Background : Protein kinases are enzymes that catalyze the phosphorylation of hydroxyl groups of tyrosine, serine, and threonine residues of proteins. Many aspects of cell life (for example, cell growth, differentiation, proliferation, cell cycle and survival) depend on protein kinase activities. Furthermore, abnormal protein kinase activity has been related to a host of disorders such as cancer and inflammation. Therefore, there is a great deal of effort directed to identifying ways to modulate protein kinase activities. In particular, many attempts have been made to identify small molecules which act as protein kinase inhibitors Various quinolinone derivatives have recently been disclosed in patents such as WO 01/28993, WO 01/29025, WO 01/62251, WO 01/62252, WO 02/22598, WO 03/20699, WO 03/37252, WO 2004/087651, and WO 2004/018419. These compounds were reported as protein kinase inhibitors. The clinical utility of these compounds has been promising, but has been partially compromised due to the relatively poor aqueous solubility and/or other drug properties. What is needed is a class of modified quinolinone based derivatives having both inhibitory activity and enhanced drug properties.

Summary : The invention is directed to quinolinone based derivatives and to their use as inhibitors of protein kinases. It is disclosed herein that hydroxy carboxy quinolinonone derivatives have enhanced and unexpected drug properties that advantageously distinguish this class of compounds over known quinolinone based derivatives having protein kinase inhibition activity. It is also disclosed herein that hydroxy carboxy quinolinone based derivatives are useful in treating disorders related to abnormal protein kinase activities such as cancer.

One aspect of the invention is directed to a compound represented by Formula (1) : R3 /=L- (CHR) n- (CH (OH)- (CHRS) Jp-COR6 X Formula N O H In Formula I, R1 is selected from the group consisting of hydrogen, halo, (C1-C6) alkyl, (C3-C8) cycloalkyl, (C1-C6) haloalkyl, hydroxy, (C1-C6) alkoxy, amino, (C1- C6) alkylamino, amide, sulfonamide, cyano, substituted or unsubstituted (C6-C10) aryl ; R2 is selected from the group consisting of hydrogen, halo, (C1-C6) alkyl, (C3-C8) cycloalkyl, (C1-C6) haloalkyl, hydroxy, alkoxy, (C1-C6) alkoxy (C1-C6) alkyl, amino, (C1-C6) alkylamino, (C6-C10) arylamino ; R3 is selected from the group consisting of hydrogen, (C1-C6) alkyl, halo, cyano; R4 is selected from the group consisting of hydrogen and (C1-C6) alkyl ; R5 is selected from the group consisting of hydrogen, (C1-C6) alkyl and hydroxyl ; R6is selected from the group consisting of hydroxyl, O-(C1-C6) alkyl, O-(C3-C8) cycloalkyl, substituted or unsubstituted O-(C6-C10) aryl, and NR7R8 ; where R7 and R3 are independently selected from the group consisting of hydrogen, (C1-C6) alkyl, (C1-C6) hydroxyalkyl, (C1-C6) dihydroxyalkyl, (C1-C6) alkoxy, (C1-C6) alkyl carboxylic acid, (C1-C6) alkyl phosphoric acid, (C1-C6) alkyl sulfuric acid, (C1-C6) hydroxyalkyl carboxylic acid, (C1-C6) alkyl amide, (C3-C8) cycloalkyl, (C5-C8) heterocycloalkyl, (C6-C10) aryl, (C5-C9) heteroaryl, (C3-C8) cycloalkyl carboxylic

acid, or R7 and R3 together with N forms a (C5-C8) heterocyclic ring either unsubstituted or substituted with one or more hydroxyls, ketones, ethers, and carboxylic acids; n and m are independently 0,1, 2, or 3; p is 1,2, or 3; X is selected from the group consisting of CR9 and N; where R9 is selected from the group consisting of hydrogen, halo, and (C1-C6) alkyl ; L is a divalent linker selected from the group consisting of-O-,-NR10-, -C(O)-NR10-, -NR10-C(O)-NR11-, -CHR10-NR11-, -CHR10-NR11-C(O)-NR12-, -S(O2)-NR10, -O-CHR10-C(O)-NR11-, and -CH2-CH2-NR10-; and R'O, R11, and R12 are independently is selected from the group consisting of hydrogen and (C1-C6) alkyl. Alternatively, this aspect of the invention may also be directed to a pharmaceutically acceptable salt, its tautomer, a pharmaceutical acceptable salt of its tautomer, or a prodrug of compounds represented by Formula (I). Key features of this aspect of the invention include the hydroxyl moiety or moieties between R4 and R5 and the carboxy moiety between R5 and R6. It is disclosed herein that these key features enhance the drug properties of the attached quinolinone pharmacophore.

As illustrated above, this first aspect of the invention may be divided into two categories. The first category includes acids and esters; the second category includes amides.

