WO2011067348A2

WO2011067348A2 - Mek inhibitor salts and solvates thereof

Info

Publication number: WO2011067348A2
Application number: PCT/EP2010/068763
Authority: WO
Inventors: Liladhar Murlidhar Waykole; Piotr H. Karpinski
Original assignee: Novartis AG
Current assignee: Novartis AG
Priority date: 2009-12-03
Filing date: 2010-12-02
Publication date: 2011-06-09
Anticipated expiration: 2012-06-03
Also published as: WO2011067348A3

Abstract

The present invention relates to sodium salt forms of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-1,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide and 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- 1,2,3,5-tetrahydro- indolizine-8-carboxylic acid cyclopropylmethoxy-amide and their corresponding solvates and polymorphs, which are useful in the treatment of a disease, disorder or syndrome associated with MEK inhibition, such as cancer, in mammals.

Description

MEK INHIBITOR SALTS AND SOLVATES THEREOF

FIELD OF THE INVENTION

The present invention relates to salt forms of 7-(4-Bromo-2-fluoro-phenylamino)-6- methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy) -amide and 7- (4-Br omo - 2-fluor o-phenyl mino)-5-oxo-l,2,3,5 -tetrahydr o-indoliz ine- 8 - carboxyl ic acid cyclopropylmethoxy-amide and their corresponding solvates and polymorphs, which are useful in the treatment of a disease, disorder or syndrome associated with MEK inhibition, such as cancer, in mammals. The invention also relates to compositions thereof and to methods of using such compositions in the treatment of cancer in mammals, especially humans.

BACKGROUND

The compound 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro- indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide (referred to herein as "Compound 1") and 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- 1,2,3, 5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide (referred to herein as "Compound 2") are potent MEK inhibitors. The crystallinity, polymorphic behavior, and hygroscopicity of the compound and its pharmaceutical salt can have a significant effect on both stability and solubility of the compound or salt. In the development of a useful and effective pharmaceutical composition, the toxicological considerations, crystallinity, monomorphism, melting point, stiochiometry, hydgroscopicity, stability in bulk, compatibility with excipients, pH of aqueous solutions, solubility in water and aqueous media, formation of solvates, morphology, handling, polymorphism behavior are all taken into consideration which is unique to a particular form of the compound and the mode of administration to a patient. Not all forms of the compound provide useful formulations, thus there is a need to identify specific forms of Compound 1 and 2 which can be used to provide a useful and stable pharmaceutical composition for use in treating diseases, conditions or disorders associated with MEK inhibition.

SUMMARY

The present invention provides sodium salts of 7-(4-Bromo-2-fluoro-phenylamino)-6- methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy) -amide and 7- (4-Br omo - 2-fluor o-phenyl amino)-5-oxo-l,2,3,5 -tetrahydr o-indoliz ine- 8 - carboxyl ic acid cyclopropylmethoxy-amide which have higher aqueous solubility, higher dissolution rate and good photo stability as compared to their corresponding free form.

In one embodiment of the present invention, a sodium salt of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide is provided. Preferably, the salt is a 1 : 1 ratio of sodium to Compound 1.

In another aspect of the present invention, a crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide, sodium salt ("Form 1 A") is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 1.

Also provided is a crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl- 5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form 1 A") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 7.5°, 10.5°, 14.2°, 15.2°, 16. P, 17.8°, 19.9°, 20.3°, 22.8°, 23.8°, 24.2°, 24.8°, 25.9°, 27.0°, 28.6°, and 30.1°.

Preferably, the crystalline form is substantially pure. The sodium to Compound 1 ratio is preferably 1: 1.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline form of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1,3,5- tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form 1 A") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 7.5°, 10.5°, 14.2°, 15.2°, 16. 1°, 17.8°, 19.9°, 20.3°, 22.8°, 23.8°, 24.2°, 24.8°, 25.9°, 27.0°, 28.6°, and 30. 1°; and a pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutical composition may optionally comprise at least one additional pharmaceutical agent described herein below (preferably, the additional pharmaceutical agent is selected from an anti-tumor or an anti-proliferative agent). A preferred additional pharmaceutical agent is a mTOR inhibitor or a PI3K inhibitor.

In yet another embodiment, the present invention provides a method of treating cancer in a mammal comprising administering to a mammal (preferably a human) in need thereof a therapeutic amount of a crystalline form of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5- oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form 1 A") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 7.5°, 10.5°, 14.2°, 15.2°, 16.1°, 17.8°, 19.9°, 20.3°, 22.8°, 23.8°, 24.2°, 24.8°, 25.9°, 27.0°, 28.6°, and 30.1°, or a pharmaceutical composition thereof. The sodium salt may be administered either simultaneously or sequentially with at least one additional pharmaceutical agent described herein below (preferably, a mTOR inhibitor or a PI3K inhibitor).

In yet another aspect of the present invention, a crystalline form of 7-(4-Bromo-2- fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide, sodium salt ("Form IB") is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 3.

Also provided is a crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl- 5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form IB") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 10.5°, 14.3°, 15. 1°, 18.5°, 19.0°, 20.2°, 21.3°, 23.1°, 24.3°, 24.8°, 25.3°, 25.8°, 26.4°, 27.2°, 28.2°, 28.4°, 29.2°, 30.0°, 31.1°, 32. P, 33.1°, 33.7°, 34.9°, and 36.3°.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline form of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1,3,5- tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form IB") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 10.5°, 14.3°, 15. 1°, 18.5°, 19.0°, 20.2°, 21.3°, 23. P, 24.3°, 24.8°, 25.3°, 25.8°, 26.4°, 27.2°, 28.2°, 28.4°, 29.2°, 30.0°, 31.1°, 32.1°, 33.1°, 33.7°, 34.9°, and 36.3°.; and a

pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutical composition may optionally comprise at least one additional pharmaceutical agent described herein below (preferably, the additional pharmaceutical agent is selected from an anti-tumor or an antiproliferative agent). A preferred additional pharmaceutical agent is a mTOR inhibitor or a PI3K inhibitor.

In yet another embodiment, the present invention provides a method of treating cancer in a mammal comprising administering to a mammal (preferably a human) in need thereof a therapeutic amount of a crystalline form of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5- oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form IB") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 10.5°, 14.3°, 15. 1°, 18.5°, 19.0°, 20.2°, 21.3°, 23.1°, 24.3°, 24.8°, 25.3°, 25.8°, 26.4°, 27.2°, 28.2°, 28.4°, 29.2°, 30.0°, 31.1°, 32.1°, 33.1°, 33.7°, 34.9°, and 36.3°., or a pharmaceutical composition thereof. The sodium salt may be administered either simultaneously or sequentially with at least one additional pharmaceutical agent described herein below (preferably, a mTOR inhibitor or a PI3K inhibitor).

The present invention also provides a sodium salt of 7-(4-Bromo-2-fluoro- phenylamino)-5-oxo- l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy- amide. Preferably, the salt is a 1 : 1 ratio of sodium to Compound 2.

In yet another aspect of the present invention, a crystalline form of 7-(4-Bromo-2- fluoro-phenylamino)- 5 -oxo- 1 , 2,3 , 5 -tetrahydro-indolizine- 8-carboxylic acid

cyclopropylmethoxy-amide, sodium salt is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 5.

Also provided is a crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt ("Form 2A") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 6.3°, 6.9°, 7.6°, 10.2°, 13.2°, 20.7°, 21.6°, 23.8°, 24.7°, 26.8°, 27.9°, 30.0°, and 32.6°.

Preferably, the crystalline form is substantially pure. The sodium to Compound 2 ratio is preferably 1 : 1.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- l,2,3, 5- tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 6.3°, 6.9°, 7.6°, 10.2°, 13.2°, 20.7°, 21.6°, 23.8°, 24.7°, 26.8°, 27.9°, 30.0°, and 32.6°; and a

pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutical composition may optionally comprise at least one additional pharmaceutical agent described herein below (preferably, the additional pharmaceutical agent is selected from an anti-tumor or an anti- proliferative agent). A preferred additional pharmaceutical agent is a mTOR inhibitor or a PI3K inhibitor.

