JP2008510783A - 4-heteroarylmethyl-substituted phthalazinone derivatives - Google Patents

Abstract

Compounds of formula (I) used to treat cancer or other diseases ameliorated by inhibition of PARP, wherein A and B together represent an optionally substituted fused aromatic ring; X is NR ^X or CR ^X R ^Y ; if X = NR ^X , n is 1 or 2, if X = CR ^X R ^Y , n is 1; R ^X is H, substituted May be selected from the group consisting of C ^1-20 alkyl, C ^5-20 aryl, C ^3-20 heterocyclyl, amide, thioamide, ester, acyl, and sulfonyl groups; R ^Y is selected from H, hydroxy, amino; Alternatively, R ^X and R ^Y may be taken together to form a spiro-C ^3-7 cycloalkyl or heterocyclyl group; R ^C1 and R ^C2 are independently from the group consisting of hydrogen and C ^1-4 alkyl When selected or X is CR ^X R ^Y , R ^C1 , R ^C2 , R ^X and R ^Y together with the carbon atom to which they are attached And may form an optionally substituted fused aromatic ring; R ¹ is selected from H and halo; and Het is (i) formula (i) (where Y ¹ is CH and N Y ² is selected from CH and N, Y ³ is selected from CH, CF and N, where only one or two of Y ¹ , Y ² and Y ³ can be N ), And (ii) a compound selected from formula (ii), wherein Q is O or S.

Description

  The present invention relates to phthalazinone derivatives and their use as medicaments. Specifically, the present invention is also known as poly (ADP-ribose) synthase and polyADP-ribosyltransferase, and is intended to inhibit the activity of the enzyme poly (ADP-ribose) polymerase-1, commonly referred to as PARP-1. It relates to the use of these compounds.

  The mammalian enzyme PARP-1 (113 kDa multidomain protein) is involved in signaling DNA damage through its ability to recognize and rapidly bind to DNA single- or double-strand breaks ( D'Amours et al., Biochem. J., 342, 249-268 (1999)).

  The family of poly (ADP-ribose) polymerases currently includes approximately 18 proteins, all of which show some degree of homology in their catalytic domains but differ in their cellular functions (Ame et al. al., Bioessays., 26 (8), 882-893 (2004)). Within this family, PARP-1 (original member) and PARP-2 have so far been the only enzymes whose catalytic activity is stimulated by the occurrence of DNA strand breaks. These enzymes are unique.

  It is now known that PARP-1 is involved in various DNA-related functions, including gene amplification, cell division, differentiation, apoptosis, DNA base excision repair, and effects on telomere length and chromosome stability (D'Adda di Fagagna, et al., Nature Gen., 23 (1), 76-80 (1999)).

Studies on the mechanism by which PARP-1 regulates DNA repair and other processes confirm its importance in poly (ADP-ribose) chain formation in the cell nucleus (Althaus, FR and Richter, C., ADP-Ribosylation of Proteins: Enzymology and Biological Significance, Springer-Verlag, Berlin (1987)). DNA-bound and activated PARP-1 utilizes NAD ⁺ to synthesize poly (ADP-ribose) on various nuclear target proteins including topoisomerase, histone and PARP itself (Rhun et al., Biochem Biophys. Res. Commun., 245, 1-10 (1998)).

  Poly (ADP-ribosyl) ation is also associated with malignant transformation. For example, PARP-1 activity is higher in isolated nuclei of SV40-transformed fibroblasts, and both leukemia cells and colon cancer cells have higher enzyme activity than the corresponding normal leukocytes and colonic mucosa. (Miwa, et al., Arch. Biochem. Biophys., 181, 313-321 (1977); Burzio, et al., Proc. Soc. Exp. Bioi. Med., 149, 933-938 (1975); And Hirai, et al., Cancer Res., 43, 3441-3446 (1983)). Recently, it has been confirmed that in malignant prostate tumors compared to benign prostate cells, a marked increase in the level of active PARP (mainly PARP-1) is associated with a higher level of genetic instability ( Mcnealy, et al., Anticancer Res., 23, 1473-1478 (2003)).

  Several low molecular weight PARP-1 inhibitors have been used to elucidate the functional role of poly (ADP-ribosyl) ation in DNA repair. In cells treated with alkylating agents, inhibition of PARP results in a significant increase in DNA strand breaks and cell killing (Durkacz, et al., Nature, 283, 593-596 (1980); Berger, NA, Radiation Research, 101, 4-14 (1985)).

  Since then, such inhibitors have been shown to promote the effects of radiation response by suppressing the repair of potentially lethal damage (Ben-Hur, et al., British Journal of Cancer , 49 (Suppl. VI), 34-42 (1984); Schlicker, et al., Int. J. Radiat. Bioi., 75, 91-100 (1999)). PARP inhibitors have been reported to be effective in radiosensitization of hypoxic tumor cells (US Pat. No. 5,032,617; US Pat. No. 5,215,738 and US Pat. No. 5,041,653). In certain tumor cell lines, chemical inhibition of PARP-1 (and PARP-2) activity is also associated with significant sensitization to very low doses of radiation (Chalmers, Clin. Oncol., 16 (1), 29-39 (2004)).

  Moreover, PARP-1 knockout (PARP-/-) animals show genomic instability in response to alkylating agents and gamma irradiation (Wang, et al., Genes Dev., 9, 509-520 (1995) Menissier de Murcia, et al., Proc. Natl. Acad. Sci. USA, 94, 7303-7307 (1997)). More recent data suggest that PARP-1 and PARP-2 possess both redundant and non-redundant functions in maintaining genomic stability, making them both interesting targets (Menissier de Murcia, et al., EMBO. J., 22 (9), 2255-2263 (2003)).

  The role of PARP-1 has also been demonstrated in certain vascular diseases, septic shock, ischemic injury and neurotoxicity (Cantoni, et al., Biochim. Biophys. Acta, 1014, 1-7 (1989) Szabo, et al., J. Clin. Invest., 100, 723-735 (1997)). Oxygen radical DNA damage that later causes DNA strand breaks recognized by PARP-1 is a major contributor to such disease states, as demonstrated by the PARP-1 inhibitor test (Cosi, et al. , J. Neurosci. Res., 39, 38-46 (1994); Said, et al., Proc. Natl. Acad. Sci. USA, 93, 4688-4692 (1996)). Recently, PARP has been demonstrated to be involved in the pathogenesis of hemorrhagic shock (Liaudet, et al., Proc. Natl. Acad. Sci. USA, 97 (3), 10203-10208 (2000)) .

  It has also been demonstrated that efficient retroviral infection of mammalian cells is blocked by inhibition of PARP-1 activity. Such inhibition of recombinant retroviral vector infection has been shown to occur in a variety of different cell types (Gaken, et al., J. Virology, 70 (6), 3992-4000 (1996)). . PARP-1 inhibitors have thus been developed for use in antiviral therapy and in cancer therapy (WO 91/18591).

  Furthermore, PARP-1 inhibition has been speculated to delay the onset of aging properties in human fibroblasts (Rattan and Clark, Biochem. Biophys. Res. Comm., 201 (2), 665-672 (1994 )). This may be related to the role PARP plays in controlling telomere function (d'Adda di Fagagna, et al., Nature Gen., 23 (1), 76-80 (1999)).

  Similarly, PARP-1 inhibitors can be used to treat inflammatory bowel disease (Szabo C., Role of Poly (ADP-Ribose) Polymerase Activation in the Pathogenesis of Shock and Inflammation, In PARP as a Therapeutic Target; Ed J. Zhang, 2002 by CRC Press; 169-204), ulcerative colitis (Zingarelli, B, et al., Immunology, 113 (4), 509-517 (2004)) and Crohn's disease (Jijon, HB, et al., Am. J Physiol. Gastrointest. Liver Physiol., 279, G641-G651 (2000).

Some of the inventors have previously described a class of 1 (2H) -phthalazinone compounds that act as PARP inhibitors (WO 02/36576). This compound has the general formula:

Where A and B together represent an optionally substituted fused aromatic ring, wherein R _C is represented by —LR _L. Many examples are of the following formula:

Where R represents one or more optional substituents.

  We now have a property that R is of a certain nature, compounds in which the phenyl group is replaced show surprising levels of inhibition of PARP activity, and / or radiation therapy and various chemotherapy of tumor cells. It has been found to show a surprising potentiation level against.

Accordingly, in a first aspect of the invention, there are provided compounds of formula (I) and isomers, salts, solvates, chemically protected forms, and prodrugs thereof:

In the formula:
A and B together represent an optionally substituted fused aromatic ring;
X can be NR ^X or CR ^X R ^Y ;
If X = NR ^X , n is 1 or 2; if X = CR ^X R ^Y , n is 1;
R ^X is selected from the group consisting of H, optionally substituted C _1-20 alkyl, C _5-20 aryl, C _3-20 heterocyclyl, amide, thioamide, ester, acyl, and sulfonyl groups;
R ^Y is selected from H, hydroxy, amino;
Or R ^X and R ^Y may be taken together to form a spiro-C _3-7 cycloalkyl or heterocyclyl group;
R ^C1 and R ^C2 are independently selected from the group consisting of hydrogen and C _1-4 alkyl, or when X is CR ^X R ^Y , R ^C1 , R ^C2 , R ^X and R ^Y are the same Together with the carbon atom which may be formed may form an optionally substituted fused aromatic ring;
R ¹ is selected from H and halo; and
Het is selected from:

Where Y ¹ is selected from CH and N, Y ² is selected from CH and N, Y ³ is selected from CH, CF and N, where one or two of Y ¹ , Y ² and Y ³ Can only be N; and

Where Q is O or S.

Thus, if X is CR ^X R ^Y , n is 1 and the compound is of formula (Ia):

If X is NR ^X and n is 1, the compound is of formula (Ib):

If X is NR ^X and n is 2, the compound is of formula (Ic):

What is possible for Het is:

  In a second aspect of the invention, there is provided a pharmaceutical composition comprising a compound of the first aspect and a pharmaceutically acceptable carrier or diluent.

  In a third aspect of the invention, there is provided the use of a compound of the first aspect in a method of treatment of the human or animal body.

In a fourth aspect of the invention, there is provided the use of a compound as defined in the first aspect of the invention in the manufacture of a medicament for:
(a) block poly (ADP-ribose) chain formation by inhibiting the activity of cellular PARP (PARP-1 and / or PARP-2);
(b) Stroke and Parkinson's disease; hemorrhagic shock; inflammatory diseases such as arthritis, inflammatory bowel disease, ulcerative colitis and Crohn's disease; multiple sclerosis; including acute and chronic treatment of secondary effects of diabetes Vascular disease; septic shock; both brain and cardiovascular ischemic injury; both brain and cardiovascular reperfusion injury; treatment of neurotoxicity; and cytoxicity or cardiovascular activity after cardiovascular surgery Acute treatment of diseases ameliorated by inhibition;
(c) Use as an adjunct in cancer therapy or use to enhance tumor cells for treatment with ionizing radiation or chemotherapeutic agents.

  Specifically, the compound defined in the first aspect of the present invention is combined with an alkylating agent such as methyl methanesulfonate (MMS), temozolomide and dacarbazine (DTIC), as well as topotecan, irinotecan, rubitecan, exatecan, lurtotecan. , Gimetecan, difuromotecan (homocamptothecin); and 7-substituted non-silathecan; 7-silylcamptothecin, topoisomerase I inhibitors such as BNP 1350; and non-camptothecin-based topoisomerase I inhibitors such as indolocarbazole, as well It can be used in anti-cancer combination therapy (or as an adjunct) with topoisomerase I and II dual inhibitors such as benzophenazine, XR 11576 / MLN 576 and benzopyridoindole. Such a combination could be given, for example, as an intravenous formulation or by oral administration depending on the preferred method of administration for a particular drug.

  In another further aspect of the invention, a PARP comprising administering to a subject in need of treatment a therapeutically effective amount of a compound as defined in the first aspect, preferably in the form of a pharmaceutical composition. Providing a treatment for diseases ameliorated by inhibition, and a therapeutically effective amount of a compound as defined in the first aspect, preferably in the form of a pharmaceutical composition, concurrently with radiation therapy (ionizing radiation) or a chemotherapeutic agent or A treatment for cancer is provided comprising the step of sequentially combining and administering to a subject in need of treatment.

  In a further aspect of the invention, the compound can be used in the manufacture of a medicament for the treatment of cancer that is deficient in DNA double-strand break (DSB) repair activity by homologous recombination (HR), or treatment of a compound Can be used in the treatment of patients with cancers that are deficient in DNA DSB repair activity by HR, including administering a therapeutically effective amount to said patient.

  The DNA DSB repair pathway by HR repairs double-strand breaks (DSB) in DNA via a homologous mechanism that rearranges continuous DNA helices (KK Khanna and SP Jackson, Nat. Genet. 27 (3): 247-254 (2001)). As components of the DNA DSB repair pathway by HR, ATM (NM_000051), RAD51 (NM_002875), RAD51L1 (NM_002877), RAD51C (NM_002876), RAD51L3 (NM_002878), DMC1 (NM_007068), XRCC2 (NM_005431), XRCC3 (NM_005432) , RAD52 (NM_002879), RAD54L (NM_003579), RAD54B (NM_012415), BRCA1 (NM_007295), BRCA2 (NM_000059), RAD50 (NM_005732), MRE11A (NM_005590) and NBS1 (NM_002485). There is nothing. Among other proteins involved in the DNA DSB repair pathway by HR are regulatory factors such as EMSY (Hughes-Davies, et al., Cell, 115, pp523-535). The HR component is also described in Wood, et al., Science, 291, 1284-1289 (2001).

  Cancers that are deficient in DNA DSB repair by HR may include one or more cancer cells that have a reduced or suppressed ability to repair DNA DSB through the pathway compared to normal cells, or It can consist of cancer cells. That is, the activity of the DNA DSB repair pathway by HR may be reduced or lost in one or more cancer cells.

  The activity of one or more components of the DNA DSB repair pathway by HR may be lost in one or more cancer cells of an individual with cancer that is deficient in DNA DSB repair by HR. The components of the DNA DSB repair pathway by HR are well characterized in the art (see, for example, Wood, et al., Science, 291, 1284-1289 (2001)), and the components described above Contains ingredients.

  In some preferred embodiments, the cancer cells can have a BRCA1 and / or BRCA2 deficient phenotype, ie, BRCA1 and / or BRCA2 activity is reduced or abolished in the cancer cells. Cancer cells having this phenotype are, for example, by mutation or polymorphism in the encoding nucleic acid, or by amplification, mutation or polymorphism in a gene encoding a regulator, such as the EMSY gene encoding a BRCA2 regulator. (Hughes-Davies, et al., Cell, 115, 523-535) or epigenetic mechanisms such as gene promoter methylation may result in loss of BRCA1 and / or BRCA2, ie BRCA1 and / or BRCA2 expression and / or activity may be reduced or disappeared in cancer cells.

  BRCA1 and BRCA2 are known tumor suppressors, and their wild-type allele is often lost in tumors of heterozygous carriers (Jasin M., Oncogene, 21 (58), 8981-93 (2002) Tutt, et al., Trends Mol Med., 8 (12), 571-6, (2002)). The association of BRCA1 and / or BRCA2 mutations with breast cancer is well characterized in the art (Radice, PJ, Exp Clin Cancer Res., 21 (3 Suppl), 9-12 (2002)) . It is also known that the amplification of the EMSY gene encoding a BRCA2 binding factor is associated with breast and ovarian cancer.

