KR20080063828A - 4-heteroarylmethyl substituted phthalazinone derivatives - Google Patents

Abstract

wherein: A and B together represent an optionally substituted, fused aromatic ring; D is selected from formula (i), where Y1 is selected from CH and N , Y2 is selected from CH and N, Y3 is selected from CH, CF and N; and formulae (ii), (A) and (B) where Q is O or S; RD is formula (C) wherein RN1 is selected from H and optionally substituted C1-10 alkyl; X is selected from a single bond, NRN2, CRC3RC4 and C=O; RN2 is selected from H and optionally substituted C1-10 alkyl; Rc3 and Rc4 are independently selected from H, R, C(=O)OR, where R is optionally substituted C1-10 alkyl, optionally substituted C5-20 aryl or optionally substituted C3-20 heterocyclyl; Y is selected from NRN3 and CRc1Rc2; RC1 and RC2 are independently selected from H, R, C(=O)OR, where R is optionally substituted C1-10 alkyl, optionally substituted C5-20 aryl or optionally substituted C3-20 heterocyclyl; RC1 and RC2 together with the carbon atom to which they are attached may form an optionally substituted spiro-fused C5-7carbocylic or heterocyclic ring; and when X is a single bond RN1 and RC2 may together with the N and C atoms to which they are bound, form an optionally substituted C5-7 heterocylic ring; and when X is CRC3RC4, RC2 and RC4 may together form an additional bond, s uch that there is a double bond between the atoms substituted by RC1 and RC3.

Description

4-heteroarylmethyl substituted phthalazinone derivative {4-HETEROARYLMETHYL SUBSTITUTED PHTHALAZINONE DERIVATIVES}

The present invention relates to phthalazinone derivatives and their use as medicaments. In particular, the present invention inhibits the activity of poly (ADP-ribose) polymerase-1, also known as poly (ADP-ribose) synthetase and poly ADP-ribosyltransferase and commonly referred to as PARP-1. It relates to the use of this compound.

Mammalian enzyme PARP-1 (113-kDa multidomain protein) has been involved in signaling DNA damage through its ability to recognize and quickly bind to DNA single or double helix damage {D'Amours , et al., Biochem . J. , 342, 249-268 (1999)].

The family of poly (ADP-ribose) polymerases now includes about 18 proteins, all of which show some level of homology in the catalytic domain but differ in cell function. Ame et al., Bioessays . , 26 (8), 882-893 (2004)]. Among these families, PARP-1 (founding member) and PARP-2 are the only enzymes whose catalytic activity is so far stimulated by the occurrence of DNA helix damage, which makes them unique in the family.

It is now known that PARP-1 is involved in various DNA-related functions, including gene amplification, cell division, differentiation, apoptosis, DNA nucleotide ablation repair, as well as effects on terminal length and chromosome stability. Adda di Fagagna, et al., Nature Gen. , 23 (1), 76-80 (1999)].

Research into the mechanism by which PARP-1 modulates DNA repair and other processes has shown that this is important in the formation of poly (ADP-ribose) chains 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 uses NAD ⁺ to synthesize poly (ADP-ribose) on a variety of nuclear target proteins, including isomerization enzymes, histones and PARP itself (Rhun, et al., Biochem . Biophys . Res . Commun . , 245, 1-10 (1998)]}

Poly (ADP-ribosyl) ation is also associated with malignant cell turnover. For example, PARP-1 activity is higher in the isolated nucleus of fibroblasts transformed with SV40, while both leukemia cells and colon cancer cells show higher enzymatic activity than the corresponding normal leukocytes and colon mucosa. Miwa, et al., Arch . Biochem . Biophys . 181, 313-321 (1977); Burzio, et al., Proc . Soc . Exp . Biol . Med . 149, 933-938 (1975); And Hirai, et al., Cancer Res . , 43, 3441-3446 (1983)]. More recently, when comparing malignant prostate tumors with benign prostate cells, it has been shown that significantly increased levels of active PARP (mainly PARP-1) are associated with high levels of gene instability (Mcnealy, et al., Anticancer Res . , 23, 1473-1478 (2003)]}

Many low molecular weight inhibitors of PARP-1 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 significant increases in DNA strand damage and cell killing (Durkacz, et al., Nature , 283, 593-596 (1980); Berger, NA, Radiation Research , 101, 4-14 (1985)].

Subsequently, these inhibitors have been found to enhance the effect of the radiation response by inhibiting the repair of potentially fatal damage. [Ben-Hur, et al., British Journal of Cancer , 49 (Suppl. VI), 34-42 (1984); Schlicker, et al., Int . J. Radiat Biol ., 75, 91-100 (1999)]. PARP inhibitors have been reported to be effective on radiosensitizing 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)].

In addition, PARP-1 knockout (PARP-/-) animals exhibit genomic instability in response to alkylating agents and γ 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 have both overlapping and non-overlapping functions in the maintenance of genomic stability, which makes them both important targets (Menissier de Murcia, et al., EMBO . J. , 22 (9), 2255-2263 (2003)].

The role of PARP-1 in certain vascular diseases, septic shock, ischemic injury and neurotoxicity has also been demonstrated. Cantoni, et al., Biochem . Biophys . Acta , 1014, 1-7 (1989); Szabo, et al., J. Clin . Invest , 100, 723-735 (1997)]. Oxygen radical DNA damage, which then results in DNA helix damage recognized by PARP-1, is a major contributor to this disease state, as evidenced by the PARP-1 inhibitor studies [Cosi, et al., J. Neurosci . Res . , 39, 38-46 (1994); Said, et al., Proc . Natl . Acad . Sci. USA , 93, 4688-4692 (1996)]. More recently, PARP has been shown to play a role in the development of hemorrhagic shock {Liaudet, et al., Proc . Natl . Acad . Sci . USA , 97 (3), 10203-10208 (2000)].

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

In addition, PARP-1 inhibition is thought to delay the expression of senescent properties in human fibroblasts (Rattan and Clark, Biochem . Biophys . Res . Comm . , 201 (2), 665-672 (1994)]. This may be related to the role of PARP in the control of terminal function [d'Adda di Fagagna, et al., Nature Gen. , 23 (1), 76-80 (1999)].

PARP inhibitors are also described in Inflammatory Bowel Disease (Szabo C., Role of Poly (ADP-Ribose) Polymerase Activation in Pathogenesis of Shock and Inflammation, In PARP as 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 previously disclosed a class of 1 (2H) -phthalazinone compounds that act as PARP inhibitors (in WO 02/36576). The compound has the formula:

In the above formulae, A and B together represent an optionally substituted fused aromatic ring, and R _c is represented by -LR _L. Many examples are of the formula:

In the above formula, R is one or more optional substituents.

Some of the present inventors have disclosed a particular class of such compounds in WO 03/093261, which has the above formula, wherein R is in the meta position, and the disclosed examples have the R group selected from:

The inventors have now found that compounds with such substituent groups as described above inhibit the synergistic action of tumor cells and / or PARP activity to radiation therapy and various chemotherapy to surprising levels. In addition, the stability of the compounds of the invention is generally improved compared to the compounds exemplified in WO 03/093261. Some of the compounds of the present invention also exhibit good solubility in both aqueous media and phosphate buffer solutions, and enhanced solubility can be used for the preparation of compounds for administration by the IV route or for pediatric oral preparations (e.g. liquid and predetermined form). Can be. Oral bioavailability of the compounds of the present invention can be enhanced.

Accordingly, a first aspect of the invention provides compounds of formula I (including isomers, salts, solvates, chemically protected forms and prodrugs of compounds of formula I):

In the above formula,

A and B together represent a fused aromatic ring optionally substituted;

D is

(i) Wherein Y ¹ is selected from CH and N, Y ² is selected from CH and N, and Y ³ is selected from CH, CF and N; And

(ii) Wherein Q is O or S

Is selected from;

R ^D is

[Wherein, R ^N1 is selected from H and optionally substituted C _{1 -10} alkyl;

X is selected from a single bond, NR ^N2 , CR ^C3 R ^C4 and C═O;

R ^N2 is selected from H and optionally substituted C _{1 -10} alkyl;

R ^C3 and R ^C4 are independently H, R, C (= O) OR (wherein, R is an optionally substituted C _{1 -10} alkyl, optionally substituted C _{5 -20} aryl or optionally substituted hetero C _{3 -20} Is a cyclic);

Y is selected from NR ^N3 and CR ^C1 R ^C2 ;

R ^C1 and R ^C2 are independently selected from H, R, C (= O) OR (wherein, R is an optionally substituted C _{1 -10} alkyl, optionally substituted C _{5 -20} aryl or optionally substituted hetero C _{3 -20} Is a cyclic);

R ^C1 and R ^C2 together with the carbon atom attached thereto may form a spiro-fused C _{5 -7} carbocyclic or heterocyclic ring optionally substituted and;

If X is a single bond, R ^N1 and R ^C2 together with the N and C atoms attached thereto an optionally substituted C _{5 -7} may form a heterocyclic ring;

When X is CR ^C3 R ^C4 , R ^C2 and R ^C4 may together form an additional bond such that a double bond is present between the atoms substituted with R ^C1 and R ^C3 .

For D the following is possible:

A second aspect of the present invention provides a pharmaceutical composition comprising the compound of the first aspect and a pharmaceutically acceptable carrier or diluent.

A third aspect of the invention provides the use of a compound of the first aspect in a method of treating a human or animal body.

The fourth aspect of the present invention

(a) prevention of poly (ADP-ribose) chain formation by inhibiting the activity of cellular PARP (PARP-1 and / or PARP-2);

(b) vascular disease; Septic shock; Ischemic damage to both the brain and cardiovascular; Reperfusion injury in both the brain and cardiovascular; Neurotoxicity, including acute and chronic treatment for stroke and Parkinson's disease; Hemorrhagic shock; Inflammatory diseases such as arthritis, inflammatory bowel disease, ulcerative colitis and Crohn's disease; Multiple sclerosis; As well as treatment of secondary effects of diabetes, as well as acute treatment of diseases that are ameliorated by cardiovascular surgery following cytotoxicity or inhibition of PARP activity;

(c) use as adjuvant in cancer therapy, or as adjuvant to increase drug efficacy on tumor cells for treatment with ionizing radiation or chemotherapy

Provided is the use of a compound as defined in the first aspect of the invention in the manufacture of a medicament.

In particular, the compounds as defined in the first aspect of the invention, together with alkylating agents such as methyl methanesulfonate (MMS), temozolomide and dacarbazine (DTIC), also contain topotecan, irinotecan, rubicatecan, exatecan , Rutothecan, zimetecan, diplomotecan (homocamptothekin) as well as 7-substituted bisilatecan; Topical isomerase 1 inhibitors such as 7-silyl campotechin, BNP 1350; And in anticancer combination therapies with noncamphorthekin topical isomerase-I inhibitors such as indolocarbazole, dual topical isomerase-I and II inhibitors such as benzophenazine, XR 11576 / MLN 576 and benzopyridoindole (or As an adjuvant). Such combinations may be provided, for example, as an intravenous preparation or by oral administration, depending on the preferred method of administration for the particular agent.

