Substituted pyridazines as EGFR and/or KRAS inhibitors

Abstract

The present invention is directed to pyridazinone compounds of formula (I) and furan compounds of formula (II), pharmaceutical compositions of compounds of formula (I) and (II), kits containing these compounds, methods of syntheses, and a method of treatment of a proliferative disease in a subject by administration of a therapeutically effective amount of a compound of formulae (I) or (II). Both classes of compounds were identified through screening of a collection of small molecule libraries. ##STR00001##

Claims

1. A method of inhibiting proliferation of a tumor cell carrying or expressing a cancer-associated epidermal growth factor receptor (EGFR) and/or Kirsten rat sarcoma viral oncogene homolog (KRAS) mutation comprising contacting the cell with a compound having the formula: ##STR00599## wherein X is oxygen or NR.sup.x; R.sup.X is hydrogen; designates a single or double bond represented by a and b; a is a double bond; b is a single bond; R.sup.1 is selected from hydrogen, aliphaticoxy, aliphaticthioxy optionally substituted with aryl or heteroaryl, thio, amino, heterocyclyl, heteroaryl, amido, cyano, and halo; R.sup.2 is selected from hydrogen, optionally substituted aliphatic, optionally substituted aliphaticoxy, optionally substituted aliphaticthioxy, heterocyclyl, optionally substituted heteroaryl, halo, and optionally substituted arylthioxy; wherein an optionally substituted group is optionally substituted with aliphatic, aliphaticoxy, aryloxy, hydroxyaliphatic, aryl, heteroaryl, acyl, or amido; R.sup.3 is selected from hydrogen, aliphatic, optionally substituted aliphaticoxy, optionally substituted acyl, optionally substituted acyloxy, and halo, wherein an optionally substituted group is optionally substituted with heteroaryl, aryl or acyl; and Ar is selected from phenyl, indanyl, benzyl, benzothiazolyl, 1,4-benzodioxanyl and pyridinyl, each optionally and independently substituted with aliphaticoxy, aliphatic, cyano, nitro, perfluoroaliphatic, or halo; or a pharmaceutically acceptable salt thereof.

2. The method according to claim 1, wherein the cancer-associated EGFR and/or KRAS mutation is selected from delE746-A750, T790M/L858R, L858R, G12C and G13C.

3. The method according to claim 2, wherein the cancer-associated EGFR mutation is selected from delE746-A750, T790M/L858R and L858R.

4. The method according to claim 2, wherein the cancer-associated Associated KRAS mutation is selected from G12C and G13C.

5. The method according to claim 1, wherein the compound is applied to a tumor cell which is resistant to erlotinib.

6. The method according to claim 1, wherein the compound is applied to a tumor cell which is resistant to gefitinib.

7. The method according to claim 1, wherein the compound is applied to a tumor cell which is resistant to both erlotinib and gefitinib.

8. The method according to claim 1, wherein R.sup.1 and R.sup.2 are the same, and R.sup.3 is selected from hydrogen, halo, and acyl.

9. The method according to claim 8, wherein R.sup.1 and R.sup.2 are both hydrogen.

10. The method according to claim 8, wherein R.sup.1 and R.sup.2 are both chloro, bromo, iodo, or fluoro.

11. The method according to claim 8, wherein R.sup.1 and R.sup.2 are both aliphaticthioxy optionally substituted with heteroaryl or aryl.

12. The method according to claim 1, wherein R.sup.1 and R.sup.2 are different, and R.sup.3 is selected from hydrogen, halo, and acyl.

13. The method according to claim 12, wherein R.sup.1 and R.sup.2 are independently selected from hydrogen, aliphaticoxy, aliphaticthioxy optionally substituted with heteroaryl or aryl, heterocyclyl, heteroaryl, and halo.

14. The method according to claim 1, wherein Ar is selected from: ##STR00600## wherein R.sup.Ar1, R.sup.Ar2, and R.sup.Ar3 are independently selected from hydrogen, methyl, and ethyl.

15. The method according to claim 1, wherein Ar is selected from: ##STR00601## wherein each R.sup.6, R.sup.7, R.sup.8, R.sup.9, and R.sup.10 is independently selected from hydrogen, aliphaticoxy, aliphatic, cyano, nitro, perfluoroaliphatic, and halo.

16. The method according to claim 15, wherein Ar is: ##STR00602## wherein R.sup.7 is selected from aliphaticoxy, aliphatic, perfluoroaliphatic, and halo.

17. The method according to claim 16, wherein R.sup.7 is selected from methoxy, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, bromo, chloro, and iodo.

18. The method according to claim 15, wherein Ar is: ##STR00603## wherein R.sup.6 and R.sup.7 are both aliphatic.

19. The method according to claim 15, wherein Ar is: ##STR00604## wherein R.sup.7 and R.sup.8 are independently selected from aliphatic, perfluoroaliphatic, and halo.

20. The method according to claim 19, wherein R.sup.7 is selected from methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, bromo, chloro, and iodo, and R.sup.8 is selected from methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, bromo, chloro, and iodo.

21. The method according to claim 15, wherein Ar is: ##STR00605## wherein R.sup.7 and R.sup.9 are both aliphatic.

22. The method according to claim 21, wherein R.sup.7 and R.sup.9 are independently selected from methyl, ethyl, n-propyl, and n-butyl.

23. The method according to claim 15, wherein R.sup.7 is not hydrogen.

24. The method according to claim 15, wherein when R.sup.8 is not hydrogen, R.sup.7 is not hydrogen.

25. The method according to claim 15, wherein when R.sup.6 is not hydrogen, R.sup.8 is hydrogen.

26. The method according to claim 1, wherein Ar is selected from benzothiazolyl, 1,4-benzodioxanyl, and pyridinyl.

27. The method according to claim 26, wherein Ar is selected from benzothiazolyl and pyridinyl.

28. The method according to claim 27, wherein Ar is pyridinyl.

29. The method according to claim 1, wherein Ar is phenyl.

30. The method according to claim 1, wherein the compound is not 4,5-dichloro-2-m-tolyl-2H-pyridazin-3-one.

31. The method according to claim 1, wherein X is oxygen.

32. The method according to claim 31, wherein the compound is selected from: ##STR00606## ##STR00607## ##STR00608## ##STR00609## ##STR00610## ##STR00611## ##STR00612## ##STR00613## or a pharmaceutically acceptable salt thereof.

33. The method according to claim 32, wherein the compound is selected from: ##STR00614## ##STR00615## ##STR00616##

34. The method according to claim 1, wherein X is NR.sup.x, and R.sup.x is hydrogen.

35. The method according to claim 34, wherein the compound is selected from: ##STR00617## or a pharmaceutically acceptable salt thereof.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1. SKI-267077 blocks DNA synthesis. H2030 cells were treated with the indicated concentration of SKI-267077 for 24 h and then the incorporation of .sup.3H-thymidinc into DNA was determined

(2) FIGS. 2A-2B. SKI-267077 induces cell cycle arrest. H2030 cells were treated with SKI-267077 for 24 h. Cell cycle progression was analyzed by FACS analysis. FIG. 2A: Percent of cells in S-phase. FIG. 2B: percent of cells in G1 phase.

(3) FIG. 3. Activation of caspase 3/7 by SKI-267077. H2030 cells were treated with 1 M SKI-267077 for 48 h and then caspase 3/7 activity measured in whole cell extracts.

(4) FIGS. 4A-4E. IC.sub.50 Curves of Primary HTS Data for SKI 104122.

(5) FIGS. 5A-5D. IC.sub.50 Curves of SKI 104122 (repeated).

(6) FIGS. 6A-6E. IC.sub.50 Curves for Resynthesized SKI 104122.

(7) FIGS. 7A-7D. IC.sub.50 Curves for Resynthesized SKI 104122

(8) FIGS. 8A-RE. IC.sub.50 Curves for Resynthesized SKI 104122.

(9) FIGS. 9A-9E. IC.sub.50 Curves for Resynthesized SKI 104122.

(10) FIGS. 10A-10E. IC.sub.50 Curves for Resynthesized SKI 104122.

(11) FIGS. 11-40. IC.sub.50 curves for active pyridazinone compounds against the H3255 cell line.

(12) FIGS. 41A-41B. SKI-267077 inhibits DNA synthesis. SKI-267077 was used to elucidate the mechanism by which this agent inhibits cell growth. Treatment of adenocarcinoma cell lines with either mutant EGFR (H1975 and H3255) resulted in a dose-dependent reduction in DNA synthesis.

(13) FIG. 42. SKI-267077 induces apoptosis. Cells were treated for 48 hours with SKI-267077 and then binding of FITC-labeled annexin V to cells determined FACS analysis was used to determine the amount of annexin V-positive cells. There was an induction of apoptosis, as measured by binding of annexin-V to the cell surface.

(14) FIG. 43A-43C. A phosphoprotein array was used to determine which signaling pathways were affected by treatment of cells with SKI-267077. To detect protein phosphorylation using phosphoprotein arrays (R and D Systems), cells were treated for 24 hours with SKI-267077 or erlotinib (Tarceva). A phosphoprotein array (R and D Systems) was used to measure the level of phosphorylation of cellular proteins. Protein phosphorylation was determined according to the manufacturer's instructions. SKI-267077 blocked phosphorylation of all three AKT isoforms (FIG. 43A) and p70 S6 kinase (FIG. 43B) (PI-3kinase signaling pathway). In addition, SKI-267077 treatment also reduced phosphorylation of the MAPK isoforms ERK1 and ERK2 (FIG. 43C) (MAPK signaling pathway). Erolotinib was used as a control in these experiments and a similar pattern of inhibition of the PI-3 kinase and MAPK pathways was observed.

DEFINITIONS

(15) Chemical Definitions

(16) Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75.sup.th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March's Advanced Organic Chemistry, 5.sup.th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3.sup.rd Edition, Cambridge University Press, Cambridge, 1987; the entire contents of each of which are incorporated herein by reference.

(17) Certain compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.

(18) Where an isomer/enantiomer is preferred, it may, in some embodiments be provided substantially free of the corresponding enantiomer, and may also be referred to as optically enriched. Thus, an optically-enriched isomer/enantiomer refers to a compound which is isolated or separated via separation techniques or prepared free of the corresponding isomer/enantiomer. Optically-enriched, as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments the compound is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments the compound is made up of at least about 95%, 98%, or 99% by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts or prepared by asymmetric syntheses. See, for example, Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, S. H., et al., Tetrahedron 33:2725 (1977); Eliel, E. L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, S. H. Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972).

(19) It will be appreciated that the compounds, as described herein, may be substituted with any number of substituents or functional moieties. In general, the term substituted whether preceded by the term optionally or not, and substituents contained in formulas of this invention, refer to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. As used herein, the term substituted is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. Furthermore, this invention is not intended to be limited in any manner by the permissible substituents of organic compounds.

(20) Examples of substituents are defined herein, and include, but are not limited to, aliphatic, carbocyclic, alkyl, alkenyl, alkynyl, aliphaticoxy, alkyloxy, alkenyloxy, alkynyloxy, aliphaticthioxy, alkylthioxy, alkenylthioxy, alkynylthioxy, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, amino, azido, hydrazino nitro, oxo, thiooxo, imino, hydroxy, thio, halo, aryloxy, arylthioxy, and arylamino; wherein any of the substituents described above and herein may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, which result in the formation of a stable moiety. These substituents recited above may be optionally substituted by additional substituents, such as oxo, thioxo, or imino, or may be optionally substituted by any of the substituents recited above, given that the combination results in a stable moiety. Exemplary substituent combinations include, for example, perfluoroaliphatic, perfluoroaliphaticoxy, aminoaliphatic, hydroxyaliphatic, thioaliphatic, arylaliphatic, heteroarylaliphatic, arylaliphaticthioxy, arylaliphaticoxy, arylaliphaticamino, heteroaliphaticoxy, heteroaliphaticthioxy, heteroaliphaticamino; Additional examples of generally applicable substituents are illustrated by the specific embodiments and in the Examples as described herein.

(21) The term stable moiety, as used herein, preferably refers to a moiety which possess stability sufficient to allow manufacture, and which maintains its integrity for a sufficient period of time to be useful for the purposes detailed herein.

(22) The term aliphatic, as used herein, includes both saturated and unsaturated, nonaromatic, straight chain (i.e., unbranched), branched, acyclic, cyclic, or polycyclic hydrocarbons, which are optionally substituted with one or more functional groups. As will be appreciated by one of ordinary skill in the art, aliphatic is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties. Thus, as used herein, the term alkyl includes straight, branched and cyclic alkyl groups. An analogous convention applies to other generic terms such as alkenyl, alkynyl, and the like. Furthermore, as used herein, the terms alkyl, alkenyl, alkynyl, and the like encompass both substituted and unsubstituted groups. In certain embodiments, as used herein, alkyl is used to indicate those alkyl groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1-6 carbon atoms. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(23) The term carbocyclic, carbocycle, or carbocycyl, as used herein, refers to a cyclic aliphatic group, such as a cycloalkyl, cycloalkenyl, and cycloalkynyl, which may be substituted or unsubstituted. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(24) The term alkyl, as used herein, refers to saturated, straight- or branched-chain hydrocarbon radicals derived from a hydrocarbon moiety containing between one and twenty carbon atoms by removal of a single hydrogen atom. In some embodiments, the alkyl group employed in the invention contains 1-10 carbon atoms. In another embodiment, the alkyl group employed contains 1-8 carbon atoms. In still other embodiments, the alkyl group contains 1-6 carbon atoms. In yet another embodiments, the alkyl group contains 1-4 carbons. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl, iso-pentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-dccyl, n-undecyl, dodecyl, and the like, which may bear one or more substitutents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(25) The term alkenyl, as used herein, denotes a monovalent group derived from a straight- or branched-chain hydrocarbon moiety having at least one carbon-carbon double bond by the removal of a single hydrogen atom. In certain embodiments, the alkenyl group employed in the invention contains 1-20 carbon atoms. In some embodiments, the alkenyl group employed in the invention contains 1-10 carbon atoms. In another embodiment, the alkenyl group employed contains 1-8 carbon atoms. In still other embodiments, the alkenyl group contains 1-6 carbon atoms. In yet another embodiments, the alkenyl group contains 1-4 carbons. Alkenyl groups include, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(26) The term alkynyl, as used herein, refers to a monovalent group derived from a straight- or branched-chain hydrocarbon having at least one carbon-carbon triple bond by the removal of a single hydrogen atom. In certain embodiments, the alkynyl group employed in the invention contains 1-20 carbon atoms. In some embodiments, the alkynyl group employed in the invention contains 1-10 carbon atoms. In another embodiment, the alkynyl group employed contains 1-8 carbon atoms. In still other embodiments, the alkynyl group contains 1-6 carbon atoms. Representative alkynyl groups include, but are not limited to, ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(27) The term aliphaticoxy, as used herein, refers to an aliphatic group, defined herein, attached to the parent molecular moiety through an oxygen atom. In certain embodiments, the aliphatic group contains 1-20 aliphatic carbon atoms. In certain other embodiments, the aliphatic group employed in the invention contain 1-8 aliphatic carbon atoms. In still other embodiments, the aliphatic group contains 1-6 aliphatic carbon atoms. In yet other embodiments, the aliphatic group contains 1-4 aliphatic carbon atoms. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, tert-butoxy, i-butoxy, sec-butoxy, neopentoxy, n-hexoxy, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the above-listed substituents, and that result in the formation of a stable moiety.

(28) The terms alkyloxy, alkenyloxy and alkynyloxy refer to an alkyl, alkenyl and alkynyl group, respectively, attached to the parent molecular moiety through an oxygen atom.

(29) The terms aliphaticthioxy, as used herein, refer to an aliphatic group attached to the parent molecular moiety through a sulfur atom. In certain embodiments, the aliphatic group contains 1-20 aliphatic carbon atoms. In certain other embodiments, the aliphatic group contains 1-10 aliphatic carbon atoms. In yet other embodiments, the aliphatic group contains 1-8 aliphatic carbon atoms. In still other embodiments, the aliphatic group contains 1-6 aliphatic carbon atoms. In yet other embodiments, the aliphatic group contain 1-4 aliphatic carbon atoms. Examples of aliphaticthioxy moieties include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

(30) The terms alkylthioxy, alkenylthioxy and alkynylthioxy refer to an alkyl, alkenyl, and alkynyl group, respectively, attached to the parent molecular moiety through a sulfur atom.