One preferred embodiment of this first category may be represented by Formula (ll) : R3 R Xi4L-(CHR4) n-(CH (oH)-(CHR5) m) p-CooR6a X R 4 Formula 11 N H In Formula II, R6a is selected from the group consisting of hydrogen, (C1-C6) alkyl, (C3-C8) cycloalkyl, and substituted or unsubstituted (C6-C10) aryl. In a preferred embodiment, X is selected from the group consisting of CH and N; R1 is selected from the group consisting of hydrogen, halo, and cyano; R2 is selected from the group consisting of hydrogen, hydroxyl, (C1-C6) alkoxy,-NH2, and-NHR13, where

R13 is (C1-C6) alkyl ; R4, R5 and R6a are hydrogen; n, and p are independently 1, or 2; m is 0 or 1; L is selected from the group consisting of-C (O)-NR11-,-NR10-C (O) - NR11-, -CHR10-NR11-C(O)-NR12-, -O-CHR10-C(O)-NR11-, -S(O2)-NR10-; where R10, R11 and R12 are independently hydrogen and (C1-C6) alkyl. Preferred examples include the following :

Other preferred examples include the following :

Other preferred examples include the following :

Other preferred examples include the following :

The second category of the first aspect of the invention is embodied by a compound, salt, tautomer, or prodrug according to claim 1 wherein R6 is NR 7 R8.

In a preferred embodiment of this second category, X is selected from the group consisting of CH and N; R1 is selected from the group consisting of hydrogen, halo, and cyano; R2 is selected from the group consisting of hydrogen, hydroxyl,

(C1-C6) alkoxy,-NH2, and-NHR13, where R13 is (C1-C6) alkyl ; R4, R5 and R6 are hydrogen; n, and p are independently 1, or 2; m is 0 or 1; L is selected from the group consisting of-C (O)-NR10-, -NR10-C (O)-NR1-, -CHR10-NR11-C (O)-NR2-,-O- CHR10-C(O)-NR11-, -S(O2)-NR10-; where R10, R11 and R12 are independently hydrogen and (C1-C6) alkyl ; and R7 an are selected from the group consisting of hydrogen, (C1-C6) alkyl, (C1-C6) hydroxyalkyl, (C1-C6) dihydroxyalkyl, (C1-C6) alkoxy, (C1-C6) alkyl carboxylic acid, (C1-C6) alkyl phosphoric acid, (C1-C6) alkyl sulfuric acid, (C1-C6) hydroxyalkyl carboxylic acid, (C1-C6) alkyl amide, (C3-C8) cycloalkyl, (C5-C8) heterocycloalkyl, (C6-10) aryl, (C5-C8) heteroaryl, (C3-C9) cycloalkyl carboxylic acid, or R7and R8 together with N form a (C5-C8) heterocyclic ring either unsubstituted or substituted with one or more hydroxyls, ketones, ethers, and carboxylic acids. Preferred examples include the following :

Other preferred examples include the following :

Other preferred examples include the following : Other preferred examples include the following : wherein R is selected from the group consisting of radicals represented by the following structures:

Provisios may apply to any of the above categories or embodiments wherein any one or more of the other above described embodiments or species may be excluded from such categories or embodiments.

Another aspect of the invention is directed to a method for the modulation of the catalytic activity of a protein kinase with a compound or salt of the first aspect of the invention. In a preferred mode, the protein kinase is selected from the group consisting of VEGF receptors and PDGF receptors.

Detailed Description : This invention discloses that hydroxy carboxy quinolinone derivatives have unexpected drug properties that advantageously distinguish them from known compounds. They are therefore useful in treating disorders related to abnormal protein kinase activities such as cancer.

It should be understood that a compound of Formula (II) may exist in its open-acid form or its cyclic-lactone form or the two forms may co-exist in solution or in vivo as illustrated below : OH OH OH O R OU R O O Open-acid form Cyclic-lactone form Furthermore, all compounds of Formula (I) have at least one asymmetric center and the stereochemistry at the asymmetric center (s) is (are) either RS, R, or S.

In addition, some of the compounds of Formula (I) may exhibit the phenomenon of tautomerism. As the chemical structures shown in the present invention can only represent one of the possible tautomeric forms, it should be understood that the invention encompasses any tautomeric form of the drawn structure. For example, any claim to compound A below is understood to include tautomeric structure B, and vice versa, as well as mixtures thereof.

Tautomerism may also result from limited rotation about a chemical single bond if there is steric hindrance and/or intra-molecular hydrogen bonding that limits the otherwise free rotation about that bond. It should be understood that the invention also encompasses any rotomers of the drawn structure.

Utility The present invention provides compounds capable of regulating and/or modulating protein kinase activities of, but not limited to, VEGFR (Vascular Endothelial Growth Factor Receptor) and/or PDGFR (Platelet-Derived Growth Factor Receptor). Thus, the present invention provides a therapeutic approach to the treatment of disorders related to the abnormal functioning of these kinases. Such disorders include, but not limited to, solid tumors such as glioblastoma, melanoma, and Kaposi's sarcoma, and ovarian, lung, prostate, pancreatic, colon and epidermoid carcinoma. In addition, VEGFR/PDGFR inhibitors may also be used in the treatment of restenosis and diabetic retinopathy.

Furthermore, this invention relates to the inhibition of vasculogenesis and angiogenesis by receptor-mediated pathways, including the pathways comprising VEGF receptors, and/or PDGF receptors. Thus the present invention provides therapeutic approaches to the treatment of cancer and other diseases which involve the uncontrolled formation of blood vessels.

Synthesis of Acid Compounds The compounds of this invention can be synthesized by following the published general procedures disclosed in WO 01/28993, WO 01/29025, WO 01/62251, WO 01/62252, WO 02/22598, WO 03/20699, WO 03/37252, WO 2004/087651, and WO 2004/018419 and in Kuethe et al. Org. Lett. (2003), 5 (21), 3975; Manley et al. Org. Lett. (2004), 6 (14), 2433; and Kumar et al. Org.

