POLYMORPHS OF A MEK INHIBITOR

FIELD OF THE INVENTION

The present invention relates to polymorphic forms of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide 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-tetrah ydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide (referred to herein as "Compound 1") is a potent MEK inhibitor. The crystallinity, polymorphic behavior, and hygroscopicity of the compound or 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 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 a crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 1 (referred to herein as "Form 1- A").

The present invention also provides a crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide ("Form 1-A") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 5.2°, 7.4°, 8.6°, 13.3°, 14.7°, 15. Γ, 17.3°, 18.8°, 20.0°, 20.8°, 22.3°, 23.7°, and 26. Γ.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 5.2°, 7.4°, 8.6°, 13.3°, 14.7°, 15. 1°, 17.3°, 18.8°, 20.0°, 20.8°, 22.3°, 23.7°, and 26. 1°; and a pharmaceutically acceptable excipient, diluent or carrier.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 5.2°, 7.4°, 8.6°, 13.3°, 14.7°, 15. P, 17.3°, 18.8°, 20.0°, 20.8°, 22.3°, 23.7°, and 26. P, or a

pharmaceutical composition thereof.

The present invention provides a crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxy- ethoxy)-amide having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 3 (referred to herein as "Form 1-B").

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-hydroxy- ethoxy)-amide ("Form 1-B") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 7.7°, 20.3°, 22.5°, 23.5°, 24.8°, 27.9° and 31.9° is provided.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 7.7°, 20.3°, 22.5°, 23.5°, 24.8°, 27.9° and 3 1.9°; and a pharmaceutically acceptable excipient, diluent or carrier.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (29) of 7.7°, 20.3°, 22.5°, 23.5°, 24.8°, 27.9° and 3 1.9°, or a pharmaceutical composition thereof. 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- hydroxy-ethoxy)-amide is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 5 (referred to herein as "Form 2").

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-hydroxy-ethoxy)-amide ("Form 2") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.7°, 15.4°, 15.8°, 21. 1°, 21.5°, 21.9°, 23. 1°, 23.5°, 26.4° and 36.6°.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.7°, 15.4°, 15.8°, 21.1°, 21.5°, 21.9°, 23. 1°, 23.5°, 26.4° and 36.6°; and a pharmaceutically acceptable excipient, diluent or carrier.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.7°, 15.4°, 15.8°, 21.1°, 21.5°, 21.9°, 23.1°, 23.5°, 26.4° and 36.6°, or a pharmaceutical composition thereof.

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-ca rboxylic acid (2-hydroxy- ethoxy)-amide is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 7 (referred to herein as "Form 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-hydroxy-ethoxy)-amide ("Form 3") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.3°, 8.5°, 13.2°, 14.6°, 19.8°, 22.6°, 23.6°, 26.2°, 31.0° and 34.8°.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.3°, 8.5°, 13.2°, 14.6°, 19.8°, 22.6°, 23.6°, 26.2°, 31.0° and 34.8°; and a pharmaceutically acceptable excipient, diluent or carrier.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 7.3°, 8.5°, 13.2°, 14.6°, 19.8°, 22.6°, 23.6°, 26.2°, 31.0° and 34.8°, or a pharmaceutical composition thereof.

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-ca rboxylic acid (2-hydroxy- ethoxy)-amide is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 9 (referred to herein as "Form 4").

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-hydroxy-ethoxy)-amide ("Form 4") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 13.0°, 13.7°, 14.5°, 23.7°, 24. F, 26.0°, 31.3°, and 32.8°.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 13.0°, 13.7°, 14.5°, 23.6°, 24.1°, 26.0°, 31.3°, and 32.8°; and a pharmaceutically acceptable excipient, diluent or carrier.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (26?) of 13.0°, 13.7°, 14.5°, 23.6°, 24. 1°, 26.0°, 31.3°, and 32.8°, or a pharmaceutical composition thereof.