(31) The term heteroaliphatic, as used herein, refers to an aliphatic moiety, as defined herein, that contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms. In certain embodiments, heteroaliphatic moieties are substituted by independent replacement of one or more of the hydrogen atoms thereon with one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(32) The term heterocyclic, or heterocyclyl, as used herein, refers to an non-aromatic, partially unsaturated or fully saturated, 3- to 10-membered ring system, which includes single rings of 3 to 8 atoms in size, and bi- and tri-cyclic ring systems which may include aromatic five- or six-membered aryl or heteroaryl groups fused to a non-aromatic ring. These heterocyclic rings include those having from one to three heteroatoms independently selected from oxygen, sulfur, and nitrogen, in which the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. In certain embodiments, the term heterocylic refers to a non-aromatic 5-, 6-, or 7-membered ring or polycyclic group wherein at least one ring atom is a heteroatom selected from O, S, and N (wherein the nitrogen and sulfur heteroatoms may be optionally oxidized), and the remaining ring atoms are carbon, the radical being joined to the rest of the molecule via any of the ring atoms. Heterocycyl groups include, but are not limited to, a bi- or tri-cyclic group, comprising fused five, six, or seven-membered rings having between one and three heteroatoms independently selected from the oxygen, sulfur, and nitrogen, wherein (i) each 5-membered ring has 0 to 2 double bonds, each 6-membered ring has 0 to 2 double bonds, and each 7-membered ring has 0 to 3 double bonds, (ii) the nitrogen and sulfur heteroatoms may be optionally oxidized, (iii) the nitrogen heteroatom may optionally be quaternized, and (iv) any of the above heterocyclic rings may be fused to an aryl or heteroaryl ring. Exemplary heterocycles include azacyclopropanyl, azacyclobutanyl, piperidinyl, piperazinyl, azocanyl, thiaranyl, thietanyl, tetrahydrothiophenyl, dithiolanyl, thiacyclohexanyl, oxiranyl, oxetanyl, tetrahydropuranyl, tetrahydropuranyl, dioxanyl, oxathiolanyl, morpholinyl, thioxanyl, tetrahydronaphthyl, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(33) The term aryl, as used herein, refer to stable aromatic mono- or polycyclic ring system having 3-20 ring atoms, of which all ring atoms are carbon, and which may be substituted or unsubstituted. In certain embodiments of the present invention, aryl refers to a mono, bi, or tricyclic C.sub.4-C.sub.20 aromatic ring system having one, two, or three aromatic rings which include, but not limited to, phenyl, biphenyl, naphthyl, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(34) The term heteroaryl, as used herein, refer to stable aromatic mono- or polycyclic ring system having 3-20 ring atoms, of which one ring atom is selected from S, O, and N; zero, one, or two ring atoms are additional heteroatoms independently selected from S, O, and N; and the remaining ring atoms are carbon, the radical being joined to the rest of the molecule via any of the ring atoms. Exemplary heteroaryls include, but are not limited to pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, pyyrolizinyl, indolyl, quinolinyl, isoquinolinyl, benzoimidazolyl, indazolyl, quinolinyl, isoquinolinyl, quinolizinyl, cinnolinyl, quinazolynyl, phthalazinyl, naphthridinyl, quinoxalinyl, thiophenyl, thianaphthenyl, furanyl, benzofuranyl, benzothiazolyl, thiazolynyl, isothiazolyl, thiadiazolynyl, oxazolyl, isoxazolyl, oxadiaziolyl, oxadiaziolyl, and the like, which may bear one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(35) The term acyl, as used herein, refers to a group having the general formula C(O)R, where R is hydrogen, halogen, hydroxy, thio, amino, aliphatic, carbocyclic, heteroaliphatic, alkyl, alkenyl, alkynyl, aryl, alkyloxy, alkylthioxy, alkylamino, dialkylamino, arylamino, diarylamino, aryl, heteroaryl, or heterocycyl. Exemplary acyl groups include aldehydes, carboxylic acids, ketones (such as an acetyl group [(CO)CH.sub.3], esters, amides, carbonates, carbamates, and ureas.

(36) The term acyloxy, as used herein, refers to an acyl group of the formula (OC(O)R), where R may be hydrogen, hydroxy, thio, amino, aliphatic, carbocyclic, heteroaliphatic, alkyl, alkenyl, alkynyl, aryl, aliphaticoxy, aliphaticthioxy, aliphaticamino, dialiphaticamino, arylamino, diarylamino, aryl, heteroaryl, or heterocycyl.

(37) The term amide, or amido, as used herein, refers to an acyl group having the general formula C(O)N(R)(R), N(H)C(O)(R), or N(R)C(O)(R), where each instance of R is, hydrogen, hydroxy, thio, amino, aliphatic, carbocyclic, heteroaliphatic, alkyl, alkenyl, alkynyl, aryl, alkyloxy, alkylthioxy, alkylamino, dialkylamino, arylamino, di arylamino, aryl, heteroaryl, or heterocycyl.

(38) The term imide, or imido, as used herein, refers to a group having the general formula C(NR)R, OC(NH)R, OC(NR)R, C(NH)R, N(H)C(NH)R, N(H)C(NR)R, N(R)C(NH)R, or N(R)C(NR)R, where each instance of R is, independently, aliphatic, heteroaliphatic, aryl, heteroaryl or heterocyclyl.

(39) The term sulfinyl, as used herein, refers to a group of the formula RS(O) where there is one double bond between the sulfur and oxygen, and where R may be aliphatic, aryl, alkoxy, hydroxy, thiol, alkylthioxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, or heterocyclyl. The term aliphaticsulfinyl refers to a sulfinyl group where R may be aliphatic, heterocyclyl, or heteroaliphatic. The term arylsulfinyl refers to a sulfinyl group where R may be aryl or heteroaryl.

(40) The term sulfonyl, as used herein, refers to an organic radical (or functional group) obtained from an sulfonic acid by the removal of the hydroxyl group. Sulfonyl groups can be written as having the general formula RS(O).sub.2, where there are two double bonds between the sulfur and oxygen, and where R may be aliphatic, heteroaliphatic, aryl, alkyloxy, hydroxy, thiol, alkylthioxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, or heterocyclic. The term aliphaticsulfonyl refers to a sulfonyl group where R may be aliphatic, heteroaliphatic, or heterocyclic. The term arylsulfonyl refers to a sulfonyl group where R may be aryl or heteroaryl. The names of sulfonyl groups typically end in -syl, such as tosyl (toluene sulfonyl, CH.sub.3C.sub.6H.sub.4SO.sub.2), mesyl (methyl sulfonyl, CH.sub.3SO.sub.2), etc.

(41) The terms aliphaticamino, dialiphaticamino, and trialiphaticamino, as used herein, refers to one, two, or three, respectively, aliphatic, heterocyclyl or heteroaliphatic groups, as previously defined, attached to the parent molecular moiety through a nitrogen atom. The term aliphaticamino refers to a group having the structure NHR wherein R is an aliphatic, heterocyclyl or heteroaliphatic group, as previously defined; and the term dialiphaticamino refers to a group having the structure NRR, wherein R and R are each independently selected from the group consisting of aliphatic, heterocyclyl or heteroaliphatic groups. The term trialiphaticamino refers to a group having the structure NRRR, wherein R, R, and R are each independently selected from the group consisting of aliphatic, heterocyclyl or heteroaliphatic groups. In certain embodiments, the R, R, or R groups contain 1-20 carbon atoms. In certain other embodiments, the R, R, or R groups contain 1-10 carbon atoms. In yet other embodiments, the R, R, or R groups contain 1-8 carbon atoms. In still other embodiments, the R, R, or R groups contain 1-6 carbon atoms. In yet other embodiments, the R, R, or R groups contain 1-4 carbon atoms. Additionally, R, R, and/or R taken together may optionally be joined to form a five to six membered ring system. Examples include, but are not limited to, methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, iso-propylamino, piperidino, piperazinyl, pyrrolidinyl, trimethylamino, and propylamino. In certain embodiments, the R, R, or R groups are substituted by independent replacement of one or more of the hydrogen atoms thereon with one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(42) The terms arylamino, and diarylamino, as used herein, refers to one, or two, respectively, aryl or heteroaryl groups, as previously defined, attached to the parent molecular moiety through a nitrogen atom. The term arylamino refers to a group having the structure NHR wherein R is an aryl or heteroaryl group, as previously defined; and the term diarylamino refers to a group having the structure NRR, wherein R and R are each independently selected from the group consisting of aryl and heteroaryl. In certain embodiments, the R or R groups are substituted by independent replacement of one or more of the hydrogen atoms thereon with one or more substituents. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(43) The term perfluoroaliphatic, as used herein, refers to an aliphatic group, as defined herein, that has only fluorine substituents. Such perfluoroaliphatic include trifluoromethyl (CF.sub.3).

(44) The term perfluoroaliphaticoxy, as used herein, refers to a perfluoroaliphatic group, as defined herein, that is attached to the parent group through an oxygen atom.

(45) The terms carboxaldehyde, or carboxyaldehyde, as used herein, refers to an acyl group of the formula CHO.

(46) The term cyano, as used herein, refers to a group of the formula (CN).

(47) The term isocyano, as used herein, refers to a group of the formula (NC).

(48) The term amino, as used herein, refers to a group of the formula (NH.sub.2).

(49) The term azido, as used herein, refers to a group of the formula (N.sub.3).

(50) The term hydrazino, as used herein, refers to a group of the formula [(N(R)N(R)(R)], wherein each instance of R, R, or R, is a hydrogen, or a substituent. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(51) The term nitro, as used herein, refers to a group of the formula (NO.sub.2).

(52) The term oxo, as used herein, refers to a group of the formula (O)

(53) The term thiooxo, as used herein, refers to a group of the formula (S).

(54) The term imino, as used herein, refers to a group of the formula (NR), wherein R is a hydrogen, or a substituent. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(55) The term hydroxy, or hydroxyl, as used herein, refers to a group of the formula (OH).

(56) The term thio, or thiol, as used herein, refers to a group of the formula (SH).

(57) The terms halo and halogen as used herein refer to an atom selected from fluorine (fluoro), chlorine (chloro), bromine (bromo), and iodine (iodo).

(58) The term aminoaliphatic, as used herein, refers to an amino group, as defined herein, attached to the parent molecular moiety through an aliphatic group.

(59) The term hydroxyaliphatic, as used herein, refers to a hydroxy group, as defined herein, attached to the parent molecular moeity through an aliphatic group.

(60) The term thioaliphatic, as used herein, refers to a thio group, as defined herein, attached to the parent molecular moiety through an aliphatic group.

(61) The term arylaliphatic, as used herein, refers to an aryl group, as defined herein, attached to the parent molecular moeity through an aliphatic group.

(62) The term heteroarylaliphatic as used herein, refers to a heteroaryl group, as defined herein, attached to the parent molecular moiety through an aliphatic group.

(63) The term aryloxy, as used herein, refers to an aryl group, as defined herein, attached to the parent molecular moiety through an oxygen atom.

(64) The term arylthioxy, as used herein, refers to an aryl group, as defined herein, attached to the parent molecular moiety through a sulfur atom.

(65) The term arylamino, as used herein, refers to an aryl group, as defined herein, attached to the parent molecular moiety through a nitrogen atom (N)(R), wherein R is a hydrogen, or a substituent. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(66) The term arylaliphaticthioxy, as used herein, refers to an arylaliphatic group, as defined herein, attached to the parent molecular moiety through a sulfur atom.

(67) The term arylaliphaticoxy, as used herein, refers to an arylaliphatic group, as defined herein, attached to the parent molecular moiety through an oxygen atom.

(68) The term arylaliphaticamino, as used herein, refers to an arylaliphatic group, as defined herein, attached to the parent molecular moiety through a nitrogen atom (N)(R), wherein R is a hydrogen, or a substituent. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(69) The term heteroaliphaticoxy, as used herein, refers to a heteroaliphatic group, as defined herein, attached to the parent molecular moiety through an oxygen atom.

(70) The term heteroaliphaticthioxy, as used herein, refers to a heteroaliphatic group, as defined herein, attached to the parent molecular moiety through a sulfur atom.

(71) The term heteroaliphaticamino, as used herein, refers to a heteroaliphatic group, as defined herein, attached to the parent molecular moiety through a nitrogen atom (N)(R)(R), wherein R and R is, independently, a hydrogen, or a substituent. Substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety.

(72) The term Ph, as used herein, refers to a phenyl group.

(73) The term Ar, as used herein, refers to an aryl or a heteroaryl group.

(74) General Terms

(75) The term neoplasia, or neoplastic, or neoplasm, as used herein, refers to the abnormal growth of tissue in a subject, and may be either benign or malignant.

(76) The presently claimed invention is directed to the treatment of neoplasia in a subject. In certain embodiments, the neoplasia is a tumor. Tn certain embodiments, the neoplasia is a benign tumor. In certain embodiments, the neoplasia is a malignant tumor (i.e., cancer).

(77) By the term tumor is meant a neoplastic growth of cells which may be either benign or malignant.

(78) The term proliferative disease as used herein refers to any disease associated with an undesired and/or abnormal proliferation of cells. The cells may be any type of cell found in the subject. The proliferation may be due to any cause (e.g., any genetic mutation, any signal). Examples of proliferative diseases include cancer, neoplasms, inflammatory diseases, autoimmune diseases, graft-vs.-host disease, diabetic retinopathy, and benign tumors.

(79) By the term cancer is meant a malignant neoplasm. Cancers are classified by the type of cell that resembles the tumor and, therefore, the tissue presumed to be the origin of the tumor; e.g., carcinoma: malignant tumors derived from epithelial cells, and represents the most common cancers, including breast, prostate, lung and colon cancer; lymphoma and leukemia: malignant tumors derived from blood and bone marrow cells; sarcoma: malignant tumors derived from connective tissue, or mesenchymal cells; mesothelioma: tumors derived from the mesothelial cells lining the peritoneum and the pleura; glioma: tumors derived from glia, the most common type of brain cell; germinoma: tumors derived from germ cells, normally found in the testicle and ovary; choriocarcinoma: malignant tumors derived from the placenta. In certain embodiments, the presently claimed invention is directed to the treatment of prostate cancer, lung cancer, breast cancer, brain cancer, bone cancer, stomach cancer, oral cancer, skin cancer (melanoma), colorectal cancer, bladder cancer, pancreatic cancer, endometrial cancer, ovarian cancer, endometrial cancer, cutaneous melanoma, leukemia, non-Hodgkin's lymphoma, Wilms' tumor, lymphomas, rhabdomyosarcoma (arising from muscle), retinoblastoma, osteosarcoma, or Ewing's sarcoma, in a subject.

(80) The term subject, as used herein, refers to any animal. In certain embodiments, the subject is a mammal. In certain embodiments, the term subject, as used herein, refers to a human (e.g., a man, a woman, or a child).

(81) The terms administer, administering, or administration, as used herein, refer to either directly administering a compound or composition to a patient, or administering a prodrug derivative or analog of the compound to the patient, which will form an equivalent amount of the active compound or substance within the patient's body.

(82) The terms treat or treating, as used herein, refers to partially or completely alleviating, inhibiting, preventing, ameliorating, and/or relieving the condition.

(83) The terms effective amount and therapeutically effective amount, as used herein, refer to the amount of a compound of the presently claimed invention that, when administered to a patient, is effective to at least partially treat a condition from which the subject is suffering. Conditions include, but are not limited to, renal cancer, bladder cancer, liver cancer, testicular cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, brain cancer, bone cancer, stomach cancer, oral cancer, skin cancer, blood cancer, leukemia, non-Hodgkin's lymphoma, Wilms' tumor, lymphomas, rhabdomyosarcoma, retinoblastoma, osteosarcoma, Ewing's sarcoma, or any other disorder as described herein. To be therapeutically effective, against tumors or cancers, as used herein, is meant reducing the neoplasm, or slowing or halting the growth or spread of the neoplasm, e.g., tumor or cancer, in a subject diagnosed or suffering from a tumor or cancer.

(84) The term pharmaceutically acceptable salts or pharmaceutically acceptable salt includes acid addition salts, that is salts derived from treating a compound of the presently claimed invention with an organic or inorganic acid such as, for example, acetic, lactic, citric, cinnamic, tartaric, succinic, fumaric, maleic, malonic, mandelic, malic, oxalic, propionic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, glycolic, pyruvic, methanesulfonic, ethanesulfonic, toluencsulfonic, salicylic, benzoic, or similarly known acceptable acids. Where a compound the presently claimed invention contains a substituent with acidic properties, for instance, phenolic hydroxyl, the term also includes salts derived from bases, for example, sodium salts.

(85) The term prodrug, as used herein, is meant a compound which is administered to a subject in an inactive (or significantly less active) form. Once administered, the prodrug is metabolised in vivo, for example, by deacylation, dephosphorylation, hydrolysis, or epimerization, into a more active compound.

(86) The term isomers, as used herein, is meant two or more organic compounds that are configurational isomers (e.g., isomers that are constitutionally identical but differ by a 3D distribution of groups in space). Configurational isomers include geometric isomers (e.g., cis, trans, E, Z) and stereoisomers (e.g., enantiomers, diastereomers, atropisomers).

