HK1116483B - Polymorphic and amorphous forms of the phosphate salt of 8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6h-azepino[5,4,3-cd]indol-6-one - Google Patents

Description

Polymorphs and amorphous forms of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one

The benefit of U.S. provisional application No.60/612,459 filed on 9/22/2004 and U.S. provisional application No.60/679,296 filed on 9/2005 is claimed and is hereby incorporated by reference in its entirety.

Technical Field

The present invention relates to novel polymorphic and amorphous forms of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one and to processes for their preparation. The invention also relates to pharmaceutical compositions comprising at least one polymorph or amorphous, and to the therapeutic or prophylactic use of such polymorphs and amorphous and compositions.

Background

The compound 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one is represented by formula 1,

it is a small molecule inhibitor of poly (ADP-ribose) polymerase (PARP). The compounds of formula 1 and their salts can be prepared by methods described in the following documents: U.S. patent No.6,495,541, PCT application No. PCT/IB2004/000915, international publication No. wo 2004/087713, and U.S. provisional patent application No.60/612,457, the disclosures of which are incorporated herein by reference in their entirety.

Currently, 18 enzymes have been identified in the PARP family by DNA sequence homology, 7 of which have been studied for biochemical and enzymatic properties: PARP-1 and PARP-2 are triggered by DNA strand breaks; PARP-3 interacts with PARP-1 and the central body; PARP-4, also known as cavern (vault) PARP (vparp), is the largest PARP and is associated with the cytoplasmic cavern; tankyrase 1 and 2(PARP-5a and 5b) is associated with telomeric proteins; the function of PARP-7(TiPARP) is not clear at present, but it is associated with T cell function and it can poly (ADP-ribosylation) histones (Ame JC, Splenlehauer C and de Murcia G.the PARP Superfamily.Bioessays 26882-893 (2004)). Pharmacological studies have shown that the compound of formula 1 is PARP-1 (K)_i1.4nM) and PARP-2 (K)_i0.17 nM). Based on the structural similarity of the amino acid sequences between these PARP enzymes, the compounds of formula 1 are also likely to bind with high affinity to other members of the family.

Enzymatic repair of single or double stranded breaks in DNA is a possible mechanism for radiation therapy or cytotoxic drugs whose resistance mechanism relies on DNA damage. Thus, a strategy to enhance the action of these agents is to inhibit DNA repair enzymes. PARP-1, the best characterized member of the PARP family, is a ribozyme that, after activation by DNA damage, causes ADP-ribose fragments to dissociate from NAD⁺Transfer to multiple receptor proteins. Depending on the extent to which DNA damage occurs, PARP-1 is activated and subsequently undergoes a poly (ADP-ribose) reaction, resulting in repair of damaged DNA or cell death. When the degree of DNA damage is moderate, PARP-1 plays a major role in the DNA repair process. Conversely, in the case of massive DNA damage, excessive activation of PARP-1 (in the case of NAD supplementation)⁺In vivo) that eventually leads to cell necrosis and apoptosis (TentoriL, Portarena I, Graziani g. potential applications of poly (ADP-ribose) polymerase (parp) inhibition. pharmacol Res 2002, 45, 73-85). This activation of PARP can also lead to AIF (apoptosis induction)Lead), triggers caspase-independent apoptotic pathways. (Hong SJ, Dawson TM and Dawson VL, Nuclear and mitochondral configurations in cell death: PARP-1 and AIF. trends in pharmaceutical Sciences 25259-264 (2004)).

Due to the dual action of PARP-1, inhibitors of this enzyme, such as 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one represented by formula 1, may have chemosensitizing (by preventing DNA repair, e.g. after anti-cancer therapy) effects, or effects in the treatment of various diseases and toxic states associated with oxidative or nitric oxide-induced stress and subsequent overactivation of PARP. Such conditions include neuropathy and neurodegenerative diseases (e.g., Parkinson ' S disease, Alzheimer ' S disease) (Love S, Barber R, Wilcock GK. incorporated Poly (ADP-ribosyl) ation of nuclear proteins in Alzheimer ' S disease. Brain1999; 122: 247-53; Mandir AS, Przedborski S, Jackson Lewis V et al. Poly (ADP-ribose) polymerase activity mediators 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) -induced physiological syndrome. Proc Natl Acad Sci USA 1999; 96: 5774-9); cardiovascular diseases (e.g., Myocardial infarction, ischemia reperfusion injury) (Pieper AA, Walles T, Wei G et al. biomedical injection in reduction dby Poly (ADP-ribose) polymerase-1 gene delivery. J Mol Med 2000; 6: 271-82; Szabo G, Bahres, Stumpf N et al. Poly (ADP-ribose) polymerase injection reduction in compression after heart disease transfer. circle Res 2002; 90: 100-6; U.S. Pat. No.6,423,705); inflammatory diseases (Szabo C, Dawson B. Roleof poly (ADP-ribose) synthesis in inflammation and ischia-reperfusion. TIPS 1998; 19: 287-98); diabetic vascular dysfunction (Sorian FG, ViragL, Szabo C. diabetic endothiolobal dysfunction: role of reactive oxidative and biochemical production and poly (ADP-ribose) polymerase activity. J. MolMed 2001; 79: 437-48); arthritis (Szabo C, Virag L, Cuzzocrea S et al, Protection against peroxide-induced fiber in the front and the Protection against level by inhibition of poly (ADP-ribose) synthsase, Proc Natl Acad Sci USA 1998; 95: 3867-72); and cisplatin-induced nephrotoxicity (Racz I, Tory K, GallyasF et al. BGP-15-a novel poly (ADP-rib) polymerase inhibitor-protectsagaint reagent of catalysis with out compounding of activity. biochem Pharmacol 2002; 63: 1099-111). Furthermore, BRCA 2-deficient tumor cells have been shown to be very sensitive to PARP-1 inhibitors only (Bryant HE, Schultz N, ThomasHD, Parker KM, Flower D, Lopez E, Kyle S, Meuth M, Curtin NJ and Helleday T. "Specific kill of BRCA2 Specific tumors with inhibition of poly (ADP-ribose) polymerase", Nature in the publication). PARP inhibitors have also been implicated in enhancing the induction of Reg gene and HGF gene expression in beta cells, thus promoting proliferation of pancreatic beta cells in the islets of langerhans (U.S. patent application publication 2004/0091453; PCT publication No. wo 02/00665). In addition, PARP inhibitors are also used in the manufacture of cosmetics, particularly after-sun creams (PCT publication No. wo 01/82877). No marketed PARP inhibitors exist at present.

Cancer remains a disease that is urgently treated. Cytotoxic chemotherapy remains the primary method of systemic treatment for most cancers, particularly advanced diseases. However, there are few cytotoxic chemotherapeutic agents or therapies that can effectively improve overall survival in patients with advanced or metastatic disease. In addition, the therapeutic window associated with cytotoxic agents is small, which results in high toxicity and less than ideal therapeutic efficacy. Therefore, chemosensitizers that can increase the efficacy of cytotoxic drugs at a suitable dose-resistant amount would meet the stringent requirements for cancer patients. U.S. provisional patent application No.60/612,458 and 60/683,006 entitled "Therapeutic Combinations Comprising Poly (ADP-Ribose) polymeraseInhibitor," which is incorporated herein by reference in its entirety, describe pharmaceutical Combinations of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one.

In order to prepare a pharmaceutical composition comprising 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one for administration to a mammal, it is desirable to manufacture such a compound in a form having physical properties suitable for reliable formulation. Accordingly, there is a need in the art to provide improved forms of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one with enhanced properties, such as improved solubility or bioavailability and stability to heat, water and light.

Summary of The Invention

In one aspect, the present invention provides six polymorphic forms and an amorphous form of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one.

In one embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form I.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph of form III.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form IV.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form V.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is substantially pure polymorph form VI.

In another embodiment, the invention provides a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is substantially pure amorphous.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form I having an X-ray powder diffraction pattern comprising peaks at diffraction angles (2 θ) of 10.9, 19.3, 22.9 and 25.0.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II having an X-ray powder diffraction pattern comprising peaks at diffraction angles (2 θ) of 11.2, 14.0, 20.1 and 23.1.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form III having an X-ray powder diffraction pattern comprising peaks at diffraction angles (2 θ) of 10.7, 11.0, 19.4, and 25.1.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form IV having an X-ray powder diffraction pattern comprising peaks at diffraction angles (2 θ) of 8.2, 16.5, 23.0, and 24.8.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form V having an X-ray powder diffraction pattern comprising peaks at diffraction angles (2 θ) of 10.8, 14.8, 21.6 and 25.8.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form VI having an X-ray powder diffraction pattern comprising peaks at diffraction angles (2 θ) of 14.8, 20.0, 22.3, and 23.5.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form I having an X-ray powder diffraction pattern including a peak at substantially the same diffraction angle (2 θ) as shown in figure 1.

In another embodiment, the present invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II having an X-ray powder diffraction pattern including a peak at substantially the same diffraction angle (2 θ) as shown in figure 4.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form III having an X-ray powder diffraction pattern including a peak at substantially the same diffraction angle (2 θ) as shown in figure 7.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form IV having an X-ray powder diffraction pattern including a peak at substantially the same diffraction angle (2 θ) as shown in figure 10.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form V having an X-ray powder diffraction pattern including a peak at substantially the same diffraction angle (2 θ) as shown in figure 13.

In another embodiment, the invention provides a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form VI having an X-ray powder diffraction pattern including peaks at diffraction angles (2 θ) substantially the same as shown in figure 18.

In another embodiment, the present invention provides 8-fluoro-2- {4- [ (methylamino) methyl group]Phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd]An amorphous form of a phosphate salt of indol-6-one, wherein the X-ray powder diffraction pattern of the amorphous form exhibits broad peaks at diffraction angles (2 theta) in the range of 4-40 ° without any peak characteristic of the crystalline form. More specifically, the amorphous substance is characterized by an X-ray powder diffraction pattern substantially the same as that shown in fig. 21. Even more specifically, the amorphous substance includes a peak (cm) shifted by substantially the same as that shown in FIG. 23^-1) Characterized by the raman spectrum of (a).