Lett. (2004), 6 (1), 7. But the following intermediates are specific to compounds of this invention and may be used in place of their respective counterparts in the published general procedures: (4R, 6R)-t-butyl-6- (2-aminoethyl)-2, 2- dimethyl-1, 3-dioxane-4-acetate, t-Butyl (3R, 5S)-6-hydroxy-3, 5-O- isopropylidene-3, 5-dihydroxyhexanoate, and 4-amino-3-hydroxy-butanoic acid.

These intermediates may be purchased from commercial sources (e. g. Fisher Scientific, Fairlawn, New Jersey, or Kaneka Corp. , Japan). Another variation from the published general procedures is that in the synthesis of compounds using (4R, 6R)-t-butyl-6-(2-aminoethyl)-2, 2-dimethyl-1, 3-dioxane-4-acetate, the protecting groups need to be removed from the final product. These variations from the published general procedures can be understood and carried out by those skilled in the art. Thus, the compounds of the present invention can be synthesized by those skilled in the art.

The compounds described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention.

Example 1: (3R, 5S)-6-{[2-(4-Amino-5-fluoro-2-oxo-1, 2-dihydro-quinolin-3- yl)-3H-benzoimidazole-5-carbonyl]-amino}-3, 5-dihydroxy-hexanoic acid, sodium salt. The preparation of the title compound was accomplished by first synthesizing compound 1-E as shown in Scheme 1 below : HCI, ethanol NH. HCI ethyl 3, 4-diaminobenzoate 0 -78- 25 OC, 4d ethanol, 75 OC, 2h o H II 1-B F F F F 2-cyano-3-fluoroaniline NH2 LiOH, H2O/THF iline \N LIHMDS,-78 OC, 2h 70 OC, 17h OH O H O O p 1-D 1-E Scheme 1 Preparation of compound 1-B. HCI in dioxane (4M, 25 mL, 100 mmol) was added to a solution of compound 1-A (50 mmol) in anhydrous ethanol (60 mmol) at-30 °C under argon. After stirring at-30 °C for 4h, the solution was sitting at 0 °C for 4d. Then, the solution was stirred at room temperature.

During this stirring, precipitation was observed. The solvents were filtered off, and the solid was washed twice using tert-butylmethyl ether. This solid was dried under high vacuum to obtain the final product 1-B (9.3g, 94%). This compound was used directly in the following steps without further characterization.

Prepratation of compound 1-C. Anhydrous THF (30 mL) and anhydrous ethanol (30 mL) were added to a flask charged with compound 1-B (2.5g, 13 mmol), and ethyl 3,4-diaminobenzoate (1.96g, 11 mmol) under argon. After stirring at 0 °C for 1 h, the solution was continuously stirred at 25 °C overnight, and LC-MS showed a complete reaction. The solvents were removed via evaporation under reduced pressure, and the resulting residue was suspended in ethyl acetate (250 mL). This solution was washed using saturated NaHCO3 (3x) and brine (3x), dried over Na2SO4, and evaporated under vacuum to get the crude product. This crude material was subjected to flash chromatography (ISCO system, 1 %-5% MeOH in DCM, 22 min.) to obtain the final product 1-C (2.85g, 95%). LC-MS: single peak at 254 nm, MH+ calcd. for C14H16N204 : 277, obtained: 277. 1H-NMR (DMSO-d6, 400 MHz), 5 12.73 (d, J= 12 Hz, 1H), 8.12 (d, J = 28 Hz, 1 H), 7.80 (dd, J = 8. 8 Hz, J = 16 Hz, 1 H), 7.60 (dd, J = 8. 4 Hz, J = 28 Hz, 1 H), 4.31 (q, J = 7. 2 Hz, 2H), 4.13 (q, J = 7. 2 Hz, 2H), 4.02 (s, 2H), 1.33 (t, J = 7. 2 Hz, 3H), 1.20 (t, J = 7. 2 Hz, 3H).

Preparation of compound 1-D. LiHMDS in THF (1. OM, 20 mL) was added to a solution of compound 1-C (2.28g, 8.24 mmol) in anhydrous THF (20 mL) under argon. After stirring at-78 °C for 1 h, 2-Cyano-3-fluoroaniline (1.2g, 8.8 mmol, dissolved in THF) was added. The solution was warmed up to 25 °C, and stirred at this temperature for another 1 h. The solvent was removed via evaporation under reduced pressure, and the resulting residue was suspended in ethyl acetate (150 mL). This solution was washed using brine (3x), dried over Na2SO4, and evaporated under vacuum to get the crude product. This crude material was subjected to flash chromatography (ISCO system, 1 %-5% MeOH in DCM, 22 min. ) to obtain the final product 1-D (2.3g, 76%). LC-MS: single peak at 254 nm, MH+ calcd. for C19H15FN403 : 367, obtained: 367. 1H-- NMR (DMSO-d6, 400 MHz), # 12.80 (s, 1H), 11.12 (s, 1H), 8.18 (b, 1H), 7.81 (t, J = 7. 2 Hz, 1 H), 7.77 (dd, J = 1. 6 Hz, J = 8. 0 Hz, 1 H), 7.67 (d, J = 8. 4 Hz, 1 H), 7.61 (b, 1 H), 7.31 (t, J = 8.8 Hz, 1 H), 4.31 (q, J = 7.2 Hz, 2H), 4.17 (s, 2H), 1.33 (t, J = 7.2 Hz, 3H).