(87) The term tautomers, as used herein, is meant two or more organic compounds generated from each other by a formal migration of a hydrogen atom, and accompanied by an exchange of valencies between a single bond and an adjacent double bond, i.e., a tautomerization reaction. Tautomers include keto-enol, amide-imidic, lactam-lactim, enamine-imine, and enamine-imine. In solutions where tautomerization is possible, a chemical equilibrium of the tautomers will be reached. The exact ratio of the tautomers depends on several factors, including temperature, solvent, and pH.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

(88) The present invention is directed to pyridazinone and furan-containing compounds, and pharmaceutically acceptable salts, prodrugs, isomers, and tautomers, thereof, pharmaceutical compositions, kits, methods of syntheses, and methods of treating proliferative diseases, such as cancer (e.g., lung cancer), in a subject by administering a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, prodrug, isomer, or tautomer, thereof.

(89) Furthermore, the compounds of the present invention may target a common effector in the oncogenic EGFR and/or KRAS pathways, and may be so identified using cell-based screens as described herein. Such compounds are effective in the treatment of cancers that are associated with EGFR and/or KRAS mutations.

(90) Compounds of Formula (I)

(91) Pyridazinone compounds of the present invention correspond to compounds of formula (I) as depicted below:

(92) ##STR00004##

(93) wherein X is oxygen or NR.sup.x;

(94) R.sup.x is hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted sulfinyl, optionally substituted sulfonyl, optionally substituted hydroxy, or optionally substituted amino;

(95) designates a single or double bond represented by a and b; with the proviso that:

(96) (i) when a is a double bond, b is a single bond, and R.sup.1, R.sup.2, and R.sup.3, are, independently, hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, halo; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; or R.sup.1 and R.sup.2 are joined to form a 5- to 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl or optionally substituted heteroaryl ring, and R.sup.3 is as defined above; or R.sup.2 and R.sup.3 are joined to form a 5- to 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl or optionally substituted heteroaryl ring, and R.sup.1 is as defined above; or

(97) (ii) when a is a single bond, b is a double bond, and R.sup.2 and the carbon directly attached to R.sup.2 form an (O), (S), or (NR.sup.y) group, wherein R.sup.y is hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, halo; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; and R.sup.1 and R.sup.3 are, independently, hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, halo; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo;

(98) Ar is a group selected from aryl, heteroaryl, aliphatic optionally substituted with aryl, aliphatic optionally substituted with heteroaryl, heteroaliphatic optionally substituted with aryl, or heteroaliphatic optionally substituted with heteroaryl, each group optionally substituted with aliphatic, heteroaliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo;

(99) and pharmaceutically acceptable salts, prodrugs, isomers, or tautomers, thereof.

(100) In certain embodiments, the compound 4,5-dichloro-2-m-tolyl-2H-pyridazin-3-one (SKI-104122), as depicted below, is specifically excluded from compounds of formula (I):

(101) ##STR00005##

4,5-dichloro-2-m-tolyl-2H-pyridazin-3-one

(102) In certain embodiments, R.sup.1 and R.sup.2 are the same, and R.sup.3 is hydrogen, halo, optionally substituted acyl, optionally substituted heterocyclic, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted hydroxy.

(103) In other embodiments, R.sup.1 and R.sup.2 are both hydrogen.

(104) In other embodiments, R.sup.1 and R.sup.2 are both chloro. In other embodiments, R.sup.1 and R.sup.2 are both bromo. In other embodiments, R.sup.1 and R.sup.2 are both iodo. In other embodiments, R.sup.1 and R.sup.2 are both fluoro.

(105) In other embodiments, R.sup.1 and R.sup.2 are both optionally substituted hydroxy. In yet other embodiments, R.sup.1 and R.sup.2 are both optionally substituted thio. In yet other embodiments, R.sup.1 and R.sup.2 are both optionally substituted amino.

(106) In yet other embodiments, R.sup.1 and R.sup.2 are both optionally substituted aliphatic. In yet other embodiments, R.sup.1 and R.sup.2 are both optionally substituted heterocylic.

(107) In yet other embodiments, R.sup.1 and R.sup.2 are both cyano.

(108) In certain embodiments, the compounds of formula (I), wherein R.sup.1 and R.sup.2 are the same, have one of the following structural formulae:

(109) ##STR00006##

(110) wherein X is oxygen or or NR.sup.x; Hal is bromo, chloro, iodo or fluoro; Het is an optionally substituted heterocycyl; each instance of R.sup.O is hydrogen, or an optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted sulfinyl, or optionally substituted sulfonyl, or two R.sup.O groups are joined to form a 5- to 6-membered optionally substituted heterocyclic ring; and each instance of R.sup.S is hydrogen, or an optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted sulfinyl, or optionally substituted sulfonyl, or two R.sup.S groups are joined to form a 5- to 6-membered optionally substituted heterocyclic ring.

(111) In certain embodiments, R.sup.1 and R.sup.2 are the different, and R.sup.3 is hydrogen, halo, optionally substituted acyl, optionally substituted heterocyclic, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted hydroxy.

(112) In certain embodiments, R.sup.1 and R.sup.2 are the different, and are selected from the group consisting of hydrogen, halo, cyano, nitro, azido, optionally substituted hydroxy, optionally substituted thio, optionally substituted amino, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl.

(113) In certain embodiments, compounds of formula (I), wherein R.sup.1 and R.sup.2 are the different, correspond to the following structural formulae:

(114) ##STR00007## ##STR00008## ##STR00009##

(115) wherein X; Hal; Het; R.sup.O; and R.sup.S are as described above; and EWG (electron withdrawing group) is acyl (e.g., carboxylic acid, carboxaldehyde, ester, amide, imide, ketone), nitro, or cyano; Aliphatic is optionally substituted aliphatic; and each instance of R.sup.N is, hydrogen, or an optionally substituted amino, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted sulfinyl, or optionally substituted sulfonyl, or two R.sup.N groups, both groups present on the same nitrogen, are joined to form a 5- to 6-membered optionally substituted heterocyclic ring, an azido group (N.sub.3), or an optionally substituted hydrazino group.

(116) In certain embodiments, the Ar group of compounds of formula (I) corresponds to an optionally substituted aryl, or an aliphatic group optionally substituted with aryl. Exemplary aryl groups include phenyl, napthyl, and biphenyl. In certain embodiments, the Ar group is an optionally substituted phenyl group.

(117) In certain embodiments, the Ar group of compounds of formula (I) corresponds to an optionally substituted heteroaryl, or an aliphatic group optionally substituted with heteroaryl. Exemplary heteroaryl groups include pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl, pyyrolizinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, indazolyl, quinolinyl, isoquinolinyl, quinolizinyl, cinnolinyl, quinazolynyl, phthalazinyl, naphthridinyl, quinoxalinyl, thiophenyl, thianaphthenyl, furanyl, benzofuranyl, thiazolynyl, isothiazolyl, thiadiazolynyl, oxazolyl, isoxazolyl, oxadiaziolyl, and oxadiaziolyl. In certain embodiments, the Ar group is an optionally substituted pyridinyl group.

(118) In certain embodiments, the Ar group of compounds of formula (I) correspond to the following structural formula:

(119) ##STR00010##

(120) wherein n is 0 or 1;

(121) W is a carbon or nitrogen atom; with the proviso that when W is nitrogen, the group directly attached to nitrogen (e.g., R.sup.6, R.sup.7, R.sup.8, R.sup.9, or R.sup.10) is an electron pair; and

(122) when W is a carbon, then the group directly attached to the carbon (e.g., R.sup.6, R.sup.7, R.sup.8, R.sup.9, or R.sup.10) is as defined herein;

(123) each instance of R.sup.6, R.sup.7, R.sup.8, R.sup.9, and R.sup.10 is, independently, hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, halo; optionally substituted with oxo, thiooxo, imino, aliphatic, heteroaliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; or

(124) R.sup.6 and R.sup.7 may be joined to form a 5- or 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl ring; R.sup.7 and R.sup.8 may be joined to form a 5- or 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl ring; R.sup.8 and R.sup.9 may be joined to form a 5- or 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl ring; or R.sup.9 and R.sup.10 may be joined to form a 5- or 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl ring; and

(125) each instance of R.sup.4 and R.sup.5 is, independently, hydrogen or optionally substituted aliphatic.

(126) In certain embodiments, the Ar group of formula (I) corresponds to the following structural formulae:

(127) ##STR00011##

(128) wherein n is 0, and R.sup.6, R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are as described above and herein.

(129) In certain embodiments, when n is 0, R.sup.7 is not hydrogen. In certain embodiments, when n is 1, R.sup.6, R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are all hydrogen. In certain embodiments, when R.sup.8 is not hydrogen, R.sup.7 is also not hydrogen. In certain embodiments, when R.sup.6 is not hydrogen, R.sup.8 is hydrogen.

(130) In certain embodiments, the Ar group of formula (I) corresponds to the following structural formulae:

(131) ##STR00012##

(132) In certain embodiments, for compounds having an Ar group corresponding to structural formula Ia-Ar, R.sup.7 is halo, optionally substituted hydroxy, optionally substituted aliphatic, or optionally substituted heteroaliphatic. In certain embodiments, R.sup.7 is bromo, chloro, iodo, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy.

(133) In certain embodiments, for compounds having an Ar group corresponding to structural formula Ib-Ar, R.sup.6 and R.sup.7 are, independently, halo, optionally substituted hydroxy, optionally substituted aliphatic, or optionally substituted heteroaliphatic. In certain embodiments, R.sup.6 is bromo, chloro, iodo, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy, and R.sup.7 is bromo, chloro, iodo, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy.

(134) In certain embodiments, for compounds having an Ar group corresponding to structural formula Ic-Ar, R.sup.7 and R.sup.8 are, independently, halo, optionally substituted hydroxy, optionally substituted aliphatic, or optionally substituted heteroaliphatic, or R.sup.7 and R.sup.8 may be joined to form a 5- or 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl ring. In certain embodiments, R.sup.7 is bromo, chloro, iodo, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy, and R.sup.8 is bromo, chloro, iodo, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy.

(135) In certain embodiments, for compounds having an Ar group corresponding to structural formula Id-Ar, R.sup.7 and R.sup.9 are, independently, halo, optionally substituted hydroxy, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, or optionally substituted heteroaryl. In certain embodiments, R.sup.7 is bromo, chloro, iodo, fluoro, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy, and R.sup.9 is bromo, chloro, iodo, fluoro, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, benzyl, or methoxy.

(136) In certain embodiments, for compounds having an Ar group of formula (I) corresponding to structural formula Ic-Ar, the group corresponds to the following structural formulae:

(137) ##STR00013##

(138) wherein H, E, and F are sulfur, oxygen, N(H), or CH.sub.2; G is carbon or nitrogen; p is 0 to 1; and R.sup.Ar1, R.sup.Ar2, and R.sup.Ar3, are, independently, hydrogen, optionally substituted heterocyclic, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heteroaliphatic, or optionally substituted aliphatic.

(139) In certain embodiments, for compounds having an Ar group of formula (I) corresponding to structural formula Ic-Ar, the group corresponds to the following structural formulae:

(140) ##STR00014##

(141) wherein each instance of R.sup.Ar1, R.sup.Ar2, and R.sup.Ar3, is, independently, hydrogen, methyl, ethyl, phenyl, or benzyl, optionally substituted with bromo, chloro, iodo, fluoro, methyl, ethyl, n-propyl, n-butyl, trifluoromethyl, or methoxy.

(142) In certain embodiments, compounds of formula (I) correspond to compounds wherein X is oxygen. Such compounds of formula (I) are exemplified in Table 1:

(143) TABLE-US-00001 TABLE 1 Compounds of formula (I) (7a) (7b) (7c) (9a) (9b) 0 (9c) (9d) (9e) (9f) (9g) (9h) (9i) (9j) (9k) (9l) 0 (9m) (9n) (12a) (12b) (12c) (12d) (12e) (14) (15a) (15b) 0 (15c) (16) (17) (20a) (20b) (21) (22) (23) (29) (30) 0 (31) 0 0 0 0 00 01 02 03 04 05 06 07 08 09 0 0 0 0 0 0

(144) In certain embodiments, compounds of formula (I) correspond to compounds 7a-7c, 9a-9n, 12a-12c, 14, 15a-15c, 16, 17, 20a-20b, 21-23 and 29-31, as depicted above in Table 1. In certain embodiments, compounds of formula (I) correspond to compounds 7a-7c, 9a-9n, 12a-12c, 14, 15a-15c, 16, 17, and 20a-20b.

(145) In certain embodiments, compounds of formula (I) correspond to compounds wherein X is NR.sup.x; and wherein R.sup.x is hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted sulfinyl, optionally substituted sulfonyl, optionally substituted hydroxy, or optionally substituted amino. In certain embodiments, X is NH. Such compounds of formula (I) are exemplified in Table 2:

(146) TABLE-US-00002 TABLE 2 Compounds of formula (I) 0

(147) In certain embodiments, compounds of formula (I) correspond to compounds wherein a is a single bond, b is a double bond, R.sup.2 and the carbon directly attached to R.sup.2 forms the group (NR.sup.y), and R.sup.y is hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted sulfinyl, optionally substituted sulfonyl, optionally substituted hydroxy, or optionally substituted amino. In certain embodiments, R.sup.y is optionally substituted amino. In certain embodiments, R.sup.y is NH.sub.2. In certain embodiments, R.sup.y is NH(CH.sub.3). Such compounds of formula (I) are exemplified in Table 3:

(148) TABLE-US-00003 TABLE 3 Compounds of formula (I) 0
Pharmaceutical Compositions of Compounds of Formula (I)

(149) The present invention is also directed to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt, prodrug, isomer or tautomer thereof, and at least one pharmaceutically acceptable excipient.

(150) Pharmaceutically acceptable excipients are well-known in the art and include, without limitation, purified water, mannitol, sorbit, silicon dioxide, terpenes (e.g., menthol), alcohols (e.g., ethanol, propylene glycol, glycerol and other similar alcohols), organic solvents (e.g., dimethylsulfoxide, dimethylformamide, dimethylacctamide), waxes, saccharides, oligosaccharides and/or polysaccharides (e.g., starch or starch fragments, corn starch, glucose, galactose, lactose, cellulose), dextrins, amino acids, gums (e.g., xanthan gum, locust bean gum, British gum) and the like, or mixtures thereof.

(151) The present invention is also directed to methods of treating a proliferative dsease in a subject by administering to a subject a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, prodrug, isomer or tautomer thereof. The present application also contemplates a method of treating a proliferative disease in a subject by administering to a subject a therapeutically effective amount of a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt, prodrug, or tautomer thereof, and a pharmaceutically acceptable excipient.

(152) In certain embodiments, the proliferative disease is neoplasia. Neoplasia, as used herein, refers to the abnormal growth of tissue in a subject, and may be either benign or malignant. In certain embodiments, the neoplasia is a tumor. In certain embodiments, the neoplasia is cancer. In certain embodiments, the cancer is renal cancer, bladder cancer, liver cancer, testicular cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, brain cancer, bone cancer, stomach cancer, oral cancer, skin cancer, blood cancer, or leukemia. In certain embodiments, the cancer is non-Hodgkin's lymphoma, Wilms' tumor, lymphomas, rhabdomyosarcoma (arising from muscle), retinoblastoma, osteosarcoma, or Ewing's sarcoma. in certain embodiments, the cancer is lung cancer. In certain embodiments, the lung cancer is human non-small cell lung cancer (adenocarcinoma, squamous cell carcinoma, and large cell carcinoma). In certain embodiments the cancer is associated with EGFR and/or KRAS mutations.

(153) In certain embodiments, compounds of formula (I) are effective against a cancer or a tumor. To be effective against tumors or cancers, as used herein, is meant reducing the mass (e.g., a tumor or cancer) or slowing or halting the growth or spread of the mass, in a subject diagnosed or suffering from a tumor or cancer. In certain embodiments, compounds of formula (I) are therapeutically effective against a cancer or a tumor with EGFR and/or KRAS mutations. In certain embodiments compounds of formula (I) are therapeutically effective against a lung cancer or a lung tumor with EGFR and/or KRAS mutations.

(154) The form of administration of a compound of the invention to a subject is not particularly restricted, and can be oral or a parenteral administration using generally employed methods. For example, administration may be enteral (by mouth, by feeding tube, rectally), parenteral by injection or infusion (e.g., intravenous, intraarterial, intramuscular, intracardiac, subcutaneous, intradermal, intrathecal, intraperitoneal), and/or parenteral other than injection (e.g., transdermal, transmucosal, or by inhalation).