In another embodiment, the present invention provides a solid form of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the solid form is a solid form comprising at least two of the following forms: I. polymorphic and amorphous forms II, III, IV, V, VI.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form I.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form III.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form IV.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form V.

In another embodiment, the present invention provides a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is substantially pure polymorph form VI.

In another embodiment, the invention provides a pharmaceutical composition comprising a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure amorphous substance.

In another embodiment, the present invention provides a pharmaceutical composition in solid form comprising a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the solid form is a pharmaceutical composition comprising at least two of the following forms: I. polymorphic or amorphous forms II, III, IV, V, VI.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form I.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form III.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form IV.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form V.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is substantially pure polymorph form VI.

In another embodiment, the invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, the method comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure amorphous form.

In another embodiment, the present invention provides a method of treating a disease condition in a mammal mediated by poly (ADP-ribose) polymerase activity, the method comprising administering to a mammal in need of treatment a therapeutically effective amount of a pharmaceutical composition comprising a solid form of a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the solid form is a pharmaceutical composition comprising at least two of the following forms: I. polymorphic and amorphous forms II, III, IV, V, VI.

In another embodiment, the present invention provides a method of treating cancer in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form I.

In another embodiment, the present invention provides a method of treating cancer in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II.

In another embodiment, the present invention provides a method of treating cancer in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form III.

In another embodiment, the present invention provides a method of treating cancer in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form IV.

In another embodiment, the present invention provides a method of treating cancer in a mammal, said method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is substantially pure polymorph form V.

In another embodiment, the present invention provides a method of treating cancer in a mammal, said method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is substantially pure polymorph form VI.

In another embodiment, the invention provides a method of treating cancer in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure amorphous substance.

In another embodiment, the present invention provides a method of treating cancer in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a solid form of a phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the solid form is a pharmaceutical composition comprising at least two of the following forms: I. polymorphic or amorphous forms II, III, IV, V, VI.

In another embodiment, the invention provides a dosage form comprising 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the dosage form is a lyophilized powder for injection, which after resuspension with sterile water for injection provides a final concentration of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one as a free base of 1.0-4.5mg/mL at a pH of 8.0-3.0.

In another embodiment, the invention provides a dosage form comprising 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the dosage form is a lyophilized powder for injection, which, after resuspension with sterile water for injection, provides a final concentration of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one as the free base of 2-3mg/mL at a pH of 5.0-6.0.

Definition of

The term "compound I" refers to the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one. The term "compound of formula 1" refers to 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, as the free base.

The term "active agent" or "active ingredient" refers to a polymorph of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one ("compound I"), or to a solid form comprising two or more polymorphs or non-crystalline forms of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (compound I).

The term "ambient temperature" refers to temperature conditions that typically occur in a laboratory, including temperature ranges of about 20-30 ℃.

The term "amorphous" refers to an amorphous form of a compound.

The term "aqueous base" refers to any organic or inorganic base. Aqueous bases include, by way of example only, metal bicarbonates such as sodium bicarbonate, potassium carbonate, cesium carbonate, and the like.

The term "aromatic solvent" refers to an organic solvent having an aromatic moiety, including, by way of example only, benzene, toluene, xylene isomers, mixtures thereof, and the like.

The term "chemical stability" refers to a type of stability in which a particular compound maintains its chemical integrity, including, but not limited to, thermal stability, light stability, and water stability.

The term "detectable amount" refers to an amount or volume unit that can be detected using conventional techniques such as X-ray powder diffraction, differential scanning calorimetry, HPLC, fourier transform infrared spectroscopy (FT-IR), raman spectroscopy, and the like.

The term "exposure to humidity" refers to the process of exposing a substance to water vapor in a humidifier, humidity cabinet, or any device capable of controlling relative humidity. This term may also describe the process of exposing a substance to ambient humidity during storage.

The term "cancer" includes, but is not limited to, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, melanoma in the skin or eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the Central Nervous System (CNS), primary CNS lymphoma, spinal tumors, brain stem glioma, pituitary adenoma, or. In another embodiment of the method, the abnormal cell growth is a benign proliferative disease including, but not limited to, psoriasis, benign prostatic hypertrophy or restenosis (restinosis).

The term "inert solvent" refers to any solvent or liquid component of a slurry that does not chemically react with other components in the solution or slurry. Inert solvents include, by way of example only, aprotic solvents such as aromatic solvents, ethyl acetate, acetone, methyl tertiary-butyl ether, dioxane, Tetrahydrofuran (THF), and the like. Protic solvents include, by way of example only, methanol, ethanol, propanol isomers, butanol isomers, and the like.

The term "mediated by poly (ADP-ribose) polymerase (PARP) activity" refers to a biological or molecular process that is regulated, modulated or inhibited by PARP activity. For certain applications, inhibition of PARP activity associated with cancer is preferred. The present invention includes methods of modulating or inhibiting PARP activity, for example, in a mammal by administering a polymorph of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (compound I) or a solid form comprising two or more polymorphs of compound I. The activity or potency of a polymorph of compound I or a solid form comprising two or more polymorphs of compound I can be measured as described, for example, in U.S. patent No.6,495,541 and U.S. provisional patent application No.60/612,458, both of which are incorporated herein by reference in their entirety.

The term "minimum amount" refers to the minimum amount of solvent required to completely dissolve a substance at a given temperature.

The term "polymorph" as used herein refers to different crystalline forms of the same compound as well as other solid state molecular forms, including pseudopolymorphs, such as hydrates (e.g., the presence of bound water in the crystal structure) and solvates (e.g., bound nonaqueous solvents) of the same compound. Different crystalline polymorphs have different crystal structures due to the different arrangement of the molecules in the crystal lattice. This results in different crystal symmetries and/or unit cell parameters, directly affecting its physical properties, such as the X-ray diffraction characteristics of the crystal or powder. For example, different polymorphs typically diffract at different series of angles and yield different intensity values. Thus, X-ray powder diffraction can be used to identify different polymorphs or solid forms containing more than one polymorph in a reproducible and reliable manner (S.Byrn et al, Pharmaceutical solutions: A Stretic Approach to Regulation Considerations, Pharmaceutical research, Vol.12, No.7, p.945-954, 1995; J.K.Haleblian and W.McCron, Pharmaceutical Applications of Polymorphism, Journal of Pharmaceutical Sciences, Vol.58, No.8, p.911-929, 1969). Crystalline polymorphs, particularly those involved in the development of suitable dosage forms, are of great importance to the pharmaceutical industry. If the polymorph does not remain unchanged during clinical or stability studies, there may be no comparability between batches of exact dosage forms used or studied. It would also be desirable to have a process for preparing a compound having a selected polymorph in high purity, since the presence of impurities may produce undesirable toxic effects when the compound is used in clinical research or commercial products. Certain polymorphs may have enhanced thermodynamic stability or may be more conveniently prepared in high purity in large quantities, and thus are more suitable for inclusion in pharmaceutical formulations. Certain polymorphs may exhibit other advantageous physical properties due to different lattice energies, such as low hygroscopicity, increased solubility, and increased dissolution rate.

The term "peak intensity" refers to the relative signal intensity within a given X-ray diffraction pattern. Factors that can affect the relative peak intensities are the sample thickness and preferred orientation (i.e., the crystalline particles are not randomly distributed).

The term "peak position" as used herein refers to the X-ray refraction position measured and observed in an X-ray powder diffraction experiment. The peak position is directly related to the unit cell size. The peaks identified by their respective peak positions have been taken from the diffraction patterns of the various polymorphic forms I, II, III, IV, V and VI of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (compound I).

The term "PEG" refers to poly (ethylene glycol). PEGs with different polymer chain lengths and thus different viscosities are commercially available. Dissolving PEG 400 in alcohol, acetone, benzene, chloroform, acetic acid, and CCl₄And water.

The term "pharmaceutically acceptable carrier, diluent or excipient" refers to a material (or materials) that can be included with a particular pharmaceutical agent to form a pharmaceutical composition and that can be solid or liquid. Exemplary solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and the like. Exemplary liquid carriers include syrup, peanut oil, olive oil, water, and the like. Similarly, the carrier or diluent may include time-delay or time-release materials known in the art, such as glyceryl monostearate or glyceryl distearate with or without a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate, and the like.

The term "pharmaceutical composition" refers to a mixture of one or more of the compounds or polymorphs described herein, or physiologically/pharmaceutically acceptable salts or solvates thereof, with other chemical components such as physiologically/pharmaceutically acceptable carriers or excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

The term "recrystallization" refers to the process of completely dissolving a solid in a first solvent under heating (if necessary), and then causing precipitation, typically by cooling the solution or by adding a second solvent that dissolves the solid less.

The term "relative humidity" refers to the ratio of the amount of water vapor in air at a given temperature to the maximum amount of water vapor that can be present at that temperature and pressure, expressed as a percentage.

The term "relative intensity" refers to the intensity value derived from the X-ray diffraction pattern of a sample. The entire ordinate range scale of the diffraction pattern is designated as value 100. Peaks whose intensity falls between about 50% and about 100% of the intensity on this scale are called very strong (vs) peaks; peaks with intensities falling between about 50% and about 25% are called strong(s) peaks. Other weaker peaks are present in the typical diffraction pattern and are also characteristic peaks for a given polymorph.

The term "slurry" refers to a solid material suspended in a liquid medium (typically water and an organic solvent).

The term "isolation" refers to a step in a synthesis in which a desired agent is separated from other undesired agents, including, but not limited to, any of the following steps: filtered, washed with additional solvent or water, dried under heat or vacuum.

The term "substantially pure" in relation to a particular polymorph of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (compound I) means that the polymorph contains less than 10 wt%, preferably less than 5 wt%, preferably less than 3 wt%, preferably less than 1 wt% of impurities, including other polymorphs of compound I. Such purity can be determined, for example, by X-ray powder diffraction.