Preparation of compound 1-E. LiOH (21 mmol) in H20 (10 mL) was added to a solution of compound 1-D (1.22g, 3.33 mmol) in THF (20 mL). The solution was heated to and stirred at 70 °C overnight. The THF was removed via evaporation under reduced pressure, and the resulting aqueous solution was acidified to pH #3 using concentrated HCI. A precipitate was formed, and the precipitation was collected by filtration, and washed using H20 several times.

The resulting solid was dried under high vacuum to give the final product 1-E (0.45g, 40%). LC-MS: single peak at 254 nm, MH+ calcd. for C17H11FN403 : 339, obtained: 339. 1H-NMR (DMSO-d6, 400 MHz), 5 13.20 (s, 1H), 6 12.70 (s, 1H), 11.62 (s, 1 H), 11.32 (s, 1 H), 8.27 (b, 1 H), 7.93 (d, J = 13.6 Hz, 1 H), 7.81 (dd, J = 1.2 Hz, J = 8. 0 Hz, 1 H), 7.70 (b, 2H), 7.57 (dd, J = 1.6 Hz, J = 8.4 Hz, 1H), 7.17 (d, J=8. 4Hz, 1H), 7.05 (dd, J=8. 0Hz, J=14. 0Hz, 1H).

Compound 1-E was converted to the title compound as shown in Scheme 2 below : Scheme 2 Preparation of ( (4R, 6S)-6-Aminomethyl-2, 2-dimethyl- [1, 3] dioxan-4-yl)-acetic acid tert-butyl ester: Triflic anhydride 1.4mL (2.36g, 8. 345mmol) was dropwise added at-78 °C to a solution of 2, 6-lutidine 1.35mL (11. 63mmol) and t-Butyl (3R, 5S) -6-hydroxy-3, 5- O-isopropylidene-3, 5-dihydroxyhexanoate 1. 981g (7.609 mmol, obtained from Kaneka Corp. ) in dichloromethane (anh. , 50mL) over 3 minutes. The mixture was stirred at-78 °C for 10 min, then placed on ice-slush bath and stirred at 0 °C for 45 min. The resulting pink mixture was transferred into ice-cooled solution of ammonia in methanol (7M solution, 200mL). The mixture was placed on ambient water bath and stirred at RT for 6 hours. The reaction mix was evaporated to dryness, the residue partitioned between ether (200mL) and aqueous potassium carbonate (6g in 200 mL of water), the aqueous phase re- extracted with ether (150mL). Combined organic extracts were dried (magnesium sulfate) and evaporated. The crude product was purified on a column of silica (125g) eluting with a mix chloroform-methanol-conc. aq. ammonia 100: 10: 1 (v/v) (1.5L) Y = 1.777g of a colorless liquid (90% th) 1H (dDMSO, 400MHz) : 4.167 (m, 1H), 3.741 (m, 1H), 2.484 (m, 2H), 2.384 (ddAB, J=15. 2Hz, 5. 1Hz, 1H), 2.201 (ddAB, J=15Hz, 7.8Hz, 1H), 1.533 (br d, J=12. 5Hz, 1 H), 1.373 (s, 9H), 1.363 (s, 3H), 1.250 (br s, 2H), 1.223 (s, 3H) Preparation of compound 2-A (n=0). The amine prepared above (0.1g, 0.39 mmol) was added to a solution of compound 1-E (0. 1g, 0.3 mmol), EDC (0.4 mmol), HOBt (0.4 mmol), and DIEA (1 mmol) in DMF (2 mL). After the solution was stirred at 25 °C for 2h, DMF was removed via evaporation under reduced pressure. The resulting residue was suspended in ethyl acetate (100 mL), washed by saturated NaHCO3 (3x) and brine (3x), and dried over Na2SO4. The ethyl acetate was removed under reduced pressure to give the crude product.

This crude material was subjected to flash chromatography (ISCO system, 1 % - 6% MeOH in DCM, 22 min. ) to obtain the final product 2-A (145mg, 85%).

LC-MS: single peak at 254 nm, MH+ calcd. for C3oH34FN506 : 580, obtained: 580. 1H-NMR (DMSO-d6, 400 MHz), # 13. 10 (d, J= 7.2 Hz, 1 H), 11.64 (d, J = 4.0 Hz, 1H), 11.35 (s, 1H), 8.45 (dt, J = 6.0 Hz, J = 29.2 Hz, 1H), 8.18 (d, J = 12.0 Hz, 1H), 7.92 (t, J= 12.0 Hz, 1H), 7.72 (m, 1H), 7.68 (m, 1H), 7.58 (m, 1H), 7.19 (d, J=8. 0Hz, 1H), 7.05 (dd, J= 8. 0 Hz, J= 14. 0 Hz, 1H), 4.19 (m, 1 H), 4.09 (m, 1 H), 3.57 (t, J = 6.0 Hz, 1 H), 3.24 (m, buried in water signal, 1 H), 2.40 (dd, J = 4.4 Hz, J = 15.2 Hz, 1H), 2.22 (dd, J = 8.- Hz, J = 152. Hz, 1H), 1.65 (m, 1H), 1.40 (s, 3H), 1.37 (s, 9H), 1.28 (s, 3H), 1.10 (m, 1H).