Another further aspect of the present invention comprises administering to a subject in need thereof a therapeutically effective amount of a compound as defined in the first aspect of the form of a pharmaceutical composition to an individual in need thereof. Treatment of diseases improved by inhibition of; And administering to the subject in need thereof a therapeutically effective amount of a compound as defined in the first aspect of the form of a pharmaceutical composition concurrently with radiation therapy (ionizing radiation) or chemotherapy. Or a combination thereof is provided sequentially.

In a further aspect of the invention, the compounds may be used in the manufacture of a medicament for the treatment of cancers lacking homologous recombination (HR) dependent DNA double strand break (DSB) repair activity or for repair of HR dependent DNA DSB. It can be used for the treatment of patients suffering from a cancer lacking HR dependent DNA DSB repair activity, comprising administering a therapeutically effective amount of a compound to a patient suffering from a cancer lacking activity.

The HR-dependent DNA DSB repair pathway repairs double helix damage (DSB) in DNA through homologous mechanisms to correct sequential DNA helices (KK Khanna and SP. Jackson, Nat. Genet. 27 (3): 247-254 (2001)]. The components of the HR dependent DNA DSB repair pathway are ATM (NM_000051), RAD51 (NM_002875), RAD51L1 (NM_002877), RAD51C (NM_002876), RAD51L3 (NM_002878), DMC1 (NM_007068), XRCC2 (NM_005431), RXC432 (NM_002879), RAD54L (NM_003579), RAD54B (NM_012415), BRCA1 (NM_007295), BRCA2 (NM_000059), RAD50 (NM_005732), MRE11A (NM_005590) and NBS1 (NM_002485). Other proteins involved in the HR dependent DNA DSB repair pathway include regulatory factors such as EMSY (Hughes-Davies, et al., Cell , 115, pp523-535). HR components are also described in Wood, et al., Science , 291, 1284-1289 (2001).

Cancers lacking HR dependent DNA DSB repair may comprise or consist of one or more cancer cells with reduced or stopped ability to repair DNA DSB through this pathway compared to normal cells. That is, the activity of the HR dependent DNA DSB repair pathway can be reduced or stopped in one or more cancer cells.

The activity of one or more components of the HR dependent DNA DSB repair pathway may be disrupted in one or more cancer cells of an individual suffering from a cancer that lacks HR dependent DNA DSB repair. Components of the HR dependent DNA DSB repair pathway are well characterized in the art (see, eg, Wood, et al., Science , 291, 1284-1289 (2001)) and include the components described above.

In some preferred embodiments, the cancer cells may have a phenotype lacking BRCA1 and / or BRCA2. That is, BRCA1 and / or BRCA2 activity is reduced or destroyed in cancer cells. Cancer cells with this phenotype may lack BRCA1 and / or BRCA2. That is, the expression and / or activity of BRCA1 and / or BRCA2 is, for example, by a mutation or polymorph in a coding nucleic acid or by amplification, mutation or in a gene encoding a regulatory factor, such as an EMSY gene encoding a BRCA2 regulatory factor. Polymorphs (Hughes-Davies, et al., Cell , 115, 523-535), or by epigenetic mechanisms such as gene promoter methylation, can be reduced or destroyed in cancer cells.

BRCA1 and BRCA2 are known tumor inhibitors that often lose wild-type alleles 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)]. Amplification of the EMSY gene encoding the BRCA2 binding factor is also known to be associated with breast and ovarian cancer.

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

In some preferred embodiments, the individual is a heterozygous for one or more variations, such as mutations and polymorphs in BRCA1 and / or BRCA2 or a modulator thereof. Detection of mutations in BRCA1 and BRCA2 is well known in the art, see, eg, EP 699 754, EP 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 amplification of the BRCA2 binding factor EMSY is disclosed in Hughes-Davies, et al., Cell , 115, 523-535.

Mutations and polymorphs associated with cancer can be detected at the nucleic acid level by detecting the presence of variant nucleic acid sequences or at the protein level by detecting the presence of a variant (ie, mutation or allelic variant) polypeptide.

Justice

The term "aromatic ring" is used herein in the conventional sense to refer to a cyclic aromatic structure, ie a cyclic structure having an unlocalized π-electron orbital.

The aromatic ring fused to the main core, ie the aromatic ring formed by -A-B-, may comprise further fused aromatic rings (eg, naphthyl or anthracenyl groups are formed). The aromatic ring (s) may comprise only carbon atoms or may comprise one or more hetero atoms including, but not limited to, carbon atoms and nitrogen, oxygen and sulfur atoms. The aromatic ring (s) preferably have 5 to 6 ring principles.

Aromatic ring (s) may be optionally substituted. When the substituents themselves comprise an aryl group, this aryl group is not considered part of the aryl group to which it is attached. Biphenyl groups, for example, are considered herein to be phenyl groups substituted with phenyl groups (aryl groups comprising a single aromatic ring). Similarly, benzylphenyl groups are considered to be phenyl groups (aryl groups containing a single aromatic ring) substituted with benzyl groups.

In a group of preferred embodiments, the aromatic group comprises a single aromatic ring optionally containing 5 or 6 ring atoms selected from carbon, nitrogen, oxygen and sulfur. Examples of such groups include, but are not limited to, benzene, pyrazine, pyrrole, thiazole, isoxazole and oxazole. 2-pyrones may also be considered aromatic rings, but are less preferred.

If the aromatic ring has six atoms, preferably at least four, or even five or all ring atoms are carbon. Other ring atoms are selected from nitrogen, oxygen and sulfur, with nitrogen and oxygen being preferred. Suitable groups include 0 hetero atoms (benzene); One nitrogen ring atom (pyridine); Two nitrogen ring atoms (pyrazine, pyrimidine and pyridazine); One oxygen ring atom (pyron); And a ring having one oxygen and one nitrogen ring atom (oxazine).

If the aromatic ring has five ring atoms, preferably at least three of the ring atoms are carbon. The remaining ring atoms are selected from nitrogen, oxygen and sulfur. Suitable rings include one nitrogen ring atom (pyrrole); Two nitrogen ring atoms (imidazole, pyrazole); One oxygen ring atom (furan); One sulfur ring atom (thiophene); One nitrogen and one sulfur ring atom (isothiazole, thiazole); And a ring having one nitrogen and one oxygen ring atom (isoxazole or oxazole).

Aromatic rings can include one or more substituent groups at any available ring position. The substituents are selected from halo, nitro, hydroxy, ether, thiol, thioether, amino, C _{1 -7} alkyl, C _{3 -20} days heterocyclyl and C _{5 -20} aryl. Aromatic rings can also include one or more substituent groups that together form a ring. In particular it may be of the formula-(CH ₂ ) _m -or -O- (CH ₂ ) _p -O-, wherein m is 2, 3, 4 or 5 and p is 1, 2 or 3 .

Spiro Fused Rings: As used herein, the term “spiro fused ring” relates to a carbocyclic or heterocyclic ring fused to the remainder of the molecule at a single carbon atom. The ring may itself be a carbocyclic ring containing only carbon ring atoms, or it may be a heterocyclic ring containing one or more hetero atoms. C _{5 -7} show examples of carbocyclic ring and heterocyclic ring herein.

A nitrogen-containing C _{5 -7} heterocyclic ring: The term As used herein "C _5-7 nitrogen-containing heterocyclic ring" of the bar to define to work in conjunction, heterocyclyl having a nitrogen ring atom or more C 1 _{5 -7} heterocyclic ring.

Alkyl: As used herein, the term "alkyl" (unless otherwise specified) has 1 to 20 carbon atoms, which may be aliphatic or cycloaliphatic and may be saturated or unsaturated (such as partially unsaturated, fully unsaturated). It relates to a monovalent moiety obtained by removing a hydrogen atom from a carbon atom of a hydrocarbon compound. Thus, the term "alkyl" includes the subclasses discussed below, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, and the like.

In the context of the group, a prefix (e.g., C _{1 -4,} C _{1 -7, -20} C _1, C _{2 -7, -7} C _3, etc.) is the number of carbon atoms, or a number range of carbon atoms, Refer. For example, the terms As used herein, "C _{1 -4} alkyl" relates to alkyl groups having 1 to 4 carbon atoms. Examples of the group of the alkyl group may have C _{1 -4} alkyl ( "lower alkyl"), C _{1 -7} alkyl, and C _{1 -20} alkyl. The first prefix may vary according to other limitations, such as for the unsaturated alkyl group the first prefix should be at least 2 and for the cyclic alkyl groups the first prefix should be at least 3.

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 thereto.

Examples of the (unsubstituted) saturated straight chain alkyl group 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 ), including but not limited to.

Examples of the (unsubstituted) saturated branched alkyl group are 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 ).

Alkenyl: The term "alkenyl" as used herein relates to an alkyl group having at least one carbon-carbon double bond. Examples of the alkenyl group is a 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 ) Do not.

Alkynyl: The term “alkynyl,” as used herein, relates to an alkyl group having at least one carbon-carbon triple bond. Examples of the alkynyl group is a 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 is not limited.

Cycloalkyl: As used herein, the term “cycloalkyl” refers to an alkyl group that is also a cyclyl group, that is, an alicyclic ring of a carbocyclic ring of a carbocyclic compound, which may be saturated or unsaturated (eg, partially unsaturated, fully unsaturated). A monovalent moiety obtained by removing a hydrogen atom from a group ring atom, and relates to a moiety having 3 to 20 carbon atoms (unless otherwise specified), including 3 to 20 ring atoms. Thus, the term "cycloalkyl" includes subclasses cycloalkenyl and cycloalkynyl. Preferably each ring has 3 to 7 ring atoms. Examples of groups of cycloalkyl groups are the C _{3 -20} cycloalkyl, C _{3 -15} cycloalkyl, C _{3 -10} cycloalkyl, C _{3 -7-cycloalkyl.}

 Examples of cycloalkyl groups are

Saturated monocyclic hydrocarbon compounds: cyclopropane (C ₃ ), cyclobutane (C ₄ ), cyclopentane (C ₅ ), cyclohexane (C ₆ ), cycloheptane (C ₇ ), methylcyclopropane (C ₄ ), dimethyl Cyclopropane (C ₅ ), methylcyclobutane (C ₅ ), dimethylcyclobutane (C ₆ ), methylcyclopentane (C ₆ ), dimethylcyclopentane (C ₇ ), methylcyclohexane (C ₇ ), dimethylcyclohexane (C ₈ ), methane (C ₁₀ );

Unsaturated monocyclic hydrocarbon compounds: cyclopropene (C ₃ ), cyclobutene (C ₄ ), cyclopentene (C ₅ ), cyclohexene (C ₆ ), methylcyclopropene (C ₄ ), dimethylcyclopropene (C ₅ ), methylcyclobutene (C ₅ ), dimethylcyclobutene (C ₆ ), methylcyclopentene (C ₆ ), dimethylcyclopentene (C ₇ ), methylcyclohexene (C ₇ ), dimethylcyclohexene (C ₈ ) ;

The saturated cyclic hydrocarbon compounds: tujan (C _10), Karan (C _10), evacuation (C _10), Vor I (C _10), Nord Curran (C _7), Nord evacuation (C _7), norbornane (C ₇ ), adamantane (C ₁₀ ), decalin (decahydronaphthalene) (C ₁₀ );

Unsaturated polycyclic hydrocarbon compounds: campene (C ₁₀ ), limonene (C ₁₀ ), pinene (C ₁₀ );

Polycyclic hydrocarbon compounds having aromatic rings: indene (C ₉ ), indane (eg 2,3-dihydro-1H-indene) (C ₉ ), tetralin (1,2,3,4-tetrahydronaphthalene ) (C _10), acenaphthene (C _12), fluorene (C _13), Fe nalren (C _13), acetoxy phenanthrene (C _15), ASEAN Transistor (C _16), kolran Transistor (C ₂₀₎

Including but not limited to.