In yet another aspect of the present invention, a crystalline hydrate of 7-(4-bromo-2- fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizi ne-8-carboxylic acid (2- hydroxy-ethoxy)-amide is provided having a X-ray diffraction spectrum substantially the same as the X-ray powder diffraction spectrum shown in FIG. 11 (referred to herein as "Hydrate"). Also provided is a hydrate crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-6- methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide ("Hydrate") having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 3.7°, 10.9°, 14.3°, 15.2°, 17.0°, 17.8°, 19.0°, 19.3°, 20.6°, 21.4°, 21.9°, 22.4, 23.0°, 23.3°, 24.6°, 25. 1°, 25.9°, 26.6°, 27.7°, 28.8°, 30.3°, 30.7°, and 32.0°.

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-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2Θ) of 3.7°, 10.9°, 14.3°, 15.2°, 17.0°, 17.8°, 19.0°, 19.3°, 20.6°, 21.4°, 21.9°, 22.4, 23.0°, 23.3°, 24.6°, 25.1°, 25.9°, 26.6°, 27.7°, 28.8°, 30.3°, 30.7°, and 32.0°; and a pharmaceutically acceptable excipient, diluent or carrier.

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-mefhyl-5- oxo-l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2(9) of 3.7°, 10.9°, 14.3°, 15.2°, 17.0°, 17.8°, 19.0°, 19.3°, 20.6°, 21.4°, 21.9°, 22.4, 23.0°, 23.3°, 24.6°, 25. P, 25.9°, 26.6°, 27.7°, 28.8°, 30.3°, 30.7°, and 32.0°, or a pharmaceutical composition thereof.

Preferably, the crystalline forms described above (Form 1-A, Form 1-B, Form 2,

Form 3, Form 4 and Hydrate) are substantially pure.

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 (2Θ) 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 "compounds of the present invention", unless indicated otherwise, refers to the crystalline forms of the free form of 7-(4-bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

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-hydroxy-ethoxy)-amide (Compound 1 "Form 1-A").

Figure 2 shows a TGA-DT A thermogram of the "Form 1-A" polymorph of 7-(4- Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

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-hydroxy-ethoxy)-amide (Compound 1 "Form 1-B").

Figure 4 shows a differential scanning calorimetry (DSC) thermogram of the "Form 1-B" polymorph of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

Figure 5 shows a powder X-ray diffraction pattern of a crystalline form of 7-(4-

Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahyd ro-indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide (Compound 1 - "Form 2").

Figure 6 shows a differential scanning calorimetry (DSC) thermogram for the "Form 2" polymorph of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrah ydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

Figure 7 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-hydroxy-ethoxy)-amide (Compound 1 - "Form 3").

Figure 8 shows a differential scanning calorimetry (DSC) thermogram of the "Form 3" polymorph of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1,3,5-tetrahydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

Figure 9 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-hydroxy-ethoxy)-amide (Compound 1 - "Form 4"). Figure 10 shows a differential scanning calorimetry (DSC) thermogram of the "Form 4" polymorph of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

Figure 11 shows a powder X-ray diffraction pattern of the crystalline hydrate form of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide (Compound 1 "Hydrate").

Figure 12 shows a TGA-DTA thermogram of the crystalline hydrate form of 7-(4- Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide.

DETAILED DESCRIPTION

The characteristic 2-theta (2Θ) values, d-spacing (A) and relative intensity (RI) for the powder X-ray diffraction (PXRD) pattern of the crystalline forms of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide (as prepared in Example 1 "Form 1-A" and "Form 1-B") are shown in Tables 1 A and IB, respectively (below). The corresponding thermograms for the crystalline form of 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrah ydro-indolizine-8- carboxylic acid (2-hydroxy-ethoxy)-amide "Form 1-A" is shown in FIG. 2 and "Form 1-B" is shown in FIG 4.

Table 1A ("Form 1-A")

Table IB ("Form 1-B")

2-Theta d-spacing

(deg°) (A) Relative intensity

The characteristic 2-theta (2Θ) values, d-spacing (A) and relative intensity (RI) for the powder X-ray diffraction (PXRD) pattern of the crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide (as prepared in Example 2 "Form 2") are shown in Table 2 (below). The corresponding DSC thermogram for the "Form 2" crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide is shown in FIG 6.

Table 2

The characteristic 2-theta (2(9) values, d-spacing (A) and relative intensity (RI) for the powder X-ray diffraction (PXRD) pattern of the crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide (as prepared in Example 3 - "Form 3") are shown in Table 3 (below). The corresponding DSC thermogram for the "Form 3" crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide is shown in FIG 8.