In yet another embodiment, the present invention provides a method of treating cancer in a mammal comprising administering to a mammal (preferably a human) in need thereof a therapeutic amount of a crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 6.3°, 6.9°, 7.6°, 10.2°, 13.2°, 20.7°, 21.6°, 23.8°, 24.7°, 26.8°, 27.9°, 30.0°, and 32.6°, or a pharmaceutical composition thereof. The sodium salt may be administered either simultaneously or sequentially with at least one additional pharmaceutical agent described herein below (preferably, a mTOR inhibitor or a PI3K inhibitor).

In another aspect of the present invention, an acetone solvate of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 7.

Also provided is a crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, acetone solvate ("2A- ACT") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (20) of 3.2°, 7.1°, 9.8°, 10.9°, 13. P, 15.4°, 17.7°, 20. P, 21.8°, 23.5°, 25.3°, 25.8° and 26.5°.

In another aspect of the present invention, an acetonitrile solvate of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt ("2A-ACN") is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 9.

In another aspect of the present invention, an ethanol solvate of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt ("2A-ETH") is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 11.

Also provided is a crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, ethanol solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 3.2°, 7.1°, 9.8°, 10.9°, 13. P, 15.4°, 17.7°, 20.1° and 21.8°.

In another aspect of the present invention, an isopropyl acetate solvate of 7-(4-bromo- 2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8- carbonyl)(cyclopropylmethoxy)amide, sodium salt ("2A-I AC") is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 13.

Also provided is a crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, isopropyl acetate solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 6.5°, 7.0°, 7.6°, 10.4°, 13.2°, 16.6°, 18.4°, 19. P, 20.9°, 21.8°, 23.8°, 24.9°, 26.2° and 26.9°. In another aspect of the present invention, an isopropanol solvate of -(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8- carbonyl)(cyclopropylmethoxy)amide, sodium salt ("2A-ISO")having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 15.

Also provided is a crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, isopropanol solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 6.6°, 8.4°, 9. P, 9.4°, 13. F, 14.2°, 15.7°, 20.3°, 24. P and 25.7°.

In another aspect of the present invention, a tetrahydrofuran solvate of -(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, -tetrahydroindolizine-8- carbonyl)(cyclopropylmethoxy)amide, sodium salt is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 17.

Also provided is a crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, tetrahydrofuran solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 3.2°, 7.2°, 7.7°, 19. 1°, 21.4°, 22.0°, 22.6°, 23. F, 23.7°, 25.3° and 26.8°.

Preferably, for each of the solvates described above, the crystalline form is substantially pure. The sodium to Compound 2 ratio is preferably 1 : 1.

In another embodiment, the present invention provides a pharmaceutical composition comprising a sodium salt of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo-l, 2,3,5- tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide or any one of the solvates thereof (i.e., 2 A- ACT, 2A-ACN, 2A-ETH, 2A-IAC. 2A-ISO or 2A-THF) and a

In yet another embodiment, the present invention provides a method of treating cancer in a mammal comprising administering to a mammal (preferably a human) in need thereof a therapeutic amount of a sodium salt of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide or any one of the solvates thereof (i.e., 2 A- ACT, 2A-ACN, 2A-ETH, 2A-IAC. 2A-ISO or 2A-THF) or a pharmaceutical composition thereof. The sodium salt may be administered either simultaneously or sequentially with at least one additional pharmaceutical agent described herein below (preferably, a mTOR inhibitor or a PI3K inhibitor).

Definitions

As used herein, the term "substantially pure" with reference to a particular polymorphic form means that the polymorphic form includes less than 10%, preferably less than 5%, more preferably less than 3%, most preferably less than 1% by weight of any other physical forms of the compound.

The term "essentially the same" with reference to X-ray diffraction peak positions means that typical peak position and intensity variability are taken into account. For example, one skilled in the art will appreciate that the peak positions (26?) will show some inter- apparatus variability, typically as much as 0.2°. Further, one skilled in the art will appreciate that relative peak intensities will show inter-apparatus variability as well as variability due to degree of crystallinity, preferred orientation, prepared sample surface, and other factors known to those skilled in the art, and should be taken as qualitative measure only.

The term "free form" refers to the compound per se without salt formation or association with a solvent (e.g., solvate).

The term "solvate" refers to a molecular complex of the compound with one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical art, which are known to be innocuous to the recipient, e.g., water, acetone, acetonitrile, ethanol, isopropanol, isopropyl acetate, tetrahydrofuran, and the like. The term "hydrate" refers to the complex where the solvent molecule is water.

The term "compounds of the present invention", unless indicated otherwise, refers to the sodium salts of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro- indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide and (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)-

(cyclopropylmethoxy)amide, the crystalline forms of 7-(4-bromo-2-fluoro-phenylamino)-6- methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy) -amide, sodium salt (Form 1 A and Form IB) and (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)-(cyclopropylmethoxy)amide, sodium salt (2A) including their corresponding hydrate or solvate(s) thereof.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows a powder X-ray diffraction pattern of the crystalline form of 7-(4- Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form 1 A").

Figure 2 shows a TGA-DTA thermogram of the "Form 1A" crystalline form of 7-(4- Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt.

Figure 3 shows a powder X-ray diffraction pattern of the crystalline form of 7-(4- Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt ("Form IB").

Figure 4 shows a TGA-DTA thermogram of the "Form IB" crystalline form of 7-(4-

Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt.

Figure 5 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt ("Form 2 A").

Figure 6 shows a TGA-DTA thermogram of the "Form 2A" crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt.

Figure 7 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, acetone solvate ("Form 2A-ACT").

Figure 8 shows a TGA-DTA thermogram of the "Form 2A- ACT" crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, acetone solvate.

Figure 9 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, acetonitrile solvate ("Form 2A-ACN").

Figure 10 shows a TGA-DTA thermogram of the "Form 2A-ACN" crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, acetonitrile solvate.

Figure 11 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, ethanol solvate ("Form 2A-ETFF '). Figure 12 shows a TGA-DTA thermogram of the "Form 2A-ETH" crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, ethanol solvate.

Figure 13 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, isopropyl acetate solvate ("Form 2A-IAC").

Figure 14 shows a TGA-DTA thermogram of the "Form 2A-IAC" crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, isopropyl acetate solvate.

Figure 15 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, isopropanol solvate ("Form 2A-ISO").

Figure 16 shows a TGA-DTA thermogram of the "Form 2A-ISO" crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, Isopropanol solvate.

Figure 17 shows a powder X-ray diffraction pattern of the crystalline form of (7-(4- bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, tetrahydofuran solvate ("Form 2A-THF").

Figure 18 shows a TGA-DTA thermogram of the "Form 2A-THF" crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt, tetrahydrofuran solvate.

DETAILED DESCRIPTION

The characteristic 2-theta (2Θ) values, d-spacing (A) and intensity (%) for the powder X-ray diffraction (PXRD) pattern of the two crystalline forms of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide, sodium salt (as prepared in Example 1) are shown in Table 1 A ("Form 1A") (below) and Table IB ("Form IB"). The corresponding TGA-DTA

thermogram for the crystalline Form 1 A and Form IB of 7-(4-Bromo-2-fluoro-phenylamino)- 6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide, sodium salt (Compound 1) are shown in FIG. 2 and FIG. 4, respectively.

Table 1A

2-Theta d-spacing Intensity

(de_g°) (A) (%) 7.5 11.771 90.7

10.5 8.458 20.6

14.2 6.237 100.0

15.2 5.835 42.5

16.1 5.485 34.1

17.8 4.985 43.0

19.9 4.460 60.2

20.3 4.377 44.9

22.8 3.903 34.1

23.8 3.733 30.2

24.2 3.667 25.8

24.8 3.586 21.8

25.9 3.443 35.9

27.0 3.295 23.8

28.6 3. 119 35.3

30.1 2.962 20.8

Table IB

The characteristic 2-theta (2Θ) values, d-spacing (A) and intensity (%) for the powder X-ray diffraction (PXRD) pattern of the crystalline form of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt ("Form 2A" as prepared in Example 2) are shown in Table 2 (below). The corresponding TGA-DTA thermogram for the crystalline form of (7- (4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)- (cyclopropylmethoxy)amide, sodium salt ("Form 2A") is shown in FIG 6.