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

In this context, the prefixes _{(e.g., C 3 -20, C 3 -7} , such as C _{5 -6)} refers to the number range of number of ring atoms, whether carbon atoms or heteroatoms, or the ring atoms. For example, the terms As used herein "C _{-6 5} day heterocyclyl" relates to a heterocyclyl group having 5 or 6 ring atoms. Examples of a heterocyclyl group the group is a C _{3 -20} days heterocyclyl, C _5-20 heterocyclyl yl, C _{3 -15} days heterocyclyl, C _{5 -15} days heterocyclyl, C _{3 -12} days heterocyclyl, C _{5 -12} heterocyclyl days, a C _{3 -10} days heterocyclyl, C _{5 -10} days heterocyclyl, C _{3 -7} days heterocyclyl, C _{5 -7} days heterocyclyl and C _{5 -6} heterocyclyl days.

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 (eg, 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 ), oxol (dihydrofuran) (C ₅ ), oxane (tetrahydropyran) (C ₆ ) , Dihydropyran (C ₆ ), pyran (C ₆ ), oxepin (C ₇ );

S _1: T is (C _3), thietane (C _4), tiolran (tetrahydro-thiophene) (C _5), Tian (tetrahydro-thiopyran) (C _6), thienyl plate (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 ), tetrahydro Oxazine (C ₆ ), dihydrooxazine (C ₆ ), oxazine (C ₆ );

N ₁ S ₁ : thiazolin (C ₅ ), thiazolidine (C ₅ ), thiomorpholine (C ₆ );

N ₂ O ₁ : oxadiazine (C ₆ );

O ₁ S ₁ : oxathiol (C ₅ ) and oxatian (thioxane) (C ₆ ); And

N ₁ O ₁ S ₁ : Oxathiazine (C ₆ ).

Examples of substituted (non-aromatic) monocyclic heterocyclyl groups include saccharides in the form of rings, such as furanose (C ₅ ) such as arabinofuranose, lysofuranose, ribofuranose and xylfuranose, And pyranose (C ₆ ) such as allopyranose, altropyranose, glucopyranose, mannofyranose, glucopyranose, aydopyranose, galactopyranose and talopyranose There are those derived from.

Spiro -C _{3 -7} cycloalkyl or heterocyclyl: The term As used herein, "spiro C _{3 -7} cycloalkyl or an alkyl heterocycle" is bonded to another ring by a single atom common to both rings C _3-7 refers to a cycloalkyl, or C _3-7 heterocyclyl ring days.

C _{5 -20} aryl: Terms As used herein, "C _{5 -20} aryl" one ring, or (e. G. Fused) having two or more rings of 5 to 20 rings (at least one of the rings is ring Im) to a 1 obtained by removing a hydrogen atom from an aromatic ring atom of a C _{5 -20} aromatic compounds of the portion with the atom. Preferably each ring has 5 to 7 ring atoms.

Ring atoms may be all carbon atoms, as in "carboxylic group-bore", in which case it can be addressed by the group group conveniently "C _{5 -20} carboxylic bore reel".

Examples of having no ring hetero atom C _{5 -20} aryl group (i. E. C _5-20 carboxylic group-bore) benzene (i.e., phenyl) (C _6), naphthalene (C _10), anthracene (C _14), phenanthrene But are not limited to those derived from trasene (C ₁₄ ) and pyrene (C ₁₆ ).

Alternatively, the ring atom may comprise one or more hetero atoms, including but not limited to oxygen, nitrogen and sulfur, such as in a "heteroaryl group". In this case, there is a group may be referred to conveniently group "C _{5 -20} heteroaryl", where "C _{5 -20"} refers to a ring atoms, whether carbon atoms or heteroatoms. Preferably, each ring has 5 to 7 ring atoms, wherein 0 to 4 are ring heteroatoms.

C _{5 -20} Examples of heteroaryl groups are furan (dioxol), thiophene (thiol group), pyrrole (azole), imidazole (1,3-diazole), pyrazole (1,2-diazole), triazole C ₅ heteroaryl groups derived from oxazoles, isoxazoles, thiazoles, isothiazoles, oxadiazoles, tetrazole and oxtriazoles; And isoxazine, pyridine (azine), pyridazine (1,2-diazine), pyrimidine (1,3-diazine; eg cytosine, thymine, uracil), pyrazine (1,4-diazine) and tree It comprises a heteroaryl, a C ₆ derived from triazine, however, and the like.

Heteroaryl groups can be bonded via carbon or hetero ring atoms.

An example of C _{5 -20} heteroaryl groups containing fused rings are benzofuran, isopropyl benzofuran, benzothiophene, derived from indole, isoindole C ₉ heteroaryl group; C ₁₀ heteroaryl groups derived from quinoline, isoquinoline, benzodiazine, pyridopyridine; Acridine and xanthene the C ₁₄ include a heteroaryl group derived from, but is not limited to this.

The alkyl, heterocyclyl and aryl groups, whether alone or as part of other substituents, may optionally be substituted by themselves with one or more groups selected from themselves and further substituents described below.

Halo: -F, -Cl, -Br and -I.

Hydroxy: -OH.

Ether: -OR [wherein, R is an ether substituent, for example, C _{1 -7} alkyl group (C _{1 -7} alkoxy group is also referred to as value), C _{3 -20} heterocyclyl group (C _{3 -20} heterocyclyl as a group acryloyloxy hereinafter), or C _{5 -20} aryl group (C _{5 -20} aryl-oxy group is also referred to), preferably C _{1 -7} alkyl group.

Nitro: -NO ₂ .

Cyano (nitrile, carbonitrile): -CN.

Acyl (keto): -C (= O) R [ wherein, R is (referred to Fig. ₁ C _-7 alkyl acyl or C _{1 -7} alkanoyl days) acyl substituent, for example H, C _{1 -7} alkyl, C _{3 -20} heterocyclyl group (also referred to as C _{3 -20} days heterocyclyl acyl), or C _{5 -20} aryl group (also referred to as a C _{5 -20} aryl acyl group), preferably C _{1 -7} alkyl group. Examples of acyl groups are —C (═O) CH ₃ (acetyl), —C (═O) CH ₂ CH ₃ (propionyl), —C (═O) C (CH ₃ ) ₃ (butyryl) and- C (= 0) Ph (benzoyl, phenone), including but not limited to.

Carboxylic Acid (carboxylic acid): -COOH.

Ester (carboxylate, carboxylic acid ester, oxycarbonyl): -C (= O) OR [ wherein, R is an ester substituent, for example, C _{1 -7} alkyl, C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably a C _{1 -7} alkyl group. Examples of ester groups include, but are not limited to, -C (= 0) OCH ₃ , -C (= 0) OCH ₂ CH ₃ , -C (= 0) OC (CH ₃ ) ₃ and -C (= 0) OPh. It is not limited.

Amido (carbamoyl, carbamyl, aminocarbonyl, carboxamide): -C (= 0) NR ¹ R ^{2 wherein} R ¹ and R ² are independently amino substituents as defined for amino groups ]. Examples of amido groups include -C (= 0) NH ₂ , -C (= 0) NHCH ₃ , -C (= 0) N (CH ₃ ) ₂ , -C (= 0) NHCH ₂ CH ₃ and -C ( In addition to ═O) N (CH ₂ CH ₃ ) ₂ , R ¹ and R ² are added to this, such as, for example, in piperinocarbonyl, morpholinocarbonyl, thiomorpholinocarbonyl and piperazinylcarbonyl. Amido groups that form a heterocyclic structure with an attached nitrogen atom include, but are not limited to.

Amino: -NR ¹ R ² [wherein, R ¹ and R ² are independently (referred to as a C ₁ -C _{1 -7 -7} alkylamino or di-alkylamino) amino substituents, for example, hydrogen, C _1-7 alkyl group , C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably H or C _{1 -7} for an alkyl group or, or "cyclic" amino group, R ¹ and R ² together with the nitrogen atom attached thereto 4 To form a heterocyclic ring having from 8 ring atoms. Examples of amino groups include, but are not limited to, -NH ₂ , -NHCH ₃ , -NHCH (CH ₃ ) ₂ , -N (CH ₃ ) ₂ , -N (CH ₂ CH ₃ ) _2, and -NHPh. Examples of cyclic amino groups include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidino, piperazinyl, perhydrodiazepynyl, morpholino and thiomorpholino. In particular, the cyclic amino group may be substituted on any of its substituents as defined herein, such as carboxy, carboxylate and amido, on its ring.

Acyl amido ^{(acylamino): -NR 1 C (= O} ) R 2 ( wherein, R ¹ is an amide substituent, for example, hydrogen, C _{1 -7} alkyl, C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably H or a C _{1 -7} alkyl, most preferably H, R ² is the acyl substituent, for example, C _{1 -7} alkyl group, a C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably It is C _{1 -7} alkyl group). Examples of acyl amide groups include, but are not limited to, -NHC (= 0) CH ₃ , -NHC (= 0) CH ₂ CH _3, and -NHC (= 0) Ph. R ¹ and R ² together form a cyclic structure, as for example in succinimidyl, maleimidyl and phthalimidyl:

^{Ureido: -N (R 1) CONR 2} R 3 ( wherein, R ² and R ³ is an amino substituent as defined for amino groups independently, R ¹ is a ureido substituent, for example, hydrogen, C _{1 -7} alkyl group, C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably hydrogen or C _{1 -7} alkyl group). Examples of ureido groups include, but are not limited to, -NHCONH ₂ , -NHCONHMe, -NHCONHEt, -NHCONMe ₂ , -NHCONEt ₂ , -NMeCONH ₂ , -NMeCONHMe, -NMeCONHEt, -NMeCONMe ₂ , -NMeCONEt _2, and -NHCONHPh Do not.

Acyloxy (reverse ester): -OC (= O) R ( wherein, R is an acyloxy substituent, for example, C _{1 -7} alkyl, C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably a C _1-7 alkyl group). Examples of acyloxy groups include -OC (= 0) CH ₃ (acetoxy), -OC (= 0) CH ₂ CH ₃ , -OC (= 0) C (CH ₃ ) ₃ , -OC (= 0) Ph , —OC (═O) C ₆ H ₄ F, and —OC (═O) CH ₂ Ph.