Preparation of compound 2-B (n=O). Aqueous HCI (4 mL, 1. OM) was added to a solution of compound 2-A (215 mg, 0.36 mmol) in MeOH (4 mL). A precipitation was observed immediately. After the suspension was stirred at 50 °C for 2.5h, the solution became clear, and LC-MS showed the reaction to remove the acetonide protection group was complete. The solvent was removed via evaporation under reduced pressure. The resulting crude material was used directly in the next step without further purification. Thus, the aqueous NaOH (5 mL, 1. OM) was added to a solution of this crude material in MeOH (7 m L). The solution was stirred at 25 °C for 1 h, and LC-MS showed the reaction to remove the t-butyl ester was complete. This solution was directly subjected to preparative HPLC to obtain the final title compound (130 mg, 72%). LC-MS: single peak at 254 nm, MH+ calcd. for the acid C23H22FN506 : 484, obtained: 484. 1HNMR (CD30D, 400 MHz), 5 8.12 (s, 1H), 7.73 (dd, J = 2. 0 Hz, J = 8. 8 Hz, 1 H), 7.63 (d, J = 8.4 Hz, 1H), 7.54 (m, 1 H), 7.14 (d, J = 8. 0 Hz, 1H), 6. 98 (dd, J= 8.0 Hz, J= 14.4 Hz, 1H), 4.19 (m, 1H), 4.05 (m, 1H), 3.55 (dd, J = 5. 2 Hz, J = 13. 2 Hz, 1 H), 3.46 (dd, J = 6. 8 Hz, J = 13. 2 Hz, 1 H), 2.41 (dd, J = 5. 6 Hz, J = 15. 2 Hz, 1 H), 2.32 (dd, J = 8. 0 Hz, J = 15. 4 Hz, 1 H), 1.73 (m, 2H).

Example 2 : (3R, 5R)-7-{[2-(4-Amino-5-fluoro-2-oxo-1,2-dihydro-quinolin-3- yl)-3H-benzoimidazole-5-carbonyl]-amino}-3,5-dihydroxy-hepta noic acid, sodium salt.

The preparation of the title compound was carried out following the procedure for Example 1.

Preparation of compound 2-A (n=1). The amine (0. 15g, 0.55 mmol) was added to a solution of compound 1-E (0.14g, 0.41 mmol), EDC (0.6 mmol), HOBt (0.6 mmol), and DIEA (1 mmol) in DMF (2 mL). After the solution was stirred at 25 °C for 2h, DMF was removed via evaporation under reduced pressure. The resulting residue was suspended in ethyl acetate (150 mL), washed by saturated NaHCO3 (3x) and brine (3x), and dried over Na2SO4. The ethyl acetate was removed under reduced pressure to give the crude product. This crude material was subjected to flash chromatography (ISCO system, 1 %-5% MeOH in DCM, 22 min. ) to obtain the product 2-A (170mg, 69%). LC-MS: single peak at 254 nm, MH+ calcd. for C31H36FN506 : 594, obtained: 594. 1H-- NMR (DMSO-d6, 400 MHz), # 13.09 (d, J = 6.4 Hz, 1 H), 11.64 (d, J = 5.2 Hz, 1H), 11.35 (s, 1H), 8.35 (m, 1H), 8.16 (d, J= 14.0 Hz, 1H), 7.92 (t, J= 12.8 Hz, 1H), 7.72 (m, 1H), 7.64 (m, 1H), 7.57 (m, 1H), 7.19 (d, J= 8.0 Hz, 1H), 7.05 (dd, J= 8. 0 Hz, J= 14. 0 Hz, 1H), 4.20 (m, 1H), 4.00 (m, 1H), 3.59 (t, J= 5.6 Hz, 1 H), 3.30 (m, buried in water signal, 1 H), 2. 38 (dd, J = 4.8 Hz, J = 15.2 Hz, 1 H), 2.22 (dd, J = 8. 0 Hz, J = 14. 8 Hz, 1 H), 1.60 (m, 3H), 1.40 (s, 3H), 1.38 (s, 9H), 1.25 (s, 3H), 1.10 (dd, J = 12.0 Hz, J = 24.0 Hz, 1 H).

Preparation of compound 2-B (n=1). Aqueous HCI (4 mL, 1. OM) was added to a solution of compound 2-A (157 mg, 0.27 mmol) in MeOH (4 mL). A precipitation was observed immediately. After the suspension was stirred at 50 °C for 0.5h, the solution became clear, and LC-MS showed the reaction to remove the acetonide protection group was complete. The solvent was removed via evaporation under reduced pressure. The resulting crude material was used directly in the next step without further purification. Thus, the aqueous NaOH (5 mL, 1. OM) was added to a solution of this crude material in MeOH (7 mL). The solution was stirred at 25 °C for 1 h, and LC-MS showed the reaction to remove the t-butyl ester was complete. This solution was directly subjected to preparative HPLC to obtain the final title compound (135 mg, 96%). LC-MS: single peak at 254 nm, MH calcd. for the acid C24H24FN506 : 496, obtained: 496. 1HNMR (CD30D, 400 MHz), 6 8.10 (s, 1 H), 7.70 (d, J = 8. 8 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 7.52 (m, 1H), 7.13 (d, J = 8. 4 Hz, 1 H), 6.97 (dd, J = 8.0 Hz, J = 13.2 Hz, 1 H), 4.16 (m, 1 H), 3.95 (m, 1 H), 3.60 (m, 1 H), 3.53 (m, 1 H), 2.35 (m, 2H), 1.88 (m, 2H), 1.72 (m, 2H).