Table 2

The characteristic 2-theta (2Θ) values, d-spacing (A) and intensity (%) for the powder X-ray diffraction (PXRD) pattern of the crystalline solvate forms of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carbonyl)-

(cyclopropylmethoxy)amide, sodium salt (as prepared in Example 2) are shown below in Table 3 (2A-ACT), Table 4 (2A-ETH), Table 5 (2A-IAC), Table 6(2A-ISO) and Table 7(2A-THF). The acetonitrile solvate (2A-ACN) is less crystalline and more amorphous in character than the other solvates. The corresponding TGA-DTA thermogram for the solvate forms of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8- carbonyl)-(cyclopropylmethoxy)amide, sodium salt are shown in figures: FIG. 8 (2 A- ACT), FIG. 10(2A-ACN), FIG. 12 (2A-ETH), FIG. 14(2A-IAC), FIG. 16(2A-ISO) and FIG. 18 (2A-THF).

Table 3

13.1 6.730 56.9

15.4 5.762 52.3

17.7 4.997 52.3

20.1 4.423 96.9

21.8 4.072 59.4

23.5 3.787 66.3

25.3 3.524 64.0

25.8 3.454 63.5

26.5 3.355 82.7

Table 4

Table 5

Table 6

9.1 9.690 100.0

9.4 9.422 66.0

13.1 6.749 62.4

14.2 6.237 68.2

15.7 5.657 49.7

20.3 4.370 55.1

24.1 3.682 35.6

25.7 3.470 42.3

Table 7

The free form of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5- tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide can be synthesized using the procedures outlined in Scheme I below or by the methods disclosed in US Publication No. 2009/0124595, incorporated herein by reference.

Scheme I

For a detailed description of the process steps outlined in Scheme I, see the Examples section below.

Alternatively, Compound 1 may be prepared using the synthesis outlined in Scheme II below.

Scheme II

For a detailed description of the process steps outlined in Scheme II, see the Examples section below.

The free base form of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- 1,2,3, 5-tetrahydro- indolizine-8-carboxylic acid cyclopropylmethoxy-amide can be synthesized using the procedures outlined in Scheme III below or by the methods disclosed in US Publication No. 2009/0124595, incorporated herein by reference.

Scheme III

In general, compounds of the invention will be administered in therapeutically effective amounts via any of the usual and acceptable modes known in the art, either singly or in combination with one or more therapeutic agents. A therapeutically effective amount may vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used and other factors. In general, satisfactory results are indicated to be obtained systemically at daily dosages of from about 0.03 to 2.5mg/kg per body weight. An indicated daily dosage in the larger mammal, e.g. humans, is in the range from about 0.5mg to about lOOmg, conveniently administered, e.g. in divided doses up to four times a day or in retard form. Suitable unit dosage forms for oral administration comprise from ca. 1 to 50 mg active ingredient.

The pharmaceutical formulations may be prepared using conventional dissolution and mixing procedures. For example, the bulk drug substance (i.e., compound of the present invention is dissolved in a suitable solvent in the presence of one or more of the excipients described below. The compound of the present invention is typically formulated into pharmaceutical dosage forms to provide an easily controllable dosage of the drug and to give the patient an elegant and easily handleable product.

Compounds of the invention can be administered as pharmaceutical compositions by any conventional route, in particular enterally, e.g., orally, e.g., in the form of tablets or capsules, or parenterally, e.g., in the form of injectable solutions or suspensions, topically, e.g., in the form of lotions, gels, ointments or creams, or in a nasal or suppository form.

Pharmaceutical compositions comprising a compound of the present invention in association with at least one pharmaceutically acceptable carrier or diluent can be manufactured in a conventional manner by mixing, granulating or coating methods. For example, oral compositions can be tablets or gelatin capsules comprising the active ingredient together with a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; b) lubricants, e.g., silica, talcum, stearic acid, its magnesium or calcium salt and/or

polyethyleneglycol; for tablets also c) binders, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and or

polyvinylpyrrolidone; if desired d) disintegrants, e.g., starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and/ or e) absorbents, colorants, flavors and sweeteners. Injectable compositions can be aqueous isotonic solutions or suspensions, and suppositories can be prepared from fatty emulsions or suspensions. The compositions may be sterilized and/ or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances. Suitable formulations for transdermal applications include an effective amount of a compound of the present invention with a carrier. A carrier can include absorbable pharmacologically acceptable solvents to assist passage through the skin of the host. For example, transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound to the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin. Matrix transdermal formulations may also be used. Suitable formulations for topical application, e.g., to the skin and eyes, are preferably aqueous solutions, ointments, creams or gels well-known in the art. Such may contain solubilizers, stabilizers, tonicity enhancing agents, buffers and preservatives.

In some therapies, it may be advantageous to administer the compounds of the invention in combination with one or more therapeutic agents (pharmaceutical combinations). For example, synergistic effects can occur with other anti-tumor or anti-proliferative agents, for example, mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors (e.g., trastuzumab, panitumumab, cetuximab, gefitinib, erlotinib, lapatinib, sorafenib, etc.), cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxics, anti-hormones, anti-androgens, an anti- angiogenesis agent, kinase inhibitor, pan kinase inhibitor or growth factor inhibitor. Suitable therapeutic agents include erlotinib, docetaxel, gemcitabine, cisplatin, carboplatin, paclitaxel, bevacizumab, trastuzumab, pertuzumab, temozolomide, taxoxifen, doxorubicin, rapamycin and lapatnib. Other suitable therapeutic agents are listed in the Physicians Desk Reference.

Preferred therapeutic agents for combination therapy include mTOR inhibitors (e.g., Rapamycin (sirolimus), TORISEL™(temsirolimus), RADOOl (everolimus),

AP23573(deforolimus), OSI-027(OSI Pharmaceuticals), compounds described in WO 06/090167; WO 06/090169; WO 07/080382, WO 07/060404; and WO08/023161): and

PI3K inhibitors (e.g., wortmannin, 17-hydroxywortmannin analogs described in WO 06/044453, 4-(2-(lH-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-l-yl)methyl)thieno- [3,2- d]pyrimidin-4-yl)morpholine, (S)- l-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4- mo holinothieno[3,2-d] yrimidin-6-yl)methyl)piperazin-l-yl)-2-hydroxypropan-l-one, 4- (2-(lH-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin- l-yl)methyl)thieno- [2,3-d]pyrimidin- 4-yl)morpholine, LY294002(2-(4-Morpholinyl)-8-phenyl-4H- l-benzopyran-4-one available from Axon Medchem), PI 103 hydrochloride (3-[4-(4-Μοφ^1ί^1ρνηάο[3',2^4,5]ΓΐΐΓθ[3,2- d]pyrimidin-2-yl]phenol hydrochloride available from Axon Medchem), PIK 75 (N'-[(1E)- (6-bromoimidazo[l,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfono- hydrazide hydrochloride available from Axon Medchem), PIK 90 (N-(7,8-dimethoxy-2,3- dihydro-imidazo[l,2-c]quinazolin-5-yl)-nicotinamide available from Axon Medchem), GDC- 0941 bismesylate (-( lH-Indazol-4-yl)-6-(4-methanesulfonyl-piperazin-lylmethyl)-4- morpholin-4-yl-thieno[3,2-d]pyrimidine bismesylate available from Axon Medchem), BEZ235 (2-Methyl-2-[4-(3-methyl-2-oxo-8-quinolin-3-yl-2,3-dihydro-imidazo[4,5- c]quinolin-l-yl)-phenyl]-propionitrile available from Axon Medchem), AS-252424 (5-[l-[5- (4-Fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione available from Axon Medchem), and TGX-221 (7-Methyl-2-(4-morpholinyl)-9-[l- (phenylamino)ethyl]-4H-pyrido-[l,2-a]pyrimidin-4-one available from Axon Medchem), XL- 765, and XL- 147.