Thiol: -SH.

Thioether (sulfide): -SR [wherein, R is a thioether substituent, for example, C _{1 -7} alkyl group (also referred to C _{1 -7} alkylthio groups), C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, is preferably a C _{1 -7} alkyl group. Examples of C _{1 -7} alkylthio include -SCH ₃ and -SCH ₂ CH _3, however, and the like.

Sulfoxide (sulfinyl): -S (= O) R ( wherein, R is a sulfoxide substituent, for example, C _{1 -7} alkyl, C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably a C _1-7 alkyl group). Examples of sulfoxide groups include, but are not limited to, -S (= 0) CH ₃ and -S (= 0) CH ₂ CH ₃ .

Sulfonyl _{(sulfone): -S (= O) 2} R ( wherein, R is a sulfone substituent, for example, C _{1 -7} alkyl, C _{3 -20} heterocyclyl group or C _{5 -20} aryl group, preferably a C _{1 -7} alkyl group. Examples of sulfone groups are -S (= O) ₂ CH ₃ (methanesulfonyl, mesyl), -S (= O) ₂ CF ₃ , -S (= O) ₂ CH ₂ CH ₃ and 4-methylphenylsulfonyl ( 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. Examples of amido groups include -C (= S) NH ₂ , -C (= S) NHCH ₃ , -C (= S) N (CH ₃ ) ₂ and -C (= S) NHCH ₂ CH ₃ , It is not limited to this.

^{Sulfone-amino-: -NR 1 S (= O)} 2 R ( wherein, R ¹ is an amino substituent as defined for amino groups, R is a sulfonic amino substituents, for example C _{1 -7} alkyl, C _{3 -20} heterocyclyl group or a C _5-20 aryl group, is preferably a C _{1 -7} alkyl group). Examples of sulfonamino groups include, but are not limited to, -NHS (= 0) ₂ CH ₃ , -NHS (= 0) ₂ Ph, and -N (CH ₃ ) S (= 0) ₂ C ₆ H ₅ .

As mentioned above, the groups that form the group described by substituents such as C _{1 -7} alkyl, C _{3 -20} days heterocyclyl and C _{5 -20} aryl may be substituted with itself. Thus, the above definition encompasses substituted substituent groups.

More preferred

The following preferences may apply to each aspect of the invention where applicable.

In the present invention, the fused aromatic ring (s) represented by -A-B- is preferably composed of only carbon ring atoms, and thus may be benzene, naphthalene, more preferably benzene. As described above, this ring may be substituted, but in some embodiments it is preferably unsubstituted.

When the fused aromatic ring represented by -AB- comprises a substituent group, it is preferably attached to an atom itself attached to the central ring β- relative to the carbon atom of the central ring. Thus, when the fused aromatic ring is a benzene ring, the preferred substitution position is represented by ^* , generally referred to as the 5 position of the phthalazinone moiety in the formula:

Substituent is preferably an alkoxy, and still is amino, halo or hydroxy group, more preferably C _{1 -7} alkoxy group (e.g., -OMe).

If the substituent is halo, it may also be present at the 6, 7 or 8 position of the phthalinone moiety, but is preferably present at the 8 position. Halo is preferably chloro or fluoro, more preferably fluoro.

Preferably, D is selected from phenylene, fluoro-phenylene, pyridylene, fluoro-pyridylene, furanylene and thiophenylene.

More preferably, D is selected from:

More preferably, D is selected from:

Most preferably, D to be.

Preferably, X is selected from a single bond, NR ^N2 and CR ^C3 R ^C4 , and Y is CR ^C1 R ^C2 .

In some embodiments, X is NR ^N2 . In this embodiment, it is also preferred that R ^C1 and R ^C2 are preferably H and R ^N2 is H. In other embodiments, X is CR ^C3 R ^C4 . In this embodiment, it is also preferred that R ^C1 and R ^C2 are preferably H and R ^C3 and R ^C4 are H.

In some embodiments, X is preferably a single bond so that R ^D is a five membered ring. In this compound, it may be preferred that at least one of R ^N1 , R ^C1 and R ^C2 is not hydrogen. In some of these preferred compounds, only one of R ^N1 , R ^C1 and R ^C2 is not hydrogen. In other of these preferred compounds, two or three of R ^N1 , R ^C1 and R ^C2 are not hydrogen.

The preferred group for R preferably ^C1 or ^C2 R comprises an unsubstituted C _{1 -4} alkyl, such as methyl, ethyl, propyl, but not limited to, methyl is preferred in some compounds.

When R ^N1 is not hydrogen, R ^C1 is preferably methyl, R ^C2 is preferably selected from H and methyl, more preferably methyl.

If R ^N1 is hydrogen, R is ^C2 is also preferred that as a hydrogen, R ^C1 is preferably a carboxy or amido substituted C _{1 -4} alkyl at its terminus (e.g., methyl). The amino substituents of the amido groups together with the N atoms attached thereto are preferably cyclic. The annular portion of the amido group is preferably a C _{5 -7} heterocyclic group containing nitrogen such as pyrrolyl Lindy carbonyl, piperazinyl, No. fur is piperazinyl, piperidinyl, morpholino, all of which are as defined above And may be further substituted as well. In particular, the substituent group is hydroxyl, substituted and unsubstituted C _{1 -4} alkyl (e.g., methyl, hydroxymethyl, methoxy-ethyl, dimethylamino-ethyl) and C _{5 -7} days heterocyclyl (e.g., piperidino N- Nil, morpholino), but is not limited thereto.

Preferred for the R ^N1 group is preferably a C _{1 -4} alkyl (e.g., methyl) substituted with a carboxy or amido esters in Fig group, and further from its end, but is not limited to this. The amino substituents of the amido groups together with the N atoms attached thereto are preferably cyclic. The annular portion of the amido group is preferably a C _{5 -7} heterocyclic group containing nitrogen such as pyrrolyl Lindy carbonyl, piperazinyl, homopiperazinyl, piperidinyl, and morpholino, all of which are as described above May be further substituted. In particular, the substituent group is hydroxyl, substituted and unsubstituted C _{1 -4} alkyl (e.g., methyl, hydroxymethyl, hydroxyethyl, methoxy-ethyl, dimethylamino-ethyl) and C _{5 -7} days heterocyclyl (e.g., N-piperidinyl, morpholino), but is not limited thereto. Where appropriate, the preferences may each be taken in combination with each other.

In some embodiments, Y is NR ^N3 , in which X is preferably C═O. In this embodiment, the R ^N1 and R ^N2 is preferably H and C _{1 -4} is selected from alkyl, more preferably both H.

Other modes included

Well-known ions, salts, solvates and protected forms of these substituents are included above. For example, a carboxylic acid refers to the (-COOH) is its anionic (carboxylate) form (-COO ^-), and, also included, as well as salts or solvates, conventional protected forms. Similarly, references to amino groups include the protonated forms of amino groups (-N ⁺ HR ¹ R ² ), salts or solvates such as hydrochloride, as well as conventional protected forms of amino groups. Similarly, the hydroxyl groups are referred to lock its anionic form (-O ^-) also includes conventional protected forms of addition, salt or solvate thereof, a hydroxyl group.

Isomers, salts, solvates, protected forms and Prodrug

Certain compounds include cis- and trans-forms, E- and Z-forms; c-, t- and r- forms; Endo- and exo-forms; R-, S- and meso-forms, D- and L-forms; d- and l- form; (+) And (-) forms; Keto-, enol- and enolate-forms; syn- and anti-forms; Flavor- and sand-shape; α- and β-forms; Axial- and horizontal-shape; Boat-, chair-, twist-, envelope- and semi- chair-forms; And combinations thereof, hereinafter referred to collectively as "isomers" (or "isomer forms"), including but not limited to one or more specific geometries, optical, enantiomers, diastereomers, epimers, stereoisomers, It may be in the form of tautomers, batches or anomers.

If the compound is present in crystalline form, it may exist in many different polymorphic forms.

Except as discussed below for tautomeric forms, structural (or constituent) isomers (i.e., isomers that differ not only by the position of atoms in space, but also in the relationship between atoms) are used herein as the term " Note that it is expressly excluded from "isomers." For example, the reference to a methoxy group, -OCH ₃ should not be construed as a reference to its structural isomer, the hydroxymethyl group, -CH ₂ OH. Similarly, reference to ortho-chlorophenyl should not be construed as reference to meta-chlorophenyl, which is a structural isomer thereof. However, refers to a class of structures may also include structurally isomeric forms being included in that class (e.g. C _{1 -7} alkyl is n- propyl and iso-propyl, and including; is butyl n-, iso-, sec And tert-butyl; methoxyphenyl includes ortho-, meta- and para-methoxyphenyl).

This exclusion does not correspond to tautomeric forms, such as keto-, enol- and enolate-forms, such as in the following tautomeric pairs: keto / enol, imine / enamine, amide / imino alcohol, amidine / amid Dine, nitroso / oxime, thioketone / enethiol, N-nitroso / hydroxyazo and nitro / aci-nitro.

The tautomer pairs illustrated below are particularly relevant to the present invention:

Note that compounds substituted with one or more isotopes are expressly included in the term "isomer". For example, H can be present in any isotopic form, including ¹ H, ² H (D) and ³ H (T); C can be in any isotopic form, including ¹² C, ¹³ C, and ¹⁴ C; O may be present in any isotopic form, including ¹⁶ O and ¹⁸ O.

Unless otherwise specified, references to particular compounds include all such isomeric forms, including (complete or partial) racemates and other mixtures thereof. Methods of preparing such isomeric forms (e.g., asymmetric synthesis) and separating (e.g., fractional crystallization and chromatography means) are known in the art or may be readily employed in a known manner by employing the methods or known methods taught herein. Is obtained.

Unless otherwise specified, references to particular compounds also include, for example, their ionic, salt, solvate and protected forms, as well as the different polymorphic forms thereof, as discussed below.

It may be convenient or desirable to prepare, purify and / or handle the corresponding salts of the active compounds, such as pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts are described in Berge, et al., "Pharmaceutically Acceptable Salts", J. Pharm . Sci. , 66, 1-19 (1977).

For example, when having the functional group in the compound is anionic, or negative ions or holy ^- may form a salt (for example, -COOH is -COO which may be a), the appropriate cation. Examples of suitable inorganic cations include other cations such as alkaline earth cations such as Na ⁺ and Al ^{3 +} and alkali metal ions such as K ^+, Ca ^{2 +} and Mg ^{2 +,} however, and the like. Examples of suitable organic cations include, but are not limited to, ammonium ions (ie NH ₄ ⁺ ) and substituted ammonium ions (eg NH ₃ R ⁺ , NH ₂ R ₂ ⁺ , NHR ₃ ⁺ , NR ₄ ⁺ ). Examples of some suitable substituted ammonium ions are those derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine , Amino acids such as phenylbenzylamine, choline, meglumine and tromethamine, as well as lysine and arginine. An example of a typical quaternary ammonium ion is N (CH ₃ ) ₄ ⁺ .