(155) Although the administered dosage of a compound of the invention will differ depending on severity of symptoms, age, gender, body weight, form of administration, and type of disease, etc., dosages may fall between about 0.01 mg/kg/day to about 100 mg/kg/day for an adult, and such dosages may be administered once or divided over several days. In certain embodiments, a therapeutically effective amount of a compound of formula (I) is between about 0.01 mg/kg/day to about 100 mg/kg/day. In certain embodiments, a therapeutically effective amount of a compound of formula (I) is at least 0.01 mg/kg/day, 0.05 mg/kg/day, 0.10 mg/kg/day, 1.0 mg/kg/day, 5.0 mg/kg/day, 10 mg/kg/day, 15 mg/kg/day, 20 mg/kg/day, 30 mg/kg/day, 40 mg/kg/day, 50 mg/kg/day, 60 mg/kg/day, 70 mg/kg/day, 80 mg/kg/day, 90 mg/kg/day, or 100 mg/kg/day.

(156) The present invention is also directed to kits, comprising at least one compound of formula (I), or pharmaceutically acceptable salts, prodrugs, isomers, or tautomers thereof. A kit of the invention can be particularly useful if it provides additional solvents, buffers, or excipients for pre-mixing before administration to a subject, or if it provides a means for oral or parental administration (e.g., syringes or graduated measurement cups). A kit of the invention can also be particularly useful if it contains additional chemotherapeutic agents for use in combination with a compound of formula (I). For instance, a physician may wish to administer to a subject one or more compounds of formula (I) in combination with one or more additional chemotherapeutic agents.

(157) Exemplary additional chemotherapeutic agents include, but are not limited to, 13-cis-Retinoic Acid, 2-CdA (2-Chlorodcoxyadcnosinc), 5-Fluorouracil (5-FU), 6-Mercaptopurine, 6-MP, 6-TG, 6-Thioguanine, Abraxane, ACCUTANE, ACTINOMYCIN-D, ADRIAMYON, ADRUCIL, AGRYLIN, ALA-CORT, Aldesleukin, Alemtuzumab, ALIMTA, Alitretinoin, ALKABAN-AQ, ALKERAN, All-transretinoic acid Alpha interferon, Altretamine, Amethopterin, Amifostine, Aminoglutethimide, Anagrelide, ANANDRON, Anastrozole, Arabinosylcytosine, Ara-C ARANESP, AREDIA, ARIMIDEX, AROMASIN, ARRANON, Arsenic trioxide, Asparaginase, ATRA, AVASTIN, Azacitidine, BCG, BCNU, Bevacizumab, Bexarotene, BEXXAR, Bicalutamidc, BiCNU, BLENOXANE, Bleomycin, Bortczomib, Busulfan, BUSULFEX, C225, Calcium Leucovorin, CAMPATH, CAMPTOSAR, Camptothecin-11, Capecitabine, CARAC, Carboplatin, Carmustine, Carmustine wafer, CASODEX, CC-5013, CCNU, CDDP, CeeNU, CERUBIDINE, Cetuximab, Chlorambucil, Cisplatin, Citrovorum Factor, Cladribine, Cortisone, COSMEGEN, CPT-11, Cyclophosphamide, CYTADREN, Cytarabine, Cytarabine liposomal, CYTOSAR-U, CYTOXAN, Dacarbazine, Dacogen, Dactinomycin, Darbepoetin alfa, Daunomycin, Daunorubicin, Daunorubicin hydrochloride, Daunorubicin liposomal, DAUNOXOME, Decadron, Decitabine, DELTA-CORTEF, DELTASONE, Denileukin diftitox, DEPOCYT, Dexamethasone, Dexamethasone acetate, Dexamethasone Sodium Phosphate, Dexasone, Dexrazoxane, DHAD, DIC, Diodex, Docetaxel, DOXIL, Doxorubicin, Doxorubicin liposomal, DROXIA, DTIC, DTIC-DOME, DURALONE, EFUDEX, ELIGARD, ELLENCE, ELOXATIN, ELSPAR, EMCYT, Epirubicin, Epoetin alfa, ERBITUXT, Erlotinib, Erwinia L-asparaginase, Estramustine, Ethyol, ETOPOPHOS, Etoposide, Etoposide Phosphate, EULEXIN, Evista, Exemestane, FARESTON, FASLODEX, FEMARA, Filgrastim, Floxuridine, FLUDARA, Fludarabine, FLUOROPLEX, Fluorouracil, Fluorouracil (cream), Fluoxymesterone, Flutamide, Folinic Acid, FUDR, Fulvestrant, G-CSF, Gefitinib, Gemcitabine, Gemtuzumab ozogamicin, GEMZAR, GLEEVEC, Gliadel wafer, GM-CSF, Goserelin granulocyte-colony stimulating factor, Granulocyte macrophage colony stimulating factor, Halotestin, Herceptin, Hexadrol, Hexylen, Hexamethylmelamine, HMM, Hycamtin, Hydrea, Hydrocort Acetate, Hydrocortisone, Hydrocortisone sodium phosphate, Hydrocortisone sodium succinate, Hydrocortone phosphate, Hydroxyurea, Ibritumomab, Ibritumomab Tiuxetan, IDAMYCIN, Idarubicin, IFEX, IFN-alpha, Ifosfamide, IL-11, IL-2, Imatinib mesylate, Imidazole Carboxamide, Interferon alfa, Interferon Alfa-2b (PEG conjugate), Interleukin-2 (t), Interleukin-11, INTRON A (interferon alfa-2b), IRESSA, Irinotecan, Isotretinoin, Kidrolase, Lanacort, L-asparaginase, LCR, Lenalidomide, Letrozole, Leucovorin, Leukeran, Leukine, Leuprolide, Leurocristine, Leustatin, Liposomal Ara-C, Liquid Pred, Lomustine, L-PAM, L-Sarcolysin, Lupron, LUPRON DEPOT, Matulane, Maxidex, Mechlorethamine, Mechlorethamine Hydrochloride, Medralone, MEDROL, Megace, Megestrol, Megestrol Acetate, Melphalan, Mercaptopurine, Mesna, Mesnex, Methotrexate, Methotrexate Sodium, Methylprednisolone, Meticorten, Mitomycin, Mitomycin-C, Mitoxantrone, M-Prednisol, MTC, MTX, Mustargen, Mustine, Mutamycin, Myleran, Mylocel, Mylotarg, NAVELBINE, Nelarabine, Neosar, Neulasta, Neumega, NEUPOGEN, NEXAVAR, Nilandron, Nilutamide, NIPENT, Nitrogen Mustard, Novaldex, Novantrone, Octreotide, Octreotide acetate, Oncospar, Oncovin, Ontak, Onxal, Oprevelkin, Orapred, Orasone, Oxaliplatin, Paclitaxel, Paclitaxel Protein-bound, Pamidronate, Panretin, Paraplatin, Pediapred, PEG Interferon, Pegaspargase, Pegfilgrastim, PEG-INTRON, PEG-L-asparaginase, PEMETREXED, Pentostatin, Phenylalanine Mustard, Platinol, Platinol-AQ, Prednisolone, Prednisone, Prelone, Procarbazine, PROCRIT, Proleukin, Prolifeprospan 20 with Carmustine implant, PURINETHOL, Raloxifene, REVLIMID, Rheumatrex, Rituxan, Rituximab, ROFERON-A (interferon alfa-2a), Rubex, Rubidomycin hydrochloride, SANDOSTATIN, Sandostatin LAR, Sargramostim, Solu-Cortef, Solu-Medrol, Sorafenib, STI-571, Streptozocin, SU11248, Sunitinib, SUTENT, Tamoxifen, TARCEVA, Targretin, TAXOL, TAXOTERE, TEMODAR, Temozolomide, Teniposide, TESPA, Thalidomide, THALOMID, TheraCys, Thioguanine, THIOGUANINE TABLOID, Thiophosphoamide, Thioplex, Thiotepa, TICE, Toposar, Topotecan, Toremifene, Tositumomab, Trastuzumab, Tretinoin, Trexall, Trisenox, TSPA, VCR, Velban, VELCADE, VePesid, Vesanoid, Viadur, Vidaza, Vinblastine, Vinblastine Sulfate, Vincasar Pfs, Vincristine, Vinorelbine, Vinorelbine tartrate, VLB, VM-26, VP-16, Vumon, XELODA, Zanosar, ZEVALIN, Zinecard, ZOLADEX Zolcdronic acid, and ZOMETA.

(158) Methods of Preparation of Compounds of Formula (I)

(159) The present invention also provides a method of synthesizing compounds of formula (I), as depicted in Scheme 1 below.

(160) ##STR00181##

(161) Compounds of formula (I) may be synthesized starting from an optionally substituted aniline (A), wherein R.sup.11 is any substitutent, as is described herein, which results in a stable substituted aniline compound (A), and q is 0-5.

(162) In step 1 (S-1), under standard hydrazine formation conditions, optionally substituted aniline (A) is converted to optionally substituted phenyl hydrazine (B).

(163) In step 2 (S-2), reaction of compound (B) with compound (C), wherein Z is, independently, hydrogen or halo (Cl, Br, I, F), generates an optionally substituted pyridazin-3-one derivative (D).

(164) Reaction of compound (D) in step 3 (S-3) with different nucleophiles can generate a wide variety of compounds with different substitutions and/or substitution patterns (for example, compounds (E), (F), and/or (G), as depicted above). In certain embodiments, R.sup.12 and R.sup.13, are, independently, optionally substituted aliphaticoxy, aliphaticthioxy, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, isocyano, amino, azido, hydrazino, hydroxy, thio, halo, aryloxy, arylthioxy, arylamino, heteroaryloxy, heteroarylamino, or heteroarylthiooxy.

(165) In certain embodiments, compound (D) is treated with a mono- or di-substituted aliphaticamine to form compounds (E), and/or (F), and/or (G), wherein R.sup.12 and R.sup.13 are, independently, N(R.sup.N).sub.2, and each instance of R.sup.N is, independently, hydrogen or optionally substituted aliphatic, heteroaliphatic, aryl, heteroaryl, or two R.sup.N groups form an optionally substituted 5- to 6-membered heteroaryl or heterocyclic ring.

(166) In certain embodiments, compound (D) is treated with an optionally substituted hydroxy group to form compounds (E), and/or (F), and/or (G), wherein R.sup.12 and R.sup.13 are, independently, O(R.sup.o), and each instance of R.sup.o is, independently, hydrogen or optionally substituted aliphatic, heteroaliphatic, aryl, heteroaryl, or two R.sup.o groups form an optionally substituted 5- to 6-membered heteroaryl or heterocyclic ring.

(167) In certain embodiments, compound (D) is treated with an optionally substituted thio group to form compounds (E), and/or (F), and/or (G), wherein R.sup.12 and R.sup.13 are, independently, S(R.sup.s), and each instance of R.sup.s is, independently, hydrogen or optionally substituted aliphatic, heteroaliphatic, aryl, heteroaryl, or two R.sup.s groups form an optionally substituted 5- to 6-membered heteroaryl or heterocyclic ring.

(168) In certain embodiments, compound (D) is treated with a Brnstead acid (such as hydrogen chloride, hydrogen bromide, hydrogen bromide, hydrogen fluoride, nitric acid, sulfuric acid, phosphoric acid, and the like), or a halogenating reagent (such as NBS, Br.sub.2, NCS, and the like) to form compounds (D), and/or (E), and/or (F), wherein R.sup.12 and R.sup.13 are, independently, hydrogen, bromo, iodo, fluoro, NO.sub.2, SO.sub.3, OP(O)(OH).sub.2 and the like.

(169) Alternatively, compounds of formula (I) can be synthesized according to the method as depicted in Scheme 2 below.

(170) In step 4 (S-4), under suitable cyclization conditions, optionally substituted phenyl hydrazine (B) reacts with a compound of formula (H) wherein Z is, independently, hydrogen or halo, to generate an optionally substituted pyridazine-3,6-dionepyridazin-3-one derivative (J). Treatment of compound (J) under suitable reducing conditions or alkylation conditions, in step 5 (S-5), generates optionally substituted pyridazin-3-one derivative (K), wherein R.sup.o is hydrogen or optionally substituted aliphatic, respectively. Reaction of compound (K) in step 6 (S-6) with different nucleophiles (as described in certain embodiments for step 3 (S-3) above) can generate a wide variety of compounds with different substitutions and/or substitution patterns (for example, compounds (L), (M), and/or (N), as depicted below).

(171) ##STR00182##
Compounds of Formula (II)

(172) The present invention is also directed to compounds of formula (II):

(173) ##STR00183##

(174) wherein each instance of R.sup.1, R.sup.2, and R.sup.3 is, independently, hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; or R.sup.1 and R.sup.2 are joined to form a 5- or 6-membered optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl ring, and R.sup.3 is as defined above;

(175) each instance of R.sup.4 and R.sup.5 is, independently, hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, hydrazino; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; or R.sup.4 and R.sup.5 are joined to form an 5- or 6-membered optionally substituted heterocyclic or optionally substituted heteroaryl ring; and

(176) J and J, together, form an oxo (O), thiooxo (S), or imino (NR.sup.N5) group, wherein R.sup.N5 is hydrogen, optionally substituted hydroxy, optionally substituted amino, optionally substituted aryl, optionally substituted sulfonyl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted aliphatic, or optionally substituted heteroaliphatic; or each instance of J and J is, independently, hydrogen, optionally substituted aliphatic or optionally substituted heteroaliphatic.

(177) In certain embodiments, the compound 5-nitro-furan-2-carboxylic acid (4-chloro-phenyl)-amide (SKI-98698), as depicted below, is specifically excluded from compounds of formula (II):

(178) ##STR00184##

(179) 5-nitro-furan-2-carboxylic acid (4-chloro-phenyl)-amide (SKI-98698)

(180) In certain embodiments, compounds of formula (II) have the structural formulae:

(181) ##STR00185##

(182) wherein each instance of R.sup.6 is, independently, hydrogen or optionally substituted aliphatic;

(183) each W is, independently, N, C(H), or C(R.sup.7); wherein each instance of R.sup.7 is, independently, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, halo; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo;

(184) is a single or double bond; and

(185) p is 0 or 1.

(186) In certain embodiments, R.sup.6 is hydrogen. In certain embodiments, R.sup.6 is alkyl. In certain embodiments, R.sup.6 is C.sub.1-6 alkyl.

(187) In certain embodiments, compounds of formula (II) have the structural formula IIa. Such compounds of formula IIa are exemplified in Table 4:

(188) TABLE-US-00004 TABLE 4 Compounds of formula (II) 0 00 01 02 03 04 05 06 07 08 09 0 0

(189) In certain embodiments, compounds of formula (II) have the structural formulae:

(190) ##STR00226##

(191) wherein Y is O, N(R.sup.12), or C(R.sup.12).sub.2;

(192) each instance of R.sup.8, R.sup.9, R.sup.10, R.sup.11, and R.sup.12, is, independently, hydrogen, aliphatic, heteroaliphatic, hydroxy, thio, amino, heterocyclic, aryl, heteroaryl, acyl, amido, imido, sulfinyl, sulfonyl, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, halo; optionally substituted with oxo, thiooxo, imino, aliphatic, carbocyclic, hydroxy, aliphaticoxy, aryloxy, thio, aliphaticthioxy, arylthioxy, heteroaliphatic, heterocyclic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, acyl, acyloxy, amido, imido, sulfinyl, sulfonyl, amino, aliphaticamino, dialiphaticamino, trialiphaticamino, arylamino, diarylamino, carboxaldehyde, cyano, isocyano, azido, hydrazino, nitro, or halo; or R.sup.8 and R.sup.9 are joined to form a 5- to 6-membered optionally substituted aryl or optionally substituted heteroaryl ring, and R.sup.10, R.sup.11, and R.sup.12 are as defined above; and

(193) m is 0 or 1.

(194) In certain embodiments, compounds of formula (II) have the structural formula IIe. Such compounds of formula IIe are exemplified in Table 5:

(195) TABLE-US-00005 TABLE 5 Compounds of formula (II) 0
Pharmaceutical Compositions of Compounds of Formula (II)

(196) The present invention is also directed to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (II), as defined herein, or a pharmaceutically acceptable salt, prodrug, isomer or tautomer thereof, and at least one pharmaceutically acceptable excipient.

(197) The present invention is also directed to methods of treating a proliferative disease in a subject by administering to a subject a therapeutically effective amount of a compound of formula (II), or a pharmaceutically acceptable salt, prodrug, isomer or tautomer thereof.

(198) In certain embodiments, the proliferative disease is neoplasia. Neoplasia, as used herein, refers to the abnormal growth of tissue in a subject, and may be either benign or malignant. In certain embodiments, the neoplasia is a tumor. In certain embodiments, the neoplasia is cancer. In certain embodiments, the cancer is renal cancer, bladder cancer, liver cancer, testicular cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, lung cancer, breast cancer, brain cancer, bone cancer, stomach cancer, oral cancer, skin cancer, blood cancer, or leukemia. In certain embodiments, the cancer is non-Hodgkin's lymphoma, Wilms' tumor, lymphomas, rhabdomyosarcoma (arising from muscle), retinoblastoma, osteosarcoma, or Ewing's sarcoma. In certain embodiments, the cancer is lung cancer. In certain embodiments, the lung cancer is human non-small cell lung cancer (adenocarcinoma, squamous cell carcinoma, and large cell carcinoma). In certain embodiments the cancer is associated with EGFR and/or KRAS mutations.