Where the compounds of the invention are administered in conjunction with other therapies, dosages of the co-administered compounds will of course vary depending on the type of co-drug employed, on the specific drug employed, on the condition being treated and so forth.

According to the methods of the invention, a compound of the present invention or a combination of a compound of the present invention and at least one additional

pharmaceutical agent is administered to a subject in need of such treatment, preferably in the form of a pharmaceutical composition. In the combination aspect of the invention, the compound of the present invention and at least one other pharmaceutical agent (described above) may be administered either separately or in the pharmaceutical composition comprising both. It is generally preferred that such administration be oral. However, if the subject being treated is unable to swallow, or oral administration is otherwise impaired or undesirable, parenteral or transdermal administration may be appropriate.

According to the methods of the invention, when a combination of a compound of the present invention and at least one other pharmaceutical agent are administered together, such administration can be sequential in time or simultaneous with the simultaneous method being generally preferred. For sequential administration, a compound of the present invention and the additional pharmaceutical agent can be administered in any order. It is generally preferred that such administration be oral. It is especially preferred that such administration be oral and simultaneous. When a compound of the present invention and the additional pharmaceutical agent are administered sequentially, the administration of each can be by the same or by different methods.

The pharmaceutical composition (or formulation) for application may be packaged in a variety of ways depending upon the method used for administering the drug. Generally, an article for distribution includes a container having deposited therein the pharmaceutical formulation in an appropriate form. Suitable containers are well-known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, ampoules, plastic bags, metal cylinders, and the like. The container may also include a tamper-proof assemblage to prevent indiscreet access to the contents of the package. In addition, the container has deposited thereon a label that describes the contents of the container. The label may also include appropriate warnings. The invention also provides for a pharmaceutical combinations, e.g. a kit, comprising a) a first agent which is a compound of the invention as disclosed herein, and b) at least one additional therapeutic agent. The kit may also include instructions for its administration.

The terms "co-administration" or "combined administration" or the like as utilized herein are meant to encompass administration of the selected therapeutic agents to a single patient, and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.

The term "pharmaceutical combination" as used herein means a product that results from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients. The term "fixed combination" means that the active ingredients, e.g. a compound of the present invention and a co-agent, are both administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredients, e.g. a compound of the present invention and a co-agent, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the 2 compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of 3 or more active ingredients.

EXAMPLES

The samples prepared in the Examples below were evaluated using the following instrumentation and procedures.

Thermogravimetric analysis/differential thermal analysis (TGA)fDTA) : TA

Instruments; Model No. 2960 SDT V3.0F and Processing Software; Universal V3 9A TA Instruments.

X-ray powder diffraction (XRPD) : Bruker AXS Discover D8, Madison, WI, USA

Hyproscopicity: Sorption/desorption isotherms were measured at 25 °C using Surface Measurement Systems Ltd's dynamic vapor sorption instrument (DVS-HT).

Polymorphism Behavior: A suspension of the drug substance in each solvent was agitated for 24 hours at 25°C and then tested by XRPD and TGA-DTA

Effect of Compaction: Approximately 100 mg of test sample was compressed with 5000 pounds in a steel die, the tablet diameter of 8 mm. Samples were analyzed by XRPD.

Effect of grinding: Approximately 100 mg of the test sample was ground in a mortar and analyzed by XRPD.

The acrynoms listed below and used in the examples have the following

corresponding meanings:

PyBOP (Benzotriazole- 1 -yl- oxy-tr is -(di methylamino)- pho sphonium

hexafluorophosphate)

HOBt Hydroxybenzotriaz ole EDCI l-Ethyl-3-(3-Dimethylaminopropyl)carbodiimide

TEA Triethylamine

BINAP 2,2'-Bis (Diphenylphosphino)- 1, Γ-Binaphthyl

Preparation of the free base of 7-(4-bromo-2-fluorophenylamino)-N-(2-hvdroxyethoxy)-6- methyl-5-oxo-l,2,3,5-tetruhydroindolizine-8-curboxamide ( Compound 1)

(Compound 1)

Step 1 : Preparation of Intermediate 7-Hydroxy-6-methyl-5-oxo- 1,2,3, 5-tetrahydro- indolizine-8-carboxylic acid ethyl ester (1-1 a):

A solution of 7-hydroxy-5-oxo- l, 2, 3, 5-tetrahydro-indolizine-8-carboxylic acid ethyl ester (SM- 1: 7.5 g, 33.6 mmol) in THF is added dropwise at 0°C to a stirred suspension of sodium hydride in THF under an inert atmosphere. The reaction mixture was allowed to warm to ambient temperature and then treated with iodomethane. The resulting reaction mixture was stirred for 2 days (TLC monitoring, 100% EtOAc) and then quenched with ice. The volatiles were removed in vacuo and the remaining aqueous phase extracted with ethyl acetate. The combined organic extracts were washed with brine and concentrated. Column chromatography of the crude product on silica gel (70% ethyl acetate in hexane) yielded 38% of the title compound.

1H NMR (DMSO-D6): δ 11.5 (s, 1H), 4.3 (q, 2H), 4.0 (t, 2H), 3.48 (t, 2H), 2.1 (q, 2H), 1.8, (s, 3H), 1.3 (t, 3H).

Step 2: Preparation of Intermediate 6-Methyl-5-oxo-7-trifluoromethanesulfonyloxy-l ,2,3,5- tetrahydro-indolizine-8-carboxylic acid ethyl ester (1-1 b):

7-hydroxy-6-methyl-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid ethyl ester (I-la: 0.75 g, 3.16 mmol) was treated with triflic anhydride (1.06 g, 3.79 mmol) after stirring for 12 hours at ambient temperature. The title compound was obtained in 40% yield after silica gel column chromatography (25% ethyl acetate in hexane). 1H NMR (DMSO-D6): 4.40 (q, 2H), 4.20 (t, 2H), 3.5 (t, 2H), 2.3-2.2 (m, 2H), 2.05 (s,3H), 1.35 ( t, 3H).

Step 3: Preparation of Intermediate 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1 ,2,3,5-tetrahydro-indolizine-8-carboxylic acid ethyl ester (I-lc):

A stirred solution of toluene (100 mL) containing 6-methyl 5-oxo-7- trifluoromethanesulfonyloxy- 1,2,3, 5-tetrahydro-indolizine-8-carboxylic acid ethyl ester (I- lb: 0.47 g, 1.26 mmol), 2-fluoro-4-bromo-aniline (0.287 g, 1.26 mmol), cesium carbonate(0.617 g, 1.89 mmol), 2,2'-bis (diphenylphosphino)- l,r-binaphthyl (BINAP)(0.6 g, 0.19 mmol), and Pd(OAc)₂ ( 0.028 g, 0. 126 mmol) was heated for 4 hours at 90°C The reaction mixture was filtered and the filtrate concentrated. The residual material was taken up in ethyl acetate and washed twice with brine, then dried (anhydrous Na₂S0₄) and concentrated. Column chromatography of the crude product on silica gel (75% ethyl acetate in hexane) afforded the title compound in 19% yield.

LC-MS purity: 96.24%, m/z 409, (M+, Br pattern).

1H NMR (DMSO-D6): 5 8.5 (s, 1H), 7.50 (d, 1H), 7.25 (d, 1H), 6.6 (t,lH), 4.2-4.0 (m, 4H), 3.45 (d, 2H), 2.2-2.1 (m, 2H), 1.70 ( t, 3H), 1.3 (t, 3H).