By "effective amount" is meant an amount of an agent that is capable of substantially inhibiting the proliferation and/or preventing de-differentiation of a eukaryotic cell (e.g., a mammalian, insect, plant, or fungal cell) and is effective for a given use (e.g., a particular therapeutic treatment).

The term "therapeutically effective amount" refers to an amount of a compound or polymorph administered that will alleviate one or more symptoms of the disease being treated to some extent. For the treatment of cancer, a therapeutically effective amount refers to an amount that has at least one of the following effects:

(1) reducing the size of the tumor;

(2) inhibit (i.e., delay, preferably stop to some extent) tumor metastasis;

(3) inhibit (i.e., retard, preferably stop, to some extent) tumor growth to some extent;

(4) to alleviate (or preferably eliminate) to some extent one or more symptoms associated with cancer.

The term "2 θ value" or "2 θ" refers to the experimentally set peak position based on X-ray diffraction experiments, and is the abscissa unit commonly used in diffraction patterns. Experimental setup required that if the reflection was diffracted when the incident beam formed an angle theta with a particular lattice plane, the reflected beam was recorded at an angle of 2 theta.

The term "treatment" refers to a method of reducing or eliminating the hyperproliferative diseases and/or their concomitant symptoms. With respect to cancer, this term simply means that the life expectancy of an individual with cancer will be extended, or one or more symptoms of the disease will be alleviated.

The term "under vacuum" refers to the typical pressure that can be obtained by a laboratory vacuum pump of the oil-or oil-free diaphragm type.

The term "X-ray powder diffraction pattern" refers to the experimentally observed diffraction pattern or parameters derived therefrom. The X-ray powder diffraction pattern is characterized by the peak position (abscissa) and the peak intensity (ordinate).

Drawings

FIG. 1 is an X-ray powder diffraction pattern of polymorph form I (hydrate A) of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (Compound I);

FIG. 2 is a chart of the infrared absorption spectrum of polymorph form I of Compound I (hydrate A);

figure 3 is a Differential Scanning Calorimetry (DSC) thermogram of polymorph form I of compound I (hydrate a), a typical thermogram showing an endotherm with an onset of 202 ℃ at a scan rate of 5 ℃/min;

FIG. 4 is an X-ray powder diffraction pattern of polymorph form II (anhydrous form) of Compound I;

FIG. 5 is a plot of the infrared absorption spectrum of polymorph form II (anhydrous form) of Compound I;

figure 6 is a Differential Scanning Calorimetry (DSC) thermogram of polymorph form II (anhydrous form) of compound I, a typical thermogram showing an endotherm with an onset of 205 ℃ at a scan rate of 5 ℃/min;

figure 7 is an X-ray powder diffraction pattern of polymorph form III of compound I (hydrate B);

FIG. 8 is a chart of the infrared absorption spectrum of polymorph form III of Compound I (hydrate B);

figure 9 is a Differential Scanning Calorimetry (DSC) thermogram for polymorph form III of compound I (hydrate B), the representative thermogram showing an endotherm with an onset of 203 ℃ at a scan rate of 5 ℃/min;

figure 10 is an X-ray powder diffraction pattern of polymorph form IV of compound I (MeOH solvate);

FIG. 11 is a plot of the infrared absorption spectrum of polymorph form IV of Compound I (MeOH solvate);

figure 12 is a Differential Scanning Calorimetry (DSC) thermogram for polymorph form IV of compound I (MeOH solvate), a typical thermogram showing an endotherm with an onset at 204 ℃ at a scan rate of 5 ℃/min;

figure 13 is an X-ray powder diffraction pattern of polymorph form V of compound I (hydrate C);

figure 14 is an aqueous slurry raman spectrum of polymorph form V of compound I (hydrate C);

figure 15 is a pH solubility plot of polymorph form II (anhydrous form) of compound I;

figure 16 is a plot of the infrared absorption spectrum of polymorph form V of compound I (hydrate C);

figure 17 is a Differential Scanning Calorimetry (DSC) thermogram for polymorph form V of compound I (hydrate C);

figure 18 is an X-ray powder diffraction pattern of polymorph form VI of compound I;

FIG. 19 is a plot of the infrared absorption spectrum of polymorph form VI of Compound I;

figure 20 is a Differential Scanning Calorimetry (DSC) thermogram for polymorph form VI of compound I;

FIG. 21 is an X-ray powder diffraction pattern of an amorphous form of Compound I;

FIG. 22 is a graph showing an infrared absorption spectrum of an amorphous compound of Compound I;

FIG. 23 is a Raman spectrum of an amorphous compound of Compound I;

fig. 24 is a Differential Scanning Calorimetry (DSC) thermogram of an amorphous of compound I.

Detailed Description

It has been surprisingly found that the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (compound I) can exist in more than one polymorphic crystalline form. These forms are useful in the formulation of products for the treatment of mammalian disease conditions mediated by poly (ADP-ribose) polymerase activity, including cancer. Each form has one or more advantages over the other forms in terms of bioavailability, stability, or manufacturability. Crystalline polymorphs of compound I have been found that may be more suitable than other polymorphs for large scale preparation and processing. Also provided are amorphous forms of compound I. Methods for preparing these polymorphs and amorphous forms in high purity are described herein. Also provided are methods of preparing each polymorph and amorphous of compound I substantially free of other polymorphs of compound I. Furthermore, the present invention provides pharmaceutical formulations of compound I comprising different polymorphs and amorphouss as described above, as well as methods of treating mammalian disease conditions mediated by poly (ADP-ribose) polymerase activity by administering such pharmaceutical formulations.

I.8-fluoro-2- {4- [ (methylamino) methyl group]Phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd]Indole Polymorphs of the phosphate salt of indole-6-one (Compound I)

The present invention provides several polymorphic crystalline forms of compound I. Each crystalline form of compound I can be characterized by one or more of the following: x-ray powder diffraction patterns (i.e., X-ray diffraction peaks at different diffraction angles (2 θ)); onset of melting point (and onset of dehydration of the hydrate form) as indicated by the endotherm of the Differential Scanning Calorimetry (DSC) thermogram; pattern of FT-IR spectrum; a raman spectrogram pattern; water solubility; photostability under high intensity light conditions at the international coordination office (ICH); and physical and chemical storage stability. For example, the polymorph samples of forms I, II, III, IV, V and VI of compound I are all characterized by the position and relative intensity of the peaks in their X-ray powder diffraction patterns. There are differences in the X-ray powder diffraction parameters for each of the polymorphic forms I, II, III, IV, V and VI of compound I. X-ray powder diffraction can therefore be used to distinguish these polymorphs of compound I.

X-ray powder diffraction pattern measurements were performed on each polymorph of the invention on a Shimadzu XRD-6000 or Bruker Discover D8X-ray diffractometer equipped with a Cu X-ray source operating at 40kV and 30mA or 40kV and 40mA, respectively. For the Shimadzu XRD-6000X-ray diffractometer, the sample was placed in a sample holder and then pressed and leveled with a glass slide. During analysis, the sample was rotated at 60rpm and analyzed at an angle of from 4 ° to 40 ° (θ -2 θ) in steps of 0.04 ° at 5 °/min or 0.02 ° at 2 °/min. If the available materials are limited, the sample is placed on a silicon plate (zero background) and analyzed without rotation. X-ray diffraction peaks, characterized by peak position and intensity values, were obtained from the X-ray powder diffraction pattern of each polymorph of compound I. For Bruker Discover D8, the samples were placed on glass slides and leveled with weighing paper. The samples were analyzed from an angle of 4 ° to 40 ° (θ -2 θ). Those skilled in the art will recognize that the peak position (2 θ) will exhibit some instrumental variation, typically about 0.1 °. Thus, when the polymorph is described by X-ray powder diffraction characteristic peaks, the peak positions (2 θ) should be understood to encompass such differences. Similarly, when the solid form of the present invention is described as having a powder X-ray diffraction pattern substantially the same as that shown in the given figures, the term "substantially the same" is intended to encompass the above-described instrumental differences in diffraction peak positions. Furthermore, those skilled in the art will recognize that relative peak intensities will also exhibit instrumental differences as well as differences due to crystallinity, preferred orientation, prepared sample surface and other factors known to those skilled in the art and thus can only be used for qualitative analysis.

Different polymorphs of compound I can also be distinguished by Differential Scanning Calorimetry (DSC). DSC measures the difference in heat energy absorbed as temperature increases between a sample solution and a suitable reference solvent. DSC thermograms are generally characterized by an endotherm (representing energy absorption) and also an exotherm (representing energy release) upon heating of the sample.

DSC (differential scanning calorimetry) thermograms were obtained over a temperature range of 25-250 ℃ using a TA instruments DSC Q1000 and a Mettler Toledo DSC821e instrument with a scanning rate of 5 ℃/min. For DSC analysis, samples were weighed and placed into airtight aluminum vessels sealed and perforated with a hole. Depending on several factors, the compounds of the invention may exhibit endotherms that differ more or less than those shown in the figures (for crystalline polymorph melts, the difference is about 0.01-5 ℃). Factors contributing to such differences include the heating rate (i.e., scan rate) at which the DSC analysis is performed, the manner in which the DSC onset temperature is defined and determined, the calibration standard used, the instrument calibration, the relative humidity, and the chemical purity of the sample. For any given sample, the instrument will be different and the endotherm observed may be different; however, if these instruments are calibrated in a similar manner, the differences are generally within the ranges defined herein.

Raman scattering spectra were obtained by using a Dispersive raman spectrometer Ramen RXN1 from Kaiser optics. The excitation light source is a 785nm external cavity stabilized diode laser. The detector is a Charge Coupled Device (CCD). Resolution was 4cm^-1。

In BrukeThe infrared spectrum was recorded on a r Vector33 FT-IR spectrophotometer. The drug samples were ground with potassium bromide and compressed into small pieces. From 4000cm^-1To 400cm^-1The patch is scanned. Marker 3400cm^-1To 500cm^-1The main peak in between.