Examples 3 and 4. The preparation of Examples 3 and 4 followed Scheme 3 shown below : Scheme 3 Preparation of Example 3. Methyl 4-amino-3-hydroxylbutyrate (0.1g, 0.7 mmol) was added to a solution of compound 1-E (0.18g, 0.53 mmol), EDC (1 mmol), HOBt (1 mmol), and DIEA (2 mmol) in DMF (4 mL). After the solution was stirred at 25 °C for 1 h, DMF was removed via evaporation under reduced pressure. The resulting residue was suspended in ethyl acetate (100 mL), washed by saturated NaHCO3 (3x) and brine (3x), and dried over Na2SO4. The ethyl acetate was removed under reduced pressure to give 240 mg crude product. A portion (60 mg) of this crude material was subjected to preparative HPLC to obtain the final product, 4- { [2- (4-Amino-5-fluoro-2-oxo-1, 2-dihydro- quinolin-3-yl)-3H-benzoimidazole-5-carbonyl]-amino}-3-hydrox y-butyric acid methyl ester (15mg, 85%). LC-MS: single peak at 254 nm, MH+ calcd. for C22H20FN505 : 454, obtained: 454. 1H-NMR (DMSO-d6, 400 MHz), S 13.11 (s, 1 H), 11.63 (s, 1 H), 11.35 (s, 1 H), 8.41 (dt, J = 5.6 Hz, J = 25.6 Hz, 1 H), 8.20 (d, J= 10.8 Hz, 1H), 7.91 (t, J= 13.2 Hz, 1H), 7.73 (s, 1H), 7.60 (m, 2H), 7.19 (d, J = 8.4 Hz, 1 H), 7.05 (dd, J = 8.0 Hz, J = 14.0 Hz, 1 H), 5.13 (d, J = 5.6 Hz, 1 H), 4.09 (m, 1 H), 3.57 (s, 3H), 3.29 (m, buried in water signal, 2H), 2.56 (dd, J = 4. 4 Hz, J = 16.- Hz, 1H), 2.31 (dd, J = 9.2 Hz, J = 14.4 Hz, 1H).

Preparation of Example 4. LiOH (1 mmol) in H20 (10 mL) was added to a solution of compound 3 (180 mg, 0.4 mmol) in MeOH (10 mL). After the suspension was stirred at 25 °C overnight, the MeOH was removed via evaporation under reduced pressure. The resulting aqueous solution was directly subjected to preparative HPLC to obtain the final pure product 4, 4- { [2- (4-Amino-5-fluoro-2-oxo-1, 2-dihydro-quinolin-3-yi)-3H-benzoimidazole-5- carbonyl]-amino}-3-hydroxy-butyric acid (135 mg, 96%). LC-MS: single peak at 254 nm, MH+ calcd. for the acid C2iHi8FNs05 : 440, obtained: 440. 1H-NMR (DMSO-d6, 400 MHz), õ 13. 05 (s, 1 H), 11.35 (s, 1 H), 8.60 (dt, J = 5.6 Hz, J = 60.0 Hz, 1H), 8.50 (s, 1H), 8.19 (d, J= 16.8 Hz, 1H), 7.90 (t, J= 12.0 Hz, 1H), 7.72 (s, 1H), 7.64 (m, 2H), 7.19 (d, J = 8.4 HZ, 1H), 7.04 (dd, J = 8. 0 Hz, J = 13.6 Hz, 1 H), 3.84 (m, 1 H), 3.24 (m, buried in water signal, 2H), 2.16 (m, 1 H), 1.97 (m, 1 H).

Synthesis of Amide Compounds The amide compounds of this invention can be readily synthesized by those skilled in the art starting from the acid compound disclosed herein.

The compounds described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention.

Examples 5 and 6. The preparation of Examples 5 and 6 followed Scheme 4 shown below : F F NEZ NEZ N H EDC, HOBt, DMF < H R HN H ---- HN H n. R OH OH O amine, 25-55 C 0 O OH OH O n=0, 1 Scheme 4 An amine (3 equiv) was added to a solution of the sodium salt of a free acid (1 equiv), EDC (5 equiv), HOBt (5 equiv), and DIEA (5 equiv) in DMF. After the solution was stirred at 25 °C overnight (stirred at 55 °C for a couple of hours if necessary), DMF was removed via evaporation under reduced pressure. The resulting residue was suspended in ethyl acetate, washed by saturated NaHCO3 (3x) and brine (3x), and dried over Na2SO4. The ethyl acetate was removed under vacuum to give the crude product. This crude material was subjected to preparative HPLC to give the final product amide, which was subsequently characterized by LC-MS and NMR spectroscopy. Example 5: 2- (4-Amino-5-fluoro-2-oxo-1, 2-dihydro-quinolin-3-yl)-3H- benzoimidazole-5-carboxylic acid ( (3R, 5R) -3,5-dihydroxy-7-oxo-7- pyrrolidin-1-yl-heptyl)-amide An amount of 86 mg (90%) product was obtained after preparative HPLC from 90 mg (0.173 mmol) of the free acid sodium salt. LC-MS: single peak at 254 nm, MH+ calcd. for C28H31FN606 : 551, obtained: 551. 1H-NMR (DMSO-d6, 400 MHz), # 13.40 (s, 1H), 11.25 (s, 1H), 8. 39 (s, 1H), 8.16 (s, 1H), 7.85 (m, 1H), 7.69 (s, 2H), 7.55 (m, 1H), 7.18 (d, J= 8. 4 Hz, 1H), 7.02 (dd, J = 8.4 Hz, J = 13.2 Hz, 1H), 4.76 (s, 1H), 4.72 (s, 1H), 4.05 (m, 1H), 3.71 (m, 1H), 3.39 (m, 2H), 3.22 (m, 2H), 3.17 (m, 2H), 2.34 (m, 2H), 1.81 (m, 2H), 1.72 (m, 4H), 1.53 (m, 2H).