Step 4: Preparation of Intermediate 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1 ,2,3,5-tetrahydro-indo!izine-8-carboxyiic acid fl-ld):

An aqueous NaOH solution (1 ml, 1 N) was added to 7-(4-bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid ethyl ester (L lc: 40 mg, 0.10 mmol) in a solvent mixture of THF:MeOH (3 : 1, v/v). The resulting reaction mixture was stirred for 3 hours at room temperature. The pH of the reaction mixture was adjusted to 1.5 with IN aqueous HC1 solution and the resulting precipitate was filtered, washed with water (20 mL) and ethyl acetate ( 10ml) to afford the title compound in 53% yield.

LC-MS purity: 99.2 %, m/z 381 (M+, Br pattern).

1H NMR (DMSO-D6): δ 13.30 (s, 1H), 9. 10 (s, 1H) 7.5 (d, 1H) 7.20 (d, 1H), 6.5 (s, 1H), 4.05 (t, 2H), 3.5 (t, 2H), 2.10 (t, 2H), 1.6 (s, 3H).

Step 5: Preparation of Intermediate 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1 ,2,3,5-tetrahvdro-indolizine-8-carboxylic acid (2-vinyloxy-ethoxy) -amide (1-le): To a stirred solution of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1,2,3,5- tetrahydro-indolizine-8-carboxylic acid (I-ld: 300 mg, 0.787 mmol) in 6 mL of dry dimethylformamide(DMF) and 2 mL of dichloromethane was added l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDCI) (165 mg, 0.865 mmol) and HOBt ( 116 mg, 0.865 mmol) at 0°C. The resulting reaction mixture was stirred for 2 hours at 0°C and then treated in succession with 0-(2-vinyloxy-ethyl)-hydroxylamine (71 mg, 0.787 mmol) and

triethylamine (158 mg, 1.57 mmol). After 12 hours, the reaction mixture was diluted with ethyl acetate and washed with saturated aqueous NaHCC>3 solution. The organic phase was dried (anhydrous Na₂S0₄) and concentrated in vacuo. The residual material was

chromato graphed on silica gel ( 1% MeOH in CHCI₃) to give 180 mg of the title compound in low purity.

LC MS: 31.7%, mfz= 466, (M+ Br pattern)

Step 6: Preparation of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl-5- oxo-1,2,3, 5-tetrahydroindolizine-8-carboxamide (Compound 1).

Crude 7-(4-bromo - 2-fluor o-phenylamino)- 6 -fluoro- 5 - oxo- 1 , 2, 3 , -tetrahy dr o - indolizine -8-carboxylic acid (2-vinyloxy-ethoxy)- amide (I-le: 180 mg, 0.386 mmol) was dissolved in ethanol containing lmL of IN HC1 and stirred for 16 hours at room temperature.

The reaction mixture was concentrated and the residue was dissolved in ethyl acetate. The organic phase was washed with brine, concentrated, and the residue was column

chromato graphed on silica gel (2 % MeOH in CHCI₃) to afford the title compound in 30% yield.

Compound 1 was also prepared using the following synthesis.

Step 1: Preparation of Intermediate Ethyl 7-chloro-6-methyl-5-oxo-l,2,3, 5-tetrahydro- indolizine-8-carboxylate (I-2a).

A four-neck 1 L round-bottomed flask equipped with a mechanical stirrer, a thermocouple, a reflux condenser, a heating/cooling capacity and a nitrogen inlet-outlet was charged with ethyl l,2,3,5-tetrahydro-7-hydroxy-6-methyl-5-oxo-8-indolizinecarboxylate (L la: 50.0 g, 210.8 mmol) and DMF (100 mL). POCI3 (25.5 mL, 274.0 mmol) was added dropwise to the mixture over 5 minutes at 30°C. The mixture was stirred at 30°C for 15 minutes after addition and a light brown solution was formed. At this point, i-PriNEt (41.9 mL, 242.4 mmol) was added dropwise to the above solution at 30°C over 5 minutes. After the addition, the reaction mixture was heated up to 83 °C and stirred at 83 °C for 8 hours. After completion of the reaction, the above mixture was cooled to room temperature and then added dropwise to brine (800 mL) at 8°C while stirring. The mixture was stirred at 8°C for 20 minutes after addition and filtered. The filter cake was washed with water (180 mL) till the washes got neutral (pH ~ 7) to yield the intermediate chloride (I-2a: 40 g, 75%) as an off white solid. [M+HJ calculated for Ci₂Hi₄ ³⁵ClN0₃, 256; found, 256.

Step 2: Preparation of Intermediate 7-C loro-6- ethyl-5-oxo-l,2,3,5-tetrahydro-indolizine- 8-carboxylic acid (I-2b).

A four-neck 12 L round bottomed flask equipped with a mechanical stirrer, a thermocouple, an addition funnel, and a nitrogen inlet/outlet was charged with intermediate ethyl ester (L2a: 750 g, 2.93 mol), THF (3 L), water ( 1.5 1) and MeOH (0.75 L) under nitrogen. LiOH.H₂0 (369.2 g, 8.8 mol) was added to the mixture. The reaction mixture was stirred at 25 °C and monitored by HPLC. After 24 hours, intermediate ethyl ester (I-2a) was consumed completely. Heptane (3 L) was then added to the reaction mixture and stirred vigorously for 30 minutes, and the top organic layer was separated. The collected bottom layer was cooled to 0°C and acidified with 3N HC1 (3.23 L, 9.69 mol) to pH 1-2. The formed precipitate was collected by filtration and washed with water till the washes got neutral (pH ~ 7). The intermediate carboxylic acid (I-2b: 633 g, 95 %) was obtained as an off white solid upon drying in a vacuum oven at 80 °C/5 torr for 16 hours. [M+H] calculated for

Ci₀Hio³ⁱClN0₃, 228; found, 228.

Step 3: Preparation of Intermediate 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1 ,2,3,5-tetrahydro-indoli∑ine-8-carboxylic acid (I-2c).

A four-neck 3 L round bottomed flask equipped with a mechanical stirrer, a thermocouple, an addition funnel, and a nitrogen inlet/outlet was charged with 7-chloro- l,2,3,5-tetrahydro-6-methyl-5-oxo-8-indolizine carboxylic acid (I-2b: 22.8 g, 0. 1 mol), 4- bromo-2-fluoroaniline (57.0 g, 0.3 mol) and tetrahydrofuran (0.3 L) under nitrogen. The suspension was stirred and cooled to— 15°C. Lithium bis(trimethylsilyl)amide (0.4 L, 1M in tetrahydrofuran, 0.4 mol) was added over 1 hour while maintaining an internal batch temperature below -10 °C throughout the addition. The resulting suspension was warmed to 22°C over 1 hour and stirred at 22°C for 4 hours till the remaining intermediate carboxylic acid (I-2b) was fully consumed as determined by HPLC analysis. The reaction mixture was cooled to 12°C and 3N hydrochloric acid (0.4 L) was added over 30 minutes while maintaining a batch temperature < 22 °C. Heptane (0.6 L) was added over 30 minutes and the mixture was stirred at room temperature over 10 hours. The suspension was filtered and the solid cake was rinsed with water (0.6 L). The solid cake was collected and slurried in 0. IN hydrochloric acid (0.35 L) at 20°C for 4 hours. The resulting slurry was filtered and the solid cake was washed with water until the washes were neutral (pH ~ 7). The solid was dried at 50°C/5 torr to afford the title compound (I-2c: 34.0 g, 89%) as an off-white solid, mp >235 °C (dec). [M+ITJ calculated for C₁₆Hi₄ ⁷⁹BrFN₂0₃, 381; found, 381.

Step 4: Preparation of 7-f (4-Bromo-2- fluorophenyl) amino] -1 ,2,3,5-tetrahydro-N-(2- hydroxyethoxy)-6-methyl-5-oxo-8-indolizinecarboxamide (Compound 1).