  Carriers of mutations in BRCA1 and / or BRCA2 are also at increased risk for ovarian, prostate and pancreatic cancer.

  In some preferred embodiments, the individual is heterozygous for one or more mutations, such as mutations and polymorphisms, in BRCA1 and / or BRCA2 or modulators thereof. Detection of mutations in BRCA1 and BRCA2 is well known in the art, for example European Patent 699 754, European Patent 705 903, Neuhausen, SL and Ostrander, EA, Genet. Test, 1, 75-83. (1992); Janatova M., et al., Neoplasma, 50 (4), 246-50 (2003). Measurement of the amplification of the BRCA2 binding factor EMSY is described in Hughes-Davies, et al., Cell, 115, 523-535).

  Mutations and polymorphisms associated with cancer are detected at the nucleic acid level by detecting the presence of a mutant nucleic acid sequence or by detecting the presence of a mutant (i.e., mutant or allelic variant) polypeptide. It can be detected at the protein level.

Definitions The term "aromatic ring", cyclic aromatic structure, that is, as used herein in the conventional sense to refer to a cyclic structure having delocalised π- electron orbitals.

  Aromatic rings fused to the main central nucleus, ie formed by -A-B-, can have additional fused aromatic rings (eg, resulting in naphthyl or anthracenyl groups). The aromatic ring may contain only carbon atoms, or may contain carbon atoms and one or more heteroatoms, including but not limited to nitrogen, oxygen and sulfur atoms. The aromatic ring preferably has 5 or 6 ring atoms.

  The aromatic ring may be substituted. If the substituent itself contains an aryl group, the aryl group is not considered to be part of the aryl group to which the substituent is attached. For example, a biphenyl group is considered herein to be a phenyl group substituted with a phenyl group (an aryl group containing a single aromatic ring). Similarly, a benzylphenyl group is considered to be a phenyl group substituted with a benzyl group (an aryl group containing a single aromatic ring).

  In one group of preferred embodiments, the aromatic group comprises a single aromatic ring having 5 or 6 ring atoms selected from carbon, nitrogen, oxygen and sulfur, which ring may be substituted . Examples of these groups include, but are not limited to, benzene, pyrazine, pyrrole, thiazole, isoxazole, and oxazole. Although 2-pyrone can be considered an aromatic ring, it is less preferred.

  If the aromatic ring has 6 atoms, preferably at least 4 or optionally 5 or all of the ring atoms are carbon. The other ring atoms are selected from nitrogen, oxygen and sulfur, with nitrogen and oxygen being preferred. Suitable groups include rings without heteroatoms (benzene), rings with one nitrogen ring atom (pyridine), rings with two nitrogen ring atoms (pyrazine, pyrimidine and pyridazine), one oxygen ring atom And a ring having one oxygen and one nitrogen ring atom (oxazine).

  If the aromatic ring has 5 ring atoms, preferably at least 3 of the ring atoms are carbon. The remaining ring atoms are selected from nitrogen, oxygen and sulfur. Suitable rings include a ring having one nitrogen ring atom (pyrrole), a ring having two nitrogen ring atoms (imidazole, pyrazole), a ring having one oxygen ring atom (furan), and one sulfur. Rings with ring atoms (thiophene), rings with one nitrogen and one sulfur ring atom (isothiazole, thiazole) and rings with one nitrogen and one oxygen ring atom (isoxazole or oxazole) Is mentioned.

The aromatic ring can have one or more substituents at any available ring position. These substituents are selected from halo, nitro, hydroxy, ether, thiol, thioether, amino, C _1-7 alkyl, C _3-20 heterocyclyl and C _5-20 aryl. Aromatic rings can similarly have one or more substituents taken together to form a ring. Specifically, they are of the formula — (CH ₂ ) _m — or —O— (CH ₂ ) _p —O—, where m is 2, 3, 4 or 5 and p is 1, 2 or 3. Can be.

  Alkyl: As used herein, the term “alkyl” is a monovalent obtained by removing a hydrogen atom from a carbon atom of a hydrocarbon compound having from 1 to 20 carbon atoms (unless otherwise specified). Which may be aliphatic or cycloaliphatic, which may be saturated or unsaturated (eg, partially unsaturated, fully unsaturated). Thus, the term “alkyl” includes the subclasses alkenyl, alkynyl, cycloalkyl, cycloalkyenyl, cycloalkynyl, and the like, discussed below.

In the context of an alkyl group, the prefix (e.g., C _1-4 , C _1-7 , C _1-20 , C _2-7 , C _3-7 etc.) indicates the number of carbon atoms, or range of carbon atoms. means. For example, the term “C _1-4 alkyl” as used herein relates to an alkyl group having 1 to 4 carbon atoms. Examples of groups of alkyl groups include C _1-4 alkyl (“lower alkyl”), C _1-7 alkyl, and C _1-20 alkyl. Note that the first prefix may vary depending on other constraints. For example, for unsaturated alkyl groups, the first prefix must be at least 2, for cyclic alkyl groups, the first prefix must be at least 3, and so forth.

Examples of (unsubstituted) saturated alkyl groups are methyl (C ₁ ), ethyl (C ₂ ), propyl (C ₃ ), butyl (C ₄ ), pentyl (C ₅ ), hexyl (C ₆ ), heptyl (C _7), octyl (C _8), nonyl (C _9), decyl (C _10), undecyl (C _11), dodecyl (C _12), tridecyl (C _13), tetradecyl (C _14), pentadecyl (C ₁₅₎ And eicodecyl (C ₂₀ ), but are not limited to these.

Examples of (unsubstituted) saturated linear alkyl groups include methyl (C ₁ ), ethyl (C ₂ ), n-propyl (C ₃ ), n-butyl (C ₄ ), n-pentyl (amyl) (C ₅ ), n-hexyl (C ₆ ) and n-heptyl (C ₇ ), but are not limited thereto.

Examples of (unsubstituted) saturated branched alkyl groups include iso-propyl (C ₃ ), iso-butyl (C ₄ ), sec-butyl (C ₄ ), tert-butyl (C ₄ ), iso-pentyl ( C ₅ ) and neo-pentyl (C ₅ ), but are not limited to these.

Alkenyl: The term “alkenyl” as used herein relates to an alkyl group having one or more carbon-carbon double bonds. Examples of alkenyl groups include C _2-4 alkenyl, C _2-7 alkenyl, C _2-20 alkenyl.

Examples of (unsubstituted) unsaturated alkenyl groups include ethenyl (vinyl, —CH═CH ₂ ), 1-propenyl (—CH═CH—CH ₃ ), 2-propenyl (allyl, —CH—CH═CH ₂ ). , Isopropenyl (1-methylvinyl, -C (CH ₃ ) = CH ₂ ), butenyl (C ₄ ), pentenyl (C ₅ ) and hexenyl (C ₆ ), but are not limited thereto. .

Alkynyl: The term “alkynyl” as used herein relates to an alkyl group having one or more carbon-carbon triple bonds. Examples of alkynyl groups include C _2-4 alkynyl, C _2-7 alkynyl, C _2-20 alkynyl.

Examples of (unsubstituted) unsaturated alkynyl groups include, but are not limited to, ethynyl (ethinyl, —C≡CH) and 2-propynyl (propargyl, —CH ₂ —C≡CH). It will never be done.

Cycloalkyl: As used herein, the term “cycloalkyl” refers to an alkyl group that is also a cyclyl group, ie, one obtained by removing a hydrogen atom from an alicyclic ring atom of a carbocyclic compound. In relation to the valent moiety, the carbocycle may be saturated or unsaturated (e.g., partially unsaturated, fully unsaturated), and this moiety contains 3 to 20 ring atoms. Including 3-20 carbon atoms (unless otherwise specified). Thus, the term “cycloalkyl” includes the subclasses cycloalkenyl and cycloalkynyl. Each ring preferably has 3 to 7 ring atoms. Examples of groups of cycloalkyl groups include C _3-20 cycloalkyl, C _3-15 cycloalkyl, C _3-10 cycloalkyl, C _3-7 cycloalkyl.

Examples of cycloalkyl groups include, but are not limited to, those derived from:
Saturated monocyclic hydrocarbon compounds:
Cyclopropane (C _3), cyclobutane (C _4), cyclopentane (C _5), cyclohexane (C _6), cycloheptane (C _7), methylcyclopropane (C _4), dimethylcyclopropane (C _5), methyl cyclobutane (C _5), dimethylcyclobutane (C _6), methylcyclopentane (C _6), dimethylcyclopentane (C _7), methylcyclohexane (C _7), dimethylcyclohexane (C _8), menthane (C _10);
Unsaturated monocyclic hydrocarbon compounds:
Cyclopropene (C _3), cyclobutene (C _4), cyclopentene (C _5), cyclohexene (C _6), methylcyclopropene (C _4), dimethyl cyclopropene (C _5), methyl-cyclo-butene (C _5), dimethyl Cyclobutene (C ₆ ), methylcyclopentene (C ₆ ), dimethylcyclopentene (C ₇ ), methylcyclohexene (C ₇ ), dimethylcyclohexene (C ₈ );
Saturated polycyclic hydrocarbon compounds:
Tsujang (C ₁₀ ), Karan (C ₁₀ ), Pinan (C ₁₀ ), Bornan (C ₁₀ ), Norcaran (C ₇ ), Norpinan (C ₇ ), Norbornane (C ₇ ), Adamantane (C ₁₀ ), Decalin ( decahydronaphthalene) (C _10);
Unsaturated polycyclic hydrocarbon compounds:
Camphene (C ₁₀ ), limonene (C ₁₀ ), pinene (C ₁₀ );
Polycyclic hydrocarbon compounds having an aromatic ring:
Indene (C ₉ ), indane (e.g., 2,3-dihydro-1H-indene) (C ₉ ), tetralin (1,2,3,4-tetrahydronaphthalene) (C ₁₀ ), acenaphthene (C ₁₂ ), fluorene (C ₁₃ ), phenalene (C ₁₃ ), acephenanthrene (C ₁₅ ), aceanthrene (C ₁₆ ), collanthrene (C ₂₀ ).

  Heterocyclyl: As used herein, the term “heterocyclyl” refers to a monovalent moiety obtained by removing a hydrogen atom from a ring atom of a heterocyclic compound, which moiety (unless otherwise specified). ) It has 3 to 20 ring atoms, 1 to 10 of which are ring heteroatoms. Preferably, each ring has 3 to 7 ring atoms, 1 to 4 of which are ring heteroatoms.

In this context, a prefix (eg, C _3-20 , C _3-7 , C _5-6 etc.) means the number of ring atoms or a range of ring atoms, whether carbon atoms or heteroatoms. For example, the term “C _5-6 heterocyclyl” as used herein relates to a heterocyclyl group having 5 or 6 ring atoms. Examples of groups of heterocyclyl groups include C _3-20 heterocyclyl, C _5-20 heterocyclyl, C _3-15 heterocyclyl, C _5-15 heterocyclyl, C _3-12 heterocyclyl, C _5-12 heterocyclyl, C _3-10 heterocyclyl, C _5-10 heterocyclyl, C _3-7 heterocyclyl, C _5-7 heterocyclyl, and C _5-6 heterocyclyl.

Examples of monocyclic heterocyclyl groups include, but are not limited to, those derived from:
N ₁ : Aziridine (C ₃ ), azetidine (C ₄ ), pyrrolidine (tetrahydropyrrole) (C ₅ ), pyrroline (for example, 3-pyrroline, 2,5-dihydropyrrole) (C ₅ ), 2H-pyrrole or 3H -Pyrrole (isopyrrole, isoazole) (C ₅ ), piperidine (C ₆ ), dihydropyridine (C ₆ ), tetrahydropyridine (C ₆ ), azepine (C ₇ );
O _1: oxirane (C _3), oxetane (C _4), oxolane (tetrahydrofuran) (C _5), Okisoru (dihydrofuran) (C _5), dioxane (tetrahydropyran) (C _6), dihydropyran (C ₆₎ , Pyran (C ₆ ), oxepin (C ₇ );
S ₁ : thiirane (C ₃ ), thietane (C ₄ ), thiolane (tetrahydrothiophene) (C ₅ ), thiane (tetrahydrothiopyran) (C ₆ ), thiepan (C ₇ );
O ₂ : dioxolane (C ₅ ), dioxane (C ₆ ), and dioxepane (C ₇ );
O ₃ : trioxane (C ₆ );
N ₂ : imidazolidine (C ₅ ), pyrazolidine (diazolidine) (C ₅ ), imidazoline (C ₅ ), pyrazoline (dihydropyrazole) (C ₅ ), piperazine (C ₆ );
N ₁ O ₁ : tetrahydrooxazole (C ₅ ), dihydrooxazole (C ₅ ), tetrahydroisoxazole (C ₅ ), dihydroisoxazole (C ₅ ), morpholine (C ₆ ), tetrahydrooxazine (C ₆ ), dihydrooxazine (C ₆ ), oxazine (C ₆ );
N ₁ S ₁ : thiazoline (C ₅ ), thiazolidine (C ₅ ), thiomorpholine (C ₆ );
N ₂ O ₁ : oxadiazine (C ₆ );
O ₁ S ₁ : oxathiol (C ₅ ) and oxathiane (thioxan) (C ₆ ); and
N ₁ O ₁ S ₁ : Oxathiazine (C ₆ ).

Substituted Examples of (non-aromatic) monocyclic heterocyclyl groups, cyclic saccharides, for example, arabinofuranose, lyxofuranose, ribofuranose and xylofuranose (Xylofuranse) furanose such as (C _5), and Aropiranosu And those derived from pyranose (C ₆ ) such as altropyranose, glucopyranose, mannopyranose, gropyranose, idopyranose, galactopyranose and talopyranose.

Spiro-C _3-7 cycloalkyl or heterocyclyl: As used herein, the term “spiro C _3-7 cycloalkyl or heterocyclyl” refers to a C connected to another ring by a single atom common to both rings. Refers to a _3-7 cycloalkyl or C _3-7 heterocyclyl ring.

C _5-20 aryl: As used herein, the term “C _5-20 aryl” refers to a monovalent moiety obtained by removing a hydrogen atom from an aromatic ring atom of a C _5-20 aromatic compound. The compound has one ring, or two or more (eg, fused) rings, 5 to 20 ring atoms, and at least one of the rings is an aromatic ring. Each ring preferably has 5 to 7 ring atoms.

The ring atoms may be all carbon atoms, as in “carboaryl groups”, in which case the groups may conveniently be referred to as “C _5-20 carboaryl” groups.

Examples of C _5-20 aryl groups having no ring heteroatoms (ie C _5-20 carboaryl groups) include benzene (ie phenyl) (C ₆ ), naphthalene (C ₁₀ ), anthracene (C ₁₄ ), phenanthrene. Examples include (C ₁₄ ) and those derived from pyrene (C ₁₆ ), but are not limited thereto.

Alternatively, the ring atoms may contain one or more heteroatoms, including but not limited to oxygen, nitrogen and sulfur, as in “heteroaryl groups”. In this case, this group may conveniently be referred to as a “C _5-20 heteroaryl” group, where “C _5-20 ” means a ring atom, whether carbon or heteroatom. Preferably, each ring has 5 to 7 ring atoms, of which 0 to 4 are ring heteroatoms.