Different polymorphs of compound I can also be distinguished by different stability and different solubility.

In one embodiment, the polymorphs of compound I of the present invention are substantially pure, i.e., each polymorph of compound I contains less than 10 wt%, such as less than 5 wt%, or such as less than 3 wt%, or even such as less than 1 wt% of impurities, including other polymorphs of compound I.

The solid forms of the invention may also comprise more than one polymorph. One skilled in the art will recognize that a crystalline form of a given compound may exist as a single substantially pure polymorph, and may also exist as a crystalline form comprising two or more different polymorphs. If the solid form comprises two or more polymorphs, the X-ray diffraction pattern will have the characteristic peaks of each polymorph of the invention. For example, a powder X-ray diffraction pattern of a solid form comprising two polymorphs will be the convolution of two X-ray diffraction patterns of the corresponding substantially pure polymorphs. For example, in one embodiment, the solid form of the invention containing the first and second polymorphs comprises at least 10% of the first polymorph. In another embodiment, the solid form comprises at least 20% of the first polymorph. Yet another embodiment comprises at least 30%, at least 40% or at least 50% of the first polymorph. One skilled in the art will recognize that many such combinations of several polymorphs in such different amounts may be present.

In the discussion that follows, X-ray diffraction and infrared absorption data are given for various polymorphs. Although the measured diffraction angles are shown as two decimal places, it is understood that the precision of the diffraction angles is ± 0.1 ° as described above.

Form IV polymorph (methanol solvate)

Form IV polymorph of compound I can be prepared by phosphorylating the compound 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one represented by formula 1 in methanol.

Polymorph form IV of compound I is physically and chemically stable at 40 ℃ for at least 3 months at 75% relative humidity.

The water solubility of polymorph form IV of Compound I was 4.5mg/mL at pH 5.4.

The X-ray powder diffraction pattern of form IV was measured as described herein, with peaks at the following approximate diffraction angles (2 θ): 8.20, 13.52, 14.88, 16.48, 18.08, 19.14, 20.26, 21.06, 22.08, 23.00, 24.80, 25.54, 26.42, 27.14, 28.36, 29.02, 29.92, 30.58, 32.48, 33.42, 34.8, 35.32, 36.22, 36.78, 37.44, 39.08. Figure 10 provides an X-ray powder diffraction pattern for form IV.

The infrared absorption spectrum of form IV was measured as described herein, at the following approximate locations (cm)^-1) Having a spectral band: 536.57, 600.07, 634.53, 657.47, 792.30, 840.23, 859.36, 873.69, 896.83, 951.68, 1021.79, 1092.70, 1135.05, 1215.91, 1259.88, 1320.12, 1352.07, 1366.62, 1418.17, 1449.57, 1468.60, 1508.23, 1578.49, 1613.42, 2366.47, 2741.05, 3020.46, 3316.12. Figure 11 provides an infrared absorption spectrum of polymorph form IV.

The DSC thermogram of form IV shown in figure 12 indicates that the endotherm begins at 204.0 ℃ at a scan rate of 5 ℃/min.

Polymorph form B.I (hydrate A)

The form I polymorph of compound I is a hydrate. Polymorph form I of compound I can be prepared by treating polymorph form IV (MeOH solvate) with water.

Polymorph form I of compound I is chemically stable at 40 ℃ for at least 3 months at 75% relative humidity, but under these conditions it converts to form III (hydrate B) after one week. Form I may be physically stable for at least 3 months at ambient conditions.

The water solubility of polymorph form I of Compound I was 2.8mg/mL at pH 5.4.

The X-ray powder diffraction pattern of form I was measured as described herein, with peaks at the following approximate diffraction angles (2 θ): 10.56, 10.88, 11.14, 11.54, 13.46, 13.90, 14.30, 15.20, 16.34, 17.12, 18.02, 19.30, 20.02, 20.72, 21.22, 21.76, 22.50, 22.94, 23.70, 24.00, 24.32, 25.02, 25.54, 26.22, 26.60, 27.20, 27.76, 29.02, 29.38, 29.74, 31.26, 31.76, 32.12, 33.52, 35.78. Figure 1 provides the X-ray powder diffraction pattern for form I.

The infrared absorption spectrum of form I was measured as described herein, at the following approximate locations (cm)^-1) Having a spectral band: 520.70, 605.60, 669.72, 705.80, 785.46, 840.18, 870.55, 895.86, 958.01, 1017.72, 1066.76, 1131.33, 1220.68, 1257.70, 1321.43, 1346.32, 1366.48, 1421.79, 1452.75, 1476.19, 1509.10, 1578.58, 1609.21. Figure 2 provides an infrared absorption spectrum of polymorph form I.

The DSC thermogram of form I shown in figure 3 indicates that the endotherm begins at 202 ℃ at a scan rate of 5 ℃/min.

Form C.II polymorph (anhydrous form)

The form II polymorph of compound I is in anhydrous form. Form II can be prepared by dehydration of form I.

The form II polymorph of compound I is stable for at least 3 months at ambient conditions. Form II polymorph of compound I can be converted to form III (hydrate B) at 75% relative humidity, after one week at 40 ℃, or after overnight at 25 ℃ at 90% relative humidity.

The water solubility of polymorph form II of Compound I was 3.0mg/mL at pH 5.4. The pH solubility of form II was studied in the pH range of 3.0-8.0. The buffer system used in this study was 50mM ammonium phosphate for pH 3.0 and 4.0 and 50mM sodium phosphate buffer for pH 5.0-8.0. The solubility results are shown in table 1 and fig. 15. It can be seen that the solubility of form II decreases as the pH increases from 3.0 to 8.0. Since form II is phosphate and phosphate buffer was used in pH solubility studies, the solubility measured in these buffer systems was lower than in water due to the co-ionic effect.

TABLE 1 pH solubility characteristics of form II

Buffer pH	Solubility (mg/mL)
		3	0.46
4	0.39
		5	0.37
6	0.32
		7	0.25
8	0.19

The X-ray powder diffraction pattern of form II was measured as described herein, with peaks at the following approximate diffraction angles (2 θ): 7.02, 11.22, 12.12, 14.00, 14.44, 15.36, 18.12, 20.12, 20.92, 22.52, 23.12, 25.28, 26.96, 28.00, 30.02, 31.40, 32.04, 33.72, 34.62, 36.66, 37.26, 39.04. Figure 4 provides the X-ray powder diffraction pattern for form II.

The infrared absorption spectrum of form II was measured as described herein, at the following approximate locations (cm)^-1) Having a spectral band: 514.55, 552.15, 595.96, 662.04, 683.16, 784.79, 842.83, 878.85, 958.55, 1017.68, 1057.72, 1129.87, 1259.81, 1320.16, 1342.76, 1367.10, 1455.18, 1508.93, 1578.68, 1610.29, 2345.53, 2375.37, 2756.52, 3015.19, 3277.71. Figure 5 provides an infrared absorption spectrum of polymorph form II.

The DSC thermogram of form II shown in figure 6 shows that the endotherm begins at 205 ℃ at a scan rate of 5 ℃/min.

Polymorph form III (hydrate B)

The form III polymorph of compound I is a hydrate. The form III polymorph of compound I can be prepared by hydration of form I or form II.

Polymorph form III of compound I is physically and chemically stable at 40 ℃ for at least 3 months at 75% relative humidity.

The water solubility of polymorph form III of Compound I was 2.7mg/mL at pH 5.4.

The X-ray powder diffraction pattern of form III was measured as described herein, with peaks at the following approximate diffraction angles (2 θ): 10.66, 10.96, 11.62, 13.54, 15.28, 16.42, 19.36, 20.80, 21.30, 21.86, 22.56, 23.28, 23.78, 24.06, 24.42, 25.12, 26.30, 26.62, 27.32, 27.84, 28.32, 29.10, 29.50, 29.80, 30.24, 31.84, 32.20, 32.58, 35.86. Figure 7 provides the X-ray powder diffraction pattern for form III.

The infrared absorption spectrum of form III was measured as described herein, at the following approximate locations (cm)^-1) Having a spectral band: 521.02, 545.63, 599.97, 672.96, 704.78, 786.38, 811.92, 839.21, 870.68, 897.67, 956.56, 1017.24, 1076.05, 1131.23, 1222.77, 1256.88, 1325.51, 1346.28, 1365.33, 1421.86, 1451.44, 1478.91, 1509.39, 1578.35, 1607.08, 2300.20, 2346.17, 2502.23, 2828.24, 3011.62, 3299.59, 3536.14. Figure 8 provides an infrared absorption spectrum of polymorph form III.

The DSC thermogram of form III shown in fig. 9 indicates that the endotherm begins at 203 ℃ at a scan rate of 5 ℃/min.

Polymorph form E.V (hydrate C)

During the stability study of form II, polymorph form V of compound I is formed when it is stored at 40 ℃ for 6 months at 75% relative humidity. Polymorph form V of compound I is physically and chemically stable at room temperature for at least 3 months.

The water solubility of polymorph form V of Compound I was 3.0mg/mL at pH 5.4.

The X-ray powder diffraction pattern of form V was measured as described herein, with peaks at the following approximate diffraction angles (2 θ): 8.64, 9.05, 10.26, 10.56, 10.84, 13.88, 14.85, 15.24, 16.24, 16.59, 17.07, 18.26, 18.56, 19.26, 19.56, 20.31, 21.16, 21.61, 22.38, 22.96, 23.40, 24.04, 24.34, 24.92, 25.46, 25.78, 26.22, 26.59, 27.10, 27.60, 27.88, 28.24, 29.03, 30.08, 30.44, 31.54, 32.08, 32.52, 36.45, 36.90, 37.14, 37.58, 37.74, 38.30, 39.00. Figure 13 provides the X-ray powder diffraction pattern for form V.

