NZ503991A

NZ503991A - Use of quinazoline derivatives as tyrosine kinase inhibitors for treating colonic polyps

Info

Publication number: NZ503991A
Application number: NZ503991A
Authority: NZ
Inventors: Philip Frost
Original assignee: American Cyanamid Co
Priority date: 1997-11-06
Filing date: 1998-11-04
Publication date: 2001-11-30
Also published as: NO20002166L; JP2001522802A; BR9814116A; HUP0004286A3; AU1308799A; AR016415A1; KR20010031813A; HUP0004286A2; WO1999024037A1; PL340800A1; ZA9810134B; NO20002166D0; IL135622A0; CN1278176A; EP1039910A1; CA2306155A1

Abstract

The use, in the manufacture of a medicament for the treatment of colonic polyps, of a compound of formula 1 wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, carboalkoxy of 2-7 carbon atoms, carboalkyl of 2-7 carbon atoms, amino, and alkanoylamino of 1- 6 carbon atoms; R and R1 are each, independently, hydrogen, halogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, or trifluoromethyl; R2 is hydrogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, trifluoromethyl; Y is a radical selected from the group consisting of formulae 2-8 wherein: R3 is independently hydrogen, alkyl of 1-6 carbon atoms, carboxy, carboalkoxy of 1-6 carbon atoms, phenyl, or carboalkyl of 2-7 carbon atoms; n = 2-4; with the proviso that each R3 of Y may be the same or different or a pharmaceutically acceptable salt thereof.

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number 503991 WO 99/24037 PCT/US98/23549 - 1 - USE OF QUINAZOLINE DERIVATIVES AS TYROSINE KINASE INHIBITORS FOR TREATING COLONIC POLYPS This invention relates to the use of certain quinazoline compounds in the 5 treatment and inhibition of colonic polyps. Colonic Polyps occur in both a familial pattern (Familial Adenomatous Polyps (FAP) and sporadically. FAP afflicts approximately 25,000 patients in the US. while it is estimated that sporadic adenomatous polyps (SAP) occur in approximately 2 million 10 people per year m the US alone. All these patients are at risk for developing adenocarcinoma of the colon. In the case of FAP, that risk is virtually 100% and these patients usually undergo a colectomy at an early age. Patients with sporadic polyps are treated with polypectomy and require periodic colonoscopic exaiflmation because of their inherent risk of developing recurrent polyps. In fact, parents and siblings of these 15 patients are also at increased risk for developing colorectal cancer. The genetic basis for FAP has been linked to the presence of mutations in the APC gene. Similar APC mutations have been found in patients with sporadic polyps. Biochemically, the APC mutation occurs in conjunction with the increased expression of cyclooxygenase enzymes, particularly COX-2. These enzymes are essential for the 20 production of prostenoids, (prostaglandin's; (PG's)) that mediate a number of functions m the bowel including motility, vascular tone, angiogenesis and mucosal protection. PG's are also purported to discourage apoptosis and this is proposed as an explanation for polyp formation. The therapy of FAP and SAP has focused on inhibiting COX enzymes. 25 Considerable evidence exists for the efficacy of COX inhibitors in reducing polyp formation These COX inhibitors are predominantly NSAID's such as chnonl, sulindac, piroxicam and etodoloc, all of which appear to be equivalent in their action. A major problem with NSAID therapy has been the development of serious side effects including peptic ulceration, and cholestatic hepatitis and renal papillary necrosis. Long 30 term therapy with NSAIDs for the treatment of polyps is therefore considered to be impractical. It has recently been proposed that the activauon and overexpression of COX-2 in adenomatous polyps is due to activation of the epidermal growth factor receptor (EGFR). EGFR sumulation by one of it's ligands - amphireguhn (AR), induces the 35 nuclear targeting of COX-2, release of PG's and subsequent mitogenesis, m polarized Printed from Mimosa 50399 1 -2- colonic epithelial cells. COX-2 inhibitors have been shown to prevent this series of events. DESCRIPTION OF THE INVENTION This invention provides the use of a compound of formula 1: R HN' 10 wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, carboalkoxy of 2-7 carbon atoms, carboalkyl of 2-7 carbon atoms, amino, and alkanoylamino of 1-15 6 carbon atoms; R and Rj are