A four-neck 12 L round bottomed flask equipped with a mechanical stirrer, a thermocouple, an addition funnel, and a nitrogen inlet/outlet was charged with 7-(4-Bromo-2- fluoro-phenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydro-indolizine-8-carboxylic acid (I-2c: 400 g, 1.05 mol), PyBOP (707 g, 1.36 mol) and tetrahydrofuran (5.2 L) under nitrogen. The suspension was stirred at 17°C for 5 minutes and triethylamine ( 176 mL, 1.26 mol) was added to the mixture. The mixture was stirred at 20°C for 30 minutes and a solution was obtained. 2-(Aminooxy)ethanol (105 g, 1.36 mol) was added to the mixture. The resulting mixture was stirred at 20°C for 30 minutes and then seeded with Compound 1 (0.3 g). The mixture was stirred at 20 °C for 2 hours and filtered. The filter cake was rinsed with tetrahydrofuran (0.8 L) and dried in a vacuum oven at 40°C/5 torr for 1 hour to yield

Compound 1 (370 g, 80%) as a white solid. [M+H] calculated for Ci₈H₁₉ ⁷⁹BrFN₃0₄, 440; found, 440.

1H NMR (DMSO-D6): δ 11.20 (s, 1H), 7.65 (s, 1H), 7.48 (d, 1H), 7.18 (d, 1H), 6.5 (t, 1H), 4.0 (t, 2H), 3.7 (t, 2H), 3.5 (s, 3H), 3.2 (t, 2H), 2. 14 (quin, 2H), 1.72 ( s, 3H). Preparation of the free form of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l, 2,3,5- tetrahvdro-indolizine-8-carboxyIic acid cyclopropylmethoxy-amide (Compound 2)

(2)

Compounds of Steps I & II below may be prepared using published methods described in J. Org. Chem., 60, 2912 (1995); and Tetrahedron, 58, 2821 (2002).

Preparation of Starting Material 5-Methoxy-3. 4-dihydro-2H-pyrrole (SM-2):

H₃C^

N

(SM-2)

Pyrrolidin-2-one (85 g, 1 mol) is added dropwise over a period of 2 hours to a stirred solution of dimethyl sulphate (126 g, 1 mol) under a nitrogen atmosphere. The reaction mixture is stirred for 16 hours at 60° C. The reaction mixture is poured onto ice and saturated potassium carbonate solution, extracted with diethyl ether (2x500mL), washed with brine, and dried (anhydrous sodium sulphate). The organic extracts are removed under reduced pressure at 20°C to give 73 g of crude 5-methoxy-3,4-dihydro-2H-pyrrole as a light yellow color liquid. This compound is used in the next step without further purification. The NMR spectrum of the title compound is consistent with the structure.

1H NMR (CDC1_3, 300 MHZ): δ 3.80 (s, 3H), 3.66 (t, 2H), 2.48 ( t, 2H), 2.08-1.95

(m,2H).

Preparation of Intermediate 7 -Hydroxy- 5 -oxo- 1 ,2,3,5-tetrahydro-indoli∑ine-8-carboxylic acid ethyl ester (I-3a) :

(1-3 a)

Triethylamine is added to a mixture of 5 -methoxy-3,4-dihydro-2H- pyrrole (SM-2: 73 g, 0.73 mmol) and 3-oxopentanedioic acid diethyl ester (200 g, 0.99 mmol) at room temperature. The resulting solution is stirred for 5 days after which the reaction mixture is filtered to give 39 g (24% yield) of 7-hydroxy-5-oxo-l,2,3,5-tetrahydro-indolizine-8- carboxylic acid ethyl ester as a white solid. The NMR spectrum of the title compound is consistent with the structure.

1H NMR (CDCI3, 300 MHZ): δ 11.4 (s, 1H), 5.80 (s, 1H), 4.40 (q, 2H), 4. 15 (t, 2H), 3.50 (t, 2H), 2.3-2.15 (m, 2H), 1.40 (t, 3H). Preparation of Intermediate 5-oxo- 7-trifluoromethanesulfonyloxy- 1 ,2,3,5-tetrahydro- indolizine-8-carboxylic acid ethyl ester (I -3b):

(I-3b)

A stirred solution of 7-hydroxy-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid ethyl ester (1-3 : 60 mg, 0.2 mmol) and triethylamine (30 mg, 0.4 mmol) in 5 mL of dichloromethane is cooled to -78°C. Triflic anhydride (91 mg, 0.32 mmol) is then added dropwise over 20 minutes and the resulting reaction mixture is stirred for 12 hours at ambient temperature with TLC monitoring (100% EtOAc). The reaction mixture is washed with aqueous sodium bicarbonate solution (4 mL) and water (4 mL). The organic layer is dried over anhydrous Na₂SC)4, concentrated and the resulting product is purified via column chromatography on silica gel (60-120 mesh) using 15% ethyl acetate in hexane as eluant to afford 40 mg (48% yield) of the title compound.

LC-MS purity: 95 %, m/z 356 (M+l).

1H NMR (CDCl_3j 300 MHZ): δ 6.15 (s, 1H), 4.40 (q, 2H), 4.20 (t, 2H), 3.58 (t, 2H), 2.32-2.2 (m, 2H), 1.40 ( t, 3H).

Preparation of Intermediate / ⁷-(4-bromo-2-fluoro-phenylamino)-5-oxo-l ,2,3,5-tetrahydro- indolizine-8-carboxylic acid ethyl ester (I-3c) :

(I-3c)

A stirred suspension of 5-oxo-7-trifluoromethanesulfonyloxy-l,2,3,5-tetrahydro- indolizine-8-carboxylic acid ethyl ester (I-3b: 2.3 g, 6.4 mmol), 2-fluoro-4-bromo-aniline (1.25 g, 6.5 mmol), cesium carbonate(3. 17 g, 9.7 mmol), BINAP (0.6 g, 0.97 mmol), and Pd(OAc)₂ (0.15 g, 0.64 mmol) in toluene (100ml) is heated at 80^'C for 16 hours. The reaction is monitored by the TLC (9: 1 CHCl₃-MeOH v/v). The reaction mixture is diluted with ethyl acetate (60ml) and filtered. The filtrate is washed with water (100ml) and the aqueous layer is re-extracted with ethyl acetate (30ml). The combined organic extracts are dried (anhydrous Na₂S0₄), concentrated, and the crude product is purified by column chromatography on silica gel (60- 120 mesh) using 0.1-0.5% MeOH in chloroform to afford 336 mg (13% yield) of the title compound.

LC-MS purity: 98 %, m/z 395, 397(M+, Br pattern).

1H NMR (DMSO-De, 300 MHZ): 6 9.48 (s, 1H), 7.70 (d, 1H), 7.49-7.39 (m, 2H), 5.32

(s, lH), 4.30 (q, 2H), 3.95 (t, 2H), 3.48 (t, 2H), 2.14-2.0 (m, 2H), 1.30 (t, 3H).

Preparation of Intermediate 7 -(4-bromo-2-fluoro-phenylamino)-5-oxo-l ,2,3,5-tetrahydro- indolizine-8-carboxylic acid (I-3d)

(I-3d)

To the solution of 7-(4-bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro- indolizine-8-carboxylic acid ethyl ester (1-3 c: 280 mg, 0.71 mmol) in, THF:MeOH (4: 1, v/v, 6ml), is added, IN aqueous LiOH solution (2 mL). The resulting mixture is stirred for 3 hours at room temperature with TLC monitoring (CHC1₃ -MeOH, 8:2). The pH of the reaction mixture is adjusted to 1 with 10% aqueous HCl solution and the resulting precipitate is filtered, washed with water (20ml) and ethyl acetate (10 mL) to afford 240 mg (92% yield) of the title compound.

LC-MS purity: 96 %, m/z 367, 369(M+, Br pattern).

1H NMR (DMSO-D_6, 300 MHZ): δ 13.40 (s, 1H), 9.90 (s, 1H) 7.70 (d, 1H), 7.45 (s,

2H), 5.35 (s, 1H), 3.95 (t, 2H), 3.49 (t, 2H), 2.15-2.0 (m, 2H).