When the compound contains a functional group that can be cationic or cationic or holy may form a salt (e.g., -NH ₂ may be that one -NH ₃ ^+), suitable anion. Examples of suitable inorganic anions include, but are not limited to, 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. Examples of suitable organic anions include, but are not limited to, those derived from 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, sulfanic acid, 2-acetyoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane Sulfonic acid, ethane disulfonic 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, carboxymethyl cellulose.

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

It may be convenient or desirable to prepare, purify and / or handle the active compounds in chemically protected form. As used herein, the term "chemically protected form" means that at least one reactive functional group is protected from an undesired chemical reaction, i.e. a protected or protected group (shielded or shielded group, or blocked or blocked) To a compound in the form of a group). By protecting the reactive functional groups, reactions involving other unprotected reactive functional groups can be carried out without affecting the protected groups, and in general, the protecting groups can be removed in the subsequent step without substantially affecting the rest of the molecule. See, eg, "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 (= 0) R], for example t-butyl ether; Benzyl, benzhydryl (diphenylmethyl) or trityl (triphenylmethyl) ether; Trimethylsilyl or t-butyldimethylsilyl ether; Or acetyl esters [-OC (= 0) CH ₃ , -OAc].

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

For example, amine groups are, for example, amides or urethanes, for example methyl amide (-NHCO-CH ₃ ); Benzyloxy amide (-NHCO-OCH ₂ C ₆ H ₅ , -NH-Cbz), t-butoxy amide [-NHCO-OC (CH ₃ ) ₃ , -NH-Boc]; 2-biphenyl-2-propoxy amide [-NHCO-OC (CH ₃ ) ₂ C ₆ H ₄ C ₆ H ₅ , -NH-Bpoc], as 9-fluorenylmethoxy amide (-NH-Fmoc) , As 6-nitroveratriryloxy amide (-NH-Nvoc), as 2-trimethylsilylethyloxy amide (-NH-Teoc), as 2,2,2-trichloroethyloxy amide (-NH-Troc), As allyloxy amide (-NH-Alloc), as 2 (-phenylsulfonyl) ethyloxy amide (-NH-Psec); Or as appropriate, N-oxides (> NO.).

For example, a carboxylic acid group is an ester such as C _{1 -7} alkyl ester (e.g., methyl ester; t- butyl ester); C _{1 -7} haloalkyl ester (e.g., alkyl esters with C ₁ to _-7 tree); Tree C _1-7 alkylsilyl -C _{1 -7} alkyl ester; As, (for example, nitrobenzyl ester, benzyl ester), or C _{5 -20} aryl -C _{1 -7} alkyl ester; Or as an amide, such as methyl amide.

For example thiol groups are thioethers (-SR), such as benzyl thioethers; It may be protected as 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 prodrug form. The term "prodrug" as used herein relates to a compound from which certain active compounds are obtained when metabolized (eg in vivo). Typically, 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 active compounds (eg, pharmacologically acceptable metabolic labile esters). During metabolism, the ester group [-C (= 0) OR] cleaves to obtain the active drug. Such esters can be formed, for example, by esterifying any of the carboxylic acid groups [—C (═O) OH] in the parent compound, if appropriate before protecting and deprotecting any other reactive groups present in the parent compound. have. Examples of such metabolically labile esters, R is C _{1 -20} alkyl (e.g., -Me, -Et); C _{1 -7-amino-alkyl} [e.g., aminoethyl; 2- (N, N-diethylamino) ethyl; 2- (4-morpholino) ethyl]; And acyloxy -C _{1 -7} alkyl (for example, acyloxy methyl, ethyl acyloxy; 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) car Carbonyloxyethyl; (4-tetrahydropyranyl) carbonyloxymethyl; and 1- (4-tetrahydropyranyl) carbonyloxyethyl).

Further suitable prodrug forms include phosphonate and glycolate salts. In particular, the hydroxy group (-OH) can be prepared as a phosphonate prodrug by reacting with chlorodibenzylphosphite and then hydrogenating to form the phosphonate group -OP (= 0) (OH) ₂ . Such groups can be cleaved by phosphatase enzymes during metabolism to obtain active drugs with hydroxy groups.

In addition, some prodrugs are activated by enzymes to give the active compound or a compound from which the active compound is obtained upon further chemical reaction. For example, the prodrug may be a sugar derivative or other glycoside conjugate, or may be an amino acid ester derivative.

Abbreviation

For convenience, many of the 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 ) Using well-known abbreviations, including but not limited to

For convenience, many of the chemical moieties are methanol (MeOH), ethanol (EtOH), iso-propanol (i-PrOH), methyl ethyl ketone (MEK), ether or diethyl ether (Et ₂ O), acetic acid (AcOH), dichloro Labeled using well-known abbreviations, including but not limited to methane (methylene chloride, DCM), trifluoroacetic acid (TFA), dimethylformamide (DMF), tetrahydrofuran (THF), and dimethyl sulfoxide (DMSO) do.

synthesis

Compounds of the present invention of Formula I may be synthesized from precursors of Formula 2:

Formula I

Formula 2

In the formulas above, OProt represents a protected hydroxy group. The various substituent groups shown may be the same as defined for the compounds of formula (I), or may be protected forms or precursors of the defined groups and further transformation may be required to reach the desired compound. Synthesis of the compounds of the present invention can proceed by removing hydroxy protecting groups using standard techniques such as base catalysis, HBTU coupling and then forming amide bonds.

Compounds of Formula 2 may be synthesized by coupling a compound of Formula 3 or Formula 4 with a compound of Formula 5 or Formula 6, respectively:

Formula 3

Formula 4

Formula 5

Formula 6

Urea bond formation reactions are carried out under standard conditions. Compounds of formulas (5) and (6) can be synthesized according to known methods, examples of which are shown below. The same applies to the compounds of the formulas (3) and (4).

Usage

The present invention provides active compounds, particularly compounds that are active in inhibiting the activity of PARP-1.

As used herein, the term "activity" refers to a compound capable of inhibiting PARP-1 activity, and may or may not exhibit some intrinsic activity, as well as a compound (drug) that specifically has intrinsic activity. Which includes both prodrugs of these compounds.

Assays that can be conveniently used to assess PARP-1 inhibition provided by certain compounds are described in the Examples below.

The invention further provides a method of inhibiting the activity of PARP-1 in a cell, comprising contacting said cell with an effective amount of an active compound, preferably in the form of a pharmaceutically acceptable composition. Such methods can be carried out in vitro or in vivo.

For example, a sample of cells can be grown in vitro and the active compound contacted with the cells, then the effect of the compound on these cells can be observed. As an example of an "effect," one can measure the amount of DNA repair that occurred at a particular time. If the active compound is found to affect the cells, it can be used as a prognostic or diagnostic marker for the efficacy of the compound in methods of treatment of patients with cells of the same cell type.

As used herein in the context of treating a condition, the term “treatment”, whether human or animal (eg, in veterinary use), generally refers to a decrease in the rate of progression, a stop in the rate of progression, a condition improvement, and Several certain therapeutic effects, including treatment, are directed to the treatments and therapies in which inhibition of pathological progress is achieved. Treatment as a prophylactic measure (ie prophylaxis) is also included.

The term "assistant" as used herein relates to the use of the active compound with known therapeutic means. Such means include ionizing radiation and / or cytotoxic therapy of drugs as used for the treatment of different cancer types. In particular, active compounds are known to increase the action of many cancer chemotherapy treatments, which include the class of local isomerases of toxins and most known alkylating agents used to treat cancer.

The active compounds can also be used as cell culture additives to inhibit PARP, for example to sensitize known chemotherapeutic agents or ionizing radiation in vitro.

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

administration

The active compound, or pharmaceutical composition comprising the active compound, can be administered orally (eg, by ingestion), whether systemic / peripheral or at a predetermined site of action; Topical (including transdermal, intranasal, ocular, buccal and sublingual); Lungs (eg, by inhalation or aeration therapy, eg using aerosol, for example through the mouth or nose); rectal; Vaginal administration; Parenteral administration, such as subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intradural, spinal canal, intraarticular, sublingual, orbital, intraperitoneal, intratracheal, subcutaneous, intraarticular, subarachnoid And parenteral administration by infusion, including intratracheal; Administration to the subject may be by any convenient route of administration, including but not limited to parenteral administration, such as by implantation of an accumulation site subcutaneously or intramuscularly.

The subject may be eukaryotic, animal, vertebrate, mammal, rodent (eg guinea pig, hamster, rat, mouse), rat (eg mouse), canine (eg dog), feline (eg cat), horse And apes (eg, horses), primates, apes (eg, monkeys or apes), monkeys (eg, silk monkeys, baboons), apes (eg, gorillas, chimpanzees, orangutans, gibbons) or humans.

Formulation

The active compound may be administered alone, but the one or more active compounds as defined above may be used in one or more pharmaceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, stabilizers, preservatives, lubricants or well known to those skilled in the art. It is preferred to provide it as a pharmaceutical composition (e.g., a formulation) comprising together with other substances and optionally other therapeutic or prophylactic agents.

Accordingly, the present invention provides a pharmaceutical composition as defined above, and at least one active compound as defined above, at least one pharmaceutically acceptable carrier, excipient, buffer, adjuvant, stabilizer or other substance as described above. Further provided is a method of preparing a pharmaceutical composition comprising mixing with.

As used herein, the term “pharmaceutically acceptable” is within the scope of sound medical judgment and used to contact an individual's (eg, human) tissue without excessive toxicity, irritation, allergic reactions or other problems or complications. It relates to compounds, materials, compositions and / or formulations which are suitable below and which have a suitable benefit / risk ratio. Each carrier, excipient, etc. must also be "acceptable" in terms of compatibility with other components of the formulation.

Suitable carriers, diluents, excipients and the like can be found in standard pharmaceutical textbooks. See, eg, "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 Excipient", 2nd edition, 1994.

The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Such methods include the step of bringing into association the active compound with the carrier which constitutes one or more accessory ingredients. Generally, formulations are prepared by uniformly and intimately associating the active compound with a liquid carrier or finely divided solid carrier or both, followed by molding the product as necessary.

Formulations include liquids, solutions, suspensions, emulsions, elixirs, syrups, tablets, lozenges, granules, powders, capsules, cachets, pills, ampoules, suppositories, pessaries, ointments, gels, pastes, creams, sprays, mists, It may be in the form of a foam, lotion, oil, cyclic, soft or aerosol.

Formulations suitable for oral administration (eg, by ingestion) may be presented as discrete units, such as capsules, cachets or tablets, each containing a predetermined amount of active compound; As a powder or granules; As a solution or suspension in an aqueous or non-aqueous liquid; Or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion; As cyclic; As a weakness; Or as a paste.