each, independendy, hydrogen, halogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, or trifluoromethyl; R2 is hydrogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, trifluoromethyl, 20 Y is a radical selected from the group consisting of R3 ^3 R3 R3 R3 • R3 S==^ R3 R3 R3 RW B fO=<3 R3 R3 , and (C(R3)2)n INTELLECTUAL PROPERTY OFFICE OF NZ. - 8 OCT 2001 received -3- R3 is independently hydrogen, alkyl of 1-6 carbon atoms, carboxy, carboalkoxy of 1-6 carbon atoms, phenyl, or carboalkyl of 2-7 carbon atoms; n = 2-4; with the proviso that each R3 of Y may be the same or different or a pharmaceutically 5 acceptable salt thereof, in the preparation of a medicament for the treatment of colonic polyps. The pharmaceutically acceptable salts are those derived from such organic and inorganic acids as: acetic, lactic, citric, tartaric, succinic, maleic, malonic, gluconic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, and similarly 10 known acceptable acids. The alkyl portion of the alkyl, alkoxy, carboalkoxy, carboalkyl, and alkanoylamino substituents include both straight chain as well as branched carbon chains, e.g. methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, n-15 pentyl or n-hexyl. Carboxy is defined as a-CO2H radical. Carboalkoxy of 2-7 carbon atoms is defined as a -CO2R" radical, where R" is an alkyl radical of 1-6 carbon atoms. Carboalkyl is defined as a -COR" radical, where R" is an alkyl radical of 1-6 carbon atoms. When used herein the term halogen refers to chlorine, bromine, iodine or fluorine. When X is substituted, it is preferred that it is mono- , di- , or tri-20 substituted, with monosubstituted being most preferred. When a compound of this invention contains an assymetric center, this invention covers the individual R and S entantiomers as well as the racemate with respect to such compound. Of the compounds of this invention, preferred members include those in which 25 R, R1, and R2 are hydrogen; and those in which R, R1, and R2 are hydrogen and X is phenyl either unsubstituted or monosubstituted with halogen or alkyl of 1-6 carbon atoms. One group of compounds within the invention are those wherein X is monosubstituted in the 3-position, preferably by a halogen, more preferably by bromine. A further group of compounds within the invention are those wherein Y is 30 -CH=CH-C02H, -CH=CH-C02Et, -CH=C(Me)2, -CH=CH-Et, -CH=CH-CH3, -CH=CH-CH=CH-CH3, -CH=CH2, -CH=CH-Ph, -CH=CH-CH3 or 2-cyclo-pentene. R3 is preferably hydrogen, methyl, ethyl, phenyl, CO2H or C02Et. WO 99/24037 PCT/US98/23549 -4- A process for the preparation of a compound of Formula 1 as defined above which comprises: a) reducing a 6-amino-4-anilinoquinazoline of Formula 6 wherein R, Ri, R2 and 5 X are as defined above r hn"x Hi Ri with: 10 i) an acid chlonde of Formula 7, wherein Y is as defined above Y-^° 7 cl 15 ii) a mixed anhydride of Formula 8, wherein Y is as defined above O r—^ OCOR4 8 , oriii) a cyclic anhydride of Formula 11 wherein each R5 is independently 20 hydrogen, phenyl, or alkyl of 1-6 carbon atoms ' $ 11° b) reacting a compound of Formula 17 wherein R, Rl, R2 and Y are as defined 25 above and Hal is any suitable halogen Printed from Mimosa WO 99/24037 PCT/US98/23549 10 15 with an aniline of Formula 18 wherein X is as defined above x-nh2 18 ore) reacung a compound of Formula 24 wherein R, Rl, R2 and Y are as defined above r2 7^n-^n(ch3)2 with an aniline of Formula 25 wherein X is as defined above. h2n-x 25 The preparation of the compounds of this invention encompassed by Formula 9 is described below m Flowsheet A where R, Ri, R2, R3, X, aind n are as defined above 20 and R4 is alkyl of 1-6 carbon atoms (preferably isobutyl). Y' is a radical selected from the group consisting of: Printed from Mimosa WO 99/24037 PCT/US98/23549 -6- R3—( ,R,3 F3 ,and R3 R's ^(C(R'3)2), n wherein each R'3 is independently alkyl of 1-6 carbon atoms, carboxy, carboalkoxy of 1-6 carbon atoms, phenyl, or carboalkyl of 2-7 carbon atoms. According to the 5 sequence of reaction outlined m flowsheet A, a 5-nitro-anthranilonitrile of Formula 2 is heated at about 100°C with or without solvent containing an excess of dimethylformamide dimethyl acetal to furnish an amidine of Formula 3. Heaung a solution of amidine 3 and the aniline 4 in acetic acid for 1 to 5 hours gives the 6-nitro-4-anilmoquinazolines of Formula 5. Reduction of the mtro