Examples of C _5-20 heteroaryl groups are furan (oxol), thiophene (thiol), pyrrole (azole), imidazole (1,3-diazole), pyrazole (1,2-diazole), triazole, oxazole, isoxazole , Thiazole, isothiazole, oxadiazole, tetrazole and oxatriazole; C ₅ heteroaryl groups; and isoxazine, pyridine (azine), pyridazine (1,2-diazine), pyrimidine (1,3-diazine; for example , Cytosine, thymine, uracil), C ₆ heteroaryl groups derived from pyrazine (1,4-diazine) and triazine, but are not limited thereto.

  A heteroaryl group may be attached via a carbon or heterocyclic atom.

Examples of C _5-20 heteroaryl groups containing fused rings include C ₉ heteroaryl groups derived from benzofuran, isobenzofuran, benzothiophene, indole, isoindole; C ₁₀ heteroaryl derived from quinoline, isoquinoline, benzodiazine, pyridopyridine Aryl groups; including but not limited to C ₁₄ heteroaryl groups derived from acridine and xanthene.

  The above alkyl, heterocyclyl, and aryl groups, whether alone or as part of other substituents, are themselves substituted with one or more groups selected from the following and further substituents: Also good.

  Halo: -F, -Cl, -Br, and -I.

  Hydroxy: -OH.

Ether: -OR, R is an ether substituent in the formula, for example, (also referred to as a C _1-7 alkoxy group) C _1-7 alkyl group (also referred to as C _3-20 heterocyclyloxy group) C _3-20 heterocyclyl group Or a C _5-20 aryl group (also referred to as a C _5-20 aryloxy group), preferably a C _1-7 alkyl group.

Nitro: -NO ₂ .

  Cyano (nitrile, carbonitrile): -CN.

Acyl (keto): -C (= O) R, wherein R is an acyl substituent, such as H, a C _1-7 alkyl group (also referred to as C _1-7 alkyl acyl or C _1-7 alkanoyl), C _{A 3-20} heterocyclyl group (also referred to as C _3-20 heterocyclyl acyl) or a C _5-20 aryl group (also referred to as C _5-20 aryl acyl), preferably a C _1-7 alkyl group. Examples of acyl groups include -C (= O) CH ₃ (acetyl), -C (= O) CH ₂ CH ₃ (propionyl), -C (= O) C (CH ₃ ) ₃ (butyryl) and -C (= O) Ph (benzoyl, phenone) can be mentioned, but is not limited thereto.

  Carboxy (carboxylic acid): -COOH.

Ester (carboxylate, carboxylic acid ester, oxycarbonyl): -C (= O) OR, wherein R is an ester substituent, such as a C _1-7 alkyl group, a C _3-20 heterocyclyl group, or C _{5- A 20} aryl group, preferably a C _1-7 alkyl group. Examples of ester groups include -C (= O) OCH ₃ , -C (= O) OCH ₂ CH ₃ , -C (= O) OC (CH ₃ ) ₃ and -C (= O) OPh. However, it is not limited to these.

Amide (carbamoyl, carbamyl, aminocarbonyl, carboxamide): -C (= O) NR ¹ R ² , wherein R ¹ and R ² are independently amino substituents, as defined for amino groups . -C Examples of amido groups _{(= O) NH 2, -C} (= O) NHCH 3, -C (= O) N (CH 3) 2, -C (= O) NHCH 2 CH 3 and -C (= O) N (CH ₂ CH ₃ ) ₂ and R ¹ and R ² together with the nitrogen atom to which they are attached, for example, piperidinocarbonyl, morpholinocarbonyl, thiomorpholinocarbonyl and pipette Examples include, but are not limited to, amide groups that form a heterocyclic structure such as radinylcarbonyl.

Amino: -NR ¹ R ² , wherein R ¹ and R ² are independently amino substituents such as hydrogen, C _1-7 alkyl groups (C _1-7 alkylamino or di-C _1-7 alkylamino Also referred to as C _3-20 heterocyclyl or C _5-20 aryl, preferably H or C _1-7 alkyl, or a “cyclic” amino group, R ¹ and R ² are Together with the attached nitrogen atoms, it forms a heterocyclic ring having 4 to 8 ring atoms. -NH ₂ Examples of amino _{groups, -NHCH 3, -NHCH (CH 3} ) 2, -N (CH 3) 2, -N (CH 2 CH 3) Although ₂ and -NHPh include, limited to It will never be done. Examples of cyclic amino groups include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidino, piperazinyl, perhydrodiazepinyl, morpholino and thiomorpholino. In particular, a cyclic amino group may be substituted on the ring by any of the substituents defined herein, such as carboxy, carboxylate and amide.

Acylamide (acylamino): -NR ¹ C (= O) R ² , wherein R ¹ is an amide substituent, such as hydrogen, a C _1-7 alkyl group, a C _3-20 heterocyclyl group, or a C _5-20 aryl group , Preferably H or a C _1-7 alkyl group, most preferably H, and R ² is an acyl substituent, such as a C _1-7 alkyl group, a C _3-20 heterocyclyl group, or a C _5-20 aryl group, preferably Is a C _1-7 alkyl group. Examples of acylamide groups include, but are not limited to, —NHC (═O) CH ₃ , —NHC (═O) CH ₂ CH ₃ and —NHC (═O) Ph. R ¹ and R ² can be taken together to form a cyclic structure, such as, for example, succinimidyl, maleimidyl and phthalimidyl:

Ureido: -N (R ¹ ) CONR ² R ³ , wherein R ² and R ³ are independently amino substituents, as defined for amino groups, and R 1 is a ureido substituent, for example, hydrogen, C _{A 1-7} alkyl group, a C _3-20 heterocyclyl group or a C _5-20 aryl group, preferably hydrogen or a C _1-7 alkyl group. Examples of ureido groups include -NHCONH ₂ , -NHCONHMe, -NHCONHEt, -NHCONMe ₂ , -NHCONEt ₂ , -NMeCONH ₂ , -NMeCONHMe, -NMeCONHEt, -NMeCONMe ₂ , -NMeCONEt ₂ and -NHCONHPh. It is not limited to.

Acyloxy (reverse ester): —OC (═O) R, wherein R is an acyloxy substituent, such as a C _1-7 alkyl group, a C _3-20 heterocyclyl group, or a C _5-20 aryl group, preferably C _{1 -7} alkyl group. Examples of the acyloxy group include -OC (= O) CH ₃ (acetoxy), -OC (= O) CH ₂ CH ₃ , -OC (= O) C (CH ₃ ) ₃ , -OC (= O) Ph, -OC (= O) but C ₆ H ₄ F and -OC (= O) CH ₂ Ph and the like, but is not limited thereto.

  Thiol: -SH.

Thioether (sulfide): -SR, wherein R is a thioether substituent, such as a _C1-7 alkyl group (also referred to as a _C1-7 alkylthio group), a _C3-20 heterocyclyl group or a _C5-20 aryl group, A C _1-7 alkyl group is preferred. Examples of C _1-7 alkylthio groups include, but are not limited to, —SCH ₃ and —SCH ₂ CH ₃ .

Sulfoxide (sulfinyl): -S (= O) R, wherein R is a sulfoxide substituent, such as a C _1-7 alkyl group, a C _3-20 heterocyclyl group, or a C _5-20 aryl group, preferably C _{1- 7 is} an alkyl group. Examples of sulfoxide groups include, but are not limited to, —S (═O) CH ₃ and —S (═O) CH ₂ CH ₃ .

Sulfonyl (sulfone): —S (═O) ₂ R, wherein R is a sulfone substituent, for example, a C _1-7 alkyl group, a C _3-20 heterocyclyl group, or a C _5-20 aryl group, preferably C _{1 -7} alkyl group. Examples of sulfone groups _{-S (= O) 2 CH 3} ( methanesulfonyl, mesyl), - S (= O) 2 CF 3, -S (= O) 2 CH 2 CH 3 and 4-methyl phenylsulfonyl ( Tosyl), but is not limited thereto.

Thioamido (thiocarbamyl): —C (═S) NR ¹ R ² , wherein R ¹ and R ² are independently amino substituents, as defined for amino groups. Include _{-C (= S) NH 2,} -C (= S) NHCH 3, -C (= S) N (CH 3) 2 and _{-C (= S) NHCH 2 CH} 3 Examples of amido groups However, it is not limited to these.

Sulfonamino: —NR ¹ S (═O) ₂ R, wherein R ¹ is an amino substituent as defined for the amino group, R is a sulfonamino substituent, such as a C _1-7 alkyl group, A C _3-20 heterocyclyl group or a C _5-20 aryl group, preferably a C _1-7 alkyl group. Examples of sulfonamino groups include, but are not limited to, —NHS (═O) ₂ CH ₃ , —NHS (═O) ₂ Ph and —N (CH ₃ ) S (═O) ₂ C ₆ H _5. Will never be done.

As noted above, the groups that form the above substituents, such as C _1-7 alkyl, C _3-20 heterocyclyl, and C _5-20 aryl, may themselves be substituted. Accordingly, the above definition covers substituted substituents.

  Where applicable, the following preferred embodiments can be applied to each aspect of the invention.

  In the present invention, the condensed aromatic ring represented by -A-B- preferably comprises only a carbon ring atom, and therefore can be benzene or naphthalene, more preferably benzene. As noted above, these rings may be substituted, but in some embodiments are preferably unsubstituted.

If the condensed aromatic ring represented by -AB- has a substituent, it is preferably bonded to the atom itself bonded to the central ring at the β-position of the carbon atom in the central ring. Thus, if the fused aromatic ring is a benzene ring, the preferred substitution position is indicated by ^{* in the following} formula:

This position is often called the 5th position of the phthalazinone moiety.

The substituent is preferably an alkoxy, amino, halo (eg, fluoro) or hydroxy group, more preferably a C _1-7 alkoxy group (eg, -OMe).

  If the substituent is halo, the group may be at the 8-position of the phthalazinone moiety.

  Preferably, Het is selected from pyridylene, fluoro-pyrdiylene, furanylene and thiophenylene.

More preferably, Het is selected from:

More preferably, Het is selected from:

R ^C1 and R ^C2 are preferably independently selected from hydrogen and C _1-4 alkyl, more preferably H and methyl. More preferably, at least one of R ^C1 and R ^C2 is hydrogen, and the most preferred option is that both are hydrogen.

When n is 2, X is NR ^X. In these embodiments, R ^X is H; optionally substituted C _1-20 alkyl (eg, _optionally substituted C _5-20 arylmethyl); _optionally substituted C _5-20 aryl; It is preferred that the ester substituent is C _1-20 alkyl; an optionally substituted ester group; an optionally substituted acyl group; an optionally substituted amide group; an optionally substituted thioamide group And preferably selected from the group consisting of optionally substituted sulfonyl groups. R ^X is H; optionally substituted C _1-20 alkyl (more preferably _optionally substituted C _1-7 alkyl, such as methyl); and the ester substituent is C _1-20 alkyl (more preferably It is preferably an optionally substituted C _1-7 alkyl (eg t-butyl), more preferably selected from the group consisting of optionally substituted ester groups.

When n is 1, X can be NR ^X or CR ^X CR ^Y.

In embodiments where X is NR ^X , R ^X is H; optionally substituted C _1-20 alkyl (eg, _optionally substituted C _5-20 arylmethyl); _optionally substituted C _{5 -20} aryl; optionally substituted acyl optionally; optionally substituted sulfonyl also be; optionally substituted amido; be selected from the group consisting of and optionally substituted thioamido groups.

  In these embodiments, Het is preferably pyridylene.

More preferably, when Het is pyridylene, R ^X is selected from the group consisting of an optionally substituted acyl; an optionally substituted sulfonyl; and an optionally substituted amide.

When Het is furanylene or thiophenylene, R ^X is optionally substituted C _1-20 alkyl (eg, optionally substituted C _5-20 arylmethyl); optionally substituted C _5- More preferably, it is selected from the group consisting of: ₂₀ aryl; optionally substituted acyl; optionally substituted sulfonyl; and optionally substituted amide.

In embodiments where X is CR ^X R ^Y , R ^Y is preferably H. R ^X is H; optionally substituted C _1-20 alkyl (eg, _optionally substituted C _5-20 arylmethyl); _optionally substituted C _5-20 aryl; _optionally substituted Good C _3-20 heterocyclyl; preferably an acyl substituent is selected from C _5-20 aryl and C _3-20 heterocyclyl (eg piperazinyl); an optionally substituted acyl; the amino group is H And optionally substituted amino, preferably selected from C _1-20 alkyl or taken together with a nitrogen atom to form a C _5-20 heterocyclic group; An optionally substituted amide, preferably selected from C _1-20 alkyl or taken together with a nitrogen atom to form a C _5-20 heterocyclic group; and the ester substituent is C it is preferably selected from _1-20 alkyl group, substituted Is preferably selected from the group consisting of which may ester group are.

In these embodiments, when Het is furylene or thiophenylene, R ^X is a C _5-20 heterocyclic group in which the amino group is selected from H and C _1-20 alkyl or together with a nitrogen atom More preferably, it is selected from optionally substituted amino, preferably forming a group, for example morpholino.

  Particularly preferred compounds include 2, 3, 5, 6, 9, 10, 12, 13, 56, 57, 58, 62, 65, 66, 67, 74 and 75.

  Where appropriate, the preferred embodiments described above can be combined with each other.

Other forms included The above include well-known ionic forms, salt forms, solvate forms, and protected forms of these substituents. For example, reference to a carboxylic acid (—COOH) also includes the anion (carboxylate) form (—COO ⁻ ), salts or solvates thereof, and the conventional protected form. Similarly, reference to an amino group includes the protonated form (—N ⁺ HR ¹ R ² ), salts or solvates of the amino group, such as the hydrochloride salt, and the conventional protected form of the amino group. Similarly, reference to a hydroxyl group also includes the anionic form (—O ⁻ ), its salts or solvates, and the conventional protected form of the hydroxyl group.

Isomers, salts, solvates, protected and prodrugs Certain compounds include cis and trans; E and Z; c, t and r; endo and exo; R, S and meso; D and L; d and l; (+) and (-); keto, enol and enolate; syn and anti; syncrin and anticlinal; α Type and beta type; axial type and equatorial type; boat type, chair type, twist type, inclusion type and half-chair type; and one or more specific geometric types, including but not limited to, optical, Type, enantiomeric form, diastereoisomeric form, epimeric form, stereoisomeric form, tautomeric form, conformational form or anomeric form, which are collectively referred to as “isomers” (or “isomers” below). It is called "body type").

  If the compound is in crystalline form, it can exist in several different polymorphic forms.

Except as noted below with respect to tautomeric forms, explicitly excluded from the term “isomer” as used herein is structural (or constitutional) isomers (ie, in space Note that the isomers differ not only in the position of the atoms but also in the connection between the atoms. For example, a reference to a methoxy group —OCH ₃ should not be construed as a reference to the structural isomer hydroxymethyl group —CH ₂ OH. Similarly, a reference to ortho-chlorophenyl should not be construed as a reference to its structural isomer meta-chlorophenyl. However, a reference to a class of structures may include structurally isomeric forms that fall within that class (eg, C _1-7 alkyl includes n-propyl and iso-propyl; butyl is n- Including butyl, iso-butyl, sec-butyl and tert-butyl; methoxyphenyl includes ortho-methoxyphenyl, meta-methoxyphenyl and para-methoxyphenyl).