(199) In certain embodiments, compounds of formula (II) are effective against a cancer or a tumor. In certain embodiments, compounds of formula (II) are effective against a cancer or a tumor associated with EGFR and/or KRAS mutations. In certain embodiments, compounds of formula (II) are effective against lung cancer or a lung tumor associated with EGFR and/or KRAS mutations.

(200) The present invention is also directed to kits, comprising at least one compound of formula (II), or pharmaceutically acceptable salts, prodrugs, isomers, or tautomers, thereof. A kit of the invention can be particularly useful if it provides additional solvents, buffers, or excipients for pre-mixing before administration, or if it provides a means for oral or parental administration (e.g., syringes or graduated measurement cups). A kit of the invention can also be particularly useful if it contains additional chemotherapeutic agents, as described herein, for use in combination with one or more compounds of formula (II).

(201) In certain embodiments, a therapeutically effective amount of a compound of formula (II) is between about 0.01 mg/kg/day to about 100 mg/kg/day. In certain embodiments, a therapeutically effective amount of a compound of formula (II) is at least 0.01 mg/kg/day, 0.05 mg/kg/day, 0.10 mg/kg/day, 1.0 mg/kg/day, 5.0 mg/kg/day, 10 mg/kg/day, 15 mg/kg/day, 20 mg/kg/day, 30 mg/kg/day, 40 mg/kg/day, 50 mg/kg/day, 60 mg/kg/day, 70 mg/kg/day, 80 mg/kg/day, 90 mg/kg/day, or 100 mg/kg/day.

(202) The present invention will be more specifically illustrated by the following examples. However, it should be understood that the present invention is not limited by these examples in any manner.

EXAMPLES

Example 1

High Throughput Screening for Novel Agents that Block Proliferation of NSCLC Cell Lines

(203) There are a variety of well-documented human lung adenocarcinoma cell lines that have been used in the cell-based screens. Characteristics of these cell lines are shown in Table 6. Some of these cell lines have gain of function mutations in either exon 2 of KRAS (H2030) or in exons 18-21 of the EGFR(H1650, H1975 and H3255). The cell lines H11-18 and H3255 express EGFR with an L858R mutation and growth of these cells is inhibited by erlotinib at much lower concentrations than is required to attenuate growth in cells with wild type EGFR (Table 6). In H1650 there is a deletion of four amino acids (E746-A750) in the EGFR. Growth of this cell line is less sensitive to erlotinib than H11-18 and H3255. This may be due to the presence of additional mutations in proteins function downstream of the EGFR. Only the L858R mutation is present in the H1975 cell line. However, we sequenced exons 18-24 of EGFR in 8 adenocarcinoma cell lines and identified a second point mutation in the kinase domain of the EGFR. This C to T change at position 2369 in exon 20 results in a T790M substitution in the receptor and is believed to confer resistance to the EGFR tyrosine kinase inhibitors erlotinib and gefitinib. Indeed, growth of the H1975 cell line is more resistant to erlotinib than the cell lines carrying mutant KRAS such as H358 and H2030 (Table 6). Introduction of double stranded siRNA specific for the EGFR deletion mutant or the L858R mutant into H1650 or H1975 and H3255 induced apoptosis, suggesting that these lung adenocarcinoma cell lines require continuous expression of the EGFR oncogene for survival.

(204) All of the adenocarcinoma cell lines with EGFR mutations shown in Table 6 express wild type KRAS. Conversely, the lines with wild type EGFR in Table 6 carry a mutation in exon 2 of KRAS, resulting in a missense amino acid substitution in codons 12 or 13. In H358 and H2030, this exon 2 mutation results in a G12C amino acid change in KRAS. In H1734, there is a G13C substitution in KRAS. introduction of a G12C-KRAS allele-specific siRNA into the H2030 cell line results in growth arrest, suggesting that this cell line is dependent on expression of mutant KRAS for growth. All adenocarcinoma cell lines with mutant KRAS in Table 6 are resistant to erlotinib and express wild type EGFR.

(205) TABLE-US-00006 TABLE 6 Cell Line EGFR KRAS IC.sub.50 (Erlotinib) 11-18 L858R WT 0.01 M H358 WT G12C 10 M H1650 DelE746-A750 WT 1 M H1734 WT G13C 7 M H1975 T790M/L858R WT 25 M H2030 WT G12C 10 M H3255 L858R WT 0.01 M HCC827 DelE746-A750 WT 0.01 M HPL1D WT WT 10 M NHBE WT WT 10 M WI-38 WT WT 10 M
Exons 18-24 of EGFR and exon 2 of KRAS were sequenced and mutations identified are shown. Sensitivity of cell proliferation to erlotinib is also shown. NHBE (normal human bronchiolar epithelial cells) are primary cells obtained from donor lungs and commercially available from Cambrex. The HPLID (human peripheral lung epithelial) cell line has been immortalized with the SV40 large T-antigen. WI-38: human lung fibroblast cell line. HPLID, NHBE and WI-38 cells will serve as controls in the chemical screens. WT: wildtype; WT*: over-expressed wildtype.
Methods

(206) Cell cycle analysis. For cell cycle analysis, cells were plated at a density of 500,000 cells per well in 6-well plates. Attached cells were treated with drugs for 24 h. Cells were then collected, washed with PBS and fixed in 70% ethanol for 1 h. For FACS analysis, fixed cells were washed with cold PBS resuspended in PBS containing 200 g/mL propidium iodide and 0.1% sodium citrate. Flow cytometry was performed on a Becton-Dickinson FACSCalibur flow cytometer and data the processed using FlowJo software.

(207) Caspase 3/7 enzyme activity assay. Apoptosis was determined by measuring the enzymatic activity of caspase 3/7 in cell homogenates. Cells were plated at a density of 10,000 cells per well in 96-well plates and treated immediately with drugs for 24-72 h. Caspase 3/7 activity will be determined in cell extracts using Z-DEVD-R110 (rhodamine 110 conjugated to the caspase substrate Z-DEVD) as substrate (Promega). The fluorescence of the cleaved substrate was measured using a micro-titer fluorescence plate reader (Ex: 499 nm, Em: 521 nm).

(208) [.sup.3H]-thymidine incorporation assay. Cells were seeded in 12-well plates at a density of 100 000 cells/well and treated with inhibitors for 24 hours. H3-thymidine (3 Ci/mL) was then added for 3 hours. Cells were washed twice with PBS, fixed with cold 10% TCA (in PBS) for 30 min then lysed with 0.5% NaOH/0.5% SDS. DNA was scraped from the wells and the amount of H3-thymidine incorporated determined by liquid scintillation counting.

(209) TABLE-US-00007 TABLE 7 Reagent/Assay Supplies Name Supplier Description Non-Small Cell Lung ATCC Human lung cancer cell lines with various Cancer Cell Line mutations in eGFR or kRas RPMI 1640 ATCC Cell Culture Growth Media, Supplement with 10% FBS Corning Cell Culture Flask Fisher 175 cm2 Flask, 600 ml capacity, sterile with Filter Caps 384 Well Cell Culture Corning Sterile Black Plate w/ Clear Bottom and Lid, Microplate tissue culture treated Staurosporine LC Laboratories Potent inhibitor of phospholipid/Ca++ dependent protein kinase and platelet aggregation, all PKC isoforms, all Akt isoforms DMSO Sigma-Aldrich Dimethyl Sulfoxide Alamar Blue Serotech Nontoxic aqueous dye for viability and proliferation measurements

(210) TABLE-US-00008 TABLE 8 Equipment used Name Description Biophile Automated Compound Plate Management System TPS-384 Compound Dispensing System from Apricot Designs CRS F3 Robot Linear Robotic Track System from Thermo System Electron Corp. Multidrop 384 Liquid dispenser for 96/384 microplates Cell Culture Incubator Incubator at 37 C., humidified, 5% CO2 FlexDrop Precision liquid handling dispenser from Perkin Elmer Wallac 1420 Victor V Multi label PMT based reader for Prompt Plate Reader Fluorescence from Perkin Elmer
Protocols

(211) Growing & Maintaining Non Small Cell Lung Cancer Cell Line. Non Small Cell Lung Cancer cell lines are grown and maintained in Corning 175 cm.sub.2 flasks in RPMI 1640 media supplemented with 10% FBS, 1% antibiotic solution, 1% additional glutamine and pyruvate. These cells have a doubling time of 1-2 days. All cell types are adherent and grow in colonies. The cells should be kept at a density between 50,000 cells/mL-500,000 cells/mL to provide for optimum growth and adequate nutrition. The cell line should be maintained at 37 C. in a humidified atmosphere with 5% CO2 in a sterile cell culture incubator.

(212) Non Small Cell Lung Cancer Proliferation Assay. To perform the assay, the Non Small Cell Lung Cancer lines are diluted in media and plated into 384 well microplates at a final density of 250 cells in 45 L. To ensure consistency and minimal pipetting error, the cells are plated using the Multidrop 384 with the following settings: Rocker Switch set at 384, Volume 45 and Column 24.

(213) Low Controls for this assay are made by treating Non Small Cell Lung Cancer cells with 5 ul of staurosporine at 250 uM in 10% DMSO and 1% SDS. [Final concentration in this assay is 25 uM staurosporine in 1% DMSO and 0.1% SDS.]

(214) High Controls for this assay are made by treating Non Small Cell Lung Cancer cells with 5 ul of 10% DMSO. [Final concentration in this assay is 1% DMSO].

(215) The remaining wells of the 384 microplate may be treated with different compounds depending on the study. Important Note: All compounds for testing in this assay should be done at the final concentration of 1% DMSO.

(216) After addition, the cells are placed in cell culture incubator at 37 C. for 48 hours for complete treatment.

(217) After drug treatment, cytotoxicity is measured by Alamar Blue reduction. 5 L of Alamar Blue is added to the 384 microplates using the Flexdrop.

(218) The cells are incubated for 48 hours at 37 C. to complete the Alamar Blue reduction.

(219) The final step in the assay is to measure the amount of Alamar Blue reduction. The cells are placed in the Wallac 1420 Victor V Plate Reader I and with the following program, DS Alamar Blue Prompt Fluo (#8). The Alamar Blue fluorescence is measured at an excitation of 530 nm and emission of 590 nm.

(220) For the NSCLC project, the cell based assay was screened against a small molecule library of approximately 200,000 compounds. The compound library is stored in our automated robotic freezer (Biophile) at 20 C. in 100% DMSO. Compound freeze and thaw cycles are kept to a minimum. Compounds for use are diluted into the appropriate concentration and plated into 384 well microplates using our custom built low volume 384-well head tool (TPS 384, Apricot Designs Inc., CA, USA). This assay was performed on a fully automated linear track robotic platform (CRS F3 Robot System, Thermo Electron Corp., Ontario, Canada) utilizing several integrated peripherals for plate handling, liquid dispensing, and readout detection. First, non-small cell lung cancer cell lines are dispensed into 384 well microplates (Corning #3712 Tissue Culture Treated Plate, Corning, N.Y., USA) with 5 L of test compounds at 100 uM in 10% DMSO for screening studies or 1 mM in 10% DMSO to 50 nM in 10% DMSO for dose response studies. The microplates are then placed at 37 C. for 48 hours to complete treatment of cells with compounds. Next, 5 ul of alamar blue was added using a liquid dispenser (FlexDrop Precision Dispenser, Perkin Elmer, Mass., USA) and incubated for 48 hours at 37 C. The alamar blue prompt fluorescence was measured on Victor V (Victor.sup.3 V, Perkin Elmer, Mass., USA) and output data files were loaded into ORIS (Oncology Research Informatics System, MSKCC, NY, USA), a screening data acquisition and analysis platform.

(221) A high throughput screening was performed using four established non small cell lung cancer cell lines (H1650, H1975, H2030, H3255) against a chemical library of 200,000 small molecules. Description of the assays is described herein. Several hits were identified inhibiting one or more cell line with the most potent one being 4,5-dichloro-2-m-tolyl-2H-pyridazin-3-one (SKI-104122).

(222) ##STR00238##

4,5-dichloro-2-m-tolyl-2H-pyridazin-3-one (SKI-104122)

(223) A substructure search was then performed against the SKI library and identified 115 derivatives of SKI-104122. Further analysis of the biological data for these derivatives reveals a coherent structure activity relationship for both active and inactive compounds with only 22 compounds identified as active during primary screening. Summary of the data is shown in Table 9. Data is expressed as percentage inhibition in the cell based assay; screening concentration of 10 M compound in 1% DMSO (v/v).

(224) TABLE-US-00009 TABLE 9 Structure SKI ID H1650 H1975 H2030 H3255 104122 48 104 101 98 0 176729 100 102 3 115 104188 13 102 4 4 224256 103 102 1 110 104288 13 102 66 46 103719 45 102 94 76 196392 87 99 13 111 104322 51 99 87 82 104004 1 98 75 90 103997 15 98 9 2 104304 52 98 14 107 0 103529 13 97 15 1 187118 101 96 7 108 202245 48 96 8 71 187116 31 90 4 27 186926 27 81 2 52 195071 71 77 9 82 199759 16 36 9 66 223337 13 5 14 61 224770 65 8 8 24 227626 15 14 0 71 0 224766 12 29 95 24 104185 9 22 2 1 104217 15 7 6 1 104325 31 12 2 4 104339 21 9 3 3 104386 12 11 6 0 104390 15 4 6 34 104415 20 15 3 9 104423 11 20 3 1 174405 13 28 9 32 0 174406 18 3 9 8 176702 29 26 14 21 103751 6 4 4 0 103868 7 18 4 3 103872 17 9 2 25 103911 7 12 5 3 103971 5 10 1 1 176963 11 5 4 17 177215 6 25 9 13 177261 14 16 8 8 0 178393 8 3 11 4 179389 3 1 8 28 180651 21 13 11 35 181612 7 11 11 11 181801 13 24 7 7 181802 2 14 5 27 181862 7 3 9 22 182721 8 9 11 4 182729 10 23 14 1 182826 13 12 11 13 0 182827 20 14 6 24 182828 25 5 8 18 182830 20 10 3 4 182831 4 5 12 18 183073 3 6 8 30 183339 15 8 12 30 185423 16 9 3 23 186017 12 6 4 13 186018 42 4 4 35 186924 7 14 1 8 00 186925 6 3 1 28 01 189015 21 29 4 32 02 189286 0 9 5 15 03 189493 2 12 8 25 04 193067 7 21 4 35 05 193326 2 1 9 49 06 194491 2 0 12 32 07 194617 9 9 4 5 08 194618 15 3 6 48 09 194841 1 15 9 35 0 195063 22 34 6 6 195066 8 3 9 9 195070 8 9 7 73 195149 4 11 5 19 195988 13 7 7 18 195989 21 35 6 27 196618 3 14 12 33 197940 11 6 2 37 199760 12 22 21 6 199761 7 25 6 41 0 199764 11 14 9 2 200052 9 5 12 18 201622 5 23 4 33 206569 2 3 7 22 208619 4 1 12 19 208693 6 7 11 0 208694 21 29 3 16 208695 14 13 11 2 208696 2 19 4 37 208697 3 16 4 31 0 208699 12 22 10 19 208701 4 25 7 7 208702 3 16 15 0 208703 2 18 9 16 208704 10 6 15 23 208705 4 20 10 0 213495 4 15 5 8 216994 1 1 4 3 222922 20 3 30 7 224358 4 9 2 14 0 224764 21 5 10 32 224765 7 1 15 12 224771 12 21 3 30 225042 18 10 6 5 227247 16 9 1 11 228706 16 11 5 25 251592 ND ND ND ND 251603 ND ND ND ND 254244 ND ND ND ND 254254 ND ND ND ND 0 254262 ND ND ND ND 254579 ND ND ND ND 254913 ND ND ND ND 255398 ND ND ND ND

Example 2

SAR of Compound SKI-104122

(225) Synthesis of Pyridazin-3(2H)-ones

(226) Based on the obtained SAR results from primary screening and the observed affinities (see Example 1), the feasibility of identifying the molecular target for SKI-104122 was investigated. To achieve this, an SAR study was undertaken to identify potential sites on the molecule for linker addition without compromising biological activity. The linker will then be attached to sepharose beads generating an affinity chromatography column to be used for identifying molecular target.

(227) The preparation of compounds screened is exemplified below.