group of 5 using a 10 reducing agent such as iron in an acetic acid-alcohol mixture at elevated temperature gives the 6-amino-4-amlinoquinazolines of Formula 6. Acylation of 6 with either an acid chloride of Formula 7 or a mixed anhydride of Formula 8 (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyridine or triethylamine gives the compounds 15 of this mvenuon represented by Formula 9. In those cases where 7 or 8 have an asymmetric carbon atom, they can be used as the racemate or as the individual R or S entantiomers in which case the compounds of this invention will be . in the racemic or R and S optically active forms, respectively. The 5-mtro-anthranilonitnles of Formula 2 needed to prepare the compounds of this invention are either already known to the art or 20 can be prepared by procedures known in the art as detailed in the following references: Baudet, Recl.Trav.Chim.Pays-Bas, 43, 710 (1924); Hartmans, Recl.Trav.Chim.Pays-Bas, 65, 468, 469 (1946) ; Taylor et al., J.Amer.Chem.Soc., 82, 6058,6063 (1960); Taylor et al., J.Amer.Chem.Soc., 82, 3152,3154 (1960); Deshpande; Seshadri, Indian J.Chem., 11 , 538 (1973); Katritzky, Alan R.; Laurenzo, Kathleen S., J.Org.Chem., 25 51 (1986); Niclas, Hans-Joachim; Bohle, Matthias; Rick, Jens-Detlev; Zeuner,Frank; Zoelch, Lothar, Z.Chem., 25(4), 137-138 (1985). Printed from Mimosa WO 99/24037 PCT/US98/23549 -7- FLOWSHEET A H °2n^AYCN (CH3)2NCH(OCH3)2 Ri DMF °2N^X^CN r2^^N' x-nh2 4 ^ ch3co2h, ' n(ch3)2 RI 3 r hn' r hn' Y—^_ Fe ^ CH3CO2H, C2H5OH OCOR4 8 thf, pyridine, or (c2hs)3n The preparation of the compounds of this invention encompassed by Formula 12 is described below in Flowsheet B wherein R, Ri, R2. X, and n are described above. Each r5 is independently hydrogen, phenyl, or alkyl of 1-6 carbon atoms. According to the reaction outlined in Flowsheet B, the 6-amino-4-anilinoquinazolines of Formula 10 (prepared as in Flowsheet A) are acylated with a cyclic anhydride of Formula 11 in an inert solvent such as tetrahydrofuran in the presence of a basic catalyst such as pyridine or triethylamine. Printed from Mimosa WO 99/24037 PCT/US98/23549 -8- FLOWSHEET B O (Z) h2n 11 o ho2c THF, pyndine, or (C2H5)3N 10 12 Representative compounds of this invention were evaluated in several standard pharmacological test procedures that showed that the compounds of this invention possess significant activity as inhibitors of protein tyrosine kinases, and are 5 antiproliferative agents. Based on the activity shown in the standard pharmacological test procedures, the compounds of this invention are therefore useful as antineoplastic agents. The test procedures used and results obtained are shown below. 10 19 is described below in Flowsheet C wherein Y', R4, and X are described above. According to the reactions outlined in Flowsheet C, 4-choro-6-nitroquinazoline, 13, (Morley, JS. and Simpson,/. Chem.. Soc., 360 (1948)) is reduced to 6-amino-4-chloroquinazoline, 14, using a reducing agent such as sodium hydrosulfite in a two phase system consisting of tetrahydrofuran and water in the presence of a small amount 15 of phase transfer catalyst. Acylation of 14 with either an acid chloride of Formula 15 or a mixed anhydride of Formula 16 (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyridine or N-methyl morpholine gives the compounds of Formula 17. In those cases where 15 or 16 have an asymmetric carbon atom, they can 20 be used as the racemate or as the individual R or S entantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. Heating a compound of Formula 17 with an aniline of Formula 18, in a inert solvent such as isopropanol, gives the compounds of this invention represented by Formula 19. The preparation of the compounds of this invention encompassed by Formula 25 Printed from Mimosa WO 99/24037 PCT/US98/23549 -9-FLOWSHEET C o2n. Na2S204i (C8H17)3NCH3+ CI" THF, H20 H2N. 13 14 or Y'-^f CI 15 OCOR4 16 THF, pyridine, or N-methyl morpholine 19 17 x—nh2 18 (CH3)2CHOH 10 The preparation of the compounds of this invention encompassed by Formula 26 is described below in Flowsheet D wherein Y', R4, and X are descnbed above. According to the reactions outlined in Flowsheet D, the nitro group of 20 (prepared as in Flowsheet A) is reduced to the corresponding amino compound 21 using a palladium catalyst and a source of hydrogen which can be hydrogen itself or cyclohexene. Acylation of 21 