  The above exclusions include, for example, the following tautomeric pairs: keto / enol, imine / enamine, amide / iminoalcohol, amidine / amidine, nitroso / oxime, thioketone / enethiol, N-nitroso / hydroxyazo and nitro It is not related to tautomeric forms, such as keto, enol and enolate forms, like / Ac-nitro.

Of particular relevance to the present invention are the tautomeric pairs shown below:

Note that specifically included in the term “isomer” are compounds with one or more isotopic substitutions. For example, H may be of any isotope type, including ¹ H, ² H (D) and ³ H (T); C is any isotope type, including ¹² C, ¹³ C and ¹⁴ C O may be of any isotopic form, including ¹⁶ O and ¹⁸ O, etc.

  Unless otherwise specified, a reference to a particular compound includes all such isomeric forms, including its (fully or partially) racemic mixtures and other mixtures. Methods for the preparation (eg, asymmetric synthesis) and separation (eg, fractional crystallization and chromatographic methods) of such isomeric forms are known in the art or as taught herein. Or can be easily obtained by adapting known methods by known methods.

  Unless specified otherwise, references to a particular compound also include, for example, its ionic, salt, solvate, and protected forms, and its different polymorphic forms, as discussed below.

  It may be convenient or desirable to prepare, purify, and / or handle a corresponding salt of the active compound, for example, a pharmaceutically-acceptable salt. Examples of pharmaceutically acceptable salts are discussed in Berge, et al., “Pharmaceutically Acceptable Salts”, J. Pharm. Sci., 66, 1-19 (1977).

For example, if the compound is anionic, or anionic in there can functional groups (e.g., -COOH may -COO ^- there may in) if you have a to form a suitable cation salt it can. Examples of suitable inorganic cations include alkali metal ions such as Na ⁺ and K ⁺ , alkaline earth cations such as Ca ²⁺ and Mg ²⁺ , and other cations such as Al ³⁺ , It is not limited to these. Examples of suitable organic cations include ammonium ions (i.e., NH ₄ ⁺⁾ and substituted ammonium ions _{^{(e.g., NH 3 R +, NH 2}} R 2 +, NHR 3 +, NR 4 +) but may be mentioned, It is not limited to these. Examples of some suitable substituted ammonium ions include ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine and tromethamine, and lysine and arginine. It is derived from the amino acid. An example of a common quaternary ammonium ion is N (CH ₃ ) ₄ ⁺ .

If the compound is cationic, or a functional group which may be cationic (e.g., -NH ₂ is capable in -NH ₃ ⁺⁾ if they have a to form a suitable anion salt it can. Examples of suitable inorganic anions include those derived from the following inorganic acids: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfurous acid, nitric acid, nitrous acid, phosphoric acid and phosphorous acid. However, it is not limited to these. Examples of suitable organic anions include the following organic acids: acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, palmitic acid, lactic acid, malic acid, pamoic acid, tartaric acid, citric acid, gluconic acid , Ascorbic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, aspartic acid, benzoic acid, cinnamic acid, pyruvic acid, salicylic acid, sulfanilic acid, 2-acetyoxybenzoic benzoic acid, fumaric acid, toluenesulfone Examples include, but are not limited to, those derived from acids, methanesulfonic acid, ethanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, valeric acid and gluconic acid. Examples of suitable polymeric anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethylcellulose.

  It may be convenient or desirable to prepare, purify, and / or handle a corresponding solvate of the active compound. The term “solvate” is used herein in the conventional sense to refer to a complex of solute (eg, active compound, salt of active compound) and solvent. If the solvent is water, the solvate may conveniently be referred to as a hydrate, such as a monohydrate, dihydrate, trihydrate, etc.

  It may be convenient or desirable to prepare, purify, and / or handle the active compound in chemically protected form. As used herein, the term “chemically protected form” refers to protecting one or more reactive functional groups from undesired chemical reactions, i.e. protected groups or protecting groups (masked groups). Or a compound in the form of a masking group or a blocked or blocking group). By protecting the reactive functional group, reactions involving other unprotected reactive functional groups can be carried out without affecting the protected group. Usually at a later stage, the protecting group can be removed without substantially affecting the rest of the molecule. See, for example, “Protective Groups in Organic Synthesis” (T. Green and P. Wuts; 3rd Edition; John Wiley and Sons, 1999).

For example, the hydroxy group is ether (-OR) or ester (-OC (= O) R), e.g. t-butyl ether; benzyl, benzhydryl (diphenylmethyl) or trityl (triphenylmethyl) ether; trimethylsilyl or t-butyldimethyl Silyl ethers; or acetyl esters (—OC (═O) CH ₃ , —OAc).

For example, an aldehyde group or a ketone group may be protected as an acetal or ketal where a carbonyl group (> C═O) is converted to a diether (> C (OR) ₂ ), for example, by reaction with a primary alcohol. it can. Aldehyde or ketone groups are readily regenerated by hydrolysis using a large excess of water in the presence of acid.

For example, an amine group is, for example, as an amide or urethane, for example, as methylamide (—NHCO—CH ₃ ); as benzyloxyamide (—NHCO—OCH ₂ C ₆ H ₅ , —NH—Cbz); t-butoxyamide (-NHCO-OC (CH ₃ ) ₃ , -NH-Boc); 2-biphenyl-2-propoxyamide (-NHCO-OC (CH ₃ ) ₂ C ₆ H ₄ C ₆ H ₅ , -NH-Bpoc) 9-fluorenylmethoxyamide (-NH-Fmoc), 6-nitroveratryloxyamide (-NH-Nvoc), 2-trimethylsilylethyloxyamide (-NH-Teoc), 2,2, As 2-trichloroethyloxyamide (-NH-Troc), as allyloxyamide (-NH-Alloc), as 2 (-phenylsulfonyl) ethyloxyamide (-NH-Psec); or, if appropriate, N-oxide It can be protected as (> NO ·).

For example, carboxylic acid groups, as esters, for example C _1-7 alkyl esters (e.g., methyl ester; t-butyl ester); C _1-7 haloalkyl ester (e.g., C _1-7 trihaloalkyl ester); tri C _{1- 7} alkylsilyl-C _1-7 alkyl ester; or as C _5-20 aryl-C _1-7 alkyl ester (eg benzyl ester; nitrobenzyl ester); or as an amide, eg as methyl amide.

For example, a thiol group can be protected as thioether (—SR), for example, benzylthioether; acetamidomethyl ether (—S—CH ₂ NHC (═O) CH ₃ ).

  It may be convenient or desirable to prepare, purify, and / or handle the active compound in the form of a prodrug. The term “prodrug” as used herein relates to a compound which upon metabolism (eg, in vivo) yields the desired active compound. In general, prodrugs are inactive or less active than the active compound, but can provide advantageous handling, administration, or metabolic properties.

For example, some prodrugs are esters of the active compound (eg, a physiologically acceptable metabolically labile ester). During metabolism, the ester group (—C (═O) OR) is cleaved to yield the active drug. Such esters can be obtained, for example, by esterification of any of the carboxylic acid groups (-C (= O) OH) in the parent compound, if appropriate, of any other reactive groups present in the parent compound. It can be formed by performing pre-protection and, if necessary, subsequent deprotection. Examples of such metabolically labile esters include R is C _1-20 alkyl (e.g. -Me, -Et); C _1-7 aminoalkyl (e.g. aminoethyl; 2- (N, N-diethylamino) ) Ethyl; 2- (4-morpholino) ethyl); and acyloxy-C _1-7 alkyl (e.g. acyloxymethyl; acyloxyethyl; e.g. pivaloyloxymethyl; acetoxymethyl; 1-acetoxyethyl; 1- (1-methoxy 1- (Methyl) ethyl-carbonyloxyethyl; 1- (benzoyloxy) ethyl; isopropoxy-carbonyloxymethyl; 1-isopropoxy-carbonyloxyethyl; cyclohexyl-carbonyloxymethyl; 1-cyclohexyl-carbonyloxy Ethyl; cyclohexyloxy-carbonyloxymethyl; 1-cyclohexyloxy-carbonyloxyethyl; (4-tetrahydropyranyloxy ) Carbonyloxymethyl; 1- (4-tetrahydropyranyloxy) carbonyloxyethyl; (4-tetrahydropyranyl) carbonyloxymethyl; and 1- (4-tetrahydropyranyl) carbonyloxyethyl) However, it is not limited to these.

Among the further suitable prodrug types are phosphonates and glycolates. Specifically, the hydroxy group (-OH) is reacted with chlorodibenzyl phosphite and then hydrogenated to form a phosphonic acid group -OP (= O) (OH) ₂ to form a phosphonic acid prodrug. can do. Such groups can be removed by the phosphatase enzyme during metabolism to yield active drugs with hydroxy groups.

  Similarly, some prodrugs are activated enzymatically to yield the active compound, or further chemical reactions yield compounds that yield the active compound. For example, the prodrug can be a sugar derivative or other glycoside complex, or can be an amino acid ester derivative.

For convenience of abbreviations , many chemical moieties are methyl (Me), ethyl (Et), n-propyl (nPr), iso-propyl (iPr), n-butyl (nBu), tert-butyl (tBu), n-hexyl ( nHex), cyclohexyl (cHex), phenyl (Ph), biphenyl (biPh), benzyl (Bn), naphthyl (naph), methoxy (MeO), ethoxy (EtO), benzoyl (Bz) and acetyl (Ac) It is expressed using well-known abbreviations that are not limited to these.

For convenience, many compounds are methanol (MeOH), ethanol (EtOH), iso-propanol (i-PrOH), methyl ethyl ketone (MEK), ether or diethyl ether (Et ₂ O), acetic acid (AcOH), dichloromethane (methylene chloride, DCM), trifluoroacetic acid (TFA), dimethylformamide (DMF), tetrahydrofuran (THF), and dimethyl sulfoxide (DMSO) are used to represent, but are not limited to, well-known abbreviations.

Synthesis In the synthesis route shown below, for convenience, the AB condensed ring is shown as a condensed benzene ring. Compounds in which the AB ring is other than benzene can be synthesized using methodologies similar to those described below by the use of appropriate alternative starting materials.

The compound of the invention is a compound of formula 1:

(Where Het is as defined above) and the compound of formula 2:

Wherein n, R ^C1 , R ^C2 and X are as defined above, and a coupling reagent system such as 2- (1H-benzotriazol-1-yl) -1 tetrafluoroborate , 1,3,3-Tetramethyluronium, 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate or (dimethylaminopropyl) ethylcarbodiimide hydrochloride It can be synthesized by reacting in the presence of a salt / hydroxybenzotriazole in the presence of a base such as diisopropylethylamine in a solvent such as dimethylacetamide or dichloromethane at a temperature ranging from 0 ° C. to the boiling point of the solvent used.

  Alternatively, the compounds of the present invention can be converted to active species using well-known methodologies, for example, active compounds such as acid chlorides or N-hydroxysuccinimide esters and conversion with compounds of formula 2 It can be synthesized by reaction of active species.

The compound of formula 1 is a compound of formula 3:

(Wherein R ¹ is as defined above) or a compound of formula 4:

(Wherein R ¹ is as defined above) or a mixture of a compound of formula 3 and a compound of formula 4 optionally with a hydrazine source, such as hydrazine hydrate or hydrazine monohydrate, such as It can be synthesized by reaction at a temperature ranging from 0 ° C. to the boiling point of the solvent used, optionally in the presence of triethylamine, optionally in the presence of a solvent, such as an industrial modified alcohol.

A compound of formula 3 or formula 4, or a mixture thereof, is a compound of formula 5:

(Wherein R ¹ is as defined above) a reagent capable of hydrolyzing the nitrile moiety, for example a solvent with sodium hydroxide, for example a temperature in the range of 0 ° C. to the boiling point of the solvent used, in the presence of water. It can synthesize | combine by reaction by.

The compound of formula 5 is a compound of formula 6:

A compound of formula 7, wherein R ¹ is as defined above:

(R _a is a C _1-4 alkyl group) in the presence of a base such as triethylamine or lithium hexamethyldisilazide in a solvent such as tetrahydrofuran at a temperature ranging from -80 ° C to the boiling point of the solvent used. It can be synthesized by reaction.

  Compounds of formula 7 can be synthesized by methods similar to those described in WO 02/26576.

Compounds of formula 4 are likewise derived from compounds of formula 7 with compounds of formula 8:

And a base such as triethylamine or lithium hexamethyldisilazide in a solvent such as tetrahydrofuran at a temperature ranging from −80 ° C. to the boiling point of the solvent used.

A compound of formula 6 wherein Het is:

Is a compound of formula 9:

Can be synthesized by oxidation of hydroxy groups with, for example, DMSO, dicyclohexylcarbodiimide (DCC) and phosphoric anhydride.

The compound of formula 9 is a compound of formula 10:

From an acetyl group using an acid such as dilute sulfuric acid in an organic solvent such as THF.

The compound of formula 10 is a compound of formula 11a and 11b:

Can be synthesized by adding acetic anhydride to a preheated solution.

Compounds of formula 11a and 11b are compounds of formula 12a and 12b, respectively:

From, for example, it can be synthesized by oxidation with m-chloroperbenzoic acid (m-CPBA) in an organic solvent such as DCM.

The compound of formula 12a is a compound of formula 13:

From the first reaction with iodomethane, followed by the dropwise addition of aqueous potassium cyanide solution to the ethanol-water solution of the first stage product.

  The compound of formula 12b can be synthesized from the compound of formula 13 by a first reaction with iodoethane followed by dropwise addition of aqueous potassium cyanide solution to the ethanol-water solution of the first stage product.

Compound of formula 8:

Wherein Het is selected from:

Is a compound of formula 14:

Can be synthesized, for example, by oxidation of alkenes with ozone in methanol and DCM (1: 1) solution of the compound of formula 14 at −78 ° C.

A compound of formula 14 wherein Het is:

Is a compound of formula 15:

From, for example, a compound of formula 16:

Can be synthesized under normal conditions.

The compound of formula 15 is a compound of formula 17:

From, for example, it can be synthesized by oxidation using potassium permanganate in an aqueous solution.

Compound of formula 14:

Where Het is:

Is a compound of formula 18:

Can be synthesized, for example, by hydrolysis of the cyano group with sodium hydroxide in methanol.

A compound of formula 18 wherein Het is:

Is a compound of formula 19:

From, for example, a compound of formula 16:

Can be synthesized under normal conditions.

The compound of formula 19 is a compound of formula 20:

Can be synthesized by reaction with sodium cyanide in an organic solvent such as DMF.

A compound of formula 18 wherein Het is:

Is a compound of formula 21:

Can be synthesized by reaction with sodium cyanide in an organic solvent such as DMF.

The compound of formula 21 is a compound of formula 20:

From, for example, a compound of formula 16:

Can be synthesized under normal conditions.

Compound of formula 8:

Where Het is:

Is a compound of formula 22:

For example, it can be synthesized by deprotection of an aldehyde group using a mixture of acetone and water together with a catalytic amount of pyridinium salt of p-toluenesulfonic acid.