(228) ##STR00354##

(229) Preparation of 1-[(1E)-But-1-en-1-yl]-3-nitrobenzene (3). n-BuLi (2.5M/hexanes, 7.7 mL, 19.46 mmol) was added drop wise to a 78 C. cooled suspension of n-propyl triphenylphosphonium bromide (2) (7.5 g, 19.46 mmol) in THF (50 mL) and the reaction was warmed to room temperature over 1 h and cooled to 78 C. A solution of 3-nitrobenzaldehyde (1) (2.94 g, 19.46 mmol) in THF (10.0 mL) was then added and the reaction mixture was gradually warmed to room temperature. After 15 h, saturated NH.sub.4Cl solution was added and diluted with water (25 mL). The organic layer was separated and aqueous layer was extracted with EtOAc (25 mL). The combined organic layers were dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The residue was purified by silica gel flash chromatography (10% EtOAc/hexanes) to afford (3) (2.5 g, 66%) as an oil. .sup.1H NMR (300 MHz, CDCl.sub.3): 1.09 (t, J=6.0 Hz, 3H), 2.25-2.42 (m, 2H), 5.75-5.90 (m, 1H), 6.39-6.49 (m, 1H), 7.40-7.64 (m, 2H), 7.95 (m, 2H).

(230) Preparation of 3-(n-butyl)-phenylamine (4). A solution of (3) (2.5 g, 14.11 mmol) in methanol (20 mL) was added to a slurry of 10% Pd/C (300 mg) in ethyl acetate (1 mL) and stirred under hydrogen atmosphere for 5 h. The catalyst was filtered and the filtrate was concentrated to afford (4) (2.0 g, 96%). .sup.1H NMR (300 MHz, CDCl.sub.3): 0.90 (t, J=6.0 Hz, 3H), 1.25-1.42 (m, 2H), 1.50-1.65 (m, 2H), 2.52 (t, J=6.0 Hz, 2H J=6.0 Hz), 3.59 (br s, 2H), 6.45-6.62 (m, 3H), 7.05 (t, J=6.0 Hz, 1H).

(231) ##STR00355##

(232) Preparation of 2-(3-butylphenyl)-4,5-dichloropyridazin-3(2H)-one (7a). A solution of NaNO.sub.2 (510 mg, 7.39 mmol) in water (2 mL) was slowly added to a cooled (15 C.) solution of (4) (1.0 g, 7.30 mmol) in Conc. HCl (5 mL) and water (5 mL). The reaction was warmed to 0 C. and slowly added to a vigorously stirred 0 C. cooled solution of SnCl.sub.2.2H.sub.2O (3.34 g) in con. HCl (15 mL). After 3 h of stirring at 0 C., crude 3-hydrazinophenyl hydrochloride (5) (300 mg) was collected by filtration and dried under vacuum. To a 90 C. stirred solution of crude 3-hydrazinophenyl hydrochloride (5) (50 mg, 0.21 mmol) in water (3 mL) was added mucochloric acid (6) (35 mg, 0.21 mmol) and stirred for 3 h. The reaction mixture was cooled to room temperature and the solid crystals were collected by filtration, washed with water (25 mL), dried under high vacuum at 40 C. to obtain (7a) (28.0 mg, 42%). .sup.1H NMR (300 MHz, CDCl.sub.3): 0.92 (t, 3H, J=6.0 Hz), 1.30-1.42 (m, 2H), 1.52-1.70 (m, 2H), 2.67 (t, 2H, J=6.0 Hz), 7.20-7.28 (m, 1H), 7.30-7.45 (m, 3H), 7.92 (s, 1H); ESI-MS m/z. 297.4 (M+1).sup.+.

(233) Preparation of 4,5-dichloro-2-(3-ethylphenyl)pyridazin-3(2H)-one (7b) was synthesized from 3-ethylaniline and mucochloric acid as described for 7a. Yield: 30 mg (53%). .sup.1H NMR (300 MHz, CDCl.sub.3): 1.27 (t, J=6.0 Hz, 3H), 2.70 (dd, J=15.0 Hz, 6.0 Hz, 2H), 7.13-7.19 (m, 1H), 7.28-7.43 (m, 3H), 7.95 (s, 1H); ESI-MS m/z. 269.0 (M+1).sup.+.

(234) Preparation of 4,5-dichloro-2-(3-methoxyphenyl)pyridazin-3(2H)-one (7c) was synthesized from 3-methoxyaniline and mucochloric acid as described for 7a. Yield: 35 mg (62%). .sup.1H NMR (300 MHz, CDCl.sub.3): 3.86 (s, 3H), 6.96-7.02 (m, 1H), 7.11-7.19 (m, 2H), 7.36-7.45 (m, 1H), 7.94 (s, 1H); ESI-MS m/z. 271.1 (M+1).sup.+.

(235) ##STR00356##

(236) Preparation of 4,5-dichloro-2-(3-methylphenyl)pyridazin-3(2H)-one (9a): Mucochloric acid (6) (6.91 g, 40.92 mmol) was added to a 100 C. solution of 3-methylphenylhydrazine (8a) (5.0 g, 40.92 mmol) in 12% aqueous HCl water (34 mL). After 5 h, the reaction mixture was cooled to room temperature and the solid crystals were collected by filtration, washed with water (250 mL) and dried under high vacuum at 40 C. to obtain (9a) as light yellow solid (10.35 g, 99%). .sup.1H NMR (300 MHz, CDCl.sub.3): 2.41 (s, 3H), 7.26 (s, 1H), 7.36-7.40 (m, 3H), 7.90 (s, 1H); ESI-MS m/z. 255.4 (M+1).sup.+.

(237) The following compounds were synthesized as described above for 9a using mucochloric acid and corresponding hydrazine.

(238) 4,5-Dichloro-2-(3-trifluoromethylphenyl)pyridazin-3(2H)-one (9b) Yield: 64%. .sup.1H NMR (300 MHz, CDCl.sub.3) 7.60-7.75 (m, 2H), 7.80-7.86 (m, 1H), 7.93 (s, 1H), 7.97 (s, 1H) ESI-MS m/z. 309.1 (M+1).sup.+.

(239) 4,5-Dichloro-2-(2,3-dimethylphenyl)pyridazin-3(2H)-one (9c) Yield: 87%. .sup.1H NMR (300 MHz, CDCl.sub.3) 2.02 (s, 3H), 2.35 (s, 3H), 7.07-7.09 (m, 1H), 7.20-7.29 (m, 2H), 7.91 (s, 1H), ESI-MS m/z. 269.1 (M+1).sup.+.

(240) 4,5-Dichloro-2-(3,4-dimethylphenyl)pyridazin-3(2H)-one (9d) Yield: 90%. .sup.1H NMR (300 MHz, CDCl.sub.3) 2.31 (s, 3H), 2.34 (s, 3H), 7.22-7.32 (m, 3H), 7.90 (s, 1H); ESI-MS m/z. 269.0 (M+1).sup.+.

(241) 4,5-Dichloro-2-(3,5-dimethylphenyl)pyridazin-3(2H)-one (9e) Yield: 86%. .sup.1H NMR (300 MHz, CDCl.sub.3) 2.37 (s, 6H), 7.07 (s, 1H), 7.15 (s, 2H), 7.90 (s, 1H); ESI-MS m/z. 269.4 (M+1).sup.+.

(242) 4,5-Dichloro-2-(2-methylphenyl)pyridazin-3(2H)-one (9f) Yield: 85%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.17 (s, 3H), 7.22-7.26 (m, 1H), 7.31-7.42 (m, 3H), 7.91 (s, 1H); ESI-MS m/z. 255.1 (M+1).sup.+.

(243) 4,5-Dichloro-2-(4-bromo-3-methylphenyl)pyridazin-3(2H)-one (9 g) Yield: 70%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.45 (s, 3H), 7.26-7.31 (m, 1H), 7.41-7.48 (m, 1H), 7.57 (d, 1H, J=6.0 Hz), 7.92 (s, 1H); ESI-MS m/z. 333.1 (M+1).sup.+ and 335.1 (M+2).sup.+.

(244) 4,5-Dichloro-2-(4-chloro-3-methylphenyl)pyridazin-3(2H)-one (9h) Yield: 71%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.43 (s, 3H), 7.29-7.38 (m, 1H), 7.44-7.59 (m, 2H), 7.94 (s, 1H); ESI-MS m/z. 291.0 (M+3).sup.+.

(245) 4,5-Dichloro-2-(2,3-dihydro-1H-inden-5yl)pyridazin-3(2H)-one (9i) Yield: 70%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.03-2.17 (m, 2H), 2.92-2.98 (m, 4H), 7.26-7.37 (m, 3H), 7.92 (s, 1H); ESI-MS m/z. 281.1 (M+1).sup.+ and 283.4 (M+3)

(246) 4,5-Dichloro-2-(2-methyl-1,3-benzothiazol-6-yl)pyridazin-3(2H)-one (9j) Yield: 15%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.96 (s, 3H), 7.59-7.69 (m, 1H), 7.95 (s, 1H), 8.08 (d, J=6.0 Hz, 1H), 8.12 (s, 1H); ESI-MS m/z. 312.4 (M+1).sup.+.

(247) 4,5-Dichloro-2-(2,3-dihydro-1,4-benzodioxin-6yl)-2H-pyridazin-3(2H)-one (9k) Yield: 75%. .sup.1H NMR (300 MHz, CDCl.sub.3): 4.26 (s, 4H), 6.93-6.98 (m, 1H), 7.02-7.12 (m, 2H), 7.88 (s, 1H); ESI-MS m/z. 299.3 (M+1).sup.+.

(248) 2-Benzyl-4,5-dichloropyridazin-3(2H)-one (91) Yield: 78%. .sup.1H NMR (300 MHz, CDCl.sub.3): 5.32 (s, 2H), 7.28-7.38 (m, 3H), 7.40-7.46 (m, 2H), 7.86 (s, 1H); ESI-MS m/z. 255.1 (M+1).sup.+.

(249) 4,5-Dichloro-2-(3-chloro-4-methylphenyl)pyridazin-3(2H)-one (9m) Yield: 75%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.44 (s, 3H), 7.32-7.35 (m, 1H), 7.40-7.44 (m, 1H), 7.61 (s, 1H), 7.93 (s, 1H); ESI-MS m/z. 298.8 (M+1).sup.+.

(250) 4,5-Dichloro-2-pyridine-2-ylpyridazin-3(2H)-one (9n) Yield: 75%. .sup.1H NMR (300 MHz, CDCl.sub.3): 7.40-7.45 (m, 1H), 7.57-7.70 (m, 1H), 7.88-7.94 (m, 1H), 7.99 (s, 1H), 8.67-8.69 (m, 1H); ESI-MS m/z. 242.1 (WO.sup.+.

(251) The following compounds were synthesized as described for 9a using mucobromic acid and corresponding hydrazine.

(252) 4,5-Dibromo-2-(3-methylphenyl)pyridazin-3(2H)-one (12a) Yield: 75%. .sup.1H NMR (300 MHz, CDCl.sub.3): 2.36 (s, 3H), 7.23-7.28 (s, 1H), 7.33-7.37 (m, 3H), 7.95 (s, 1H); ESI-MS m/z. 343.0 (M+1).sup.4.

(253) 4,5-Dibromo-2-(3-trifluoromethylphenyl)pyridazin-3(2H)-one (12b) Yield: 70%. .sup.1HNMR (300 MHz, CDCl.sub.3): 7.59-7.67 (m, 2H), 7.84-8.01 (m, 3H), ESI-MS m/z. 397.0 (M+1).sup.+.

(254) 4,5-Dibromo-2-pyridine-2-ylpyridazin-3(2H)-one (12c) Yield: 75%. .sup.1H NMR (300 MHz, CDCl.sub.3): 7.41-7.46 (m, 1H), 7.55-7.83 (m, 1H), 7.87-7.95 (m, 1H), 8.00 (s, 1H), 8.65-8.67 (m, 1H); ESI-MS m/z. 330.1 (M+1).sup.+.

(255) 4,5-Dibromo-2-(3-methoxyphenyl)pyridazin-3(2H)-one (12d) Yield: 48%. .sup.1H NMR (300 MHz, CDCl.sub.3): 3.83 (s, 3H), 6.95-6.99 (m, 1H), 7.11-7.16 (m, 2H), 7.28-7.43 (m, 1H), 7.93 (s, 1H); ESI-MS m/z. 358.9 (M+1).sup.+.

(256) 4,5-Dibromo-2-(3-(1-methoxy)ethoxyphenyl)pyridazin-3(2H)-one (12e) Yield: 65%. .sup.1H NMR (300 MHz, CDCl.sub.3): 3.51 (s, 3H), 3.74-3.82 (m, 2H), 4.13-4.19 (m, 2H), 6.98-7.02 (m, 1H), 7.15-7.18 (m, 2H), 7.34-7.43 (m, 1H), 7.92 (s, 1H); ESI-MS m/z. 403.1 (M+1).sup.+.

(257) ##STR00357##

(258) Preparation of 5-Chloro-4-(diethylamino)-2-(3-methylphenyl) pyridazin-3(2H)-one (14) and 4-Chloro-5-(diethylamino)-2-(3-methylphenyl)pyridazin-3(2H)-one (15a). A mixture of (9a) (250 mg, 0.95 mmol) and N,N-diethylamine (143 mg, 1.95 mmol) in dioxane (3 mL) was heated at 100 C. for 10 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The products were separated by flash silica gel column chromatography.

(259) 5-Chloro-4-(diethylamino)-2-(3-methylphenyl)pyridazin-3(2H)-one (14): Oil (30.0 mg); R.sub.f=0.45 (25% EtOAc/hexaness); .sup.1H NMR (300 MHz, CDCl.sub.3): 1.22 (t, J=6.0 Hz, 6H), 2.40 (s, 3H), 3.50 (q, J=6.0 Hz, 4H), 7.17-7.20 (m, 1H), 7.26-7.37 (m, 3H), 7.72 (s, 1H). ESI-MS m/z. 292.3 (M+1).sup.+.

(260) 4-Chloro-5-(diethylamino)-2-(3-methylphenyl)pyridazin-3(2H)-one (15a): Solid (120.0 mg); R.sub.f=0.35 25% EtOAc/hexaness); .sup.1H NMR (300 MHz, CDCl.sub.3): 1.31 (t, J=6.0 Hz, 6H), 2.39 (s, 3H), 3.55 (q, J=6.0 Hz, 4H), 7.15-7.18 (m, 1H), 7.28-7.40 (m, 3H), 7.70 (s, 1H) ESI-MS m/z. 292.4 (M+1).sup.+.

(261) ##STR00358##

(262) Preparation of 5-Azido-4-chloro-2-(3-methylphenyl pyridazin-3(2H)-one (15b). This compound was prepared as described above for (15a) from (9a) and sodium azide using ethanol (2 mL) and water (2 mL). Yield 55%. .sup.1HNMR (300 MHz, CDCl.sub.3): 2.41 (s, 3H), 7.18-7.26 (m, 1H), 7.32-7.41 (m, 3H), 7.74 (s, 1H) ESI-MS m/z. 235.9 (M+1).sup.+.

(263) ##STR00359##

(264) Preparation of 4-Chloro-5-ethoxy-2-(3-methylphenyl)pyridazin-3(2H)-one (15c). This compound was prepared as described above for (15a) from (9a) and ethanol (5 mL). Yield 75%. .sup.1H NMR (300 MHz, CDCl.sub.3): 1.56 (t, J=6.0 Hz, 6H), 2.42 (s, 3H), 4.40 (q, J=6.0 Hz, 4H), 7.20-7.29 (m, 1H), 7.33-7.40 (m, 3H), 7.92 (s, 1H) ESI-MS m/z. 265.0 (M+1).sup.+.

(265) ##STR00360##

(266) Preparation of 4,5-Diiodo-2-(3-methylphenyl)pyridazin-3(2H)-one (16) and 5-iodo-2-(3-methylphenyl)pyridazin-3(2H)-one (17). A mixture of (9a) (700 mg, 2.74 mmol) and 57% aqueous HI (10 mL) was heated at 100 C. After 12 h, the reaction was cooled, diluted with water (30 mL) and extracted with ethyl acetate (225 mL). The organic extracts were washed with water (220 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The pure products were separated on silica gel column using 20% ethyl acetate/hexanes.

(267) 4,5-Iodo-2-(3-methylphenyl)pyridazin-3(2H)-one (16): 200.0 mg, R.sub.f=0.40; .sup.1H NMR (300 MHz, CDCl.sub.3): 2.40 (s, 3H), 7.17-7.26 (m, 1H), 7.32-7.37 (m, 3H), 7.95 (s, 1H) ESI-MS m/z. 439.1 (M+1).sup.+.

(268) 5-Iodo-2-(3-methylphenyl)pyridazin-3(2H)-one (17): 75.0 mg, R.sub.f=0.20; .sup.1H NMR (300 MHz, CDCl.sub.3): 2.41 (s, 3H), 7.09-7.26 (m, 1H), 7.33-7.38, 7.60 (s, 1H), 8.04 (s, 1H). ESI-MS m/z. 313.1 (M+1).sup.+.