with either an acid chloride of Formula 22 or a mixed anhydride of Formula 23 (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyndine or N-methyl morpholine gives the compounds of Formula 24. In those cases where 22 or 23 have an asymmetric carbon atom, they can be used as the racemate or as the individual R or S entantiomers in which case the compounds of this invention Printed from Mimosa WO 99/24037 PCT/US98/23549 - 10- will be in the racemic or R and S optically active forms, respectively Heating a compound of Formula 24 with an aniline of Formula 25, in a inert solvent such as acetic acid gives the compounds of this invention represented by Formula 26. FLOWSHEET D o2n CN N^N(CH3)2 cyclohexene CH3OH, Pd/C h2nY^V°N ^^vN<:KN(CH3)2 2d ° /? r-4 or Y'-< ^ci OCOR4 22 23 THF, pyridine, or (C2H5)3N H 'Y. N nx cn 21 N(CH3)2 24 h2n-x 25 acetic acid H VYN o HN' J 26 The ability of the compounds of this invention to treat or inhibit colonic polyps 10 was demonstrated in an ]n vivo standard pharmacological test procedure as descnbed below. The compound of Example 9 was evaluated in this procedure, which emulates familial adenomatous polyps (FAP) in humans, as a representative compound of this invention. The Min mouse used in this test procedure, currently the best available model for FAP, is a strain which has lost both copies of the APC gene. These animals 15 develop multiple intestinal polyps (Adenomas) that ultimately progress to form adenocarcinomas. The polyps that develop in Min mice express EGFR and have activated COX-2. NSAID's such as sulindac and etodoloc can reduce (but not eradicate) intestinal polyp formulation in these animals indicating that COX-2 and the Printed from Mimosa WO 99/24037 PCT/US98/23549 - 11 - ultimate production of PG's is likely responsible for these effects. The following briefly describes the procedure used and the results obtained in this standard pharmacological test procedure. 5 The compound of Example 9 was blended with a standard murine chow and animals were given ad libitum access to the food. Based on estimated food consumption, the compound of Example 9 was added at a concentration commensurate with animals ingesting 20 mg/kg/day. At day 30, 4 treated + 4 control (chow alone) animals were sacrificed and assessed for polyp number. All control animals had greater 10 than 30 polyps in their bowel, while the treated animals had none. Identical results were observed at 60 days - when 15 animals/group were assessed. The control animals had greater than 30 (larger) polyps while the treated animals had none. These data demonstrate that the compounds of this invention effectively inhibit 15 polyp formation in animals having mutations in their APC genes. Based on the results obtained in this standard pharmacological test procedure, the compounds of this invention are useful in treating or inhibiting the formauon of colonic polyps. The compounds of this invention may formulated neat or may be combined with 20 one or more pharmaceutically acceptable earners for administration. For example, solvents, diluents and the like, and may be administered orally in such forms as tablets, capsules, dispersible powders, granules, or suspensions containing, for example, from about 0.05 to 5% of suspending agent, syrups containing, for example, from about 10 to 50% of sugar, and elixirs containing, for example, from about 20 to 50% ethanol, 25 and the like, or panenterally in the form of sterile injectable solution or suspension containing from about 0.05 to 5% suspending agent in an isotonic medium. Such pharmaceutical preparations may contain, for example, from about 0.05 up to about 90% of the active ingredient in combination with the carrier, more usually between about 5% and 60% by weight. 