The compound of formula 22 is a compound of formula 23:

From a strong base such as lithium diisopropylamide (LDA) followed by the addition of CO ₂ . This reaction may be performed, for example, in THF at −78 ° C., in which case CO ₂ is added as dry ice.

The compound of formula 23 is a compound of formula 24:

For example, by deprotection of the aldehyde group by reaction with ethylene glycol (e.g., 1.5 equivalents) in the presence of a catalytic amount of para-toluenesulfonic acid in toluene under reflux in a Dean-Stark apparatus. it can.

The compound of formula 24 is a compound of formula 25:

Can be synthesized by reaction with a strong base such as butyllithium followed by addition of DMF. This reaction may be performed, for example, at −78 ° C.

Compound of formula 8:

Wherein Het is selected from:

Is commercially available or can be readily synthesized.

  Compounds of formula 1 can also be synthesized by methods analogous to those described above, in which the nitrile moiety in all formulas is replaced with other moieties capable of producing carboxylic acids, such as ester or carboxamide moieties.

A compound of the invention where X is NH has the formula 26:

^Where n, R ^C1 , R ^C2 and R ¹ are as defined above. These compounds can be used to create a library of compounds of the invention described below.

X is NR ^X and R ^X is an acyl moiety, thus formula 27:

Wherein n, R ^C1 , R ^C2 and R ¹ are as defined above and R ^C3 is ^optionally substituted C _1-20 alkyl, C _5-20 aryl and C _3-20 heterocyclyl the compounds of the present invention can be represented by selected from the group) consisting of the compounds of formula 26, wherein R ^C3 COX (wherein R ^C3 is as previously defined, X is an appropriate leaving Optionally in the presence of a base such as pyridine, triethylamine or diisopropylethylamine, optionally in the presence of a solvent such as dichloromethane, ranging from 0 ° C. to the boiling point of the solvent used. It can be synthesized by reaction at temperature.

Compounds of formula R ^C3 COX are commercially available or can be synthesized by methods reported in the chemical literature.

A compound of formula 27 is likewise a coupling reagent system of a compound of formula 26 with a compound of formula R ^C3 CO ₂ H, wherein R ^C3 is as defined above, eg tetrafluoroboric acid. 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium, 2- (1H-benzotriazol-1-yl) -1,1,3,3 hexafluorophosphate Range from 0 ° C. to the boiling point of the solvent used in the presence of tetramethyluronium or (dimethylaminopropyl) ethylcarbodiimide hydrochloride / hydroxybenzotriazole, in the presence of a base such as diisopropylethylamine, in a solvent such as dimethylacetamide or dichloromethane It can synthesize | combine by reaction at the temperature of.

Compounds of formula R ^C3 CO ₂ H are commercially available or can be synthesized by methods reported in the chemical literature.

X is NR ^X and R ^X is an amide or thioamide moiety, thus formula 28:

Wherein n, R ^C1 , R ^C2 and R ¹ are as defined above, Y is O or S, R ^N3 is an optionally substituted C _1-20 alkyl, C _5- The compounds of the present invention that can be represented by the formula R ^N3 NCY (wherein Y and R ^N3 are as defined above), are represented by the formula 26, selected from the group consisting of ₂₀ aryl and C _3-20 heterocyclyl: In the presence of a solvent such as dichloromethane at a temperature ranging from 0 ° C. to the boiling point of the solvent used.

Compounds of formula R ^N3 NCY are commercially available or can be synthesized by methods reported in the chemical literature.

X is NR ^X and R ^X is a sulfonyl moiety, thus formula 29:

Wherein n, R ^C1 , R ^C2 and R ¹ are as defined above, R ^S1 is ^optionally substituted C _1-20 alkyl, C _5-20 aryl and C _3-20 heterocyclyl. Of the compound of formula 26 with a compound of formula R ^S1 SO ₂ Cl (wherein R ^S1 is as defined above). It can be synthesized by reaction at a temperature ranging from 0 ° C. to the boiling point of the solvent used, optionally in the presence of a base such as pyridine, triethylamine or diisopropylethylamine, in the presence of a solvent such as dichloromethane.

Compounds of formula R ^S1 SO ₂ Cl are commercially available or can be synthesized by methods reported in the chemical literature.

X is NR ^X and R ^X is selected from the group consisting of optionally substituted C _1-20 alkyl or C _3-20 heterocyclyl, thus formula 30:

Wherein n, R ^C1 , R ^C2 and R ¹ are as defined above, R ^C4 and R ^C5 are H, ^optionally substituted C _1-20 alkyl, C _5-20 aryl, C Each independently selected from the group consisting of _3-20 heterocyclyls, or taken together to form an optionally substituted C _3-7 cycloalkyl or heterocyclyl group). The compound of the invention is a reducing agent of a compound of formula 26 with a compound of formula R ^C4 COR ^{C5 where} R ^C4 and R ^C5 are as defined above, eg sodium cyanoborohydride or triacetoxy It can be synthesized by reaction in the presence of sodium borohydride, in the presence of a solvent such as methanol, and optionally in the presence of an acid catalyst such as acetic acid, at a temperature ranging from 0 ° C. to the boiling point of the solvent used.

Compounds of formula R ^C4 COR ^C5 are commercially available or can be synthesized by methods reported in the chemical literature.

Uses The present invention provides active compounds, specifically compounds that are active in inhibiting the activity of PARP.

  The term “activity” as used herein relates to a compound capable of inhibiting PARP activity, specifically a compound (drug) having an intrinsic activity and a prodrug of such a compound. As such, it includes both prodrugs that themselves exhibit little or no intrinsic activity.

  One assay that can be conveniently used to assess PARP inhibition conferred by a particular compound is described in the examples below.

  The present invention further provides a method of inhibiting the activity of PARP in a cell comprising contacting the cell with an effective amount of an active compound, preferably in the form of a pharmaceutically acceptable composition. . Such methods can be practiced in vitro or in vivo.

  For example, a sample of cells can be grown in vitro, the active compound can be contacted with the cells, and the effect of the compound on the cells can be observed. As an example of “effect”, the amount of DNA repair performed in a certain time can be measured. If the active compound is found to affect the cell, the cell can be used as a prognostic or diagnostic marker for the efficacy of the compound in a method of treating a patient having cells of the same cell type.

  The term `` treatment '' as used herein in the context of treating a condition, whether it relates to humans or animals (e.g., for veterinary use), refers to any desired therapeutic effect, e.g., progression of the condition. It is broadly related to the treatments and therapies in which inhibition is achieved, including slowing progression, stopping progression, improving the condition, and curing the condition. Treatment as a prophylactic measure (ie prevention) is also included.

  The term “adjuvant” as used herein relates to the use of active compounds in conjunction with known therapeutic means. Such means include cytotoxic measures of drugs and / or ionizing radiation used in the treatment of various cancer types. Specifically, the active compounds are known to enhance the action of several cancer chemotherapeutic methods, including most of the known alkylating agents used in treating cancer and toxics of topoisomerases. Yes.

  The active compounds can also be used as cell culture additives to inhibit PARP, for example to increase the sensitivity of cells to known chemotherapeutic agents or ionizing radiation therapy in vitro.

  The active compounds can also be used as part of an in vitro assay, for example, to determine whether a candidate host is likely to benefit from treatment with a compound of interest.

The administered active compound or pharmaceutical composition comprising the active compound, whether administered systemically / peripherally or at the desired site of action, orally (e.g., by oral ingestion); topical (e.g., (E.g., intradermal, intranasal, ocular, buccal and sublingual); pulmonary (e.g. by aerosol, e.g. by inhalation or ventilation through the mouth or nose); rectal; vagina; , Subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intradural, spinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal By any convenient route of administration including, but not limited to, parenteral routes of administration by injection, such as subcutaneous, intraarticular, intrathecal and intrasternal; for example, subcutaneously or intramuscularly It can be administered by an implant of a depot preparation.

  Subjects are eukaryotes, animals, vertebrates, mammals, rodents (e.g. guinea pigs, hamsters, rats, mice), murines (e.g. mice), canines (e.g. dogs), felines (e.g. cats), Horses (e.g. horses), primates, simians (e.g. monkeys or apees), monkeys (e.g. marmoset, baboons), tailless monkeys (e.g. gorillas, chimpanzees, orangutans, gibbons) or humans can do.

While the pharmaceutically active compound can be administered alone, at least one active compound as defined above can be combined with one or more pharmaceutically acceptable carriers, adjuvants, excipients, diluents It is provided as a pharmaceutical composition (e.g., a formulation) comprising a bulking agent, buffer, stabilizer, preservative, lubricant or other material well known to those of skill in the art and optionally other therapeutic or prophylactic agents It is preferable.

  Accordingly, the present invention further provides a pharmaceutical composition as defined above, and at least one active compound as defined above in one or more pharmaceutically acceptable forms as described herein. A method of making a pharmaceutical composition is provided comprising mixing with a carrier, excipient, buffer, adjuvant, stabilizer or other material.

  As used herein, the term `` pharmaceutically acceptable '' refers to a subject (e.g., without excessive toxicity, irritation, allergic reactions, or other problems or complications within the scope of appropriate medical judgment). It relates to compounds, materials, compositions and / or dosage forms suitable for use in contact with human tissues and corresponding to a reasonable risk-to-effect ratio. Each carrier, excipient, etc. must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.

  Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical texts. For example, `` Handbook of Pharmaceutical Additives '', 2nd Edition (eds. M. Ash and I. Ash), 2001 (Synapse Information Resources, Inc., Endicott, New York, USA), `` Remington's Pharmaceutical Sciences '', 20th edition, pub Lippincott, Williams & Wilkins, 2000; and “Handbook of Pharmaceutical Excipients”, 2nd edition, 1994.

  The formulations may conveniently be provided in unit dosage form and may be prepared by any method well known in the pharmaceutical art. Such methods include the step of associating the active compound with the carrier that constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active compound with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product.

  Formulation is liquid, liquid, suspension, emulsion, elixir, syrup, tablet, lozenge, granule, powder, capsule, cachet, pill, ampoule, suppository, suppository , Ointments, gels, pastes, creams, sprays, mists, foams, lotions, oils, boluses, lozenges or aerosols.

  Formulations suitable for oral administration (eg, by oral ingestion) include capsules, cachets or tablets, each as a discrete unit containing a predetermined amount of the active compound; as a powder or granules; aqueous or non-aqueous liquid As a solution or suspension in; or as an oil-in-water emulsion or water-in-oil emulsion; as a bolus; as a lozenge; or as a paste.

  A tablet may be made by conventional means, eg, compression or molding, optionally with one or more accessory ingredients. A compressed tablet contains the active compound in a free flowing form such as a powder or granules in an appropriate machine, optionally with one or more binders (e.g., povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethylcellulose). Bulking agents or diluents (eg lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (eg magnesium stearate, talc, silica); disintegrants (eg sodium starch glycolate, crosslinked povidone, crosslinked) Sodium carboxymethyl cellulose); surfactant or dispersant or wetting agent (e.g. sodium lauryl sulfate); and preservatives (e.g. methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid) and compress Can be prepared. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. Tablets may be optionally coated or scored and provide, for example, hydroxypropylmethylcellulose in varying proportions to provide a sustained or controlled release of the active compound in the tablet to provide the desired release characteristics You may mix | blend as follows. The tablets may optionally be enteric coated to provide release in intestinal parts other than the stomach.

  Formulations suitable for topical (e.g., transdermal, intranasal, ocular, buccal and sublingual) ointments, creams, suspensions, lotions, powders, solutions, pastes, They can be formulated as gels, sprays, aerosols or oils. Alternatively, the formulation may comprise a patch or dressing such as a bandage or adhesive bandage impregnated with the active compound and optionally one or more excipients or diluents.

  Some formulations suitable for topical administration in the mouth include flavored bases, usually sucrose and lozenges containing the active compound in acacia or tragacanth; gelatin and glycerin, or inert such as sucrose and acacia Perfumes containing the active compound in a base; and mouthwashes containing the active compound in a suitable liquid carrier.

  Formulations suitable for topical administration to the eye also include eye drops wherein the active compound is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active compound.

  Formulations suitable for nasal administration, wherein the carrier is a solid, are administered so that the key agent is inhaled, i.e. by rapid inhalation through the nasal passage from a container of the powder held tightly to the nose, e.g. about Includes a coarse powder having a particle size in the range of 20 to about 500 microns. Suitable formulations in which the carrier is a liquid for administration, eg, as a nasal spray, aerosol as a nasal spray or by nebulizer, include an aqueous or oily solution of the active compound.

  Formulations suitable for administration by inhalation include aerosols from pressurized packs using a suitable high pressure gas such as dichlorodifluoromethane, trichlorofluoromethane, dichoro-tetrafluoroethane, carbon dioxide or other suitable gas. Includes those provided as propellants.

  Formulations suitable for topical administration via the skin include ointments, creams and emulsions. When formulated in an ointment, the active compound may optionally be utilized with either a paraffinic or water-miscible ointment base. Alternatively, the active compound may be formulated in a cream with an oil-in-water cream base. Optionally, the aqueous phase of the cream base may include, for example, at least about 30% by weight of a polyhydric alcohol, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol, and mixtures thereof. And alcohols having two or more hydroxyl groups. Topical formulations may desirably include compounds that facilitate absorption or penetration of the active compound through the skin or other affected areas. Examples of such skin penetration enhancers include dimethyl sulfoxide and related analogs.

  When formulated as a topical emulsion, the oil phase may optionally contain only an emulsifier (also known as an emulgent), or the oil phase may be with a fat or oil or with both fat and oil. A mixture of at least one emulsifier. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferable to contain both an oil agent and fat. Together, the emulsifier with or without stabilizer constitutes a so-called emulsifying wax, and the waxy substance together with the oil and / or fat constitutes a so-called emulsifying ointment base that forms the oil-based dispersed phase of the cream formulation.

  Suitable emergency and emulsion stabilizers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate. The selection of a suitable oil or fat for the formulation is based on achieving the desired cosmetic properties. This is because the solubility of the active compound in most oils likely to be used in pharmaceutical emulsion formulations may be very low. Thus, the cream should preferably be a non-sticky, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Diisoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acid, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a mixture of branched esters known as Crodamol CAP Linear or branched monobasic or dibasic alkyl esters can be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and / or liquid paraffin or other mineral oils can be used.

  Formulations suitable for rectal administration can be presented as suppositories with a suitable base comprising, for example, cocoa butter or a salicylate.

  Formulations suitable for vaginal administration include vaginal suppositories, tampons, creams, gels, pastes, foams or the like, which contain, in addition to the active compound, a carrier as is known in the art. It can be provided as a spray formulation.

  Formulations suitable for parenteral administration (e.g., by injection, including cutaneous, subcutaneous, intramuscular, intravenous and intradermal injection) include antioxidants, buffers, preservatives, stabilizers, Aqueous and non-aqueous isotonic and pyrogen-free sterile injections that can contain solutes that make the fungus and preparation isotonic with the intended recipient's blood; Including aqueous and non-aqueous sterile suspensions, as well as liposomes or other particulate systems designed to target the compound to blood components or one or more organs. Examples of isotonic media suitable for use in such formulations include sodium chloride injection, Ringer's solution, or lactated Ringer's injection. Usually, the concentration of the active compound in solution is from about 1 ng / ml to about 10 μg / ml, for example from about 10 ng / ml to about 1 μg / ml. The formulation can be dispensed in unit dose or multi-dose sealed containers, such as ampoules and vials, in a lyophilized (lyophilized) state that requires the addition of a sterile liquid carrier, such as water for injection, just prior to use. Can be saved. Extemporaneous injections and suspensions can be prepared from sterile powders, granules, and tablets. The formulations can be in the form of liposomes or other particulate systems designed to target the active compound to blood components or one or more organs.