(269) ##STR00361##

(270) Preparation of 4,5-Dichloro-1-(3-methylphenyl)-1,2-dihydropyridazin-3,6-dione (19a). 3,4-Dichloromaleic anhydride (18a) (2.0 g, 12.27 mmol) was added to a solution 3-methylphenylhydrazine (5) (1.5 g, 12.27 mmol) in 20% aqueous HCl (12 mL) at 100 C. and stirred for 3 h. The reaction mixture was cooled to room temperature and diluted with water (15 mL) and extracted with ethyl acetate (320 mL). The combined extracts was dried, and concentrated in vacuo to afford a residue that was purified on silica gel column (20% EtOAc/hexanes) to afford (19a) (1.35 g, 40%). .sup.1H NMR (300 MHz, DMSO-D.sub.6): 2.36 (s, 3H), 7.22-7.25 (m, 1H), 7.33-7.41 (m, 3H); ESI-MS m/z. 271.2 (M-F1).sup.1.

(271) Preparation of 4,5-Dichloro-6-methoxy-2-(3-methylphenyl)-pyridazin-3,6-dione (20a). A mixture of (19a) (1.5 g, 5.5 mmol) dimethyl sulfate (1.6 g, 6.9 mmol) and K.sub.2CO.sub.3 (2.28 g, 16.5 mmol) in acetone (20 mL) was stirred at reflux for 12 h. The reaction was cooled and the solids were filtered-off. The filtrate was concentrated and dissolved in ethyl acetate (30 mL), washed with water (215 mL), dried and concentrated under vacuum. The crude compound was purified on silica gel column using 25% ethyl acetate/hexanes to afford (20a) (1.8 g 76%). .sup.1H NMR (300 MHz, CDCl.sub.3): 2.24 (s, 3H), 3.96 (s, 3H), 7.17-7.28 (m, 1H), 7.34-7.53 (m, 3H); ESI-MS m/z. 285.3 (M+1).sup.+.

(272) 6-Methoxy-2-(3-methylphenyl)-pyridazin-3,6-dione (20b) This compound was prepared as described for (20a) using 19b [Data for 19b: Yield: 73%; .sup.1H NMR (300 MHz, DMSO-Ds): 2.35 (s, 3H), 6.99-7.03 (m, 1H), 7.19 (d, 2H, J=6.0 Hz), 7.28-7.42 (m, 3H), 11.32 (s, 1H); ESI-MS m/z. 203.4 (M-1-1).sup.+.] Data for 20b: Yield: 82% .sup.1H NMR (300 MHz, CDCl.sub.3): 2.43 (s, 3H), 3.90 (s, 3H), 7.17-7.27 (m, 2H), 7.35-7.42 (m, 2H); ESI-MS m/z. 217.1 (M+1).sup.+.

(273) ##STR00362##

(274) Preparation of tert-Butyl-3-{{4,5-dichloro-1-(3-methylphenyl)-6-oxo-1,6-dihydropyridazin-3-yl]oxy}propylcarbamate (21) A mixture of (19a) (1.5 g, 5.5 mmol), tert-butyl 3-bromopropylcarbamate (1.6 g, 6.9 mmol) and K.sub.2CO.sub.3 (2.28 g, 16.5 mmol) in acetone (20 mL) was refluxed for 12 h. The reaction was cooled and the solids were filtered-off. The filtrate was concentrated and dissolved in ethyl acetate (30 mL), washed with water (215 mL), dried and concentrated in vacuo. The crude compound was purified on silica gel column using 25% EtOAc/hexanes to afford (21) (1.8 g 76%). .sup.1H NMR (300 MHz, CDCl.sub.3): 1.43 (s, 9H), 1.97-2.05 (m, 2H), 2.41 (s, 3H), 3.30-3.36 (m, 2H), 4.29-5.32 (m, 2H), 4.82 (bs, 1H), 7.19-7.42 (m, 4H); ESI-MS m/z. 428.3 (M+1).sup.+.

(275) Preparation of 6-(3-Aminopropoxy)-4,5-dichloro-2-(3-methylphenyl)pyridazin-3(2H)-one hydrochloride (22) To a solution of (21) (1.8 g, 4.20 mmol) was added 4 N HCl in dioxane (5.25 mL) and stirred at room temperature. After 15 h, the solid was filtered and washed with hexanes (225 mL) and dried in vacuo to afford (22) (1.32 g, 86%) as an off-white solid. .sup.1H NMR (300 MHz, DMSO-D.sub.6): 1.99-2.10 (m, 2H), 2.37 (s, 3H), 2.85-2.95 (2H), 4.20-4.36 (m, 2H), 7.15-7.27 (m, 1H), 7.39-7.59 (m, 3H), 8.03 (br s, 3H); ESI-MS m/z. 328.4 (M+1).sup.+.

(276) Preparation of 4-[(3-{[4,5-Dichloro-1-(3-methylphenyl)-6-oxo-1,6-dihydropyridazin-3-yl]oxy}propyl)amino]-4-oxobutanoic acid (23) A mixture of (21) (1.30 g, 3.57 mmol) and succinic anhydride (357 mg, 3.57 mmol) in dichloromethane (10 mL) and pyridine (1.5 mL) was stirred at room temperature for 24 h. The mixture was concentrated and re-dissolved in ethyl acetate (20 mL) washed with 1 N HCl (25 mL) and with water (210 mL). The organic solution was dried over Na.sub.2SO.sub.4 and concentrated and the obtained syrup was stirred with 50% hexanes/CH.sub.2Cl.sub.2. The solid was collected by filtration and dried at 40 C. to afford (23) (1.4 g, 91%) as off-white solid. .sup.1H NMR (300 MHz, DMSO-D.sub.6): 1.84-1.90 (m, 2H), 2.26-2.30 (m, 2H), 2.37-2.43 (m, 5H), 3.18 (m, 2H), 4.20 (t, J=6.0 Hz, 2H) 7.24-7.26 (m, 1H), 7.30-7.41 (m, 3H), 7.90-7.96 (m, 1H), 12.12 (s, 1H); ESI-MS m/z. 428.3 (M+1).sup.+.

(277) ##STR00363## ##STR00364##

(278) Preparation of tert-Butyl 3-(3-nitrophenoxy)propylcarbamate (25). A mixture of 3-nitrophenol (24) (7.0 g, 50.3 mmol), tert-butyl 3-bromopropylcarbamate (14.35 g, 60.3 mmol) and K.sub.2CO.sub.3 (13.90 g, 100.63 mmol) in acetone (150 mL) was stirred at reflux for 12 h. The reaction was cooled and the solids were filtered-off. The filtrate was concentrated, dissolved in EtOAc (250 mL), washed with water (215 mL), dried and concentrated in vacuo to afford (25) (14.25 g 95%) that was used without further purification. .sup.1H NMR (300 MHz, CDCl.sub.3): 1.46 (s, 9H), 1.99-2.07 (m, 2H), 3.26-3.47 (m, 2H), 4.10 (t, J=6.0 Hz, 2H), 4.84 (bs, 1H), 7.20-7.24, 7.43 (t, J=6.0 Hz, 1H), 7.71 (s, 1H), 7.79-7.83 (m, 1H); ESI-MS m/z. 297.3 (M+1).sup.+.

(279) Preparation of 3-(3-Nitrophenoxy)propan-1-amine hydrochloride (26). To a solution of (25) (14.25 g, 47.80 mmol) in dichloromethane (150 mL) was added 4 N HCl in dioxane (60 mL) and stirred at room temperature. After 7 h, the reaction was diluted with hexanes (200 mL). The solid was filtered, washed with hexanes (225 mL) and dried under vacuum to afford (26) (11.0 g, 98%) as off-white solid. ESI-MS m/z. 197.3 (M+1).sup.+.

(280) Preparation of 4-{[3-(3-Nitrophenoxy)propyl]amino}-4-oxobutanoic acid (27). A mixture of (26) (11.0 g, 47.11 mmol) and succinic anhydride (4.71 g, 47.11 mmol) in dichloromethane (75 mL) and pyridine (18.63 g, 235.5 mmol) was stirred at room temperature for 24 h. The mixture was concentrated, re-dissolved in ethyl acetate (150 mL) and washed with 2 N HCl (250 mL). The precipitated solid product was collected by filtration and the filtrate was dried and concentrated to afford crude product. The combined solid was washed with 50% EtOAc/hexanes (25 mL) and dried under vacuum to afford (27) (13.5 g, 96%). .sup.1H NMR (300 MHz, DMSO-D.sub.6): 1.80-1.92 (m, 2H), 2.27-2.31 (m, 2H), 2.36-2.43 (m, 2H), 3.19 (m, 2H), 4.09 (t, J=6.0 Hz, 2H) 7.38-7.41 (m, 1H), 7.54 (t, J=6.0 Hz, 3H), 7.67 (s, 1H), 7.78-7.81 (m, 1H), 7.94 (t, J=6.0 Hz, 1H); ESI-MS m/z. 297.3 (M+1).sup.+.

(281) Preparation of 4-{[3-(3-Aminophenoxy)propyl]amino}-4-oxobutanoic acid (28). To a slurry of 10% Pd/C (1.75 g) in EtOAc (400 mL) was added (27) (13.5 g, 45.5 mmol) and stirred under hydrogen atmosphere (30 Psi). After 5 h, the catalyst was filtered-off and filtrate was concentrated to afford (28) (11.75, 96%) that was used for next reaction without further purification. ESI-MS m/z. 267.4 (M+H).sup.+.

(282) Preparation of tert-Butyl 3-[3-(4,5-dibromo-6-oxopyridazin-1(6H)-yl)phenoxy]propylcarbamate (29). A solution of NaNO.sub.2 (1.29 g, 18.77 mmol) in water (3 mL) was slowly added to a cooled (0 C.) solution of (28) (5.0 g, 18.77 mmol) in conc. HCl (15 mL) and water (5 mL). After 30 min, the reaction mixture was added to cooled (0 C.) slurry of SnCl.sub.2.2H.sub.2O (10.72 g, 47.5 mmol) in conc. HCl (10 mL) with vigorous stirring. The resulting mixture was stirred for 3 h at 0 C. and crude 4-{[3-(3-hydrazinophenoxy)propyl]amino}4-oxobutanoic acid hydrochloride was collected by filtration and dried under vacuum (6.5 g). To a stirred solution of the crude acid (6.5 g, 18.35 mmol) in water (10 mL) at 90 C. was added mucobromic acid (4.73 g, 18.35 mmol). After 3 h, the reaction mixture was cooled to room temperature, basified with 1 N NaOH (pH=6.0) and treated with (Boc).sub.2O (5.0 g, 22.9 mmol) for overnight at room temperature. The reaction was extracted with EtOAc (350 mL) and organic solution was washed with water (225 mL) and dried over Na.sub.2SO.sub.4 and concentrated under vacuum. The obtained residue was purified on silica gel using 25% EtOAc/hexanes to afford (29) (1.8 g, 19.0% 3 steps). .sup.1H NMR (300 MHz, CDCl.sub.3): 1.46 (s, 3H), 1.94-2.06 (m, 2H), 3.31 (t, J=6.0 Hz, 2H), 4.04 (t, J=6.0 Hz, 2H), 4.75 (br s, 1H), 6.94-6.98 (m, 1H), 7.11-7.17 (m, 2H), 7.35-7.42 (m, 1H), 7.93 (s, 1H); ESI-MS m/z 502.1 (M+1).sup.+.

(283) Preparation of 2-[3-(3-Aminopropoxy)phenyl]-4,5-dibromopyridazin-3(2H)-one hydrochloride (30). To a solution of (29) (1.75 g, 3.4 mmol) in dichloromethane (30 mL) was added 4 N HCl in dioxane (5 mL) and stirred at room temperature. After 7 h, the reaction was diluted with hexanes (20 mL). The solid was filtered, washed with hexanes (215 mL) and dried under vacuum to afford (30) (1.40 g, 94%) as off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): 1.96-2.10 (m, 2H), 2.94 (t, J=6.0 Hz, 2H), 4.10 (t, J=6.0 Hz, 2H), 7.05-7.13 (m, 2H), 7.43 (t, J=6.0 Hz, 1H), 8.26 (s, 1H); ESI-MS m/z 402.1 (M+1).sup.+.

(284) Preparation of 4-({3-[3-(4,5-Dibromo-6-oxopyridazin-[(6H)-yl)phenoxy]propyl}amino)-4-oxobutanoic acid (31) A mixture of (30) (1.30 g, 2.95 mmol) and succinic anhydride (310.0 mg, 3.10 mmol) in dichloromethane (15 mL) and pyridine (3.5 mL) was stirred at room temperature for 15 h. The mixture was concentrated, re-dissolved in EtOAc (30 mL) and washed with 1 N HCl (310 mL) and then with water (215 mL). The organic solution was dried (Na.sub.2SO.sub.4) and concentrated to afford crude product. The obtained solid was stirred with 50% EtOAC/hexanes (10 mL), collected by filtration and dried under vacuum to afford (31) (1.2 g, 87%). .sup.1H NMR (300 MHz, DMSO-Ds): 1.81-1.86 (m, 2H), 2.30 (t, J=6.0 Hz, 2H), 2.4 (t, J=6.0 Hz, 2H), 3.19 (m, 2H), 4.00 (m, 2H) 7.01-7.03 (m, 1H), 7.07-7.10 (m, 2H), 7.40 (t, J=3.0 Hz, 1H), 7.93 (t, J=3.0 Hz, 1H), 8.26 (s, 1H), 12.05 (br s, 1H); ESI-MS m/z. 502.3 (M+1).sup.+.

Example 3

SAR of compound SKI-104122

(285) Screening of pyridazin-3(2H)-ones

(286) We undertook a SAR study to identify potential sites on the compound SKI-104122 for linker addition without compromising biological activity. Exemplary syntheses of compounds screened are detailed above in Example 2. The screening results of these compounds are summarized in Tables 10, 11, and 13 below. Compounds were tested in dose response studies from 10 M to 5 nM against several non small cell lung cancer lines: H358, H827, H1118, H1650, H1734, H1975, H2030, and H3255, and were also studied in a 72 hour cytotoxicity assay against the NHBE and WI-38 cell lines. Additionally, Tables 12 and 14 summarize similar screenings of gefitinib (IRESSA, Astra-Zeneca) and erlotinib (TARCEVA, OSI Pharmaceuticals, Genentech).

(287) In FIGS. 4(A-E) to 10(A-E), IC.sub.50 curves are shown for all experiments performed on SKI 104122 including primary HTS and resynthesis. FIGS. 11 to 40 depict IC.sub.50 curves for active pyridazinone compounds against the H3255 cell line.