The infrared absorption spectrum of form V was measured as described herein, at the following pointsSimilar position (cm)^-1) Having a spectral band: 955.28, 1019.70, 1045.84, 1067.25, 1092.06, 1104.99, 1133.50, 1260.13, 1320.27, 1366.85, 1418.85, 1450.75, 1470.01, 1579.05, 1613.39, 1632.61, 2761.48, 3024.44, 3278.09, 3312.93. Figure 16 provides an infrared absorption spectrum of polymorph form V.

The DSC thermogram for form V, shown in figure 17, has an endotherm at 199.40 ℃ and desolvation peaks at 57.29 ℃ and 110.73 ℃, respectively.

Polymorph form F.VI

Polymorph form VI of compound I can be prepared by obtaining an aqueous slurry of form II and heating at 100 ℃ overnight. As shown in FIG. 14, transformation began at 80 ℃ and was complete after overnight storage at 100 ℃.

The X-ray powder diffraction pattern of form VI was measured as described herein, with peaks at the following approximate diffraction angles (2 θ): 8.44, 8.71, 14.78, 15.10, 15.73, 16.06, 16.24, 16.9, 17.2, 19.99, 22.32, 22.60, 22.94, 23.49, 23.84, 24.55, 25.30, 25.48, 27.74, 26.02, 27.47, 27.84, 28.10, 28.40, 34.02, 35.12, 35.54, 35.88. Figure 18 provides an X-ray powder diffraction pattern of polymorph form VI.

The infrared absorption spectrum of form VI was measured as described herein, at the following approximate locations (cm)^-1) Having a spectral band: 535.82, 786.11, 841.00, 954.18, 1020.17, 1133.96, 1216.98, 1260.79, 1320.11, 1367.35, 1418.66, 1450.88, 1470.60, 1508.44, 1579.24, 1613.51, 2410.94, 2760.82, 3025.77, 3277.18. Figure 19 provides an infrared absorption spectrum of polymorph form VI.

The DSC thermogram for form VI shown in figure 20 has an endotherm at 219.68 ℃ and desolvation peaks at 88.42 ℃ and 112 ℃, respectively.

G. Amorphous substance

The amorphous material can be prepared by freeze-drying an aqueous solution of compound I.

The X-ray powder diffraction pattern of the amorphous is characterized by broad peaks from 4-40 ° typical of amorphous without the sharp peaks characteristic of any crystalline. Fig. 21 provides the X-ray powder diffraction pattern of the amorphous.

The infrared absorption spectrum of the amorphous (shown in FIG. 22) was measured as described herein, at the following approximate locations (cm)^-1) Having a spectral band: 433. 505, 518, 596, 609, 664, 674, 705, 746, 785, 856, 896, 937, 955, 1020, 1066, 1106, 1132, 1217, 1260, 1319, 1349, 1367, 1419, 1452, 1472, 1508, 1579, 2300, 2349, 2407, 2830, 3031, 3256.

The Raman spectrum of the amorphous substance shown in FIG. 23 includes Raman shift peaks (cm)^-1) The position of the displacement peak is about 1068, 1323, 1350, 1371, 1453, 1556, 1581, 1616.

The DSC thermogram of the amorphous shown in fig. 24 is characterized by the absence of isolated peaks.

Pharmaceutical compositions of the invention

The active agents of the present invention (i.e., the polymorphic forms of compound I described herein or solid forms comprising two or more such polymorphic forms or amorphous forms) may be formulated into pharmaceutical compositions suitable for medical use in mammals. Any suitable route of administration may be employed to provide an effective dose of any of the polymorphic forms I, II, III, IV, V, VI and amorphous form of Compound I to a patient. For example, oral formulations or parenteral formulations and the like may be used. Dosage forms include capsules, tablets, dispersions, suspensions and the like, e.g., enteric-coated capsules and/or tablets, capsules and/or tablets containing enteric-coated particles of compound I. In all dosage forms, the polymorphic forms I, II, III, IV, V, VI and amorphous forms of compound I may be combined with other suitable ingredients. The compositions may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy. The pharmaceutical compositions of the present invention comprise a therapeutically effective amount of the active agent, one or more inert pharmaceutically acceptable carriers, optionally any other therapeutic ingredients, stabilizers, and the like. The carrier must be pharmaceutically acceptable, by which is meant that the carrier is compatible with the other ingredients of the formulation and does not adversely affect the recipient thereof. Such compositions may also include diluents, buffers, binders, disintegrants, thickeners, lubricants, preservatives (including antioxidants), flavoring agents, taste masking agents, inorganic salts (e.g., sodium chloride), antibacterial agents (e.g., benzalkonium chloride), sweeteners, antistatic agents, surfactants (e.g., polysorbates available from BASF such as "TWEEN 20" and "TWEEN 80", and Pluronic block copolymers such as F68 and F88), sorbitan esters, lipids (e.g., phospholipids such as lecithin and other phosphatidylcholines, phosphatidylethanolamines, fatty acids and fatty esters, steroids (e.g., cholesterol)), and chelating agents (e.g., EDTA, zinc, and other suitable cations). Other pharmaceutical excipients and/or additives suitable for use in the compositions of the present invention are listed in "Remington: the Science & practice of Pharmacy, 19 th edition, Williams & Williams, (1995), "Physician's desk Reference", 52 th edition, Medical Economics, Montvale, NJ (1998) and "Handbook of Pharmaceutical Excipients", 3 rd edition, Ed.A.H.bbe, Pharmaceutical Press, 2000. Compositions in which the active agents of the invention are formulated include those suitable for oral, rectal, topical, nasal, ocular or parenteral administration (including intraperitoneal, intravenous, subcutaneous or intramuscular injection).

The amount of active agent in the formulation may vary depending on various factors, including dosage form, condition being treated, target patient population and other considerations, and is generally readily determined by one skilled in the art. A therapeutically effective amount should be that amount necessary to modulate, regulate or inhibit the PARP enzyme. In fact, the amount may vary widely depending on the particular active agent, the severity of the condition being treated, the patient population, formulation stability, and the like. The compositions generally comprise from about 0.001 wt% to about 99 wt% of the active agent in any amount, preferably from about 0.01 wt% to about 5 wt% of the active agent, more preferably from about 0.01 wt% to about 2 wt% of the active agent, the amount also depending on the relative amounts of excipients/additives contained in the composition.

The pharmaceutical compositions of the present invention are administered in conventional dosage forms prepared by combining a therapeutically effective amount of the active agent as an active ingredient with one or more suitable pharmaceutical carriers according to conventional methods. These methods may include mixing, granulating and compressing or dissolving the ingredients to form the desired formulation.

The pharmaceutical carrier used may be solid or liquid. Exemplary solid carriers include sugars (e.g., lactose, sucrose, mannitol, or sorbitol), talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and the like. Exemplary liquid carriers include syrup, peanut oil, olive oil, water, and the like. Similarly, the carrier may include time-delay or time-release materials known in the art, such as glyceryl monostearate or glyceryl distearate with or without a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate, and the like.

Various pharmaceutical forms may be used. Thus, if a solid carrier is used, the formulation may be tableted, filled into hard gelatin capsules as a powder or granules, or in the form of round or diamond shaped tablets. The amount of solid carrier can vary, but generally should be from about 25mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampoule or vial, or an anhydrous liquid suspension.

To obtain a stable water-soluble dosage form, compound I can be dissolved in an aqueous solution of an organic or inorganic acid (e.g., a 0.3M solution of succinic or citric acid). If a soluble salt form is not available, the active agent may be dissolved in a suitable co-solvent or combination of co-solvents. Examples of suitable co-solvents include, but are not limited to, ethanol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerol, and the like, at concentrations ranging from 0-60% of the total volume. The compositions may also be in the form of a solution of compound I in a suitable aqueous medium, for example water or isotonic saline or dextrose solution.

It will be appreciated that the actual dosage of active agent used in the compositions of the invention may vary depending upon the particular crystalline form employed, the particular composition formulated, the mode and site of administration, the host, and the condition being treated. The optimal dosage of an agent under given conditions can be determined by one skilled in the art using routine dosing assays in combination with assay data. For oral administration, an exemplary daily dosage of about 0.001-1000mg/kg body weight, more preferably about 0.001-50mg/kg body weight, is generally employed, and the course of treatment is repeated at appropriate intervals. Prodrug administration is generally measured at a weight level that is chemically equivalent to the weight level of the fully active form. In practicing the present invention, the most suitable route of administration and the size of the therapeutic dose will depend on the nature and severity of the condition being treated. The dose and dose frequency may also vary according to the age, weight and response of the individual patient. In general, suitable oral dosage forms may include a total daily dosage in the range of 5 to 250mg, which may be administered in a single dose or divided equally into multiple doses. The preferred dosage range is 10-80 mg. In general, suitable parenteral dosage forms may include a total daily dosage in the range of 5 to 200mg, which may be administered in a single dose or divided equally into multiple doses. The preferred dosage range is 10-100 mg.

The compositions of the present invention may be manufactured in accordance with generally known processes for the manufacture of pharmaceutical compositions, for example using conventional techniques, such as mixing, dissolving, granulating, dragee-making, finely grinding, emulsifying, encapsulating, entrapping or lyophilizing. Pharmaceutical compositions may be formulated in conventional manner using one or more physiologically acceptable carriers, which may be selected from excipients and auxiliaries, in order to facilitate processing of the active compounds into preparations which can be used as medicaments.

For oral administration, the compounds may be formulated by combining the active agent with pharmaceutically acceptable carriers known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, gels, syrups, slurries, suspensions and the like, for oral administration to a patient to be treated. Pharmaceutical preparations for oral use can be obtained using solid excipients in admixture with the active agents, optionally grinding the resulting mixture and, after adding suitable auxiliaries, treating the mixture of granules, if desired, to obtain tablets or dragee cores. Suitable excipients include: fillers, such as sugars including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, gums, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof (e.g., sodium alginate).

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinylpyrrolidone, Carbopol gel, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablets or sugar coatings for identification or to describe different active agent combinations.

Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient in admixture with fillers (e.g. lactose), binders (e.g. starches) and/or lubricants (e.g. talc or magnesium stearate) and, optionally, stabilizers. In soft capsules, the active agent may be dissolved or suspended in a suitable liquid (e.g., fatty oil, liquid paraffin, or liquid polyethylene glycol). In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may be in the form of tablets or lozenges formulated in conventional manner.

For nasal or inhaled administration, the compounds for use according to the invention are suitably delivered in the form of a spray presentation from a pressurised pack or nebuliser, together with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. When a pressurized spray is employed, the dosage unit may be determined by providing a valve for delivering a metered amount. Gelatin capsules and kits for use in an inhaler or insufflator or the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

The active agent may be formulated for parenteral administration by injection, for example by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added protective agent. The compositions may be in the form of suspensions, solutions or emulsions in, for example, an oily or aqueous carrier, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration include suspensions of the active agent and may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils (e.g. sesame oil) or synthetic fatty acid esters (e.g. ethyl oleate or triglycerides) or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the active agent, so that highly concentrated solutions can be prepared.

For administration to the eye, the active agent may be delivered in a pharmaceutically acceptable ophthalmic vehicle to maintain the compound in contact with the surface of the eye for a sufficient period of time to allow the compound to penetrate the cornea and internal regions of the eye, including, for example, the anterior chamber, the posterior chamber, the vitreous body, aqueous humor, vitreous humor, cornea, iris/ciliary body, lens, choroid/retina and sclera. The pharmaceutically acceptable ophthalmic carrier may be, for example, an ointment, a vegetable oil or an encapsulating material. The compounds of the invention may also be injected directly into the vitreous and aqueous humor or subconjunctival (subtenon) injections.

Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use. The compounds may also be formulated, for example, as rectal or vaginal compositions, e.g., suppositories or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides.

In addition to the above formulations, the polymorphs can also be formulated as depot preparations. Such long acting formulations may be administered by implantation (e.g. subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the polymorph can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives (e.g., as a sparingly soluble salt).

In addition, sustained release systems may be used to deliver the active agent, for example using semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. There are a variety of sustained release materials known to those skilled in the art. Sustained release capsules can release the compound over a period of weeks up to over 100 days depending on their chemical nature. Depending on the chemical nature and biological stability of the therapeutic agent, additional protocols for protein stabilization may be used.

The pharmaceutical compositions may also contain suitable solid and gel phase carriers or excipients. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starch, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

Methods of use of polymorphs of the invention

The polymorphic forms of compound I of the present invention are useful for modulating the activity of poly (ADP-ribose) polymerase (PARP). More specifically, this polymorph is suitable for use as a chemosensitizer, which may enhance the efficacy of radiation therapy or cytotoxic drugs (whose mechanism is dependent on DNA damage). Such drugs include, but are not limited to, temozolomide (SCHERING), irinotecan (irinotecan) (PFIZER), topotecan (GLAXOSMITHKLINE), cisplatin (cisclin) (BRISTOL MEYERS SQUIBB; AMPHARM PARTNERS; BEDFORD; GENSIA SICOR PHARMS; PHARMACHEMIE) and doxorubicin hydrochloride (AM PHARM PARTNERS; BEDFORD; GENSIA; SICOR PHARMS; PHARMACHEMIE; ADRIA; ALZA).

The polymorphic forms of compound I of the present invention are also useful for enhancing the induction of Reg gene and HGF gene expression in beta cells, thus promoting proliferation of pancreatic beta cells of islets of langerhans and inhibiting apoptosis of these cells. Furthermore, the polymorphic forms of compound I of the present invention are suitable for use in the manufacture of cosmetics, such as, for example, after-sun creams.

The agents of the invention are generally administered in therapeutically effective amounts in the form of pharmaceutical compositions to treat diseases by modulating or regulating PARP. By "effective amount" is meant an amount of an agent sufficient to effect treatment of a disease mediated by the activity of one or more PARP enzymes when administered to a mammal, including a human, in need of such treatment. Thus, a therapeutically effective amount of a compound of the present invention is an amount sufficient to modulate, regulate or inhibit the activity of one or more PARP enzymes, such that a disease condition mediated by such activity is alleviated or alleviated. An effective amount of a given compound will vary depending upon factors such as the disease condition and its severity and the characteristics and conditions (e.g., weight) of the mammal in need of treatment, but can nevertheless be determined by one skilled in the art by routine methods. By "treating" is meant at least alleviating the disease condition in a mammal (including a human being) that is at least partially affected by the activity of one or more PARP enzymes, and includes: preventing the occurrence of a disease condition in a mammal, particularly when the mammal is found to be predisposed to having the disease condition but has not yet been diagnosed; modulating and/or inhibiting the disease condition; and/or alleviating the disease condition. Exemplary disease conditions include cancer.

The activity of the polymorph of the invention as a modulator of PARP activity may be determined by any method known to those skilled in the art, including in vivo and/or in vitro assays. Examples of suitable assays for determining activity include those described in the following references: U.S. patent No.6,495,541 and U.S. provisional patent application No.60/612,458, which are incorporated herein by reference in their entirety.

The present invention also relates to therapeutic methods for treating disease conditions mediated by PARP activity, such as cancer and various diseases and toxic states associated with oxidative or nitric oxide-induced stress and subsequent overactivation of PARP. Such conditions include, but are not limited to, neuropathy and neurodegenerative diseases (e.g., parkinson's disease, senile dementia), cardiovascular diseases (e.g., myocardial infarction, ischemia reperfusion injury), diabetic vascular dysfunction, cisplatin-induced nephrotoxicity. The methods of treatment of the present invention comprise administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising any one of the polymorphs or a pharmaceutical composition as discussed above.

The present invention also relates to a combination therapy method for treating disease conditions mediated by PARP activity comprising administering to a mammal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising any one of the polymorphs or a pharmaceutical composition as discussed above, and a therapeutically effective amount of one or more substances selected from the group consisting of anti-tumor agents, anti-angiogenic agents, signal transduction inhibitors and antiproliferative agents. Such materials include those disclosed in PCT publications WO 00/38715, WO 00/38716, WO 00/38717, WO00/38718, WO 00/38719, WO 00/38730, WO 00/38665, WO 00/37107 and WO 00/38786, which are hereby incorporated by reference in their entirety.

Examples of antineoplastic agents include temozolomide (SCHERING), irinotecan (PFIZER), topotecan (GLAXO SMITHKLINE), cisplatin (BRISTYLMEYERS SQUIBB; AM PHARM PARTNERS; BEDFORD; GENSIAOR PHARMS; PHARMACHEMIE), and doxorubicin hydrochloride (AM PHARMPARTNERS; BEDFORD; GENSIA; SICOR PHARMS; PHARMACHEMIE; ADSIRIA; ALZA).

Other examples of antineoplastic agents include mitotic inhibitors, such as, for example, vinorelbine,vinblastine derivatives such as vindesine and vincristine, colchicine allochosine, halichondrine, N-benzoyltrimethyl-methyl ether colchicic acid, dolastatin 10, maystasine, rhizoxin, taxanes, such as paclitaxel (paclitaxel), docetaxel (Taxotere), 2' -N- [3- (dimethylamino) propyl ] taxane]Glutarate (paclitaxel derivative), thiocolchicine, trityl cysteine, teniposide, methotrexate, azathioprine, fluorouricil, cytarabine, 2' -difluorodeoxycytidine (gemcitabine), doxorubicin and mitomycin. Alkylating agents, for example carboplatin (carboplatin), oxoplatin (oxipulin), isoplatinum, ethyl ester of N-acetyl-DL-sarcosine-L-leucine (Asaley or Asalex), 1, 4-cyclohexadiene-1, 4-dicarbamic acid, 2, 5-bis (1-aziridinyl) -3, 6-dioxo-diethyl ester (diazaquinone), 1, 4-bis (methanesulfonyloxy) butane (bisfan or leucosin), zourecin (chlorotocin), cromethasone (clomesone), cyanomorpholino doxorubicin, cyclodiosone, dianhydrogalactitol, fluoropolypane (fluorodomopan), hepsulfam, mitomycin C, hydroxyanthrone mitomycin C, mitoxantrone, 1- (2-chloroethyl) -4- (3-chloropropyl) -piperazine dihydrochloride, polipidium chloride, polipidium bromide, propidium bromide, chloropicrin C, mitomycin C, mitoxanilide, bromopicloratadine, and the like, Methyl mitomycin, spirohydantoin mustard, tirofoximon (teroxirone), platine, thiotepa, triethylenemelamine, uracil mustard, bis (3-methanesulfonyloxypropyl) amine hydrochloride, mitomycin, nitrosourea agents such as cyclohexyl-chloroethylnitrosourea, methylcyclohexyl-chloroethylnitrosourea, 1- (2-chloroethyl) -3- (2, 6-dioxo-3-piperidinyl) -1-nitrosourea, bis (2-chloroethyl) nitrosourea, procarbazine, dacarbazine, nitrogen mustard related compounds such as methylchloroethylamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, estramustine sodium phosphate and strptozoin. DNA antimetabolites, e.g. 5-fluorouracil, arabinocytosine, hydroxyurea, 2- [ (3-hydroxy-2-pyridyl) methylene]Hydrazinothiosemicarbazide, deoxyfluorouracil, 5-hydroxy-2-formylpyridine, thiosemicarbazone, alpha-2' -deoxy-6-thioguanine, aphidicolin glycinate, 5-azadeoxycytidine, beta-thioguanineRibonucleosides, cyclocytidine, guanazol, inosine glycinedial, macbecin II, pyrazoloimidazole, cladribine (cladribine), pentostatin (pentostatin), thioguanine, mercaptopurine, bleomycin, 2-chlorodeoxyadenosine, thymidylate synthase inhibitors such as raltitrexed and pemetrexed disodium, clofarabine (clofarabine), fluorouracil, and fludarabine (fludarabine). DNA/RNA antimetabolites, e.g., L-aranorubine (alanosine), 5-azacytidine, acivicin (acivicin), aminopterin and derivatives thereof, e.g., N- [ 2-chloro-5- [ [ (2, 4-diamino-5-methyl-6-quinazolinyl) methyl]Amino group]Benzoyl radical]-L-aspartic acid, N- [4- [ [ (2, 4-diamino-5-ethyl-6-quinazolinyl) methyl ] amino]Amino group]Benzoyl radical]-L-aspartic acid, N- [ 2-chloro-4- [ [ (2, 4-diaminopterinyl) methyl]Amino group]Benzoyl radical]-L-aspartic acid, soluble becker antifolate, methamphetamine dichloride, brequinar (brequinar), ftoraf, dihydro-5-azacytidine, methotrexate, N- (phosphonoacetyl) -L-aspartic acid tetrasodium salt, pyrazolofuran, trimetrexate (trimetrexate), plicamycin (plicamycin), actinomycin D, cryptophycin, and analogs such as cryptophycin-52, or one of the preferred antimetabolites such as N- (5- [ N- (3, 4-dihydro-2-methyl-4-oxoquinazolinyl-6-methyl) -N-methylamino-disclosed in, for example, European patent application No.239362]-2-thienyl) -L-glutamic acid; a growth factor inhibitor; a cell cycle inhibitor; intercalating antibiotics, such as doxorubicin and bleomycin; proteins, such as interferon; and anti-hormonal agents, e.g. anti-oestrogens such as Nolvadex^TM(tamoxifen), or e.g. antiandrogens such as Casodex^TM(4 '-cyano-3- (4-fluorophenylsulfonyl) -2-hydroxy-2-methyl-3' - (trifluoromethyl) propenanilide). Such combination therapy may be achieved by administering the individual therapeutic components simultaneously, sequentially or separately.