Tablets may be prepared by conventional methods, such as by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may include one or more binders (eg, povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethyl cellulose); Fillers 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); Surfactants or dispersants or wetting agents (eg, sodium lauryl sulfate); And compressing the active compound in a suitable machine in a free flowing form such as powder or granules, optionally mixed with a preservative (eg methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid). . 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 optionally be coated or scored and formulated to sustain or controlled release of the active compound using, for example, hydroxypropylmethyl cellulose in various ratios to provide the desired release profile. Tablets may optionally be enteric coated to release in parts of the intestine other than the stomach.

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

Formulations suitable for topical administration to the oral cavity include lozenges comprising the active compound in a flavored base, usually sucrose and acacia or tragacanth; Pastilles comprising the active compound in an inert medicine such as gelatin or glycerin, or sucrose and acacia; And mouthwashes comprising 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, in which the carrier is a solid, are coarse particles having a particle size such as in the range of about 20 to about 500 microns, which are administered by nasal inhalation, ie, by rapid inhalation through the nasal cavity from a powder container close to the nose. It includes. Suitable carriers include aqueous or oily oils of the active compounds when the carrier is, for example, a nasal spray, a liquid for administration by nasal drops or by aerosol administration by nebulizer.

Formulations suitable for administration by inhalation include those provided as an aerosol spray from a pressurized pack using a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichloro-tetrafluoroethane, carbon dioxide or other suitable gas. do.

Formulations suitable for topical administration through the skin include ointments, creams and emulsions. When formulated into ointments, the active compounds may optionally be used with paraffinic or water miscible ointment drops. Alternatively, the active compounds may be formulated into creams with oil-in-water cream dispensers. If necessary, the water phase of the cream main agent may contain at least two hydroxyl groups, such as at least about 30% w / w of a polyhydric alcohol, ie propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof. Having alcohols. Topical formulations may preferably include compounds that enhance absorption or penetration of the active compound through the skin or other affected areas. Examples of such dermal penetration enhancers are dimethylsulfoxide and related analogs.

When formulated as a topical emulsion, the oil phase may optionally comprise only an emulsifier (also known as an emulgent), or one or more emulsifiers comprising fats or oils, or both fats and oils. It may include a mixture of. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferred to include both oils and fats. The emulsifier (s) comprising the stabilizer (s) together constitute the so-called emulsifying wax, and the wax together with the oil and / or fat constitutes the so-called emulsifying ointment preparation which forms the oily dispersed phase of the cream formulation.

Suitable emulsifiers and emulsion stabilizers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate. The oils or fats suitable for the formulation are chosen based on the achievement of certain cosmetic properties, since the solubility of the active compounds in the majority of oils that may be used in pharmaceutical emulsion formulations may be very low. Thus, the cream should preferably be a non-greasy, non-colored and washable product with adequate firmness to prevent leakage from tubes or other containers. Straight or branched mono or dibasic alkyl esters such as diisoadiate, isocetyl stearate, propylene glycol diesters of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate Blends of branched esters known as 2-ethylhexyl palmitate or Crodamol CAP can be used, the last three being the preferred esters. These may be used alone or in combination according to predetermined characteristics. 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 provided as suppositories, including suitable main medicaments, including, for example, cocoa butter or salicylate.

Formulations suitable for vaginal administration can be provided as pessaries, tampons, creams, gels, pastes, foams or spray formulations contained in addition to the active compound, such as carriers as are known in the art.

Formulations suitable for parenteral administration (eg, by infusion, including skin, subcutaneous, intramuscular, intravenous and intradermal) include antioxidants, buffers, preservatives, stabilizers, bacteriostatic agents and the blood and preparations of the intended recipients isotonic. Sterile injectable solutions, both aqueous and non-aqueous, isotonic and pyrogen-free, which may contain solutes such that; And aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickeners, and other particulate systems designed for applying the compound to liposomes or blood components or one or more organs. Examples of suitable isotonic vehicles for use in such formulations are sodium chloride injection, Ringer's solution or lactate Ringer's injection. Typically, the concentration of active compound in the solution is from about 1 ng / ml to about 10 μg / ml, such as from about 10 ng / ml to about 1 μg / ml. The formulations may be provided in unit dose or multi-dose sealed containers, such as ampoules and vials, and may be stored under lyophilized (freeze drying) conditions requiring only the addition of a sterile liquid carrier, such as water for injection, immediately before use. Term injections and suspensions can be prepared from sterile powders, granules and tablets. The formulations may be in the form of liposomes or other particulate systems designed for applying the compound to blood components or one or more organs.

Dosage

It will be appreciated that the appropriate dosage of the active compound, and composition comprising the active compound, may vary from patient to patient. Determination of the optimal dosage will generally involve balancing the therapeutic benefit levels against any risks or deleterious side effects of the treatment of the present invention. The dosage level chosen is the activity of the particular compound, route of administration, time of administration, rate of release of the compound, duration of treatment, other drugs, compounds and / or materials used in combination, and the age, sex, weight, condition, general health of the patient. It may vary depending on various factors, including but not limited to condition and previous medical history. Generally, the dosage will be to achieve local concentration at the site of action that achieves the desired effect without causing substantially harmful or deleterious side effects, but the amount and route of administration will ultimately be determined by the physician.

In vivo administration can be carried out continuously or intermittently (eg, in divided doses at appropriate intervals) at one dose during the course of treatment. The most effective means and methods of determining the dosage are well known to those skilled in the art and will vary depending on the agent used for the therapy, the purpose of the therapy, the target cell to be treated and the individual to be treated. The treating physician may make single or multiple administrations while selecting dosage levels and patterns.

In general, suitable dosages of the active substance range from about 100 μg to about 250 mg for 1 kg of the subject's body weight per day. If the active compound is a salt, ester, prodrug or the like, the amount to be administered is calculated on the basis of the parent compound and thus the actual working weight increases proportionally.

General Experiment Method

Preparative ( preparative ) HPLC

Samples were purified with a Waters mass induction purification system using a Waters 600 LC pump, Genesis AQ 120A 4 μm 500 mm × 4.6 mm column, and a Micromass ZQ mass spectrometer operating in a cationic electrospray ionization mode. Mobile phases A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile) were used in the following gradients:

Flow rate: 2.0 ml / min

analysis HPLC - MS

Analytical HPLC was routinely performed using a Spectra System P4000 pump and a Jones Genesis C18 column (4 μm, 50 mm × 4.6 mm). Mobile phases A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile) were used in the gradient as described below. Detection was performed with a TSP UV 6000LP detector in PDAs with 254 nm UV and 210-600 nm range. The mass spectrometer is a Waters ZMD LC-MS system No. 2 operating in electrospray ionization mode. LD352.

Flow rate: 1.0 ml / min

NMR

¹ H NMR and ¹³ C NMR were typically recorded using Bruker DPX 300 spectrometer at 300 MHz and 75 MHz, respectively. Chemical shifts were reported in ppm on a δ balance against tetramethylsilane internal standard. Unless otherwise specified, all samples were dissolved in DMSO-d ₆ .

Example  One

Wherein X is a single bond or NH

Compound 1 was synthesized as described in Example 23 of WO 03/093261, which is incorporated herein by reference.

(a) 4- (4-fluoro-3-isocyanato-benzyl) -2H-phthalazin-1-one (2)

4- (3-amino-4-fluoro-benzyl) -2H-phthalazin-1-one (1) in anhydrous DCM (1.6 L) and triethylamine (4.62 mL, 40.86 mmol) (4.0 g, 14.8 To a suspension of mmol) was added dropwise a solution of triphosgene (2.75 g, 9.28 mmol) in anhydrous DCM (327 mL) and stirred at room temperature for 70 minutes. The reaction mixture was then concentrated to dryness in vacuo to yield a gray solid. A single peak (the yield deemed quantitative) was obtained in the LC-MS analysis and no purification was performed. m / z (LC-MS, ESP), RT = 4.49 min, (M + MeOH) 328.0.

(b) {3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -ureido} -amino ethyl ester (4)

To a solution of the appropriate amino ester (3) (0.026 g, 0.17 mmol) in anhydrous DCM (16.7 mL) was triethyl amine (2 μL, 0.170 mmol) and 4- (4-fluoro-3-isocyanato-benzyl ) -2H-phthalazin-1-one (2) (0.05 g, 0.17 mmol) was added. The reaction mixture was stirred for 2 hours, then washed with water (2 × 15 mL), dried over MgSO ₄ , filtered and concentrated to give the corresponding ureido ester. Corresponding ureido esters were used without the need for purification.

(c) 3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -substituted imidazolidine-2,4-dione (5)

(i) Base Catalysis Method (5a-5e, 5g-5i)

Suitable {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -ureido ester in anhydrous dimethylacetimide (0.5 mL) ( 4) (0.065 mmol) was added sodium hydroxide (2.6 mg, 0.065 mmol) and heated to 50 ° C. for 30 minutes. The reaction mixture was then diluted with DCM (2 mL) and washed with brine (2.5 mL). Crude samples were submitted to preparative HPLC.

(ii) HBTU coupling method (5f, 5j)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -ureido ester (4) in ethanol (0.5 mL) (0.065) sodium hydroxide (2.6 mg, 0.065 mmol) was added and stirred at room temperature for 30 minutes. Hydrochloric acid (0.065 mmol) was then added to neutralize the reaction mixture and concentrated to dryness in vacuo.

The resulting solid was then suspended in DCM (0.5 mL), N'N'diisopropylethylamine (10.5 μl, 0.06 mmol) and 2- (1H-benzotriazol-1-yl) -1,1, Treated with 3,3-tetramethyluronium hexafluorophosphate (0.024 g, 0.06 mmol). The reaction mixture was stirred for 18 hours and then submitted to preparative HPLC.

Example  2

Wherein X is a single bond or NH

Compound 6 was synthesized as described in Example 1 of WO 03/093261, which is incorporated herein by reference.

(a) 4- (3-isocyanato-benzyl) -2H-phthalazin-1-one (7)

To a solution of 4- (3-amino-benzyl) -2H-phthalazin-1-one (6) (1.0 g, 4.0 mmol) in anhydrous DCM (415 mL) and triethylamine (1.5 mL, 10.9 mmol). A solution of triphosgene (0.73 g, 2.5 mmol) in anhydrous DCM (85 ml) was added dropwise over 30 minutes. After an additional 2 hours, the reaction mixture was concentrated to dryness to give a pale yellow powder. A single peak (the yield deemed quantitative) was obtained in the LC-MS analysis and no purification was performed. m / z (LC-MS, ESP), RT = 3.90 min, (M + MeOH) 310.0.

(b) {3- [5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -ureido} -amino ethyl ester (8)

To a solution of the appropriate amino ester (0.026 g, 0.17 mmol) in anhydrous DCM (16.7 mL) was triethylamine (24 μl, 0.170 mmol) and 4- (3-isocyanato-benzyl) -2H-phthalazine- 1-one (7) (0.05 g, 0.17 mmol) was added. The reaction mixture was stirred for 2 hours, then washed with water (2 × 15 mL), dried over MgSO ₄ , filtered and concentrated to give the corresponding ureido ester. Corresponding ureido esters were used without the need for further purification.