(288) TABLE-US-00010 TABLE 10 Summary of SKI-104122 against H1650, H1975, H2030, H3255, NHBE and WI-38 cell lines H1650 H1975 H2030 H3255 NHBE WI-38 Structure SKI ID Dose Response (uM) CytoTox (uM) 104122 104122 104122 104122 104122 104122 104122 0.04 0.97 1.24 2.71 3.80 1.18 1.88 0.01 0.14 0.14 0.31 0.28 0.29 0.29 0.001 0.30 0.27 0.58 0.65 0.82 0.45 0.04 0.38 0.64 1.33 0.38 0.53 0.84 0.80 ND 2.64 ND 3.40 1.59 1.97 ND ND 3.70 ND 6.06 6.86 8.95 ND = No Data, Assay Not Performed with this Cell Line

(289) TABLE-US-00011 TABLE 11 Summary of Pyridazinone Compounds against H1650, H1975, H2030, H3255, NHBE, and WI-38 Cell Lines H1650 H1975 H2030 H3255 NHBE WI-38 Structure SKI ID Dose Response (uM) CytoTox (uM) 257017 2.74 0.47 1.08 2.95 5.17 7.52 257020 2.10 0.27 0.56 0.92 3.18 6.78 257019 2.40 0.29 0.53 1.23 3.24 7.02 257018 2.42 0.35 0.89 1.63 3.89 7.35 0 176729 1.44 0.16 0.34 0.73 2.76 3.42 257015 1.14 0.16 0.39 0.62 2.38 3.03 257016 2.73 0.32 0.65 1.32 3.29 6.43 257014 3.18 0.29 0.55 1.36 3.29 6.93 257021 >10 >10 >10 >10 >10 >10 257022 >10 >10 >10 >10 >10 >10 257023 >10 >10 >10 >10 >10 >10 257024 >10 >10 >10 >10 >10 >10 257025 >10 >10 >10 >10 >10 >10 257029 >10 >10 >10 >10 >10 >10 0 257028 2.04 0.65 1.26 1.76 5.24 7.52 267061 3.52 0.37 0.72 1.34 1.70 3.91 267064 2.83 0.71 0.88 2.47 4.04 >10 267065 >10 1.89 >10 >10 >10 >10 267062 2.48 0.46 0.72 1.50 1.85 6.44 267063 5.82 1.93 2.75 3.89 5.34 >10 267059 >10 >10 >10 >10 >10 >10 267060 3.92 0.69 1.09 2.27 3.44 >10 267066 1.69 0.19 1.41 1.35 1.74 5.37 267067 4.47 0.45 3.04 3.64 >10 >10 0 267068 2.33 0.25 1.23 1.20 1.56 3.96 267069 >10 2.16 6.83 6.50 7.32 >10 267070 >10 0.91 5.18 3.10 >10 >10 267071 0.74 0.22 1.22 0.81 1.61 2.61 267072 0.54 0.20 1.42 1.00 1.43 2.17 267073 0.80 0.09 0.76 0.66 1.33 0.89 267074 >10 >10 >10 >10 >10 >10 267075 >10 >10 >10 >10 >10 >10 267076 >10 >10 >10 >10 >10 >10 267077 0.56 0.05 1.27 1.11 1.10 1.74 00 267082 >10 >10 >10 >10 >10 >10 01 267083 1.06 0.23 0.57 0.35 1.85 2.60 02 267084 1.51 0.23 0.61 0.38 2.56 2.33 03 267085 1.93 0.50 0.99 0.71 2.88 5.51 04 267086 >10 6.76 >10 >10 >10 >10 05 267090 1.29 0.20 0.41 0.50 1.43 1.19 06 273632 >10 >10 >10 >10 >10 >10 07 273633 >10 >10 >10 >10 >10 >10 08 273634 1.36 0.44 0.67 1.11 2.94 4.53

(290) TABLE-US-00012 TABLE 12 Iressa and Tarceva against H1650, H1975, H2030, H2030, H3255, NHBE, and WI-38 H1650 H1975 H2030 H3255 NHBE WI-38 Structure SKI ID Original ID Dose Response (uM) CytoTox (uM) 09 267079 Iressa >10 >10 >10 <1.0 ND ND 0 267080 Tarceva >10 >10 >10 <1.0 ND ND 267079 Iressa >10 >10 >10 <0.01 5.44 >10 267080 Tarceva >10 >10 >10 0.02 >10 >10 267079 Iressa >10 6.74 4.55 <0.01 8.27 6.96 267080 Tarceva >10 >10 >10 0.02 >10 >10

(291) TABLE-US-00013 TABLE 13 Summary of Pyridazinone Compounds against H358, H827, H1118, and H1734 H358 H827 H1118 H1734 Structure SKI ID Dose Response (uM) 104122 0.15 0.09 0.11 2.21 104122 ND ND ND 1.73 104122 0.21 ND ND 2.17 104122 0.10 ND ND 1.17 104122 0.12 ND ND 1.17 0 257017 0.74 0.71 0.49 3.20 257020 0.22 0.30 0.34 1.83 257019 0.24 0.32 0.29 2.01 257018 0.55 0.50 0.44 2.12 176729 0.12 0.17 0.20 0.61 257015 0.16 0.17 0.22 0.87 257016 0.35 0.29 0.38 3.37 257014 0.31 0.24 0.27 4.11 257021 >10 >10 >10 >10 257022 >10 >10 >10 >10 0 257023 >10 >10 >10 >10 257024 >10 >10 >10 >10 257025 >10 >10 >10 >10 257029 >10 ND ND >10 257028 0.94 ND ND 2.42 267061 0.24 ND ND 2.41 267064 0.47 ND ND 1.21 267065 1.24 ND ND >10 267062 0.24 ND ND 1.75 267063 0.76 ND ND 3.25 0 267059 >10 ND ND >10 267060 0.29 ND ND 1.60 267066 0.42 ND ND 1.45 267067 1.38 ND ND 2.57 267068 0.43 ND ND 2.12 267069 4.76 ND ND 6.86 267070 0.97 ND ND 2.69 267071 0.21 ND ND 0.80 267072 0.15 ND ND 0.51 267073 0.07 ND ND 0.72 0 267074 >10 ND ND >10 267075 >10 ND ND >10 267076 >10 ND ND >10 267077 0.14 ND ND 0.52 267082 >10 ND ND >10 267083 0.13 ND ND 1.31 267084 0.13 ND ND 1.63 267085 0.30 ND ND 1.94 267086 >10 ND ND >10 267090 0.06 ND ND 1.10 0 273632 >10 ND ND >10 273633 >10 ND ND >10 273634 0.24 ND ND 0.93

(292) TABLE-US-00014 TABLE 14 Iressa and Tarceva against H358 and H1734 H358 H827 H1118 H1734 Structure SKI ID Original ID Dose Response (uM) 267079 Iressa >10 ND ND 3.01 267080 Tarceva >10 ND ND >10 267079 Iressa >10 ND ND 4.48 267080 Tarceva >10 ND ND >10 267079 Iressa 7.33 ND ND 2.67 267080 Terceva >10 ND ND >10

Example 4

High Throughput Screening for Novel Agents that Block Proliferation of NSCLC Cell Lines

(293) We have performed high throughput screening campaigns using four established non small cell lung cancer cell lines (H1650, H1975, H2030, H3255) against a chemical library of 200,000 small molecules. Description of the assays is described herein. We have identified several hits inhibiting one or more cell line with the most potent one being 5-nitro-furan-2-carboxylic acid (4-chloro-phenyl)-amide (SKI-98698). We then performed a substructure search against the library and identified 51 close derivatives of SKI-98698 (Table 15). Further analysis of the biological data for these derivatives reveals a coherent structure activity relationship for both active and inactive compounds with only 8 compounds identified as active during primary screening (Table 16). Data is expressed as percentage inhibition in the cell based assay; screening concentration of 10 M compound in 1% DMSO (v/v).

(294) ##STR00469##

5-nitro-furan-2-carboxylic acid (4-chloro-phenyl)-amide (SKI-98698)

(295) TABLE-US-00015 TABLE 15 pre-SAR summary for substituted furan derivatives R1650 R1975 H2030 H3255 Structure SKI ID PRIMARY HTS [%] 0 87200 103 67 23 6 98697 103 70 62 83 98698 101 74 17 20 71762 104 74 101 100 71762 104 74 101 100 64228 87 82 93 91 71763 87 61 82 77 64228 87 82 93 91 60998 10 19 1 1 61302 13 16 2 2 0 66598 8 8 0 1 66776 13 2 3 2 72456 12 9 8 7 75072 13 0 5 1 76739 13 9 3 5 76839 20 3 6 2 77021 16 11 1 4 77942 14 2 6 7 77953 12 1 6 0 78107 14 14 4 6 0 78158 20 2 6 4 78535 41 2 26 8 79434 4 7 0 4 82456 6 15 2 2 83809 22 10 2 2 84487 27 5 5 3 87246 8 3 9 5 88935 4 10 1 2 91753 3 22 5 1 92553 2 3 5 1 00 93911 4 22 3 1 01 95436 2 12 2 3 02 97216 1 4 4 3 03 97253 2 15 5 3 04 98705 8 14 8 9 05 100403 13 13 3 2 06 100483 1 8 1 1 07 100507 3 15 7 1 08 100588 5 14 5 4 09 100652 12 2 6 3 0 100792 5 9 43 1 101070 2 18 5 1 102031 2 11 1 3 103710 33 17 5 4 103960 62 41 73 68 104003 3 18 0 5 104039 61 69 13 6 105503 10 13 7 0 107911 8 15 2 7 108321 38 7 6 2 0 109007 6 3 3 1

(296) TABLE-US-00016 TABLE 16 SAR summary for substituted phenylfuran-2-carboxamide derivatives 1650 1975 2030 3255 1650 1975 2030 3255 NHBE Structure SKI ID CONFIRMATION [%] IC.sub.50 (uM) 87200 112 95 107 103 0.29 0.44 1.01 0.69 ND 98697 136 70 101 130 0.44 1.09 1.06 0.91 29.40 98698 133 57 98 118 0.38 0.63 1.12 0.95 21.36 71762 135 59 101 122 0.43 1.24 1.43 0.95 15.77 71762 106 76 103 97 0.41 1.23 1.46 1.01 14.75 64228 111 70 90 97 1.60 2.24 1.12 2.54 13.20 71763 136 74 97 135 1.39 3.54 3.79 2.98 24.90 64228 107 97 105 97 2.31 5.46 6.23 7.51 ND

(297) SK1-267077 inhibits DNA synthesis. We used SKI-267077, a derivative of SKI-104122 to elucidate the mechanism by which this family of compounds caused growth arrest of human lung adenocarcinoma cell lines. H2030 cells were treated with increasing concentrations of SKI-267077 for 24 hours and then incorporation of .sup.3H-thymide into DNA determined (FIG. 1). As shown in FIG. 1, SKI-267077 caused a dose-dependent decrease in DNA synthesis.

(298) SKI-267077 induces cell cycle arrest at the G1 phase. Cell cycle progression in the presence of SKI-267077 was determined by FACS analysis of H2030 cells that were treated for 24 hours. SKI-267077 treatment resulted in a reduction of the percent of cells in the S phase (FIG. 2A) of the cell cycle and a concomitant increase in the percent of cells in the induce G1 phase (FIG. 2B).

(299) SKI-267077 stimulates the enzymatic activity of caspase 3/7. Treatment of H2030 with SKI-267077 for 48 hours lead to an increase in the activity of caspase 3/7 (FIG. 3).

Example 5

Dose Response Studies

(300) Dose response studies against 34 compounds from the pyridazinone and phenylfuran-carboxamide class were conducted. The 34 compounds were run in a 12 point doubling dilution from 10 uM to 5 nM for IC50 determination against 15 non small cell lung cancer lines representing various EGFR and KRAS mutations (see Table 17). In addition, Tarceva was included in these dose response studies.

(301) TABLE-US-00017 TABLE 17 Non Small Cell Lung Cancer Cell Lines Mutation Cell line EGFR KRAS P53 PTEN H1118 L858R WT K164X H820 T790M/DelE746-A750 WT H1650 DelE746-A750 WT No protein H1975 T7980M/L858R WT H3255 L858R WT WT PC9 DclE746-A750 WT A549 WT G12S H23 WT G12C I246M H358 WT G12C R273H H460 WT Q61H H1734 WT G13C H2030 WT G12C H2122 WT G12C G262V WT H2444 WT G12V HPL1D* WT WT *SV40-immortalized human lung epithelian cells

(302) The dose response studies were done with the following standard procedure. First, the cells were counted and dispensed at 250 cells/well into 384-well microplates with compound. The cells were incubated for 48 hours at 37 C to complete treatment, and then alamar blue was added for an additional 48 hours. Finally, alamar blue reduction was measured and the data was loaded into ORIS, our data screening acquisition platform. Table 18 provides a summary of these experiments.

(303) TABLE-US-00018 TABLE 18 Compound Metric Summary of 15 NSCLC Lines (IC.sub.50 in uM) Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 104122 0.34 0.18 1.41 0.29 0.87 0.65 1.45 0.21 0 257014 0.45 0.24 1.64 0.36 1.50 0.83 1.68 0.26 257015 0.37 0.22 0.92 0.32 0.76 0.71 1.20 0.23 257016 0.54 0.37 2.49 0.47 1.53 1.13 2.63 0.32 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 104122 0.41 1.56 2.14 0.29 1.15 0.24 0.28 257014 0.68 2.01 2.28 0.41 1.16 0.34 0.38 257015 0.43 1.30 1.62 0.31 1.26 0.30 0.30 257016 0.78 2.55 3.22 0.51 2.07 0.41 0.48 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 257017 0.83 0.47 1.67 0.76 3.16 2.00 2.13 0.31 257018 0.54 0.34 1.69 0.54 1.86 1.57 2.27 0.24 257019 0.94 0.67 4.15 0.89 2.01 1.98 4.20 0.69 0 257020 0.58 0.45 2.58 0.58 1.35 1.55 2.87 0.46 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 257017 1.70 3.68 3.20 1.38 2.16 1.05 1.14 257018 1.13 3.51 2.51 0.90 2.16 0.85 1.30 257019 0.79 4.09 4.65 0.80 4.38 0.82 0.62 257020 0.62 2.93 2.62 0.57 2.21 0.40 0.41 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 176729 0.31 0.28 0.80 0.31 0.66 0.90 1.19 0.23 257028 1.10 0.77 3.04 1.18 2.38 2.40 3.76 0.68 267060 1.66 1.48 3.95 1.27 2.29 3.49 5.31 1.07 267061 0.40 0.26 1.96 0.31 1.20 0.85 1.76 0.28 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 176729 0.32 1.26 1.22 0.32 1.50 0.21 0.25 0 257028 1.83 5.23 3.87 1.79 4.04 1.17 1.93 267060 1.39 4.26 5.23 1.11 6.00 1.26 0.56 267061 0.46 1.63 3.04 0.36 1.20 0.37 0.29 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 267062 0.67 0.45 1.91 0.47 1.51 1.51 2.81 0.39 267063 6.41 6.24 >10 6.57 8.35 8.89 >10 5.47 267064 >10 6.72 >10 7.58 >10 >10 >10 5.75 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 267062 0.66 2.88 2.89 0.54 2.55 0.43 0.38 267063 6.66 >10 >10 6.43 >10 5.28 7.04 267064 7.65 >10 >10 7.80 >10 6.41 4.72 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 267065 >10 >10 >10 >10 >10 >10 >10 6.96 0 267066 2.73 2.80 >10 2.23 4.61 5.87 7.52 2.04 267067 >10 8.25 >10 >10 >10 >10 >10 5.68 267068 0.48 0.32 2.26 0.37 1.23 0.88 1.93 0.29 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 267065 >10 >10 >10 >10 >10 6.37 6.66 267066 2.81 6.56 8.85 2.34 8.72 1.53 1.66 267067 >10 >10 >10 9.19 >10 6.92 7.01 267068 0.78 1.97 2.83 0.47 1.59 0.41 0.42 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 267069 3.21 2.57 >10 3.60 >10 6.21 7.39 2.53 267070 >10 >10 >10 >10 >10 >10 >10 >10 267071 0.41 0.27 1.10 0.36 0.76 0.86 1.62 0.26 0 267072 1.21 0.82 2.09 0.93 1.64 2.59 3.42 0.84 267073 0.10 0.10 0.79 0.16 0.45 0.42 0.31 0.07 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 267069 6.66 >10 >10 4.01 9.04 4.13 5.25 267070 >10 >10 >10 >10 >10 >10 >10 267071 0.40 1.43 1.72 0.37 1.26 0.31 0.24 267072 0.85 2.76 3.19 0.85 4.61 1.05 0.51 267073 0.14 0.42 1.23 0.12 0.78 0.13 0.11 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 267077 3.73 2.68 >10 2.08 8.75 >10 >10 1.72 267083 0.27 0.20 1.08 0.24 0.65 0.74 1.07 0.21 267084 0.23 0.18 1.04 0.21 0.71 0.68 0.84 0.17 0 267085 1.03 0.41 3.16 0.80 2.13 2.24 3.27 0.72 267090 0.15 0.41 1.42 0.16 0.70 0.47 0.54 0.08 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 267077 3.22 >10 >10 3.55 >10 4.68 4.95 267083 0.30 0.91 1.36 0.26 1.15 0.28 0.22 267084 0.27 0.89 1.50 0.22 1.12 0.23 0.18 267085 1.33 3.49 2.99 0.81 2.34 0.53 0.68 267090 0.21 0.79 2.47 0.17 0.89 0.22 0.17 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 273634 0.57 0.29 0.51 0.41 0.99 0.97 1.65 0.38 98697 0.56 3.53 0.60 4.57 2.72 2.17 1.81 0.91 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 273634 0.69 1.47 1.23 0.47 1.42 0.40 0.35 0 98697 1.20 2.20 4.42 1.18 5.20 4.79 >10 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 98698 0.53 1.83 0.47 2.63 1.77 1.58 1.33 0.60 71762 0.38 3.61 0.63 4.23 2.41 1.89 1.46 0.83 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 98698 0.81 1.62 3.47 0.91 3.09 2.43 >10 71762 1.27 1.85 4.88 0.99 4.83 3.80 >10 Structure SKI ID H1118 H820 H1650 H1975 H3255 PC9 A549 H23 87200 4.33 >10 7.49 >10 >10 8.67 >10 4.55 Tarceva 267080 >10 >10 >10 >10 0.03 >10 >10 >10 Structure SKI ID H358 H460 H1734 H2030 H2122 H2444 HPL1D 87200 7.17 >10 >10 8.51 >10 >10 >10 Tarceva 267080 >10 >10 >10 >10 >10 >10 >10

Other Embodiments

(304) The foregoing has been a description of certain non-limiting preferred embodiments of the invention. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.