Anti-angiogenic agents include MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase II) inhibitors. Examples of COX-II inhibitors that may be used include CELEBREX^TM(alecoxib), valdecoxib, and rocholNon-coxib (rofecoxib). Examples of matrix metalloproteinase inhibitors that may be used are disclosed in WO96/33172 (published 24.10.1996), WO 96/27583 (published 7.3.1996), European patent application No.97304971.1 (filed 8.7.1997), European patent application No.99308617.2 (filed 29.10.1999), WO 98/07697 (published 26.2.1998), WO 98/03516 (published 29.1.1998), WO 98/34918 (published 13.8.1998), WO 98/34915 (published 13.8.1998), WO 98/33768 (published 6.8.1998), WO 98/30566 (published 16.7.1998), European patent application 606,046 (published 13.7.1994), European patent publication 931,788 (published 28.7.1999), WO 90/05719 (published 31.1990), WO 99/52910 (published 21.10.1999), WO 99/52889 (published 21/10/1999), WO 99/29667 (published 17/6/1999), PCT International application No. PCT/IB98/01113 (filed 21/7/1998), European patent application No.99302232.1 (filed 25/3/1999), British patent application No.9912961.1 (filed 3/6/1999), U.S. provisional patent application No.60/148,464 (filed 12/8/1999), U.S. patent 5,863,949 (granted 26/1999), U.S. patent 5,861,510 (granted 19/1999), and European patent publication 780,386 (published 25/6/1997), all of which are incorporated herein by reference. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity for inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and/or MMP-9 relative to other matrix metalloproteinases (i.e., MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Examples of MMP inhibitors include AG-3340, RO32-3555, RS 13-0830 and the following compounds: 3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl ] - (1-hydroxycarbamoyl-cyclopentyl) -amino ] -propionic acid; 3-exo-3- [4- (4-fluoro-phenoxy) -benzenesulfonylamino ] -8-oxa-bicyclo [3.2.1] octane-3-carboxylic acid hydroxyamide; (2R, 3R)1- [4- (2-chloro-4-fluoro-benzyloxy) -benzenesulfonyl ] -3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 4- [4- (4-fluoro-phenoxy) -benzenesulfonylamino ] -tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl ] - (1-hydroxycarbamoyl-cyclobutyl) -amino ] -propionic acid; 4- [4- (4-chloro-phenoxy) -benzenesulfonylamino ] -tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3- [4- (4-chloro-phenoxy) -benzenesulfonylamino ] -tetrahydro-pyran-3-carboxylic acid hydroxyamide; (2R, 3R)1- [4- (4-fluoro-2-methyl-benzyloxy) -benzenesulfonyl ] -3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl ] - (1-hydroxycarbamoyl-1-methyl-ethyl) -amino ] -propionic acid; 3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl ] - (4-hydroxycarbamoyl-tetrahydro-pyran-4-yl) -amino ] -propionic acid; 3-exo-3- [4- (4-chloro-phenoxy) -benzenesulfonylamino ] -8-oxa-bicyclo [3.2.1] octane-3-carboxylic acid hydroxyamide; 3-endo-3- [4- (4-chloro-phenoxy) -benzenesulfonylamino ] -8-oxa-bicyclo [3.2.1] octane-3-carboxylic acid hydroxyamide; 3- [4- (4-fluoro-phenoxy) -benzoylamino ] -tetrahydro-furan-3-carboxylic acid hydroxyamide; and pharmaceutically acceptable salts, solvates and hydrates thereof.

Examples of signal transduction inhibitors include agents that can inhibit EGFR (epidermal growth factor receptor) responses, such as EGFR antibodies, EGF antibodies, and molecules that are EGFR inhibitors; VEGF (vascular endothelial growth factor) inhibitors; and erbB2 receptor inhibitors, e.g. organic molecules or antibodies that bind to the erbB2 receptor, e.g. HERCEPTIN^TM(Genentech，Inc.of SouthSan Francisco，California，USA)。

EGFR inhibitors are described, for example, in WO 95/19970 (published 27.7.1995), WO 98/14451 (published 9.4.1998), WO 98/02434 (published 22.1.1998), and U.S. Pat. No. 5,747,498 (granted 5.5.1998). EGFR inhibitors include, but are not limited to, monoclonal antibody C225 and anti-EGFR 22Mab (Imclone systems incorporated of New York, New York, USA), the compound ZD-1839(AstraZeneca), BIBX-1382(Boehringer Ingelheim), MDX-447 (Mercax of Annandale, New Jersey, USA), and OLX-103(Merck & Co. of Whitehouse State, New Jersey, USA) VRCTC-310(Ventech research) and EGF fusion toxins (Seragen incorporated of Hopkinton, Massachusetts).

VEGF inhibitors, such as SU-5416 and SU-6668(Sugen Inc. of South san Francisco, Calif., USA), may also be combined or co-administered with the compositions of the present invention. VEGF inhibitors are described in, for example, WO 99/24440 (published 20/5/1999), PCT International application PCT/IB99/00797 (published 3/5/1999), WO 95/21613 (published 17/8/1995), WO 99/61422 (published 2/12/1999), US patent 5,834,504 (granted 10/11/1998), WO 98/50356 (published 12/11/1998), US patent 5,883,113 (granted 16/3/1999), US patent 5,886,020 (granted 23/1999), US patent 5,792,783 (granted 11/8/1998), WO 99/10349 (published 4/1999), WO 97/32856 (published 12/1997), WO 97/22596 (published 26/1997), WO98/54093 (published 3/1998), WO 98/02438 (published 22/1998), WO 99/16755 (published 8/1999), and WO 98/02437 (published 22/1998) Cloth), the entirety of which is hereby incorporated by reference. Other examples of certain specific VEGF inhibitors are IM862(Cytran inc. of Kirkland, Washington, USA); anti-VEGF monoclonal antibody bevacizumab (Genentech, Inc. of South San Francisco, California); and angiozyme, synthetic nucleic acid enzymes from Ribozyme (Boulder, Colorado) and Chiron (Emeryville, California).

ErbB2 receptor inhibitors, such as GW-282974(GlaxoWellcome plc), as well as monoclonal antibodies AR-209(Aronex Pharmaceuticals Inc. of The Woodlands, Texas, USA) and 2B-1(Chiron), may be administered in combination with The compositions of The present invention. Such erbB2 inhibitors include those described in WO 98/02434 (published 22/1/1998), WO99/35146 (published 15/7/1999), WO 99/35132 (published 15/7/1999), WO 98/02437 (published 22/1/1998), WO 97/13760 (published 17/4/1997), WO 95/19970 (published 27/7/1995), U.S. Pat. No. 5,587,458 (issued 24/12/1996) and U.S. Pat. No. 5,877,305 (issued 2/3/1999), the entire contents of which are incorporated herein by reference. erbB2 receptor inhibitors useful in the present invention are also described in U.S. provisional application No.60/117,341 filed on 27.1.1999 and U.S. provisional application No.60/117,346 filed on 27.1.1999, both of which are incorporated herein by reference in their entirety.

Other antiproliferative agents which may be used include inhibitors of farnesyl protein transferase and inhibitors of the tyrosine kinase PDGFr receptor, including the compounds disclosed and claimed in the following U.S. patent applications: 09/221946 (filed 28/12/1998), 09/454058 (filed 2/12/1999), 09/501163 (filed 9/2/2000), 09/539930 (filed 31/3/2000), 09/202796 (filed 22/5/1997), 09/384339 (filed 26/8/1999), and 09/383755 (filed 26/8/1999), as well as the compounds disclosed and claimed in the following U.S. provisional patent applications: 60/168207 (filed on 30/11/1999), 60/170119 (filed on 10/12/1999), 60/177718 (filed on 21/1/2000), 60/168217 (filed on 30/11/1999), and 60/200834 (filed on 1/5/2000). All of the above-mentioned patent applications and provisional patent applications are incorporated herein by reference in their entirety.