30 The effective dosage of active ingredient employed may vary depending on the particular compound employed, the mode of administration and the severity of the condition being treated. However, in general, satisfactory results are obtained when the compounds of the invention are administered at a daily dosage of from about 0.5 to about 1000 mg/kg of animal body weight, optionally given in divided doses two to four 35 times a day, or in sustained release form. For most large mammals the total daily dosage is from about 1 to 1000 mg, preferably from about 2 to 500 mg. Dosage forms Printed from Mimosa WO 99/24037 PCT/US98/23549 - 12- suitable for internal use comprise from about 0.5 to 1000 mg of the active compound in intimate admixture with a solid or liquid pharmaceutically acceptable earner. This dosage regimen may be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily or the dose may be 5 proportionally reduced as indicated by the exigencies of the therapeuuc situation. These active compounds may be administered orally as well as by intravenous, intramuscular, or subcutaneous routes. Solid earners include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include sterile water, polyethylene glycols, non-ionic surfactants and edible oils such as corn, 10 peanut and sesame oils, as are appropnate to the nature of the active ingredient and the particular form of administration desired Adjuvants customanly employed in the preparation of pharmaceutical compositions may be advantageously included, such as flavonng agents, coloring agents, preserving agents, and antioxidants, for example, vitamin E, ascorbic acid, BHT and BHA. 15 The preferred pharmaceutical compositions from the standpoint of ease of preparation and administration are solid compositions, particularly tablets and hard-filled or liquid-filled capsules. Oral administration of the compounds is preferred. In some cases it may be desirable to administer the compounds directly to the airways in the form of an aerosol. 20 These active compounds may also be administered parenterally or intrapentoneally. Solutions or suspensions of these active compounds as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxy-propylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary 25 conditions of storage and use, these preparation contain a preservative to prevent the growth of microorganisms. f • ■*• ;The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and 30 must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bactena and fungi. The earner can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and 35 vegetable oils. ;Printed from Mimosa ;WO 99/24037 ;PCT/US98/23549 ;- 13- ;The preparation of representative examples of the compounds of this invention is described below. ;A 40.8 g portion of 5-nitro-anthranilonitnle and 40 ml of N, N-dimethylformamide dimethyl acetal were heated on a steam bath for 2 hours. The solvents were removed at reduced pressure and the residue was taken up in methylene 10 chloride. After passing this solution through Magnesol the solvent was removed. After washing with ether 50.8 g of N'-(2-cyano-4-nitrophenyl)-N,N-dimethyl-formamidine was obtained. ;A solution of 23.74 ml of 3-bromo aniline and 40 5 g N'-(2-cyano-4-nitrophenyl)-N,N-dimethylformamidine in 100 ml of glacial acetic acid was stirred and heated in an oil bath at 148°C for 1.5 hours. On cooling, filtration of the resulting solid 20 gives a quantitative yield of N-(3-bromophenyl)-6-nitro-4-quinazolinamine: mp = 267-270°C; mass spectrum (m/e): 345. ;25 <• * ' • '■' A mixture of 34.5 g of N-(3-bromophenyl)-6-nitro-4-quinazolinamine and 16.8 g of iron powder in 150 ml of ethanol and 150 ml of glacial acetic acid was heated in an oil bath at 120°C for 2 hours. After filtration of the solid, solid sodium carbonate was added to the filtrate giving a solid. This was filtered, and the solid was extracted with 30 methanol. The extracts were treated with charcoal and evaporated to a solid. After washing the solid with ether 27.5 g of N-(3-bromophenyl)-4,6-quinazolindiamine was obtained: mass spectrum (m/e): 315. 5 Example 1 N'-f2-Cvano-4-nitrophenvl')-N.N-dimethvlformamidine 15 Example 2 N-(3-BromoDhenvl1-6-nitro-4-quinazoIinanune Example 3 N-(3-BromophenvlV4.6-quinazolindiamine Printed from Mimosa WO 99/24037 PCT/US98/23549 - 14-Example 4 4-rr4-r(,3-Bromophenvl1aminol-6-quinazolinvnaminol-4-oxo-(Z')-2-butenoic acid A 15 ml portion of pyridine was added to 1.6 g of N-(3-broinophenyl)-4,6-5 quinazolindiamine and 0.6 g of maleic anhydride. After stirring overnight, the solvents were removed on the rotary evaporator. The solid was taken up in about 400 ml of hot ethanol and the insoluble matenal filtered to give 0.33 g of 4-[[4-[(3-Bromophenyl)ammo]-6-quinazolinyl]amino]-4-oxo-(Z)-2-butenoic acid: mass spectrum (m/e): M+H 413, 415. Example 5 4-IT4-IY3-Bromophenvl')aniinol-6-quinazolinvnamino1-4-oxo-(E')-2-butenoic acid, ethvl ester 15 A solution of N-(3-bromophenyl)-4,6-quinazolindiamme in 15 ml of pyridine was cooled in an ice bath and a solution of 1.22 g of ethyl fumaryl chloride in 10 ml of methylene chloride was added dropwise. After stirring for 1.5 hours, the reaction was allowed to come to room temperature. The solvents were removed at reduced pressure 20 and the residue was treated with water. The red solid was filtered and extracted into hot acetone. After filtration of the insoluble matenal, the filtrate was concentrated to give 0 45 g of 4-[[4-[(3-Bromophenyl)amino]-6-qumazolinyl]amino]-4-oxo-(E)-2-butenoic acid, ethyl ester: mp = 259-263°C, mass spectrum (m/e): M+H 441, 443. 