Table 3

2-Theta d-spacing

(deg°) (A) Relative intensity

The characteristic 2-theta (2(9) values, d-spacing (A) and relative intensity (RT) for the powder X-ray diffraction (PXRD) pattern of the crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide (as prepared in Example 4 "Form 4") are shown in Table 4 (below). The corresponding DSC thermogram for the "Form 4" crystalline form of 7-(4-Bromo-2-fluoro- phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indolizine-8-ca rboxylic acid (2-hydroxy- ethoxy)-amide is shown in FIG 10.

Table 4

The characteristic 2-theta (2Θ) values, d-spacing (A) and relative intensity (RI) for the powder X-ray diffraction (PXRD) pattern of the hydrate crystalline form of 7-(4-Bromo-2- fluoro-phenylamino)-6-methyl-5-oxo- l,3,5-tetrahydro-indolizine-8-carboxylic acid (2- hydroxy-ethoxy)-amide (as prepared in Example 5 "Hydrate") are shown in Table 5 (below). The corresponding thermogram for the "Hydrate" crystalline form of 7-(4-Bromo- 2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetrahydro-indoli zine-8-carboxylic acid (2- hydroxy-ethoxy)-amide is shown in FIG 12.

Table 5

Compound 1 (7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo-l,3,5-tetra hydro- indolizine-8-carboxylic acid (2-hydroxy-ethoxy)-amide) can be synthesized using the procedures outlined in Schemes I or 2 below or by the methods disclosed in PCT Publication No. WO 2008/082846 or US Publication No. 2009/0275606, incorporated herein by reference.

BINAP,

Pd OAc) ₂

Scheme I

For a detailed description of the process steps outlined in Scheme I, see the Examples section below. Those skilled in the art will appreciate that alternative conditions and reagents may be used in the synthetic scheme outlined above.

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. Those skilled in the art will appreciate that alternative conditions and reagents may be used in the synthetic scheme outlined above.

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]pyrirrlidin-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^^ηάο[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-i midazo[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]-th iazolidine-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.

Differential scanning calorimetry (DSC) : T A Instruments: DSC Q1000 V23.5 Build 72 and Processing Software: Universal V4 3 A TA Instruments. Temperature increase rate = 10°C/minute with a range of -40°C to 300°C.

Thermogravimetric analysis/differential thermal analysis (TGA)/DTA) TA

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

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

Examples 1 through 4 illustrate crystalline forms of 7-(4-bromo-2- fluorophenyl amino )-N-( 2-hydroxyethoxy )- 6 -methyl- 5-oxo-l,2,3,5 -tetrahydroindol izine- 8- carboxamide.

Example 1

Preparation of 7-f(4-Bromo-2-fluorophenyl)amino]-l,2,3,5-tetrahydro-N-(2-hy droxyethoxy)- 6-methyl-5-oxo-8-indolizinecarboxamide (Compound 1- Forml-A) :

(Compound 1)

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

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 (I-l b):

7-hydroxy-6-methyl-5-oxo-l,2,3,5-tetrahydro-indolizine-8-car boxylic 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 Na ₂ S04) 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): δ 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-indoli∑ine-8-carboxylic acid (I- Id):

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- l,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 NaHC(¾ solution. The organic phase was dried (anhydrous Na ₂ S0 ₄ ) and concentrated in vacuo. The residual material was

chromato graphed on silica gel ( 1% MeOH in CHC1 ₃ ) to give 180 mg of the title compound in low purity.

LC MS: 31.7%, m/z= 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 CHCI ₃ ) to afford the title compound in 30% yield.

Isolation of crystalline form— "Form I -A "

Compound 1 from step 6 was purified by column chromatography using silica gel of mesh size of 60- 120. A 1% Methanol in chloroform solution was used as the eluting system. The product obtained was triturated with methanol. Form 1 contains a fraction of solvent which evaporates at 89.7°C. Melting point = 141.57°C.