Final Step: Preparation of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l ,2,3,5-tetrahydro- indolizine-8-carboxylic acid cyclopropylmethoxy- amide (Compound 2):

To a stirred solution of 7-(4-bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro- indolizine-8-carboxylic acid (1-3 d: 140 mg, 0.38 mmol) in 10ml of dry DMF are added EDCI (220 mg, 1.14 mmol) and HOBt (160 mg, 1.14 mmol). The reaction mixture is stirred for 30 minutes and then treated with O-cyclopropylmethylhydroxylamine (141 mg, 1. 14 mmol) and TEA (232 mg, 1.14 mmol). The resulting reaction mixture is stirred for 16 hours with TLC monitoring (MeOH-CHCL 2:8, v/v). The reaction mixture is diluted with ethyl acetate (20ml) and washed with saturated aqueous NH₄CI solution (25ml), saturated aqueous NaHCOs solution (25ml), and brine (25ml). The combined organic extracts are dried (anhydrous a₂S0₄) and concentrated. The residual material is purified by column chromatography on silica gel (1% MeOH in CHC1₃) to afford the title compound in 36% yield.

LC-MS purity: 97%, m/z 436, 438 (M+, Br Pattern).

1H NMR (DMSO-De, 300 MHZ): δ 11.20 (s, 1H), 8.25 (s, 1H), 7.65 (d,lH), 7.45-7.3 ( m, 2H), 5.38 (s, US), 3.91 (t, 2H), 3.72 (d, 2H), 3.25 (t, 2H), 2. 15-2.0 (m, 2H), 1.18-1.02 ( m, 1H), 0.6-0.5 (m, 2H), 0.31-0.25 (m, 2H).

Example 1 illustrates the preparation of 7-(4-bromo-2-fluorophenylamino)-N-(2- hydroxyethoxy)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carboxamide, sodium salt in its crystalline form.

Example 1

Preparation of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl-5-oxo- 1 ,2,3,5-tetrahydroindoli∑ine-8-carboxamide, sodium salt (Form 1A):

( 1A)

The free form of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl- 5-oxo- l,2,3,5-tetrahydroindolizine-8-carboxamide (4.9 g) was suspended in 50 mL of propan-2-ol. The slurry was stirred for 5 minutes under nitrogen at 25°C. To the slurry was added 3N aqueous sodium hydroxide over 10 minutes. After stirring for 30 minutes a clear solution resulted. The solution was stirred for 1 hour and filtered for clarification. The clear filtrate was concentrated at 25-30°C at 30 mm vacuum to approximately 10 mL volume. The solids precipitated. The mixture was stirred for 6 hours at 25°C. The solid was collected by suction filtration and washed with approximately 5 mL of propan-2-ol and then dried at 55°C for 16 hours to obtain 4.68 g (yield 92.1%) of colorless crystalline salt. The salt is a crystalline solid having >99% purity and a 1: 1 acid:base stoichiometry (N:Na). The

TGA/DT A data indicated that it decomposes slowly above approximately 100°C. Unlike the corresponding free form, the salt ( 1A) has a very high solubility in aqueous media as well as in alcohols and aqueous-organic solvent mixtures.

The crystalline sodium salt exhibited polymorphic behavior: two polymorphic forms (Forms A and B) were identified (see, FIG. 1 and FIG. 3). The crystalline salt is non- hygroscopic: absorbs 1.6% moisture at 85% relative humidity and retained up to 10% moisture upon desorption to 25% relative humidity with loss of crystallinity. The salt form lost crystallinity upon compression but retained crystallinity upon grinding. The crystalline salt had very high solubility in the aqueous media as well as in alcohols and aqueous-organic solvent mixtures. It had only 0.37% of residual solvent/moisture content.

The X-ray diffraction pattern for the Sodium Salt crystal form (Form 1 A) shown in Figure 1 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 180 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -2.01 mm

Y: -9.92 mm

Z: -7.58 mm

Preparation of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl-5-oxo- 1 ,2,3,5-tetrahydroindolizine-8-carboxamide, sodium salt (Form IB):

A suspension of 50 mg of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)- 6-methyl-5-oxo-l,2,3,5-tetrahydroindolizine-8-carboxamide, sodium salt in 1 mL of acetonitrile was agitated for 24 hours at 25 °C and filtered. The solids were air dried and tested by XRPD and TGA-DT The same crystalline form is also produced when absolute ethanol or 2-propanol is used as the solvent in place of acetonitrile. The X-ray diffraction pattern for the Sodium Salt crystal (Form IB) shown in Figure 3 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -2.15 mm

Y: -7.59 mm

Z: -7. 16 mm

Example 2 illustrates the preparation of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt in its crystalline form.

Example 2

Preparation of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indolizine-8- carboxylic acid cyclopropylmethoxy-amide, sodium salt (2A):

(2A)

The free form of 7-(4-Bromo-2-fluoro-phenylamino)-5-oxo- 1,2,3, 5-tetrahydro- indolizine-8-carboxylic acid cyclopropylmethoxy-amide (Compound 2: 5.0 g) was suspended in 50 mL of propan-2-ol. The slurry was stirred for 5 minutes under nitrogen at 25°C. To the slurry was added 3N aqueous sodium hydroxide over 10 minutes. After stirring for 30 minutes a clear solution resulted. The solution was stirred for 1 hour and filtered for clarification. The clear filtrate was concentrated at 25-30 °C at 30 mm vacuum to ~ 10 mL volume. The solids came out. The mixture was stirred for 6 hours at 25°C. The solid was collected by suction filtration and washed with ~5 mL of propan-2- ol. It was dried at 55°C for 16 hours to obtain 5.0 g (yield 95.41%) of colorless crystalline solid.

Salt (2A) is a crystalline solid having > 99% purity and has a 1: 1 acid:base stiochiometry (N-Na). The TGA data indicated that it decomposes at approximately 232°C. Unlike the corresponding free form, the salt has a very high solubility in aqueous media as well as in alcohols and aqueous- organic solvent mixtures.

The X-ray diffraction pattern for the Salt 2A crystalline form (Form A) shown in Figure 5 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 180 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -1.21 mm

Y: 10.87 mm

Z: -7.58 mm

Salt (2A) formed an amorphous solvate with acetonitrile. It also formed crystalline solvates with acetone, absolute ethanol, isopropyl acetate, propan-2-ol, and tetrahydrofuran. The crystalline solvates of the sodium salt (2A) were prepared with acetone, ethanol, isopropyl acetate, isopropanol, and tetrahydrofuran using the following general procedure. A suspension of—50 mg of salt in 1 mL of solvent was agitated for 24 hours at 25°C and filtered. The solids were air dried and tested by XRPD and TGA-DTA.

7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy- amide, sodium salt , acetone solvate (2A-ACT):

The X-ray diffraction pattern for the Acetone solvate (2A-ACT)) shown in Figure 7 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -14.68 mm

Y: -33.66 mm

Z: -7.87 mm

7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indoli∑ine-8-carboxylic acid cyclopropylmethoxy- amide, sodium salt , Acetonitrile solvate (2A-ACN):

The X-ray diffraction pattern for the Acetonitrile solvate (2A- ACN)) shown in Figure

9 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3° Theta: 5.95"

Chi: 5.67°

Phi: 339.68°

X: -14.17 mm

Y: -48.13 mm

Z: -7.99 mm

7-(4-Bromo-2-fiuoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indoli∑ine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt , Ethanol solvate (2A-ETH):

The X-ray diffraction pattern for the Ethanol solvate (2A-ETH) shown in Figure 11 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -1.47 mm

Y: -47.66 mm

Z: -7.46 mm

7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt , Isopropyl acetate solvate (2A-IAC):

The X-ray diffraction pattern for the Isopropyl acetate solvate (2A-I AC) shown in Figure 13 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020° Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -7.78 mm

Y: 5.13 mm

Z: -7.35 mm

7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt , Isopropanol solvate (2A-ISO):

The X-ray diffraction pattern for the Isopropanol solvate (2A-ISO) shown in Figure 15 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -6.52 mm

Y: -28.15 mm

Z: -7.35 mm

7-(4-Bromo-2-fluoro-phenylamino)-5-oxo-l,2,3,5-tetrahydro-indolizine-8-carboxylic acid cyclopropylmethoxy-amide, sodium salt , Tetrahydofuran solvate (2A-THF) :

The X-ray diffraction pattern for the Tetrahydrofuran solvate (2A-THF) shown in Figure 17 was generated using the following settings: Type : 2 Theta alone

Start: 3°

End: 39.100°

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

2-Theta: 3°

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -7.20 mm

Y: -44.10 mm

Z: -7.35 mm

Claims

CLAIMS What is claimed is:

1. Sodium salt of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6- methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carboxamide.