25 Example 6 N-r4-[Y3-BromoDhenvl1aminol-6-quinazolinvn-3-methvi-2-butenamide A solution of 1.58 g of N-(3-bromophenyl)-4,6-quinazolindiamine in 15 ml of pyridine was cooled in an ice bath and a solution of 0 67 ml of 3,3-dimethylaciyloyl 30 chlonde in 7 ml of ether was added dropwise. After stirring and cooling for 2 hours, the solvents were removed at reduced pressure. The residue was treated with water and the resulting solid was recrystallized from methyl cellusolve to give 0.97 g of N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-3-methyl-2-butenamide: mp = 300-301°C, mass spectrum (m/e): 396, 398. Printed from Mimosa WO 99/24037 PCT/US98/23549 - 15- Example 7 N-r4-rn-Bromophenvl')aniino1-6-quinazohnvll-fE')-2-butenamide A solution of 1.6 g of N-(3-bromophenyl)-4,6-quinazolindiamine in 15 ml of 5 pyridine was cooled m an ice bath and a solution of 0.57 ml of trans-crotonoyl chloride in 6 ml of ether was added dropwise. After stirring and cooling for 2 hours, the solvents were removed at reduced pressure. The residue was treated with water and the resulting solid recrystallized from n-butanol to give 0.69 g of N-[4-[(3-bromophenyl)amino]-6-quinazohnyl]-(E)-2-butenamide: mp = 153-160°C, mass 10 spectrum (m/e): M+H 383,385. Example 8 N-r4-rG-BromophenvDaminol-6-quinazohnvll-2-methvI-2-Dropenamide 15 A solution of 1.6 g of N-(3-bromophenyl)-4,6-quinazolindiamine in 15 ml of pyridine was cooled in an ice bath and a solution of 0.59 ml of methaciyoyl chloride in 6 ml of ether was added dropwise. After stirring and cooling for 2 hours, the solvents were removed at reduced pressure. The residue was treated with water and the 20 resulting solid was taken up in n-butanol (warming). Addition of ether to the cooled solution gives 0.44 g of N-[4-[(3-bromophenyl)ammo]-6-quinazolinyl]-2-methyl-2-propenamide: mp = 40-245°C, mass spectrum (m/e): M+H 383, 385. Example 9 25 N-r4-IY3-Bromophenvl1a'minol-6-quinazolinvn-2-butvnamide A solution of 0.50 g of 2-butynoic acid in 10 ml of tetrahydrofuran was cooled in an ice bath. A 0.79 ml portion of isobutyl chloroformate followed by a 0.66 ml portion of N-methyl morpholine were added. After about 1 minute a solution of 1.6 g 30 of N-(3-bromophenyl)-4,6-quinazoIindiamine in 10 ml of pyridine was added. The reaction was allowed to come to room temperature and stir overnight. The solvents were removed at reduced pressure and the solid was recrystallized from n-butanol to give 1.07 g of N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide: mass spectrum (m/e): 381, 383. 35 Printed from Mimosa WO 99/24037 PCT/US98/23549 - 16-Example 10 4-ff4-r(3-Bromophenvl1aminol-6-qumazolinvllamino1-4-oxo-(E1-2-butenoic acid A 2.5 ml portion of 10 N aqueous sodium hydroxide was added to 2.3 g of 4-[[4-[(3-5 bromophenyl)amino]-6-quinazolinyl]amino]-4-oxo-(E)-2-butenoic acid ethyl ester (Example 5) in 25 ml of ethanol. After stirring for an hour, 2.1 ml of concentrated hydrochloric acid was added, and the reaction was stirred an additional 2 hours. The resulting solid was recrystallized from n-butanol to give 0.97 g of 4-[[4-[(3-Bromophenyl)amino]-6-quinazolinyl]amino]-4-oxo-(E)-2-butenoic acid: mass spectrum 10 (m/e): M+H 413. Example 11 N-r4-rC3-BromophenvPamino1-6-quinazolinvn-2.4-hexadienamide 15 A solution of 0.67g of 2,4-hexadienoic acid in 10 ml of tetrahydrofuran was cooled in an ice bath. A 0.79 ml portion of isobutyl chloroformate followed by a 0.66 ml portion of N-methyl morpholine were added. After about 1 minute a solution of 1.6 g of N-(3-bromophenyI)-4,6-quinazolindiamine in 10 ml of pyridine was added. The 20 reaction was allowed to come to room temperature and stir overnight. The solvents were removed at reduced pressure and the solid was recrystallized to give 1.0 g of