The X-ray diffraction pattern for the "Form 1-A" crystal of Compound 1 shown in Figure 1 was generated using the following settings:

Type : 2 Theta alone

Start: 3°

End: 39.100 ,0

Step: 0.020°

Step time: 60 seconds

Temperature: 25 °C

Time Started: 0 second

0

2-Theta: 3 ^'

Theta: 5.95°

Chi: 5.67°

Phi: 339.68°

X: -7.52 mm Y: -20.73 mm

Z: -7.79 mm

Isolation of crystalline form— "Form 1-B "

When the Form 1-A crystalline material was heated to 150°C and then cooled to room temperature, a second crystalline form ("Form 1-B") was produced and isolated. Melting point = 163-165°C.

The X-ray diffraction pattern for the "Form 1-B" crystal of Compound 1 shown in Figure 3 was generated using the following settings:

Type: 2 Theta alone

Start: 6.400°

End: 39.100°

Step: 0.020°

Step time: 240.1 seconds

Temperature: 25°C (Room)

Time Started: 0 second

2-Theta: 6.400°

Theta: 7.725°

Chi: 1.74°

Phi: 276.55°

X: -14.2 mm

Y: 20.4mm

Z: -4.4mm

Example 2

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

7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl -5-oxo- 1,2,3,5- tetrahydroindolizine-8-carboxamide containing Form 1-A (2.5 mg) was suspended in 100 μL of ethylacetate. The suspension was warmed to 60°C. A clear solution was obtained upon heating. After 24 hours, the solution temperature was reduced to room temperature

(approximately 25 °C). The observed precipitate was kept at room temperature for 48 hours under constant shaking. The precipitate was then isolated by filtration. Alternatively, the crystalline form may be prepared by substituting ethyl acetate with 2-butanone or absolute ethanol.

Melting point = 170-172°C. The powder X-ray diffraction pattern is shown in FIG. 5 and the differential scanning calorimetry (DSC) thermogram is shown in FIG. 6. Form 2 is an ansolvated form. Since this form is found to be the prevailing form in a series of crystallization experiments, it is considered the more thermodynamically stable form.

The X-ray diffraction pattern for the "Form 2" crystal of Compound 1 shown in Figure 5 was generated using the following settings:

Type: 2 Theta alone

Start: 6.400°

End: 39.100°

Step: 0.020°

Step time: 120.1 seconds

Temperature: 25°C (Room)

Time Started: 0 seconds

2-Theta: 6.400°

Theta: 7.725°

Chi: 1.74°

Phi: 276.55°

X: -12.8 mm

Y: -26.7mm

Z: 6.2mm

Example 3

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

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-indolizinecarb oxylate Q la: 50.0 g, 210.8 mmol) and DMF (100 mL). POCl ₃ (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, ;-Pr ₂ NEt (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+H] calculated for Ci ₂ Hi ₄ ³⁵ ClN0 ₃ , 256; found, 256. Step 2: Preparation of Intermediate 7-Chloro-6-methyl-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 L) and MeOH (0.75 L) under nitrogen. LiOH.H ₂ 0 (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

CioHio ³⁵ ClN0 ₃ , 228; found, 228. Step 3: Preparation of Intermediate 7-(4-Bromo-2-fluoro-phenylamino)-6-methyl-5-oxo- 1 ,2,3,5-tetrahvdro-indolizine-8-carboxylic acid (l-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+H] calculated for C ₁₆ Hi ₄ ⁷⁹ BrFN ₂ (¾, 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 I).

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-l,2,3,5-tetrahydro-indoli zine-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 Ci ₈ H _1E , ⁷⁹ BrFN ₃ 0 ₄ , 440; found, 440.

1H NMR (DMSO-D6): 5 11.20 (s, 1H), 7.65 (s, lH), 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).