Example 6: 2- (4-Amino-5-fluoro-2-oxo-1, 2-dihydro-quinolin-3-yl)-3H- benzoimidazole-5-carboxylic acid ((2S, 4R) -2, 4-dihydroxy-6-oxo-6- pyrrolidin-1-yl-hexyl)-amide An amount of 33 mg (89%) product was obtained after preparative HPLC from 35 mg (0.069 mmol) of the free acid sodium salt. LC-MS: single peak at 254 nm, MH+ calcd. for C27H29FN606 : 537, obtained: 537. 1H-NMR (DMSO-d6, 400 MHz), 5 11.20 (s, 1 H), 8.30 (s, 1 H), 8. 18 (s, 1 H), 7.80-7. 60 (m, 3H), 7.52 (m, 1H), 7.17 (d, J=8. 8Hz, 1H), 6.99 (dd, J=8. 0Hz, J=13. 2Hz, 1H), 4.95 (s, 1H), 4.78 (s, 1H), 4.09 (m, 1H), 3.81 (m, 1H), 3.40 (m, 2H), 3.26 (m, 2H), 3.11 (m, 2H), 2.35 (m, 2H), 1.82 (m, 2H), 1.72 (m, 2H), 1.61 (m, 2H).

Example 7. Further amide derivatives of Examples 1 and 2.

Following the above procedures or other known procedures, the following amides can be made.

Example 8. Amide derivatives of Example 4.

Following known procedures, the following amide derivatives of Example 4 can be made by those skilled in the art.

Examples 9-15: Examples 9-15 are illustrated by the general structures below. Example 9: Compounds 9a-I where R=OH can be made by those skilled in the art based on known procedures.

Example 10: Compounds 10a-I where R=diethylamine can be made by those skilled in the art based on known procedures.

Examples 11: Compounds 11a-I where R=dimethylamine can be made by those skilled in the art based on known procedures.

Examples 12: Compounds 12a-I where R=pyrrolidine can be made by those skilled in the art based on known procedures.

Examples 13: Compounds 13a-I where R=morpholine can be made by those skilled in the art based on known procedures.

Examples 14: Compounds 14a-I where R=ethylamine can be made by those skilled in the art based on known procedures.

Examples 15: Compounds 15a-I where R=cyclopropylamine can be made by those skilled in the art based on known procedures.