N-[4-[(3-bromophenyl)ammo]-6-quinazolinyl]-2,4-hexadienamide: mp = 258-260°C. 25 TMf Example 12 N-r4-rC3-Bromophenvnaminol-6-quinazolinvn-2-cvclopenteneamide A solution of 0.43 g of 2-cyclopeiitenoic acid in 5ml of tetrahydrofuran was cooled in an ice bath. A 0.49 ml portion of isobutyl chloroformate followed by a 0.41 30 ml portion of N-methyl morpholine were added. After about 1 minute a solution of 1.0 g of N-(3-bromophenyl)-4,6-quinazolindiamine in 10 ml of pyridine was added. The reaction was allowed to come to room temperature and stir overnight. Another 0.5 equivalents of mixed anhydride was added. The mixture was stirred for 5 hours. The solvents were removed at reduced pressure and the solid was purified by 35 Printed from Mimosa WO 99/24037 PCT/US98/23549 - 17- chromatography on silica gel to give 0.30 g of N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-cyclopenteneamide: mass spectrum (m/e): 409 (M+H, EI). Example 13 5 N-r4-r(,3-Bromophenvl'>anunol-6-quinazolinvll-2-propenamide A solution of 2.0 g of N-(3-bromophenyl)-4,6-quinazolindiamine in 10 ml of pyridine was cooled m an ice bath and a solution of 0.61 ml of acryoyl chloride m 30 ml of ether was added dropwise at 0°C. After stirring at room temperature for 3.5 10 hours, the solvents were removed at reduced pressure. The residue was purified by chromatography to give 0.2 g of N-[4-[(3-bromophenyl)amino]-6-quinazolmyl]-2-propenamide: mass spectrum (m/e): M+H 369. 15 Example 14 N-r4-IY3-Bromophenvl1)amino1-6-quinazolinvll-f3-phenvl-2-propvnamide') A solution of 0.93 g of 3-phenyl-2-propynoic acid in 10ml of tetrahydrofuran was cooled in an ice bath. A 0.82 ml portion of isobutyl chloroformate followed by a 20 0.69 ml portion of N-methyl morpholine were added After about 1 minute a solution of 1.0 g of N-(3-bromophenyl)-4,6-quinazolindiamme in 7 ml of pyridine was added. The reacuon at 0°C for 1 hr.. The solvents were removed at reduced pressure and the solid was purified by chromatography on silica gel to give 001 g of N-[4-[(3-bromophenyl)amino]-6-quinazohnyl]-(3-phenyl-2-propynamide): mass spectrum (m/e): 25 443.2, 445.2 (M+H, electrospray). . Example 15 f " 6-amino-4-chloroqumazoline 30 A mixture consisting of 3.25 g of 4-chloro-6-nitroquinazoline, 10.8 g of sodium hydrosulfite, and 0.3 g of the phase transfer catalyst (CsHi7)3NCH3+ CI- in 97 ml of tetrahydrofuran and 32 ml of water was stirred rapidly for 2 hours. The mixture was diluted with ether and the organic layer was separated. The organic solution was washed with brine and then dried over magnesium sulfate. The solution 35 was passed through a small column of silica gel. The solvent was removed at 30°C at reduced pressure giving 6-amino-4-chloroquinazoline which is used in the next step without additional purification. Printed from Mimosa WO 99/24037 PCT/US98/23549 - 18- Example 16 r4-chloro-6-quinazolinvll-2-butvnamide 5 A solution of 1.64 g of 2-butynoic acid in 46 ml of tetrahydrofuran was cooled in an ice bath. A 2.34 ml portion of isobutyl chloroformate followed by a 4.13 ml portion of N-methyl morpholine were added. After about 10 minutes, this was poured into a solution of 6-amino-4-chloroquinazoline in 46 ml tetrahydrofuran. This mixture was stirred at room temperature for 2 hours. The mixture was poured into a mixture of 10 brine and saturated sodium bicarbonate and extracted with ether. The ether solution was dried over magnesium sulfate and filtered. The solvent was removed giving £4-chloro-6-quinazolinyl]-2-butynamide as colored oil that was used m the next step without additional purification. IS Example 17 N-r4-r(3-BromophenvI'>aminol-6-quinazolmvll-2-butvnamide A solution consisting of 1.76 g of £4-chloro-6-qumazolinyl]-2-butynamide and 1.23 g of 3-bromo aniline was refluxed under an inert atmosphere in 23 ml of 20 isopropanol for 40 minutes. The mixture was cooled to room temperature and 200 ml of ether was added giving 0.4 g of N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide as the hydrochloride salt. Neutralizing with sodium bicarbonate solution, extracting with ethyl acetate, removal of the solvent, and recyrstallization from 1-butanol gives N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide as the free 25 base. Example 18 N'-(4- Amino-2-cvanophenvlVN.N -dimethvlformamidine A solution of 6.0 g (27.5 mmol) of N'-(2-cyano-4-nitrophenyl)-N,N-30 dimethylformamidine, 33.9 g (41.8 ml, 412.4 mmol) of cyclohexene, and 0.6 g of 10% Pd/C in 360 ml of methanol was refluxed for 4 hrs. The hot mixture was filtered through Celite. Solvent was removed and the residue was recrystallized from chloroform-carbon tetrachloride giving 4.9 g (95%) of the title compound as a light gray crystalline solid, mass spectrum (m/e)-188.9 (M+H, electrospray). Printed from Mimosa </div>