It will be appreciated that the appropriate dose of the active compound and the composition comprising the active compound may vary from patient to patient. The determination of the optimal dose will generally involve a balance of the degree of therapeutic effect on any risk or toxic side effects of the treatment of the present invention. The selected dosage level depends on the activity of the particular compound, the route of administration, the time of administration, the excretion rate of the compound, the duration of treatment, the other drugs, compounds and / or materials used in combination, and the patient's age, gender, weight, It will depend on a variety of factors, including but not limited to condition, general health and previous medical history. In general, the dosage should be at the site of action, achieving a local concentration that achieves the desired effect without causing substantially harmful or toxic side effects, but the amount and route of administration of the compound ultimately It will be at the discretion of the doctor.

  Administration in vivo can be performed in a single dose throughout the treatment period, continuously or intermittently (eg, in divided doses at appropriate intervals). Methods of determining the most effective means of administration and dosage are well known to those of skill in the art and will vary with the formulation used for therapy, the purpose of the therapy, the target cell being treated, and the subject being treated. . Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician.

  In general, a suitable dose of active compound is in the range of about 100 μg to about 250 mg per kilogram of subject body weight per day. When the active compound is a salt, ester, prodrug, etc., the amount administered will be calculated based on the parent compound, so the actual weight used will increase proportionally.

Example
General experimental method <br/> Preparative HPLC
Samples were purified using a Waters mass spectrometer direct purification system using a Waters 600 LC pump, a Waters Xterra C18 column (5 μm 19 mm x 50 mm) and a Micromass ZQ mass spectrometer operated in positive ion electrospray ionization mode did. Gradient mobile phase A (0.1% formic acid in water) and mobile phase B (0.1% formic acid in acetonitrile) were used at a flow rate of 20 ml / min, with 5% to 100% B held for 7 minutes over 7 minutes. .

Analytical HPLC-MS
Analytical HPLC was typically performed with a Spectra System P4000 pump and a Jones Genesis C18 column (4 μm, 50 mm × 4.6 mm). Mobile phase A (0.1% formic acid in water) and mobile phase B (acetonitrile solution) were used at a flow rate of 2 ml / min from 5% B for 1 minute to 98% B after 5 minutes and used for a gradient of 3 minutes. Detection was performed with a TSP UV 6000LP detector at a UV of 254 nm and a PDA range of 210-600 nm. The mass spectrometer was a Finnigan LCQ operated in positive ion electrospray mode.

NMR
¹ H NMR and ¹³ C NMR were usually recorded using Bruker DPX 300 spectrometer at 300 MHz and 75 MHz, respectively. Chemical shifts are reported in parts per million (ppm) of the δ scale relative to the internal standard tetramethylsilane. Unless otherwise stated, all samples were dissolved in DMSO-d ^6.

Synthesis of key intermediates
(i) Synthesis of (3-oxo-1,3-dihydro-isobenzofuran-1-yl) phosphonic acid dimethyl ester

Dimethyl phosphite (22.0 g, 0.2 mol) was added dropwise to a solution of sodium methoxide (43.0 g) in methanol (100 ml) at 0 ° C. 2-Carboxybenzaldehyde (21.0 g, 0.1 mol) was then added in portions to the reaction mixture as a slurry in methanol (40 ml) while maintaining the temperature below 5 ° C. The resulting pale yellow solution was warmed to 20 ° C. over 1 hour. Methanesulfonic acid (21.2 g, 0.22 mol) was added dropwise to the reaction and the resulting white suspension was evaporated in vacuo. The white residue was quenched with water and extracted with chloroform (3 × 100 ml). The combined organic extracts were washed with water (2 × 100 ml), dried over MgSO ₄ and evaporated in vacuo to give (i) a white solid (32.0 g, 95%, purity 95%). This was then used in the next step without further purification.

(ii) Synthesis of aldehyde intermediate
6-Formyl-pyridine-2-carboxylic acid (iia)

  (a) 1 equivalent of potassium permanganate was added to an aqueous solution of 6-bromo-2-methyl-pyridine. The solution was heated to reflux for 2 hours. The reaction was followed and potassium permanganate was added until no starting material remained. After cooling, the solution was filtered and the solution was acidified to pH = 3. The precipitate was filtered and dried.

  (b) DMA / water 9/1 solution of 6-bromo-2-pyridine-carboxylic acid, 1.5 equivalents of vinylboronic acid dibutyl ester and 1.2 equivalents of potassium carbonate is degassed for 20 minutes, then 0.06 equivalents of palladium tetrakis is added and suspended. The suspension was degassed for an additional 30 seconds and the suspension was heated at 170 ° C. in a microwave for 25 minutes. The resulting suspension was filtered through a pad of silica gel and the filtrate was concentrated.

  (c) A methanol / DCM 1/1 solution of 6-vinyl-pyridine-2-carboxylic acid was cooled to −78 ° C., and ozone was bubbled through the solution until the solution turned blue. Next, excess ozone was removed through a nitrogen stream, and 1.5 equivalents of methyl sulfide was added. The solution was allowed to warm to room temperature and concentrated. The product (iia) was then purified by silica flash chromatography.

6-formyl-pyridine-2-carbonitrile (iib)

(a) Iodomethane (204 ml, 3.1 mol) was added dropwise to 2-picoline-N-oxide (100 g, 0.9 mol) at 25 ° C. over 1 hour. The reaction was left for 12 hours. The reaction mixture was filtered, washed with Et ₂ O and dried in a vacuum oven for 3 hours to give the product, which was used in the next step without purification.

(b) An aqueous solution of KCN (112.0 g, 1.7 mol in 250 ml of H ₂ O) was added dropwise to the ethanol-H ₂ O (7: 3) solution of the previous product at 0 ° C. over 180 minutes. The reaction was left to stir for an additional 30 minutes. The reaction was then warmed to 25 ° C. and extracted into 200 ml dichloromethane and a further 4 × 100 ml. The combined organic layers were washed with 200 ml saturated brine, dried over MgSO ₄ , filtered and evaporated to give a dark red liquid (51.0 g) which crystallized upon standing for 12 hours. The solid was filtered, washed with cold hexane (2 × 50 ml) and air dried. The solid was then purified by column chromatography (silica 70 g, hexane: ethyl acetate) to give the product as a white solid (7.0 g, 7%). m / z [M + 1] ⁺ 119 (purity 98%).

(c) m-CPBA (16.2 g, 0.14 mol) was added to a solution of the previous stage product (52.0 g, 0.15 mmol) in DCM (50 ml) and the reaction was left stirring for 12 hours. Na ₂ S ₂ O ₃ (21.5 g) was added and the reaction mixture was left to stir for an additional 30 minutes. The reaction was then filtered and washed with saturated NaHCO ₃ (2 × 30 ml), brine (2 × 30 ml), dried over MgSO ₄ , filtered and evaporated to give the product as a white solid (13.6 g, 73.8%), which was used in the next step without purification.

(d) The previous product (13.5 g, 101.3 mmol) was added to a preheated solution of acetic anhydride (60 ml) at 120 ° C. and the reaction was refluxed for 90 minutes. Then 60 ml of ethanol was carefully added to the reaction mixture, refluxed for another 10 minutes and cooled to 25 ° C. The reaction was added to water (100 ml) and neutralized with NaHCO ₃ (50 g). The reaction was extracted into diethyl ether (2 × 30 ml). The combined organic layers were washed with water (2 × 20 ml), dried over MgSO ₄ , filtered and evaporated to give a brown oil that was purified by column chromatography (hexane: ethyl acetate). The product was obtained as a yellow oil (5.4 g, 30%). m / z [M + 1] ⁺ 177 (purity 30%).

(e) 1 NH ₂ SO ₄ (6 ml) was added to a solution of the previous stage product (5.4 g, 30.6 mmol) in tetrahydrofuran (15 ml) and the reaction was refluxed for 18 hours. The reaction was cooled, poured into water (150 ml), neutralized with NaHCO ₃ and extracted into DCM (3 × 50 ml). The combined organic layers were washed with 100 ml saturated brine, dried over MgSO ₄ , filtered and evaporated to give the product as a brown solid that was used in the next step without purification. m / z [M + 1] ⁺ 134 (71% purity).

(f) The product from the previous step and N, N′-dicyclohexylcarbodiimide (19.3 g, 93.0 mmol) was added to a mixture of DMSO (22 ml) and anhydrous H ₃ PO ₄ (1.4 g) and the reaction was stirred for 1.5 hours. I left it alone. The reaction was filtered and washed with diethyl ether (2 × 30 ml) and water (2 × 30 ml). The reaction layer was separated and the organic layer was washed with saturated brine (2 × 30 ml), dried over MgSO ₄ , filtered and evaporated to give (iib) as a yellow solid which was not purified. Used in the next stage.

2-formyl-isonicotinic acid (iic)

  (a) 1 equivalent of potassium permanganate was added to an aqueous solution of 2-bromo-4-methyl-pyridine. The solution was heated to reflux for 2 hours. The reaction was followed and potassium permanganate was added until no starting material remained. After cooling, the solution was filtered and the solution was acidified to pH = 3. The precipitate was filtered and dried.

  (b) 2-bromo-isonicotinic acid, vinylboronic acid dibutyl ester 1.5 equivalents, potassium carbonate 1.2 equivalents in DMA / water 9/1 solution was degassed for 20 minutes, then palladium tetrakis 0.06 equivalents were added and the suspension was The mixture was further degassed for 30 seconds and the suspension was heated in a microwave oven at 170 ° C. for 25 minutes. The resulting suspension was filtered through a pad of silica gel and the filtrate was concentrated.

  (c) A methanol / DCM 1/1 solution of 2-vinyl-isonicotinic acid was cooled to −78 ° C. and ozone was bubbled through the solution until the solution turned blue. Next, excess ozone was removed through a nitrogen stream, and 1.5 equivalents of methyl sulfide was added. The solution was allowed to warm to room temperature and concentrated. The product (iic) was then purified by silica flash chromatography.

2-formyl-isonicotinonitrile (iid)

(a) Iodoethane (265 ml, 3.3 mol) was added dropwise to 2-picoline-N-oxide (100 g, 0.9 mol) at 25 ° C. over 1 hour. The reaction was left for 12 hours. The reaction mixture was filtered, washed with Et ₂ O and dried in a vacuum oven for 3 hours to give the product, which was used in the next step without purification.

(b) An aqueous solution of KCN (52.0 g, 0.8 mol in 100 ml of H ₂ O) was added dropwise to the ethanol-H ₂ O (7: 3) solution of the previous product at 50 ° C. over 110 minutes. The reaction was left to stir for an additional 30 minutes. The reaction was then warmed to 25 ° C. and extracted into 200 ml dichloromethane and a further 4 × 100 ml. The combined organic layers were washed with 200 ml saturated brine, dried over MgSO ₄ , filtered and evaporated to give a dark red liquid (51.0 g). This procedure was repeated to give a total of 102.0 g of reaction mixture which was purified by column chromatography (silica 360 g, hexane: ethyl acetate) to give the product as a white solid (10.6 g, 10%) Got as. m / z [M + 1] ⁺ 119 (purity 98%).

(c) m-CPBA (27.7 g, 80.2 mmol) was added to a solution of the previous stage product (8.6 g, 72.8 mmol) in DCM (30 ml) and the reaction was left stirring for 12 hours. Na ₂ S ₂ O ₃ (10.0 g, 16.0 mmol) was added and the reaction mixture was left to stir for an additional 30 minutes. The reaction was then filtered and washed with saturated NaHCO ₃ (2 × 30 ml), brine (2 × 30 ml), dried over MgSO ₄ , filtered and evaporated in vacuo to give the product as a white solid ( 6.5 g, 67%), which was used in the next step without purification.

(d) The previous product (8.0 g, 60.0 mmol) was added to a preheated solution of acetic anhydride (30 ml) at 120 ° C. and the reaction was refluxed for 90 minutes. Then 30 ml of ethanol was carefully added to the reaction mixture, refluxed for another 10 minutes and cooled to 25 ° C. The reaction was added to water (100 ml) and neutralized with NaHCO ₃ (50 g). The reaction was extracted into diethyl ether (2 × 30 ml). The combined organic layers were washed with water (2 × 20 ml), dried over MgSO ₄ , filtered and evaporated in vacuo to give a brown oil that was purified by column chromatography (hexane: ethyl acetate). The product was obtained as a yellow solid (4.0 g, 38%). m / z [M + 1] ⁺ 177 (purity 96%).

(e) 1 NH ₂ SO ₄ (16 ml) was added to a solution of the previous stage product (2.7 g, 15.6 mmol) in tetrahydrofuran (25 ml) and the reaction was refluxed for 18 hours. The reaction was cooled, poured into water (150 ml), neutralized with NaHCO ₃ and extracted into DCM (3 × 50 ml). The combined organic layers were washed with 100 ml saturated brine, dried over MgSO ₄ , filtered and evaporated in vacuo to give the product as a yellow solid (1.4 g, 67%) which was purified without further purification. Used at the stage.

(f) The product from the previous stage (1.4 g, 10.2 mmol) and N, N′-dicyclohexylcarbodiimide (6.2 g, 30.0 mmol) were added to a mixture of DMSO (22 ml) and anhydrous H ₃ PO ₄ (0.45 g). The reaction was left stirring for 1.5 hours. The reaction was filtered and washed with diethyl ether (2 × 30 ml) and water (2 × 30 ml). The reaction layer was separated and the organic layer was washed with saturated brine (2 × 30 ml), dried over MgSO ₄ , filtered and evaporated in vacuo to give (iid) as a yellow solid which was purified. It was used in the next stage.

(iii) Coupling of aldehyde intermediate (ii) with (3-oxo-1,3-dihydro-isobenzofuran-1-yl) -phosphonic acid dimethyl ester (i)
(a) Synthesis of 2- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -isonicotinic acid (iiia)

(3-Oxo-1,3-dihydro-isobenzofuran-1-yl) -phosphonic acid dimethyl ester (i) (0.18 g, 0.77 mmol) and 2-formyl-isonicotinic acid (iic) (0.12 g, 0.77 mmol) ) In tetrahydrofuran (10 ml) was added triethylamine (0.32 ml, 2.8 mmol). The reaction mixture was stirred at 50 ° C. for 4 hours and then allowed to cool to room temperature. Tetrahydrofuran was evaporated to half its volume, then 1 N HCl solution was added to pH 3. Water was added until the solid was no longer crushed by the solution. The white solid was filtered, washed with water, then hexane, and recrystallized from acetonitrile. Amount: 0.9 g, m / z [M + 1] ⁺ 268 (purity 90%).

  Except that the reaction was carried out at room temperature for 16 hours instead of 4 hours at 50 ° C., all the aldehydes containing carboxylic acid functional groups were converted to (3-oxo-1,3-dihydro-iso Coupled to benzofuran-1-yl) -phosphonic acid dimethyl ester.