(c) 3- [5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -substituted imidazolidine-2,4-dione (9)

Suitable {1- [3- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenylcarbamoyl] -substituted} -carbamic acid in anhydrous dimethylacetimide (0.5 mL) To the ester (0.065 mmol) was added sodium hydroxide (2.6 mg, 0.065 mmol) and heated to 50 ° C. for 0.5-6 h. The reaction mixture was then diluted with DCM (2 mL) and washed with brine (2.5 mL). Crude samples were submitted to preparative HPLC.

Example  3

(a) carbonylmethyl-amino acetic acid bis (tert-butylsilyl ester) (11)

A suspension of (carboxymethyl-amino) -acetic acid (10) (0.6 g, 4.51 mmol) in bis (trimethylsilyl) -trifluoroacetamide (45 mL, 169 mmol) was heated until complete dissolution (about 1 time). The reaction mixture was then concentrated under vacuum to give a yellow oil (1.6 g). The oil was used without the need for further purification.

(b) {3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl-2A-dioxo-imidazolidine-1- Japanese-acetic acid (12)

Triethylamine (61) in a suspension of 4- (4-fluoro-3-isocyanato-benzyl) -2H-phthalazin-1-one (2) (4.51 mmol) in dioxane anhydrous (180 mL). Μl, 4.51 mmol) was added followed by carbonylmethyl-amino acetic acid bis (tert-butylsilyl ester) (11) (1.25 g, 4.51 mmol). The suspension was stirred for 48 hours at room temperature.

The reaction mixture was concentrated to dryness, then diluted with water (150 mL) and the pH was adjusted to 10 using sodium hydroxide solution (about 2N 10 mL). The aqueous phase was then extracted with ethyl acetate (2 × 20 mL). The pH of the aqueous phase was acidified to 1 with 2N HCl and extracted with ethyl acetate (2 × 100 mL). The latter ethyl acetate layer was then combined, dried over MgSO ₄ , filtered and concentrated in vacuo to afford a yellow solid. No purification was performed with a 92% peak in the LC-MS analysis (1.0 g, 57%). m / z (LC-MS, ESN), RT = 3.47 min, (MH) 409.0.

(c) {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -2,4-dioxo-imidazolidine -1-yl} -acetic acid amide (13)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -2,4-dioxo- in anhydrous DCM (0.5 mL) A suitable amine (0.150 mmol) was added to a suspension of imidazolidin-1-yl} -acetic acid (12) (0.05 g, 0.122 mmol) with N'N'diisopropylethylamine (0.150 mmol) and 2- (1H- Add with benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate (0.057 g, 6 0.150 mmol) and stir at room temperature for 7 hours. The reaction mixture was stirred for 18 hours and then submitted to preparative HPLC.

Example  4

(a) 2-methyl-4-nitro-pyridin-1-ol (15)

2-Picoline-N-oxide 14 (21.9 g, 0.2 mmol) was added to concentrated sulfuric acid (76 mL) while maintaining the temperature below 40 ° C. The mixture was cooled to 5 ° C. and fuming nitric acid (60 mL) was added dropwise over 15 minutes while maintaining the reaction temperature below 10 ° C. After the addition was complete, the reaction mixture was heated to 100 ° C. for 1.5 hours. The reaction was then cooled to room temperature, poured over ice (about 500 g) and the resulting greenish yellow solution was neutralized by addition of sodium carbonate (about 205 g over 1 hour). The mixture was then filtered and washed with water (2 L). The solid cake was then dissolved in DCM (500 mL) and dried over Na ₂ SO ₄ . Hexane (100 mL) was added to the DCM solution to give a yellow suspension which was filtered and dried. A single peak in LC-MS analysis (25.40 g, 82.5%) was taken as crude for the next step without purification.

(b) 4-nitro-pyridin-2-ylmethyl ester (16)

To methyl acetic anhydride (60 mL, 0.635 mol) under 110 ° C. was added partial portions of 2-methyl-4-nitro-pyridin-1-ol (15) (17.1 g, 111 mmol) over 1 minute. After about 2 minutes the solution began to darken and was kept heated at 100-120 ° C. for 30 minutes and then cooled to 80 ° C. Ethanol (60 mL) was added and the solution was concentrated in vacuo. The residue was slurried in water (140 mL) and the mixture was neutralized with NaHCO ₃ (about 40 g). The aqueous liquid was extracted with ethyl acetate (4 × 100 mL) and the combined extracts were washed with brine (2 × 100 mL), dried over Na ₂ SO ₄ and concentrated to crude oil under vacuum. The material was subjected to flash chromatography using hexanes: TBME (2: 1) as eluent. After the brown oil was isolated, a single peak was obtained by LC-MS analysis (6.44 g, 30%); m / z (LC-MS, ESP), RT = 3.65 min, (M + H) 197.

(c) (4-nitro-pyridin-2-yl) -methanol (17)

To a solution of 4-nitro-pyridin-2-ylmethyl ester (16) (0.49 g, 2.5 mmol) in methanol (5 mL) was added a solution of sodium hydroxide (2N, 1.6 mL) and the reaction was stirred for 30 minutes. After stirring, it was concentrated in vacuo. The residue was diluted with water (10 mL) and extracted with ethyl acetate (2 × 10 mL). The combined extracts were washed with brine (10 mL) and concentrated in vacuo to afford a crude oil which solidified upon standing. LC-MS analysis gave a single peak (0.13 g 33%); m / z (LC-MS, ESP), RT = 2.67 min, (M + H) 155.

(d) 4-nitro-pyridine-2-carbaldehyde (18)

Dimethyl sulfoxide (0.593 mL, 8.37 mmol) was added dropwise to a cooled solution of oxalyl chloride (0.533 g, 4.18 mmol) in DCM (2 mL) at −78 ° C. under a nitrogen atmosphere. After 30 minutes while maintaining the temperature at -78 ° C, (4-nitro-pyridin-2-yl) -methanol (17) (0.129, 0.837 mmol) in DCM (2 mL) was added dropwise. After 2 hours, the mixture was warmed to -55 ° C. Triethylamine (1.74 mL, 12.55 mmol) was then added and the mixture was allowed to warm to room temperature over 2 hours. Then brine (10 mL) was added and the mixture was extracted with DCM (4 × 10 mL). The combined organics were then dried over MgSO ₄ and concentrated to oil under vacuum. The material was used directly without the need for further purification in anticipation of quantitative conversion.

(e) 3- (4-nitro-pyridin-2-ylmethylene) -3H-isobenzofuran-1-one (19)

To a cooled solution of (4-oxo-3,4-dihydro-phthalazin-1-yl) -phosphonic acid dimethyl ester (0.202 g, 0.837 mmol) in THF (4 mL) at 10 ° C. under nitrogen atmosphere. -Nitro-pyridine-2-carbaldehyde (0.837 mmol) was added. Triethylamine (0.174 mL, 1.25 mmol) was added dropwise over 10 minutes, followed by stirring at room temperature for 18 hours. The yellow suspension is poured into water (9 mL) and filtered, the solid is washed with water (2 × 2 mL), hexane (2 × 2 mL) and then with diethyl ether (2 × 2 mL) and under vacuum Drying yielded 0.73 g of the desired product. Two peaks were obtained in the LC-MS analysis (0.13 g, 18% 2 steps), requiring no further purification; m / z (LC-MS, ESP) 1 RT = 4.26 min (M + H) 269, & RT = 4.52 min (M + H) 269.

(f) 4- (4-amino-pyridin-2-ylmethyl) -2H-phthalazin-1-one (20)

A suspension of 3- (4-nitro-pyridin-2-ylmethylene) -3H-isobenzofuran-1-one (0.67 g, 2.5 mmol) in hydrazine monohydrate (0.25 g, 5.0 mmol) was heated to 90 ° C. for 1 hour. Heated to and cooled to room temperature. Ethanol (5 mL) was added followed by ammonium chloride (0.16 g, 3.0 mmol) and iron powder (0.28 g, 5.0 mmol) and the mixture was heated to 90 ° C. for 3 h. The reaction mixture was filtered hot through celite and washed with ethyl acetate (20 mL). The filtrate was concentrated in vacuo, then diluted with ethyl acetate (10 mL), washed with brine (10 mL), dried over magnesium sulfate and dried under vacuum to afford the title compound as a pale yellow powder. Single peak in LC-MS analysis (0.20 g, 31% yield), requiring no further purification; m / z (LC-MS, ESP), RT = 1.58 (M + H) 253;

(g) 1,5,5-substituted-3- [2- (4-oxo-3,4-dihydro-phthalazin-1-yl methyl) -pyridin-4-yl] -imidazolidine -2,4-dione (23)

To a solution of the appropriate amino ester isocyanate (0.026 g, 0.17 mmol) in anhydrous DCM (16.7 mL) was triethyl amine (2 μL, 0.170 mmol) and 4- (4-amino-pyridin-2-ylmethyl) -2H-prop Talazine-1-one (21) (0.05 g, 0.17 mmol) was added. The reaction mixture was stirred for 8 hours and concentrated in vacuo to give the corresponding ureido ester. Anhydrous dimethylacetimide (0.5 mL) was added followed by sodium hydroxide (2.6 mg, 0.065 mmol) and the mixture was heated to 50 ° C. for 30 minutes. The reaction mixture was then diluted with DCM (2 mL) and washed with brine (2.5 mL). Crude samples were submitted to preparative HPLC.

(xi) 1,5,5-substituted-3- [2- (4-oxo-3,4-dihydro-phthalazin-1-yl methyl) -pyridin-4-yl] -imidazolidine Alternative route to -2,4-dione (23)

(a) 4- (4-isocyanato-pyridin-2-ylmethyl) -2H-phthalazin-1-one (22)

4- (4-amino-pyridin-2-ylmethyl) -2H-phthalazin-1-one (21) (0.40 g, 1.48 in anhydrous DCM (160 mL) and triethylamine (0.46 mL, 4.09 mmol) To a suspension of mmol) was added dropwise a solution of triphosgene (0.27 g, 0.93 mmol) in dry DCM (30 mL). The reaction was stirred at rt for 6 h. The reaction mixture was then concentrated to dryness in vacuo to give a gray solid. A single peak (the yield deemed quantitative) was obtained in the LC-MS analysis and no purification was performed. m / z (LC-MS, ESP), RT = 3.79 min, (M + MeOH) 329.0.

(b) Triethyl amine (24 μl, 0.170 mmol) and 4- (4-isocyanato-pyridin-2-ylmethyl in a solution of the appropriate amino ester (0.026 g, 0.17 mmol) in dry DCM (16.7 mL). ) -2H-phthalazin-1-one (22) (0.05 g, 0.18 mmol) was added. The reaction mixture was stirred for 8 hours and concentrated in vacuo to afford the corresponding crude ureido ester. Anhydrous dimethylacetimide (0.5 mL) was added followed by sodium hydroxide (2.6 mg, 0.065 mmol) and heated to 50 ° C. for 30 minutes. The reaction mixture was then submitted to preparative HPLC.