Examples 16-315: Still further amide examples are shown in the following table: F NH2 N 0 F NHz N - I N N H u"H CORE O I N 0 H HO fi0 R I N O H H° /R "CORE II O H H 0 H 0 ° o CORE 11 CORE III OH O I \ \ \ \ N CORE IV R H fiOHO R I (J p H O R CORE V o"CORE VI Ex# Core R Ex# Core R Ex# Core R 16 I a 66 II a 116 III a 17 I b 67 II b 117 III b 18 I c 68 II c 118 III c 19 I d 69 II d 119 III d 20 I e 70 II e 120 III e 21 I f 71 II f 121 III f 22 I g 72 II g 122 III g 23 I h 73 II h 123 III h 24 I i 74 II i 124 III i 25 I j 75 H j 125 III j 26 I k 76 II k 126 III k 27 I 1 77 H 1 127 III 1 28 I m 78 II m 128 III m 29 I n 79 II n 129 III n 30 I o 80 H o 130 in o Ex# Core R Ex# Core R Ex# Core R 31 I p 81 II p 131 III p 32 I q 82 II q 132 III q 33 I r 83 II r 133 III r 34 I s 84 II s 134 III s 35 I t 85 II t 135 III t 36 I u 86 II u 136 III u 37 I v 87 II v 137 III v 38 I w 88 II w 138 III w 39 I x 89 II x 139 III x 40 I y 90 II y 140 III y 40 I z 91 II z 141 III z 42 I aa 92 II aa 142 III aa 43 I ab 93 II ab 143 III ab 44 I ac 94 II ac 144 III ac 45 I ad 95 II ad 145 III ad 46 I ae 96 II ae 146 III ae 47 I af 97 II af 147 III af 48 I ag 98 II ag 148 III ag 49 I ah 99 II ah 149 III ah 50 I ai 100 II ai 150 III ai 51 I aj 101 II aj 151 in aj 52 I ak 102 II ak 152 III ak 53 I al 103 II al 153 III al 54 I am 104 II am 154 III am 55 I an 105 II an 155 III an 56 I ao 106 II ao 156 III ao 57 I ap 107 II ap 157 III ap 58 I aq 108 II aq 158 III aq 59 I ar 109 II ar 159 III ar 60 I as 110 II as 160 III as 61 I at 111 II at 161 III at 62 I au 112 II au 162 III au 63 I av 113 II av 163 III av 64 I aw 114 II aw 164 III aw 65 I ax 115 II ax 165 III ax Ex# Core R Ex# Core R Ex# Core R <BR> <BR> <BR> <BR> <BR> <BR> 166 IV a 216 V a 266 VI a 167 IV b 217 V b 267 VI b 168 IV c 218 V c 268 VI c 169 IV d 219 V d 269 VI d 170 IV e 220 V e 270 VI e 171 IV f 221 V f 271 VI f 172 IV g 222 V g 272 VI g 173 IV h 223 V h 273 VI h 174 IV i 224 V i 274 VI i 175 IV j 225 V j 275 VI j 176 IV k 226 V k 276 VI k 177 IV 1 227 V 1 277 VI 1 178 IV m 228 V m 278 VI m 179 IV n 229 V n 279 VI n 180 IV o 230 V o 280 VI o 181 IV p 231 V p 281 VI p 182 IV q 232 V q 282 VI q 183 IV r 233 V r 283 VI r 184 IV s 234 V s 284 VI s 185 IV t 235 V t 285 VI t 186 IV u 236 V u 286 VI u 187 IV v 237 V v 287 VI v 188 IV w 238 V w 288 VI w 189 IV x 239 V x 289 VI x 190 IV y 240 V y 290 VI y 191 IV z 241 V z 291 VI z 192 IV aa 242 V aa 292 VI aa 193 IV ab 243 V ab 293 VI ab 194 IV ac 244 V ac 294 VI ac 195 IV ad 245 V ad 295 VI ad 196 IV ae 246 V ae 296 VI ae 197 IV af 247 V af 297 VI af 198 IV ag 248 V ag 298 VI ag 199 IV ah 249 V ah 299 VI ah 200 IV ai 250 V ai 300 VI ai 201 IV aj 251 V aj 301 VI aj 202 IV ak 252 V ak 302 VI ak 203 IV al 253 V al 303 VI al 204 IV am 254 V am 304 VI am 205 IV an 255 V an 305 VI an 206 IV ao 256 V ao 306 VI ao 207 IV ap 257 V ap 307 VI ap 208 IV aq 258 V aq 308 VI aq 209 IV ar 259 V ar 309 VI ar 210 IV as 260 V as 310 VI as 211 IV at 261 V at 311 VI at Ex# Core R Ex# Core R Ex# Core R 212 IV au 262 V au 312 VI au 213 IV av 263 V av 313 VI av 214 IV aw 264 V aw 314 VI aw 215 IV ax 265 V ax 315 VI ax In the above table, R is selected from the following radicals : These amide examples 16-315 can be made by those skilled in the art following the above procedure and/or known procedures.

Example 316 : (3R, 5S)-3, 5-Dihydroxy-6-[2-(2-oxo-1, 2-dihydro-quinolin-3-yl)- 1 H-indol-5-yloxy]-hexanoic acid The title compound can be made by those skilled in the art following known procedures.

Example 317 : 3-Hydroxy-4-[2-(2-oxo-1, 2-dihydro-quinolin-3-yl)-1H-indol-5- yloxy]-butyric acid The title compound can be made by those skilled in the art following known procedures.

VEGFR Biochemical Assay The compounds were assayed for biochemical activity by Upstate Ltd at Dundee, United Kingdom, according to the following procedure. In a final reaction volume of 25 pI, KDR (h) (5-10 mU) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.33 mg/ml myelin basic protein, 10 mM MgAcetate and [y- 33P-ATP] (specific activity approx. 500 cpm/pmol, concentration as required).

The reaction is initiated by the addition of the MgATP mix-After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of 5 lli of a 3% phosphoric acid solution. 10 ßli of the reaction is then spotted onto a P30 filtermat and washed three times for 5 minutes in 75 rnM phosphoric acid and once in methanol prior to drying and scintillation counting.

Compounds of the present invention were tested in this assay and exhibited IC50 between 1-5,000 nM.

Cellular Assay : HUVEC: VEGF induced proliferation The compounds were assayed for cellular activity in the VEGF induced proliferation of HUVEC cells. HUVEC cells (Cambrex, CC-2517) were maintained in EGM (Cambrex, CC-3124) at 37°C and 5% CO2. HUVEC cells were plated at a density 5000 cells/well (96 well plate) in EGM. Following cell attachment (1hour) the EGM-medium was replaced by EBM (Cambrex, CC- 3129) + 0. 1 % FBS (ATTC, 30-2020) and the cells were incubated for 20 hours at 37°C. The medium was replaced by EBM +1% FBS, the compounds were serial diluted in DMSO and added to the cells to a final concentration of 0- 5,000 nM and 1% DMSO. Following a 1 hour pre-incubation at 37°C cells were stimulated with 10ng/ml VEGF (Sigma, V7259) and incubated for 45 hours at 37°C. Cell proliferation was measured by BrdU DNA incorporation for 4 hours and BrdU label was quantitated by ELISA (Roche kit, 16472229) using 1 M H2SO4 to stop the reaction. Absorbance was measured at 450nm using a reference wavelength at 690nm.