2. A crystalline form of 7-(4-bromo-2-fluorophenylamino)-N-(2- hydroxyethoxy)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carboxamide, sodium salt having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 1.

3. The crystalline form of 7-(4-bromo-2-fluorophenylamino)-N-(2- hydroxyethoxy)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carboxamide, sodium salt having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.5°, 10.5°, 14.2°, 15.2°, 16.1°, 17.8°, 19.9°, 20.3°, 22.8°, 23.8°, 24.2°, 24.8°, 25.9°, 27.0°, 28.6°, and 30. 1°.

4. A crystalline form of 7-(4-bromo-2-fluorophenylamino)-N-(2- hydroxyethoxy)-6-methyl-5-oxo- 1,2,3, -tetrahydroindolizine-8-carboxamide, sodium salt having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 3.

5. The crystalline form of 7-(4-bromo-2-fluorophenylamino)-N-(2- hydroxyethoxy)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8-carboxamide, sodium salt having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 10.5°, 14.3°, 15. 1°, 18.5°, 19.0°, 20.2°, 21.3°, 23. P, 24.3°, 24.8°, 25.3°, 25.8°, 26.4°, 27.2°, 28.2°, 28.4°, 29.2°, 30.0°, 31.1°, 32.1°, 33.1°, 33.7°, 34.9°, and 36.3°.

6. The crystalline form of Claims 2, 3, 4 or 5 where said crystalline form is substantially pure.

7. A pharmaceutical composition comprising a sodium salt of 7-(4-bromo-2- fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide in accordance with any one of the preceding claims; and a pharmaceutically acceptable excipient, diluent or carrier.

8. The pharmaceutical composition of Claim 7 further comprising at least one additional pharmaceutical agent selected from an anti-tumor or an anti-proliferative agent.

9. A method of treating cancer in a mammal comprising administering to a mammal in need thereof a therapeutic amount of a sodium salt of 7-(4-bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxyethoxy)-amide in accordance with Claims 1 through 6, or a pharmaceutical composition thereof.

10. The method of Claim 9 further comprising administering at least one additional pharmaceutical agent selected from an anti-tumor or an anti-proliferative agent.

11. The method of Claim 10 where said at least one additional pharmaceutical agent is a mTOR inhibitor or a PI3K inhibitor.

12. The method of Claim 10 or 11 wherein said at least one additional

pharmaceutical agent and said sodium salt of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5- oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide are administered simultaneously.

13. The method of Claim 10 or 11 wherein said at least one additional

pharmaceutical agent and said sodium salt of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5- oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide are administered sequentially.

14. The use of a sodium salt of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5- oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxyethoxy)-amide according to Claims 1 through 6 in the treatment of cancer.

15. Sodium salt of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo-l,2,3,5- tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide.

16. A crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 5.

17. The crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 6.3°, 6.9°, 7.6°, 10.2°, 13.2°, 20.7°, 21.6°, 23.8°, 24.7°, 26.8°, 27.9°, 30.0°, and 32.6°.

18. The crystalline form of Claims 16 or 17 where said crystalline form is substantially pure.

19. An acetone solvate of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo-

1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy )ami de, sodium salt .

20. A crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, acetone solvate having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 7.

21. The crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, acetone solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (29) of 3.2°, 7.1°, 9.8°, 10.9°, 13.1°, 15.4°, 17.7°, 20.1°, 21.8°, 23.5°, 25.3°, 25.8° and 26.5°.

22. The crystalline form of Claims 20 or 21 where said crystalline form is substantially pure.

23. An acetonitrile solvate of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy) ami de, sodium salt .

24. An ethanol solvate of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt.

25. A crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, ethanol solvate having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 11.

26. The crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, ethanol solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 3.2°, 7.1°, 9.8°, 10.9°, 13. Γ, 15.4°, 17.7°, 20. Γ and 21.8°.

27. The crystalline form of Claims 25 or 26 where said crystalline form is substantially pure.

28. An isopropyl acetate solvate of 7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt.

29. A crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, isopropyl acetate solvate having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 13.

30. The crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt, isopropyl acetate solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 6.5°, 7.0°, 7.6°, 10.4°, 13.2°, 16.6°, 18.4°, 19.1°, 20.9°, 21.8°, 23.8°, 24.9°, 26.2° and 26.9°.

31. The crystalline form of Claims 29 or 30 where said crystalline form is substantially pure.

32. An isopropanol solvate of -(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy )ami de, sodium salt .

33. A crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo- 1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy )ami de, sodium salt, isopropanol solvate having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 15.

34. The crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- 1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy )ami de, sodium salt, isopropanol solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 6.6°, 8.4°, 9. 1°, 9.4°, 13.1°, 14.2°, 15.7°, 20.3°, 24. Γ and 25.7°.

35. The crystalline form of Claims 33 or 34 where said crystalline form is substantially pure.

36. An tetrahydrofuran solvate of -(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide, sodium salt.

37. A crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo-

1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy )am de, sodium s lt, tetrahydrofuran solvate having a X-ray diffraction spectrum substantially the same as the X- ray powder diffraction spectrum shown in FIG. 17.

38. The crystalline form of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5-oxo-

1,2,3,5 -tetrahydr oindol izine- 8 -carbonyl) (cycl opropylmethoxy )ami de, sodium salt, tetrahydrofuran solvate having a powder X-ray diffraction pattern comprising peaks at diffraction angles (20) of 3.2°, 7.2°, 7.7°, 19. 1°, 21.4°, 22.0°, 22.6°, 23. P, 23.7°, 25.3° and 26.8°.

39. The crystalline form of Claims 37 or 38 where said crystalline form is substantially pure.

40. A pharmaceutical composition comprising a sodium salt of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8- carbonyl)(cyclopropylmethoxy)amide or a solvate thereof in accordance with Claims 15 through 39; and a pharmaceutically acceptable excipient, diluent or carrier.

41. The pharmaceutical composition of Claim 40 further comprising at least one additional pharmaceutical agent selected from an anti-tumor or an anti-proliferative agent.

42. A method of treating cancer in a mammal comprising administering to a mammal in need thereof a therapeutic amount of a sodium salt of (7-(4-bromo-2- fluorophenylamino)-6-methyl-5-oxo- 1,2,3, 5-tetrahydroindolizine-8- carbonyl)(cyclopropylmethoxy)amide or a solvate thereof in accordance with Claims 15 through 39, or a pharmaceutical composition thereof.

43. The method of Claim 42 further comprising administering at least one additional pharmaceutical agent selected from an anti-tumor or an anti-proliferative agent.

44. The method of Claim 43 where said at least one additional pharmaceutical agent is a mTOR inhibitor or a PI3K inhibitor.

45. The method of Claim 43 or 44 wherein said at least one additional pharmaceutical agent and said sodium salt of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5 oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide or said solvate thereof are administered simultaneously.

46. The method of Claim 43 or 44 wherein said at least one additional pharmaceutical agent and said sodium salt of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5 oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide or said solvate thereof are administered sequentially.

47. The use of a sodium salt of (7-(4-bromo-2-fluorophenylamino)-6-methyl-5- oxo-l,2,3,5-tetrahydroindolizine-8-carbonyl)(cyclopropylmethoxy)amide or said solvate thereof according to Claims 15 through 39 in the treatment of cancer.