The compounds synthesized according to the above protocol are:
6-formyl-pyridine-2-carboxylic acid (iia) to 6- [3-oxo-3H-isobenzofuran-1-ylidenemethyl] -pyridine-2-carboxylic acid (iiib)
From 5-formyl-furan-2-carboxylic acid to 5- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -thiophene-2-carboxylic acid (iiic): m / z [M + 1] ⁺ 287 ( (Purity 92%)
From 5-formyl-thiophene-2-carboxylic acid, 5- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -furan-2-carboxylic acid (iiid): m / z [M + 1] ⁺ 257 ( (Purity 90%)
(b) Alternative synthesis of 2- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -isonicotinic acid (iiia)

  (i) Triethylamine (2.2 ml, 15 mmol) was added to (3-oxo-1,3-dihydro-isobenzofuran-1-yl) -phosphonic acid dimethyl ester (i) (2.4 g, 10.2 mmol) and (iid) ( 10.2 mmol) in tetrahydrofuran (10 ml). The reaction mixture was stirred at 25 ° C. for 12 hours and concentrated in vacuo to give 2- [3-oxo-3H-isobenzofuran- (1E, Z) -ylidenemethyl] -isonicotinonitrile (iii-int) as a red solid This was used in the next step without purification.

  (ii) 2- [3-Oxo-3H-isobenzofuran- (1E, Z) -ylidenemethyl] -isonicotinonitrile (iii-int) (10.2 mmol), water (50 ml) and potassium hydroxide pellets (1.7 g, 30.7 mmol) was refluxed for 16 hours. The reaction was cooled to 25 ° C. and washed with dichloromethane (2 × 30 ml). Thereafter, the aqueous layer was concentrated in vacuo, and the resulting solid (iiia) was used in the next step without further purification.

  All aldehydes containing nitrile groups were coupled to (3-oxo-1,3-dihydro-isobenzofuran-1-yl) -phosphonic acid dimethyl ester and then hydrolyzed to carboxylic acids using the above conditions. .

The compound synthesized according to the above protocol is: 6-formyl-pyridine-2-carbonitrile (iib) to 6- [3-oxo-3H-isobenzofuran-1-ylidenemethyl] -pyridine-2 -Carboxylic acid (iiib); step (a) gave a yellow solid: m / z [M + 1] ⁺ 249 (purity 98%); step (b) gave a dark yellow oil: m / z [M + 1] ⁺ 286 (83% purity).

(c) 4- (3-Oxo-3H-isobenzofuran-1-ylidenemethyl) -pyridine-2carbonitrile (iii'e)

  To a solution of 4-formyl-pyridine-2-carbonitrile (4.89 g, 37.0 mmol) in anhydrous THF (200 mL) was added dimethyl ((3-oxo-1,3-dihydro-isobenzofuran-1-yl) -phosphonate. Ester] phosphonic acid (9.2 g, 37.0 mmol) was added followed by triethylamine (5.1 mL, 37.0 mmol) and the reaction was stirred at room temperature for 18 hours. The reaction mixture was then filtered and the isolated solid was washed with dry THF (2 × 25 mL) and dried in vacuo. Two peaks (geometric isomer) by LC-MS analysis, (7.0 g, 76%); m / z (LC-MS, ESP), rt = 4.19 min, (M + H) = 249 and rt = 4.36 Min, (M + H) = 249. This material was used without the need for purification.

(iv) Conversion of isobenzofuran compounds to phthalazinone compounds
(a) Synthesis of 5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -furan-2-carboxylic acid (iva)

  To a suspension of 2- [3-oxo-3H-isobenzofuran-ylidenemethyl] isonicotinic acid (iiia) (87 mg, 0.32 mmol) in water (2 ml), hydrazine monohydrate (33 mg, 0.64 mmol) Was added and the mixture was heated at reflux for 5 hours. The solution was concentrated to half its volume and acidified with 1 N HCl solution to pH 3. The white solid was filtered, washed with water and dried. Amount: 32 mg.

m / z [M + 1] ⁺ 282 (purity 42%).

  The phthalazinone core was formed on all compounds according to the protocol described above.

The compounds synthesized according to the above protocol are:
6- [3-oxo-3H-isobenzofuran-1-ylidenemethyl] -pyridine-2-carboxylic acid (iiib) to 6- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -pyridine- 2-carboxylic acid (ivb): m / z [M + 1] ⁺ 282 (purity 48%);
From 5- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -thiophene-2-carboxylic acid (iiic) to 5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -thiophene- 2-carboxylic acid (ivc): m / z [M + 1] ⁺ 287 (purity 60%);
From 5- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -furan-2-carboxylic acid (iiid) to 5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -furan- 2-carboxylic acid (ivd): m / z [M + 1] ⁺ 271 (94% purity).

(b) Synthesis of 4- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -pyridine-2-carboxylic acid (ive)

  4- (3-Oxo-3H-isobenzofuran-1-ylidenemethyl) -pyridine-2carbonitrile (iii'e) (3.72 g, 15.0 mmol) was added to water (100 mL) and hydrazine monohydrate (1.5 g 30.0 mmol) was added. The reaction mixture was then heated to 100 ° C. for 6 hours and then cooled to room temperature. The white suspension was filtered and washed with diethyl ether (2 × 20 ml). This material was then vacuum dried. Major peak by LC-MS analysis (7.0 g, 76%); m / z (LC-MS, ESN), rt = 3.54 min (M + H) = 261.

  Concentrated hydrochloric acid (5 mL) was added to a solution of the obtained material (2.36 g, 9.0 mmol) in ethanol (10 mL). The reaction mixture was then heated to 70 ° C. for 18 hours and then cooled to 5 ° C., and the resulting white suspension was filtered, water (2 × 5 mL) followed by diethyl ether (2 × 20 mL). ). A beige solid was isolated and became the main peak in LC-MS analysis (2.40 g, 94%); m / z (LC-MS, ESP), RT = 3.49 min, (M + H) = 282; and (2M + H) = 563. This material was used without the need for purification.

(v) Addition of piperazine group
(a) Synthesis of 4- [6- (piperazin-1-carbonyl) -pyridin-2-ylmethyl] -2H-phthalazin-1-one (vb)

6- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -pyridine-2-carboxylic acid (ivb) (0.3 g, 1.1 mmol), triethylamine (0.3 ml, 2.1 mmol), tert-butyl- Dimethylformamide of 1-piperazinecarboxylic acid (0.23 g, 1.3 mmol) and tetraethylammonium 2- (1H-benzotriazol-1-yl) -1,1,3,3-hexafluorophosphate (0.5 g, 1.3 mmol) (10 ml) The mixture was stirred for 18 hours. The reaction mixture was precipitated by the addition of water (50 ml) and air dried. The white precipitate was dissolved in ethanol (3 ml), 12 M hydrochloric acid (6 ml) was added to the solution and the reaction was stirred for 30 minutes. The reaction was then concentrated in vacuo, redissolved in water (10 ml) and washed with dichloromethane (2 × 10 ml). The aqueous layer was basified with ammonium hydroxide and extracted into dichloromethane (2 × 10 ml). The combined organic layers were dried over MgSO _4, filtered, expected product were evaporated to (va) was obtained as a pink solid (0.23 g, 73%). m / z [M + 1] ⁺ 250 (purity 96%).

  All compounds were coupled to piperazine-1-carboxylic acid tert-butyl ester and the protecting group removed according to the protocol described above.

The compounds synthesized according to the above protocol are:
5- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -furan-2-carboxylic acid (iva) to 4- [4- (piperazin-1-carbonyl) -pyridin-2-ylmethyl] -2H-phthalazin-1-one (va): m / z [M + 1] ⁺ 250 (purity 96%);
From 5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -thiophene-2-carboxylic acid (ivc) to 4- [5- (piperazin-1-carbonyl) -thiophen-2-ylmethyl] -2H-phthalazin-1-one (vc): m / z [M + 1] ⁺ 339 (80% purity);
5- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -furan-2-carboxylic acid (ivd) to 4- [5- (piperazin-1-carbonyl) -furan-2-ylmethyl] -2H-phthalazin-1-one (vd): m / z [M + 1] ⁺ 355 (84% purity).

(b) Synthesis of 4- [2- (piperazin-1-carbonyl) -pyridin-4-ylmethyl] -2H-phthalazin-1-one (ve)

  To a solution of 4- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -pyridine-2-carboxylic acid (ive) (0.563 g, 2/0 mmol) in anhydrous DCM (30 mL) was added 1- Piperazine carboxylate tert-butyl (0.45 g, 2.4 mmol) and hexafluorophosphate O-benzotriazole-N, N, N ′, N′-tetramethyluronium (0.91 g, 2.4 mmol) were added. The mixture was stirred for 5 minutes before adding N ′, N′-diisopropylethylamine (0.42 mL, 2.4 mmol). After 30 minutes of stirring at room temperature, the reaction mixture was filtered and concentrated in vacuo. The resulting oil was chromatographed with EtOAc: MeOH 9: 1 (rf of 0.23) to isolate a white solid. LC-MS analysis gave a single peak (0.71 g, 79%) and did not require further purification. m / z (LC-MS, ESP), RT = 3.75 min. (M + H) = 450.

  4 M hydrogen chloride (3.25 mL, 13.0 mmol) was added to the dioxane solution of the obtained compound (0.60 g, 1.35 mmol). After 15 minutes, the solvent was removed in vacuo and 7 N ammonia in methanol (3 mL, 15.0 mmol) was added. The resulting cream colored precipitate was filtered. The filtrate was concentrated in vacuo to give a sticky gum (0.31 g yield 89%). LC-MS analysis showed 93% purity and no further purification was attempted. m / z (LC-MS, ESP), RT = 2.86 min. (M + H) = 350.

  Appropriate acid chloride or sulfonyl chloride (0.24 mmol) in 4- [4- (piperazin-1-carbonyl) -pyridin-2-ylmethyl] -2H-phthalazin-1-one (va) in dichloromethane (2 ml) Added to. Then Hunig's base (0.4 mmol) was added and the reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (a) Appropriate acid chloride or sulfonyl chloride (0.24 mmol) with 4- [4- (piperazin-1-carbonyl) -pyridin-2-ylmethyl] -2H-phthalazin-1-one (vb) in dichloromethane (2 ml) was added to the solution. Then Hunig's base (0.4 mmol) was added and the reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (b) Appropriate isocyanate (0.24 mmol) was added to a solution of 4- [4- (piperazin-1-carbonyl) -pyridin-2-ylmethyl] -2H-phthalazin-1-one (vb) in dichloromethane (2 ml). It was. The reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (c) 6- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -pyridine-2-carboxylic acid (iva) (0.3 g, 1.1 mmol), triethylamine (0.3 ml, 2.1 mmol), appropriate Amine (1.3 mmol) and 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate (0.5 g, 1.3 mmol) in dimethylformamide (10 ml) The mixture was stirred for 18 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (a) Appropriate acid chloride (0.24 mmol) in 4- [5- (piperazin-1-carbonyl) -furan-2-ylmethyl] -2H-phthalazin-1-one (vd) in dichloromethane (2 ml) Added to. Then Hunig's base (0.4 mmol) was added and the reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (b) Add the appropriate isocyanate (0.24 mmol) to a solution of 4- [5- (piperazin-1-carbonyl) -furan-2-ylmethyl] -2H-phthalazin-1-one (vd) in dichloromethane (2 ml). It was. The reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (c) 5- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -furan-2-carboxylic acid (ivd) (1.1 mmol), triethylamine (0.3 ml, 2.1 mmol), the appropriate amine ( 1.3 mmol) and 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium (0.5 g, 1.3 mmol) hexafluorophosphate in a dimethylformamide (10 ml) mixture. Stir for hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (a) Appropriate acid chloride (0.24 mmol) in 4- [5- (piperazin-1-carbonyl) -thiophen-2-ylmethyl] -2H-phthalazin-1-one (vc) in dichloromethane (2 ml) Added to. Then Hunig's base (0.4 mmol) was added and the reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (b) Add the appropriate isocyanate (0.24 mmol) to a solution of 4- [5- (piperazin-1-carbonyl) -thiophen-2-ylmethyl] -2H-phthalazin-1-one (vc) in dichloromethane (2 ml). It was. The reaction was stirred at room temperature for 16 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (c) 5- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -thiophene-2-carboxylic acid (ivc) (1.1 mmol), triethylamine (0.3 ml, 2.1 mmol), the appropriate amine ( 1.3 mmol) and 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate (0.5 g, 1.3 mmol) in dimethylformamide (10 ml) Stir for hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (a) Appropriate acid chloride (0.13 mmol) to 4- [2- (piperazin-1-carbonyl) -pyridin-4-ylmethyl] -2H-phthalazin-1-one (ve) (0.045 g, 0.13 mmol) To a solution of anhydrous DCM (1.0 mL). Next, N ′, N′-diisopropylethylamine (47 μL, 0.26 mmol) was added, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

  (b) 4- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -pyridine-2-carboxylic acid (ive) (0.037 g, 0.13 mmol) in anhydrous dimethylacetamide (1 mL) The appropriate secondary amine (0.14 mmol) and hexafluorophosphate O-benzotriazole-N, N, N ′, N′-tetramethyluronium (0.060 g, 0.16 mmol) were added. The mixture was stirred for 5 minutes before adding N′N′-diisopropylethylamine (0.47 μL, 0.26 mmol) and stirred overnight at room temperature. The reaction mixture was then purified by preparative HPLC.

The synthesized compounds are shown below.

(c)

  Compound 64 was deprotected with concentrated HCl in an organic solvent to give compound 63. Rt 3.21 min, M + 1 364.

5 alternative synthesis

(a) 2-hydroxymethyl-isonicotinonitrile Concentrated sulfuric acid (5 mL) was added to a stirred solution of 4-cyanopyridine (10.4 g, 100.0 mmol) in methanol (100 mL) and water (50 mL) at room temperature. A slight fever was observed. After 10 minutes, iron sulfate heptahydrate (910 mg, 3.0 mmol) was added and the reaction immediately turned dark yellow / orange. After sonication of the reaction mixture under nitrogen for 20 minutes, hydroxylamine-O-sulfuric acid (11.3 g, 100.0 mmol) was added in one portion. A slight exotherm was observed after 10 minutes and the reaction was maintained under a nitrogen atmosphere.

After 2 hours, additional hydroxylamine-O-sulfuric acid (11.3 g, 100.0 mmol) was added along with concentrated sulfuric acid (5 mL) and iron (II) sulfate heptahydrate (910 mg, 3.0 mmol). After an additional 3 hours of stirring, the reaction was neutralized by the addition of sodium carbonate (18.0 g 200 mmol). The mixture is then diluted with water (100 ml) and filtered to remove the crimson / brown precipitate, the filtrate is concentrated to dryness, and the resulting solid is dissolved in water (50 mL), Extracted with EtOAc (5 × 100 ml). The combined organics were dried over MgSO ₄ and concentrated in vacuo to give 6.1 g of a crude gray solid.