Claims

<div class="application article clearfix printTableText" id="claims"> WO 99/24037 PCT/US98/23549 - 19-Example 19 N-r3-Cvano-4-rrCdimethvlamino'lmethvlenelaminol phenvll-2-butvnainide To a solution of 2.01 g (23.9 mmol) of 2-butynoic acid and 2.9 ml (22.3 mmol) isobutyl chloroformate in 30 ml tetrahydrofuran was stirred at 0°C under 5 nitrogen as 2.42 g (2.63 ml, 22.3 mmol) of N-methyl morpholine was added over 3 min. After stirring for 15 min., a solution of N'-(4-amino-2-cyanophenyl)-N,N-dimethylformamidine and 1.6 g (1.75 ml, 15.9 mmol) of N-methyl morpholine in 25 ml tetrahydrofuran was added over 4 min. The mixture was stirred 30 min. at 0°C and 30 min. at room temperature. The mixture was diluted with 70 ml of ethyl acetate and 10 poured into a mixture of brine and saturated sodium bicarbonate. The organic layer was dried (MgS04) and filtered through a pad of silica gel. The solvent was removed and the residue was stirred with 50 ml of ether. The suspended solid was collected to give 3.61 g (89%) of an off-white solid, mass spectrum (m/e): 255.0 (M+H, electrospray). 15 Example 20 N-r4-rC3-Bromophenvl1aminol-6-quinazolinvll-2-butvnamide A solution of 3.0 g (11.8 mmol) of N-[3-cyano-4-[[(dimethylamino)-methylene]amino] phenyl]-2-butynamide and 2.23 g (12.98 mmol) of 3-bromo aniline 20 in 18 ml of acetic acid was refluxed gently with stirnng under nitrogen for 1 hr 15 min.. The mixture was cooled in an ice bath and a solid mass formed. The solid was collected by filtration and washed with ether-acetonitnle 1:1 to give a yellow solid which was recrystallized from ethanol giving 2.51 g of N-[4-[(3-bromophenyl)amino]-6-qumazolinyl]-2-butynamide: mass spectrum (m/e): 381,383. Printed from Mimosa 50398 WE CLAIM -20-

1. Use of a compound of the formula R HN"X O R 1 wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, carboalkoxy of 2-7 carbon atoms, carboalkyl of 2-7 carbon atoms, ammo, and alkanoylamino of 1-6 carbon atoms; R and Rj are each, independently, hydrogen, halogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, or trifluoromethyl; r2 is hydrogen, alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, trifluoromethyl; Y is a radical selected from the j»roup consisting of R3 is independently hydrogen, alkyl of 1-6 carbon atoms, carboxy, carboalkoxy of 1-6 carbon atoms, phenyl, or carboalkyl of 2-7 carbon atoms; n = 2-4; with the proviso that each R3 of Y may be the same or different or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for the treatment of colonic polyps. intellectual property office of nz. ' 8 SEP 2001 RECEIVED 50 -21-

2. Use of a compound as claimed in Claim 1 wherein R, R„ and R2 are hydrogen or a pharmaceutically acceptable salt thereof.

3. Use of a compound as claimed in Claim 1 or Claim 2 wherein X is phenyl unsubstituted or substituted with halogen or alkyl of 1-6 carbon atoms.

4. Use of a compound as claimed in any one of Claims 1 to 3 wherein the compound is one of the following: N-[4-[(3-Bromophenyl)amino]-6-quinazohnyl]-2-butynamide, N-[4-[(3-Bromophenyl)amino]-6-quinazolinyl]-2-methyl-2-propenamide, N-[4-[(3-Bromophenyl)amino]-6-quinazolinyl]-2,4-hexadienamide, N-[4-[(3-Bromophenyl)amino]-6-quinazolinyl]-(E)-2-butenamide, N-[4-[(3-Bromophenyl)aminq]-6-quinazolinyl]-3-methyl-2-butenamide, 4-[[4-[(3-Bromophenyl)amino]-6-quinazolinyl]amino]-4-oxo-(Z)-2-butenoic acid, 4-[[4-[(3-Bromophenyl)amino]-6-quinazolinyl]arruno]-4-oxo-(E)-2-butenoic acid, 4-[[4-[(3-Bromophenyl)amino]-6-quinazohnyl]amino]-4-oxo-(E)-2-butenoic acid, ethyl ester, N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-cyclopentenamide, N-[4-[(3-Bromophenyl)amino]-6-quinazolinyl]-2-propenamide, N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-(3-phenyl-2-propynamide) or a pharmaceutically acceptable salt of any of these.

5. The use of Claim 1 substantially as herein described with reference to any example thereof. intellectual property office of n z. 1 8 SEP 2001 D fi A c 11< n ■> </div>