The compositions of the invention may also be used with other agents useful in the treatment of abnormal cell growth or cancer, including, but not limited to, agents capable of enhancing anti-tumor immune responses, such as CTLA4 (cytotoxic lymphocyte antigen 4) antibodies and other agents capable of blocking CTLA4, as well as anti-proliferative agents, such as other farnesyl protein transferase inhibitors. Specific CTLA4 antibodies useful in the present invention include those disclosed in U.S. provisional patent application 60/113,647 (filed 12/23 of 1998), which is incorporated herein by reference in its entirety.

The disclosures of all references are incorporated herein by reference in their entirety.

Examples

The following examples serve to further illustrate the processes for the preparation of the various polymorphs of the present invention (i.e., polymorphic forms I, II, III, IV, V, VI and amorphous form of compound I), but are not intended to limit the scope of the invention as defined herein or claimed in the claims. All temperatures used are in degrees Celsius and all parts and percentages are by weight unless otherwise indicated.

Example 1: preparation and characterization of polymorph form IV of Compound I (methanol solvate)

Form IV polymorph of compound I was prepared by the following method. A500 mL round bottom flask was charged with the compound 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one represented by formula 1 (1.65g, 5.10mmol, 1.0 equiv.) and methanol (200 mL). The mixture was stirred until a clear solution was obtained (about 10 minutes). A0.5M solution of phosphoric acid in methanol (11.0mL, 5.87mmol, 1.15 equiv., prepared by dissolving 0.7g of 85% phosphoric acid in 11.0mL of methanol) was added. The resulting mixture was stirred at ambient temperature for 30 minutes. The resulting solid was filtered and dried at 45 ℃ to give polymorph form IV of the phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one (1.43 g).

Figure 10 is an X-ray powder diffraction pattern of polymorph form IV of compound I. Figure 11 is an infrared absorption spectrum of polymorph form IV. Polymorph form IV of compound I was further characterized by differential scanning calorimetry (figure 12).

Example 2: preparation and characterization of polymorph form I of Compound I (hydrate A)

Polymorph I of compound I was prepared by the following method. A 50mL round bottom flask was charged with polymorph form IV (methanol solvate) of compound I slurried with 10mL water (1.0g) and stirred at ambient temperature for 18-24 hours. The resulting solid was filtered and dried at 45 ℃ to give polymorph form I of compound I (0.67 g). The product was analyzed by NMR without methanol.

Figure 1 is an X-ray powder diffraction pattern of polymorph form I of compound I. Figure 2 is an infrared absorption spectrum of form I polymorph. Form I polymorph of compound I was further characterized by differential scanning calorimetry (figure 3).

Example 3: preparation and characterization of polymorph form II (anhydrous form) of Compound I

Form II was prepared as follows:

(a) heating form I overnight at 60 deg.C;

(b) maintaining form I overnight under vacuum at room temperature; or

(c) Form I is maintained at 25 ℃ for more than 5 hours at a relative humidity of 0%.

Figure 4 is an X-ray powder diffraction pattern of polymorph form II of compound I. Figure 5 is an infrared absorption spectrum of polymorph form II. Form II polymorph of compound I was further characterized by differential scanning calorimetry (figure 6).

Storing form II at 2-8 deg.C in the presence of a desiccant.

Example 4: preparation and characterization of polymorph form III of Compound I (hydrate B)

The form III polymorph of compound I was prepared as follows:

(a) exposing form I to a relative humidity of 90% for more than 5 hours at 25 ℃;

(b) exposing form I to 75% relative humidity for 1 week at 40 ℃; or

(c) Form II was exposed to 75% relative humidity overnight at 40 ℃.

Figure 7 is an X-ray powder diffraction pattern of polymorph form III of compound I. Figure 8 is an infrared absorption spectrum of polymorph form III. Form III polymorph of compound I was further characterized by differential scanning calorimetry (figure 9).

Example 5: preparation and characterization of polymorph form V of Compound I (hydrate C)

Form V polymorph of compound I was formed when stability studies were performed on form II after 6 months at 40 ℃ at 75% relative humidity. Polymorph form V of compound I is physically and chemically stable at room temperature for at least 3 months.

The water solubility of polymorph form V of Compound I was 3.0mg/mL at pH 5.4.

Figure 13 provides an X-ray powder diffraction pattern for polymorph form V of compound I. Figure 16 is an infrared absorption spectrum of polymorph form V. The DSC thermogram for form V has an endotherm at 199.40 ℃ and desolvation peaks at 57.29 ℃ and 110.73 ℃, respectively (fig. 17).

Example 6: preparation and characterization of polymorph form VI of Compound I

Polymorph form VI of compound I can be prepared by obtaining an aqueous slurry of form II and heating at 100 ℃ overnight. As shown in FIG. 14, transformation began at 80 ℃ and was complete after overnight storage at 100 ℃.

The VI polymorph has the characteristics described above. Figure 18 is an X-ray powder diffraction pattern of form VI of compound I. Figure 19 is an infrared absorption spectrum of form VI polymorph of compound I. Figure 20 is a Differential Scanning Calorimetry (DSC) thermogram for polymorph form VI of compound I.

Example 7: use of polymorph form II (anhydrous form) of Compound I for the preparation of pharmaceutical compositions

A. Complete composition

The following provides the use of polymorph form II (anhydrous form) of compound I for the preparation of lyophilized powder for injection at 12 mg/vial (as free base) for clinical use.

The pharmaceutical product is first formulated as a solution of compound I for lyophilization. The quantitative composition of the compound I solution used for lyophilization is shown in table 2.

TABLE 2 Compound I solution for lyophilization, 3mg/mL (as free base)

The quantitative unit compositions of the lyophilized powders of compound I for injection are shown in table 3.

TABLE 3 Dry powder for injection, 12 mg/vial (as free base)

B. Amount of excess

The clinical composition of the lyophilized powder of compound I for injection, 12 mg/vial (as free base), contains a theoretical excess (overage) of 0.45 mg/vial (as free base). This excess compensates for the solid volume in the vial after resuspension with 6mL sterile water for injection (SWFI) and ensures delivery of 2.02mg/mL (as free base) of the drug solution.

C. Container with a lid

The components of the packaging system for the lyophilized powder for injection of compound I are listed below:

D. development of pharmaceutical principles for dosage form selection

Lyophilized powders for injection are conventional dosage forms for administration. The clinical formulation comprises mannitol as a bulking agent and an osmolality adjusting agent. Resuspend drug product with 6mL SWFI to give a clear, hypotonic 2.02mg/mL (as free base) solution. The resuspended drug product will be diluted with an acceptable isotonic sterile diluent for infusion.

The clinical drug product was originally designed to be resuspended with 4mL of SWFI to give a clear, isotonic 3mg/mL (as free base) solution. During the drug product stability evaluation, the haze/turbidity in the solution was observed and studied. Haze/turbidity is caused by drug crystallization of the drug polymorph (hydrate B). The water solubility of polymorph form III (hydrate B) was 2.7mg/mL at pH 5.4, thus very close to the originally desired resuspension concentration of the drug product (3 mg/mL). The SWFI resuspension volume was changed from 4mL to 6mL to ensure complete dissolution of the drug. The final concentration of the resulting drug product solution was 2.02mg/mL (as the free base), suitably lower than the aqueous solubility of polymorph form III (hydrate B).

E. Clinical manufacturing formula, manufacturing process, and in-process control and assembly process

The manufacturing process of lyophilized powder for injection (12 mg/vial (as free base)) of compound I is listed below. The current clinical batch size is 9.3L per manufacturing stage. The formulation was made the same as the clinical composition (see tables 2 and 3).

a) Adding about 75% of the total amount of water for injection (WFI) to the mixing vessel;

b) adding the required amount of mannitol while stirring and allowing it to dissolve completely;

c) heating the WFI/mannitol solution to about 58 deg.C, adding the desired amount of Compound I drug substance, and mixing until completely dissolved;

d) the solution was brought to final volume (weight) with WFI, mixed for 10 minutes, and the solution was cooled to room temperature;

e) equal aliquots were taken for in-process control testing (i.e., appearance, pH, density and UV analysis);

f) the bulk solution for lyophilization was sterile filtered through 0.45 μm and 0.22 μm membrane filters and 4.15mL (including 0.15mL excess) was aseptically filled into 10mL/20mm type I amber glass vials;

g) freeze-drying the filled vial with the partially inserted stopper;

h) at the end of the freeze-drying cycle, the vials were back-filled with nitrogen and stoppers at room temperature under a low vacuum;

i) sealing the freeze-dried vial with an aluminum end cap;

j) the vials were refrigerated.

Example 8: preparation and characterization of amorphous Compound I

An amorphous form of compound I was prepared by dissolving polymorph form II of compound I (anhydrous form) in sterile water for injection at a concentration of 4.46 mg. 2mL of this solution was filled into 10mL clean type I vials and freeze dried in an FTS LyoStar freeze dryer (S/N LSACC 3). The freeze-drying cycle is described below.

The product was frozen to-50 ℃ and then vacuum dried at-30, -20 ℃ and-15 ℃ for 12 hours, respectively, to complete the first drying step. The vacuum pressure was maintained at 200 mtorr. The product was further dried at 25 ℃ under vacuum of 200mtorr for 24 hours, completing the second drying step.

A white/pale yellow lyophilized cake of compound I amorphous was obtained. The amorphous compound I can be resuspended in 2mL sterile water for injection to give a clear yellow solution.

Claims (3)

1. A crystalline phosphate salt of 8-fluoro-2- {4- [ (methylamino) methyl ] phenyl } -1,3,4, 5-tetrahydro-6H-azepino [5,4,3-cd ] indol-6-one, wherein the salt is a substantially pure polymorph form II having an X-ray powder diffraction pattern with peaks at diffraction angles (2 Θ) of 7.02, 11.2, 12.12, 14.0, 20.1, and 23.1.
2. 2. A pharmaceutical composition comprising a salt of claim 1.
3. 3. Use of a therapeutically effective amount of a salt according to claim 1 in the manufacture of a medicament for treating a disease mediated by poly (ADP-ribose) polymerase activity in a patient in need of treatment.