Step 5: Recrystallization and isolation of Form 3 crystalline material:

A four-neck 5 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 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl -5- oxo-l,2,3,5-tetrahydroindolizine-8-carboxamide from above (226 g, 0.51 mol) and methanol (2.2 L) under nitrogen. The suspension was heated to 64°C and stirred at 64°C for 10 minutes to form a clear solution. The mixture was filtered while hot through a

Celite® pad and the filter cake was rinsed with hot (> 55 °C) methanol (0.3 L). The filtrate was heated to 64°C and stirred at 64°C for 10 minutes to form a solution. The mixture was allowed to cool to 20°C over 1 hour. The mixture was concentrated to a volume of 0.8 L under vacuum with batch temperature below 20°C. The mixture was filtered and the solid cake was washed with methanol (0. 1 L) and dried in a vacuum oven at 60°C/5 torr for 16 hours to give crystalline Compound 1 Form 3 (200 g, 88%), mp 163~165°C. [M+H] calculated for Ci ₈ Hi9 ⁷ ¾rFN30 ₄ , 440; found, 440

The X-ray diffraction pattern for the "Form 3" crystal of Compound 1 shown in Figure 7 was generated using the following settings:

Type: 2 Theta alone

Start: 6.400°

End: 39.100°

Step: 0.020°

Step time: 240.1 seconds

Temperature: 25°C (Room)

Time Started: 0 seconds

2-Theta: 6.400°

Theta: 7.725°

Chi: 1.74°

Phi: 276.55°

X: -13.8 mm

Y: 20.0mm

Z: 6.2mm

Example 4

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

A 22 L 4-necked round- bottomed flask, equipped with mechanical stirrer, thermocouple, reflux condenser, heating/cooling capacity, nitrogen inlet-outlet under a nitrogen atmosphere at ambient temperature, was charged with 7-(4-bromo-2- fluorophenyl amino )-N-( 2-hydroxyethoxy )- 6 -methyl- 5-oxo-l,2,3,5 -tetrahydroindol izine- 8- carboxamide from step 4 of Example 3 (598 g, 1.36 mol) and 200 proof ethanol (10.8 L). The suspension was heated to 78° C and stirred at 78°C for 10 minutes to form a clear solution. The mixture was filtered while hot through a Celite ^® pad and the filter cake was rinsed with hot (> 60 °C) 200 proof ethanol (1.2 L). The filtrate was heated to 78°C and stirred at 78°C for another 10 minutes. The mixture was allowed to cool to 40°C over a minimum of 30 minutes. After seeding with 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6- methyl-5-oxo- l,2,3,5-tetrahydroindolizine-8-carboxamide (0.5 g), the mixture was stirred gently for 6 hours at 40°C during which precipitation occurs. The mixture was cooled to 20°C over a minimum of 1 hour and stirred at 20°C for 12 hours. The mixture was then concentrated to 4.18 L in vacuo at 40°C (bath temperature )/20°C (process tem erature)/30 torr. The product was filtered and the filter cake rinsed with 200 proof ethanol (0.2 L). The product was dried in a vacuum oven at 40°C/5 torr for 3 hours to give a white crystalline solid (Form 4: 518 g, 87%), mp 195-196 °C. [M+H] calc'd for Ci ₈ Hi9 ⁷⁹ BrFN ₃ 0 ₄ , 440; found, 440.

Substitution of the absolute ethanol with 2-propanol or isopropyl acetate also produced the "Form 4" crystal of Compound 1.

The X-ray diffraction pattern for the "Form 4" crystal of Compound 1 shown in Figure 9 was generated using the following settings:

Type: 2 Theta alone

Start: 6.400°

End: 39.100°

Step: 0.020°

Step time: 240.1 seconds

Temp. : 25 °C (Room)

Time Started: 0 seconds

2-Theta: 6.400°

Theta: 7.725°

Chi: 1.74°

Phi: 276.55°

X: -18.3mm mm

Y: 9.1mm

Z: -4.6mm

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

40 mg of 7-(4-bromo-2-fluorophenylamino)-N-(2-hydroxyethoxy)-6-methyl -5-oxo- l,2,3,5-tetrahydroindolizine-8-carboxamide was agitated in 1 mL of water for 24 hour at 25 ^U C. The suspension was filtered, air-dried for 12 hours and the solids were tested by XRPD and TGA-DTA. The crystalline material is primarily the dihydrate.

The X-ray diffraction pattern for the crystalline form of 7-(4-bromo-2- fluorophenyl amino )-N-( 2-hydroxyethoxy )- 6 -methyl- 5 -oxo- 1 , 2, 3 , 5 -tetrahydroindol izine- 8- carboxamide hydrate, 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: -2.06 mm

Y: -7.41 mm

Z: -7.31 mm