The material is then subjected to flash chromatography with eluent Hex: EtOAc 8: 1 to remove unreacted 4-cyanopyridine and then the polarity is increased to Hex: EtOAc 2: 1 to give the desired product. Was isolated as a fluffy white solid (rf 0.5, Hex / EtOAc 2: 1). Single peak by LC-MS analysis (3.3 g, 24.6%), m / z (LC-MS, ESP), RT = 1.70 min, (M + H) = 135.0. ¹ H NMR (300 MHz) 8.72 (1H, dd, J 0.9, 6.0 Hz), 7.78 (1H, m), 7.71 (1H, dt, J 0.9, 6.0 Hz), 5.65 (1H, t, J 6.9Hz- OH), 4.61 (1H, d, J 6.9 Hz); ¹³ C NMR (100 MHz), 163.72, 149.85, 123.60, 121.69, 119.77, 116.99, 63.70.

(b) 2-Formyl-isonicotinonitrile (iid)
DMSO (21.2 mL) was added dropwise over 20 minutes to a cooled solution of oxalyl chloride (13.2 mL, 150 mmol) in anhydrous DCM (86 mL) at −78 ° C. under a nitrogen atmosphere. The mixture was stirred at -78 ° C for 15 min before 2-hydroxymethyl-isonicotinonitrile (4.0 g, 30 mmol) dissolved in anhydrous DCM (60 mL) was added dropwise to the reaction mixture over 5 min. did. The reaction was stirred at −78 ° C. for 2 hours while maintaining a nitrogen atmosphere. A white solid precipitate formed, the temperature was raised to (-55 ° C), triethylamine (6.15 mL, 450 mmol) was added dropwise over 15 minutes, the cooling bath was removed and the mixture was allowed to warm to room temperature over 2 hours. Allowed to warm. The mixture was diluted with DCM (400 mL) and washed with brine (2 × 50 mL). The aqueous phase was extracted with DCM (3 × 50 mL). The organic layers were combined and concentrated in vacuo. A pale yellowish white solid was isolated and used without further purification. Single peak in LC-MS analysis (yield interpreted as quantitative), m / z (LC-MS, ESP), RT = 2.53 min, (M + H) = 133.0.

(c) 2- (3-Oxo-3H-isobenzofuran-1-ylidenemethyl) -isonicotinonitrile (iii-int)
[(3-Oxo-1,3-dihydro-isobenzofuran-1-yl) -phosphonic acid dimethyl ester] A cooled suspension of phosphonic acid (i) (8.0 g, 33.0 mmol) in THF (400 mL) (about 0 To the mixture was added crude 2-formyl-isonicotinonitrile (iid) (30.0 mmol) followed by triethylamine (6.2 mL, 33.0 mmol). The mixture was stirred at 0 ° C. for 30 minutes and then allowed to warm overnight.

  The reaction mixture was then evaporated in vacuo and the resulting solid was then washed with ethyl acetate (2 × 50 mL), methanol (1 × 15 mL) then diethyl ether (2 × 20 mL). Two peaks in LC-MS analysis, the desired product; m / z (LC-MS, ESP), RT = 4.27 min, (M + H) = 249.0, and impurity peak (approximately 40%) RT = 4.47 min M + H 194) was detected. This material was used without the need for purification.

(d) 2- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -isonicotinonitrile Unpurified 2- (3-oxo-3H-isobenzofuran-1-ylidenemethyl) -isonicotino Nitrile (iii-int) (about 4.0 mmol) was suspended in water (30 mL) and hydrazine monohydrate (2 mL). The reaction mixture was then heated to 90 ° C. for 90 minutes and then cooled to room temperature. The resulting suspension was filtered and washed with methanol (5 mL), water (10 ml) and then ethyl ether (2 × 30 mL). A pale yellow solid was isolated and became a single peak in LC-MS analysis. (0.61 g, 26.7% over 3 steps); m / z (LC-MS, ESP), RT = 3.48 min, (M + H) = 263.0.

(e) 2- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -isonicotinic acid (iva)
To a suspension of 2- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -isonicotinonitrile (0.54 g, 2.06 mmol) in absolute ethanol (2.5 mL) was added concentrated hydrochloric acid (1.3 mL). added. The reaction mixture was then heated to 70 ° C. overnight. The reaction was then cooled to 5 ° C. and then the white suspension was filtered and washed with water (2 × 5 mL) followed by diethyl ether (2 × 20 mL). A bright yellow solid was isolated and became a single peak in LC-MS analysis (0.49 g, 88%); m / z (LC-MS, ESP), RT = 3.08 min, (M + H ) = 282; and (2M + H) = 563.

(f) 4- [4- (4-Cyclohexanecarbonyl-piperazin-1-carbonyl) -pyridin-2-ylmethyl-2H-phthalazin-1-one (5)
2- (4-Oxo-3,4-dihydro-phthalazin-1-ylmethyl) -isonicotinic acid (iva) (0.25 g, 0.89 mol) in dimethylacetamide (2 mL) to a stirred solution of cyclohexyl-piperazine 1-yl-methanone (0.20 g, 1.0 mmol) was added followed by HBTU (0.38 g, 1.0 mmol) and diisopropylethylamine (0.35 mL, 20.0 mmol) and stirred. The reaction mixture was then concentrated in vacuo and the resulting oil was subjected to flash chromatography with eluent EtOAc / MeOH 9: 1. (Rf of 0.3) The title compound was isolated as a white solid. Single peak in LC-MS analysis, (0.18 g, 56%); m / z (LC-MS, ESP), RT = 4.07 min, (M + H) = 460; ¹ H NMR (300 MHz) 12.57 (1H, S -NH), 8.56 (1H, d, J 5.1 Hz), 8.26 (1H, dd, J 1.5, 8.1 Hz), 7.95-7.80 (3H, m), 7.38 (1H, S), 7.25 ( 1H, d, J 5.7 Hz), 4.51 (2H, S), 3.56-3.17 (8H, m), 2.58 (1H, m), 1.71-1.61 (5H, m), 1.38-1.22 (5H, m).

In order to evaluate the inhibitory action of the compounds, IC ₅₀ values were measured using the following assay (Dillon, et al., JBS., 8 (3), 347-352 (2003)).

Mammalian PARP isolated from HeLa cell nucleus extract in Z-buffer (25 mM Hepes (Sigma); 12.5 mM MgCl ₂ (Sigma); 50 mM KCl in 96-well FlashPlatesTM (NEN, UK) (Sigma); 1 mM DTT (Sigma); 10% glycerol (Sigma) 0.001% NP-40 (Sigma); pH 7.4) and various concentrations of the inhibitor were added. All compounds were diluted in DMSO to a final assay concentration of 10-0.01 μM, resulting in a DMSO final concentration of 1% per well. The total assay volume per well was 40 μl.

After 10 minutes incubation at 30 ° C., the reaction was initiated by the addition of 10 μl of reaction mixture containing NAD (5 μM), ³ H-NAD and 30mer double stranded DNA-oligo. In order to calculate% enzyme activity, designated positive and negative reaction wells were performed in combination with compound wells (unknown). The plate was then shaken for 2 minutes and incubated at 30 ° C. for 45 minutes.

  Following incubation, the reaction was quenched by the addition of 50 μl of 30% acetic acid to each well. The plate was then shaken for 1 hour at room temperature.

  Plates were transferred to TopCount NXT ™ (Packard, UK) for scintillation counting. The value recorded is the count per minute (cpm) after counting each well for 30 seconds.

The% enzyme activity for each compound is then calculated by the following formula:
% Inhibition = 100− [100 × (unknown cpm−negative average cpm) / (positive average cpm−negative average cpm)].

IC ₅₀ values (concentration at which 50% of enzyme activity is inhibited) were calculated. IC ₅₀ values are measured in different concentration ranges, usually ranging from 10 μM to 0.001 μM. Such IC ₅₀ values are used as comparative values to confirm increased compound potency.

All compounds tested had an IC ₅₀ of less than 1 μM.

The following compounds have an IC ₅₀ of less than 0.1 μM: 1-7, 9-14, 17-18, 21, 24, 26-29, 35, 50-52, 56-75.

The compound enhancement factor (PF ₅₀ ) is calculated as the ratio of the IC ₅₀ of the control cell growth divided by the IC ₅₀ of the cell growth plus the PARP inhibitor. Growth inhibition curves for both control and compound treated cells are in the presence of the alkylating agent methyl methanesulfonate (MMS). Test compounds were used at a fixed concentration of 0.2 micromolar. The concentration of MMS ranged from 0 to 10 μg / ml. Cell proliferation was assessed by the sulforhodamine B (SRB) assay (Skehan, P., et al., (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J. Natl. Cancer Inst. 82, 1107-1112). 2,000 HeLa cells were seeded in each well of a flat bottom 96-well microtiter plate in a volume of 100 μl and incubated at 37 ° C. for 6 hours. Cells were replaced with medium alone or medium containing PARP inhibitor at a final concentration of 0.5, 1 or 5 μM. Before adding MMS at a series of concentrations (typically 0, 1, 2, 3, 5, 7, and 10 μg / ml) to either untreated cells or PARP inhibitor-treated cells, Grow for 1 hour. Growth inhibition by PARP inhibitors was evaluated using cells treated with PARP inhibitors alone.

  Cells were left for a further 16 hours before the medium was changed and the cells were allowed to grow for an additional 72 hours at 37 ° C. The medium was then removed and the cells were fixed with 100 μl of ice cold 10% (w / v) trichloroacetic acid. Plates were incubated at 4 ° C. for 20 minutes and then washed 4 times with water. Subsequently, the cells in each well were stained with 100 μl of a 1% acetic acid solution of 0.4% (w / v) SRB for 20 minutes and then washed 4 times with 1% acetic acid. The plate was then dried at room temperature for 2 hours. The dye from the stained cells was lysed by adding 100 μl of 10 mM Tris Base to each well. The plate was gently shaken and allowed to stand at room temperature for 30 minutes before measuring the optical density at 564 nM with a Microquant microtiter plate reader.

All compounds tested had at least 1 PF ₅₀ at 200 nM. The following compounds had at least 2 PF ₅₀ at 200 nM: 2, 3, 5, 6, 9, 10, 12, 13, 56, 57, 58, 62, 65, 66, 67, 74, 75.

Claims (23)

Formula (I):

[Where:
A and B together represent an optionally substituted fused aromatic ring;
X can be NR ^X or CR ^X R ^Y ;
If X = NR ^X , n is 1 or 2; if X = CR ^X R ^Y , n is 1;
R ^X is selected from the group consisting of H, optionally substituted C _1-20 alkyl, C _5-20 aryl, C _3-20 heterocyclyl, amide, thioamide, ester, acyl, and sulfonyl groups;
R ^Y is selected from H, hydroxy, amino;
Or R ^X and R ^Y may be taken together to form a spiro-C _3-7 cycloalkyl or heterocyclyl group;
R ^C1 and R ^C2 are independently selected from the group consisting of hydrogen and C _1-4 alkyl, or when X is CR ^X R ^Y , R ^C1 , R ^C2 , R ^X and R ^Y are the same Together with the carbon atom which may be formed may form an optionally substituted fused aromatic ring;
R ¹ is selected from H and halo; and
Het is selected from:

(Where Y ¹ is selected from CH and N, Y ² is selected from CH and N, Y ³ is selected from CH, CF and N, where one or two of Y ¹ , Y ² and Y ³ Only one can be N)
and

(Where Q is O or S)]
Or an isomer, salt, solvate, chemically protected form, or prodrug thereof.   2. The compound according to claim 1, wherein the fused aromatic ring represented by -A-B- consists only of carbon ring atoms.   The compound according to claim 2, wherein the condensed aromatic ring represented by -A-B- is benzene. The compound according to any one of claims 1 to 3, wherein R ^C1 and R ^C2 are hydrogen. n is 2, X is NR ^X , and R ^X is H; _optionally substituted C _1-20 alkyl; _optionally substituted C _5-20 aryl; _optionally substituted ester A group; an optionally substituted acyl group; an optionally substituted amide group; an optionally substituted thioamide group; and an optionally substituted sulfonyl group; 5. The compound according to any one of 4. n is 1, X is NR ^X , and R ^X is H; _optionally substituted C _1-20 alkyl; _optionally substituted C _5-20 aryl; _optionally substituted acyl 5. A compound according to any one of claims 1 to 4, selected from the group consisting of: optionally substituted sulfonyl; optionally substituted amide; and optionally substituted thioamide group. 7. The compound of claim 6, selected from the group consisting of Het is pyridylene and R ^X is an optionally substituted acyl; an optionally substituted sulfonyl; and an optionally substituted amide. Het is furanylene or thiophenylene and R ^X is optionally substituted C _1-20 alkyl; optionally substituted C _5-20 aryl; _optionally substituted acyl; optionally substituted 7. A compound according to claim 6 selected from the group consisting of sulfonyl; and optionally substituted amides. n is 1, X is CR ^X R ^Y , R ^Y is H, R ^X is H; _optionally substituted C _1-20 alkyl; _optionally substituted C _5-20 aryl An _optionally substituted C _3-20 heterocyclyl; an _optionally substituted acyl; an optionally substituted amino; an optionally substituted amide; and an optionally substituted ester group The compound according to any one of claims 1 to 4, which is selected. Het is furanylene or thiophenylene, R ^X is selected from optionally substituted amino, the amino group is selected from H and C _1-20 alkyl, or together with a nitrogen atom, C _{5 10.} A compound according to claim 9, forming a _-20 heterocyclic group.   11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 and a pharmaceutically acceptable carrier or diluent.   11. A compound according to any one of claims 1 to 10 for use in a method of treatment of the human or animal body.   11. The production of a medicament for preventing poly (ADP-ribose) chain formation by inhibiting the activity of cellular PARP (PARP-1 and / or PARP-2), according to any one of claims 1-10. Use of the compound.   Vascular disease; Septic shock; Ischemic injury; Reperfusion injury; Neurotoxicity; Hemorrhagic shock; Inflammatory disease; Multiple sclerosis; Treatment of secondary effects of diabetes; Cytotoxicity after cardiovascular surgery ) Or the use of a compound according to any one of claims 1 to 10 in the manufacture of a medicament for the acute treatment of diseases ameliorated by inhibition of the activity of PARP.   11. Use of a compound according to any one of claims 1 to 10 in the manufacture of a medicament for use as an adjunct in cancer therapy or for enhancing tumor cells for treatment with ionizing radiation or a chemotherapeutic agent.   Use of a compound according to claims 1 to 10 in the manufacture of a medicament for use in the treatment of a cancer deficient in the DNA DSB repair pathway by HR in an individual.   17. Use according to claim 16, wherein the cancer comprises one or more cancer cells whose ability to repair DNA DSB by HR is reduced or suppressed compared to normal cells.   18. Use according to claim 17, wherein the cancer cells have a BRCA1 or BRCA2 deficient phenotype.   Use according to claim 18, wherein the cancer cells are deficient in BRCA1 or BRCA2.   20. Use according to any one of claims 16 to 19, wherein the individual is heterozygous for a mutation in a gene encoding a component of the DNA DSB repair pathway by HR.   21. Use according to claim 20, wherein the individual is heterozygous for a mutation in BRCA1 and / or BRCA2.   The use according to any one of claims 16 to 21, wherein the cancer is breast cancer, ovarian cancer, pancreatic cancer or prostate cancer.   23. Use according to any one of claims 16 to 22, wherein the treatment further comprises administration of ionizing radiation or a chemotherapeutic agent.