Example  5

(a) 4-nitro-thiophene-2-carbaldehyde (25)

Concentrated sulfuric acid (15 mL) was added to concentrated nitric acid (15 mL), the mixture was cooled to -10 ° C, and 2-thiophenecarboxyaldehyde (24) (10.0 g) was maintained at -5 to 0 ° C. , 89.2 mmol) was added dropwise over 15 minutes. The mixture was stirred for an additional 1 hour, then poured into water (200 mL) and extracted with DCM (3 × 100 mL). The combined organic layer was dried over MgSO ₄ and concentrated in vacuo. The crude oil was flash chromatographed using hexane: ether (10: 1) as eluent. The desired isomer (rf 0.32 in 2: 1 hexanes: ether) was collected. Single peak in LC-MS analysis (11.5 g, 82% yield), no further purification needed; m / z (LC-MS, ESP), RT = 3.55 min. no isolation at ms was observed; ¹ H NMR (300 MHz, D ₆ -DMSO) 10.0 (1H, s), 9.2 (1H, s), 8.6 (1H, s).

(b) 3- (4-nitro-thiophen-2-ylmethylene) -3H-isobenzofuran-1-one (26)

4-nitro-thiophene in a solution of (4-oxo-3,4-dihydro-phthalazin-1-yl) -phosphonic acid dimethyl ester (3.85 g, 15.9 mmol) in THF (30 mL) under nitrogen atmosphere. 2-Carbaldehyde (25) (2.5 g, 15.9 mmol) was added. Triethylamine (2.2 mL, 15.9 mmol) was added dropwise over 10 minutes and then stirred at room temperature for 4 hours. The yellow suspension was poured into water (80 mL) and filtered, the solid was washed with water (15 mL) and hexane (10 mL), then with diethyl ether (10 mL) and dried under vacuum to give 4.1 g. The desired product was obtained. Two peaks were obtained in the LC-MS analysis (4.1 g, 94%), so no further purification was necessary; m / z (LC-MS, ESP), RT = 4.71 min (M + H) 274, & RT = 4.84 min (M + H) 274.

(c) 4- (4-nitro-thiophen-2-ylmethyl) -2H-phthalazin-1-one (27)

A suspension of 3- (4-nitro-thiophen-2-ylmethylene) -3H-isobenzofuran-1-one (26) (1.95 g, 5.17 mmol) in hydrazine monohydrate (0.6 mL, 12.0 mmol) was added to 1 After heating to 90 ° C. for hours, it was cooled to room temperature. The mixture was filtered and the filtered cake was washed with water (3 × 40 mL), hexane (2 × 40 mL) and dried at 50 ° C. under vacuum. Two peaks were obtained in the LC-MS analysis (1.0 g, 61%), so no further purification was necessary; m / z (LC-MS, ESP), RT = 4.05 min (M + H) 289, & RT = 3.71 min (M + H) 289.

(d) 4- (4-amino-thiophen-2-ylmethyl) -2H-phthalazin-1-one (28)

Water (18) in a suspension of 4- (4-nitro-thiophen-2-ylmethyl) -2H-phthalazin-1-one (27) (0.90 g, 5.71 mmol) in ethanol (25 mL) under a nitrogen blanket. ML) was added dropwise a solution of ammonium chloride (0.31 g, 5.7 mmol). The resulting solution was stirred for 5 minutes, then iron powder (0.64 g, 11.5 mmol) was added in portions. The yellow mixture was stirred for 3 h at 80 ° C. The reaction mixture was hot filtered through celite and washed with ethyl acetate (80 mL). After concentration in vacuo, diluted with ethyl acetate (20 mL), washed with brine (20 mL), dried over magnesium sulfate and dried under vacuum to afford the title compound as a light yellow powder. A single peak was obtained in the LC-MS analysis (0.68 g, 84% purity), so no further purification was necessary; m / z (LC-MS, ESP), RT = 2.94 (M + H) 258.

(e) substituted {3- [5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -thiophen-3-yl] -ureido} -acetic acid ethyl ester (29 )

Isocyanate suitable for suspension of 4- (4-amino-thiophen-2-ylmethyl) -2H-phthalazin-1-one (28) (75 mg, 0.29 mmol) in DCM (1.5 mL) under nitrogen atmosphere. 0.380 mmol) was added. After stirring for 3 hours, the reaction was quenched with water and the resulting precipitate was filtered and dried. The isolated solid was used without the need for purification.

(f) {1- [3- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) phenylcarbamoyl substituted imidazolidine-2,4-dione (30)

Suitable {3- [5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -thiophen-3-ylJ-ureido}-in anhydrous dimethylacetimide (0.5 mL)- To the acetic acid ethyl ester 29 (0.065 mmol) was added sodium hydroxide (2.6 mg, 0.065 mmol) and heated to 50 ° C. for 0.5-6 h. The reaction mixture was then diluted with DCM (2 mL) and washed with brine (2.5 mL). Crude samples were submitted to preparative HPLC.

Example  6

A: R ^C2 = H

B: R ^C2 = Me

a) 2- (tert-butoxycarbonylmethyl-amino) -propionic acid methyl ester (33A)

To a solution of tert-butyl chloroacetate (31) (4.5 g, 29.7 mmol) in anhydrous DMF (50 mL) was added Hunig's base (6.28 mL, 36.0 mmol) followed by potassium carbonate (5.24 g, 38.0). mmol) was added. After the mixture was heated to 90 ° C., DL-alanine methyl ester (32A) (4.55 g, 37.2 mmol) dissolved in DMF (10 mL) was added dropwise over 1 hour. Heating was continued for an additional 1 hour and the reaction was cooled to room temperature. The reaction mixture was then diluted with water (70 mL) and diluted with DCM (3 × 50 mL). The combined organics were then dried over sodium sulphate and concentrated to dryness in vacuo. The resulting oil was purified by flash chromatography with Rf of 0.19 using hexane: ethyl acetate (5: 1) as eluent, stained blue with 'Anisaldehyde stain ^* . 3.5 g of a colorless oil were obtained. It was expected to be 100% pure and sent to the next step without any further analysis.

^* 'Anisaldehyde stain "Recipe in absolute ethanol was added to the 135 ㎖ p- anisaldehyde in 5 ㎖ concentrated sulfuric acid, glacial acetic acid and 1.5 ㎖ 3.7 ㎖ of. The solution was then stirred vigorously to make it uniform.

b) {3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo-imi Dazolidin-1-yl} -acetic acid tert-butyl ester (35A)

Of 4- (3-isocyanato-benzyl) -2H-phthalazin-1-one (2) (3.12 mmol) in anhydrous DCM (80 mL) in the presence of activated 4 ′ molecular sieve (about 5 g). To the suspension was added 2- (tert-butoxycarbonylmethyl-amino) -propionic acid methyl ester (33A) (1.07 g, 4.7 mmol). The reaction was then stirred at ambient temperature for 48 hours. HPLC analysis suggested that there was a mixture of cyclic and acyclic ureas in a 1: 1 ratio. The mixture was filtered and concentrated in vacuo. The crude oil was then flash chromatographed using hexane: ethyl acetate (1: 1), Rf = 0.09 as eluent. A single peak was obtained in the LC-MS analysis (1.3 g, 95% purity), so no further purification was necessary; m / z (LC-MS, ESP), RT = 3.66 min (M + H) 481-Compound cyclized on column.

c) {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo-imi Dazolidin-1-yl} -acetic acid (36A)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4 in dry DCM (20 mL) Trifluoroacetic acid (2 mL, about 27 mmol) was added dropwise over 2 minutes to a solution of -dioxo-imidazolidin-1-yl} -acetic acid tert-butyl ester (35A) (1.3 g, 2.7 mmol). It was. After stirring overnight, the reaction mixture was concentrated to dryness and treated by flash chromatography. Eluent was 1% acetic acid in ethyl acetate. The title compound was isolated as a white solid. A single peak was obtained in the LC-MS analysis (0.680 mg, 100% purity), so no further purification was needed; m / z (LC-MS, ESN), RT = 2.95 min (M-H) 423

d) synthesis of library compounds

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4- in DCM (5 mL) In a suspension of dioxo-imidazolidin-1-yl} -acetic acid (36A) (145 mg, 0.34 mmol), HBTU (0.32 g, 0.85 mmol), Hunig base (0.85 mmol) and the appropriate amine (0.85 mmol) Was added. After the reaction mixture was stirred for 18 hours, the resulting compound was purified by preparative HPLC.

e) {3- [2-fluoro-5- (4-oxo-3,4-dihydro) by an alternative route using 2-amino-2-methyl-propionic acid methyl ester instead of DL-alanine methyl ester -Phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4-dioxo-imidazolidin-1-yl} -acetic acid (36B), followed by part d) Coupling to the appropriate amine as discussed in. The resulting compound was purified by preparative HPLC.

* Long method: Waters ZQ LC-MS system No. 2 operating in electrospray ionization mode. LAA 254; Mobile phase A: 0.1% formic acid in water; Mobile phase B: 0.1% formic acid in acetonitrile; Column: Genesis C18 4 μm 50 × 4.6 mm

gradient:

Flow rate: 2.0 ml / min

f) 1- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl]-[1,3,5] triazinane-2,4 , 6-trion (38)

Ethyl allophanate (52 mg, 0.34 g) in a suspension of 4- (3-isocyanato-benzyl) -2H-phthalazin-1-one (2) (0.37 mmol) in anhydrous DMA (1 mL) Was added. The suspension was heated to 160 ° C. for 1 hour and then cooled to room temperature to give a dark brown suspension. The solid was filtered and sampled, which showed about 80% purity RT = 3.56 min. The material was then purified by preparative HPLC purification.

Long method (see above)

g) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo -Imidazolidin-1-yl} -acetamide (40a)

(i) {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo- Imidazolidin-1-yl} -acetic acid methyl ester (39)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4- in methanol (4 mL) To a solution of dioxo-imidazolidin-1-yl} -acetic acid (36A) (0.04 g, 0.094 mmol) was added concentrated sulfuric acid (1 mL), stirred at 70 ° C. for 90 minutes and then cooled to room temperature. I was. The reaction mixture was then concentrated in vacuo, diluted with water (10 mL) and extracted with ethyl acetate (2 × 10 mL). The organic layer was then dried over sodium sulphate and concentrated in vacuo to give a colorless oil. Obtaining a single peak in LC-MS (52 mg, 95% purity), no further purification was needed; m / z (LC-MS, ESP), RT = 3.24 min (M + H) 439;

(ii) {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5 methyl-2,4-dioxo-imi Dazolidin-1-yl} -acetic acid (40a)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo-imidazoli Din-1-yl} -acetic acid methyl ester (39) (0.03 g, 0.068 mmol) was dissolved in a solution of 7N ammonia in methanol (2 mL, 14 mmol) and placed in a sealed tube. The reaction was then heated to 60 ° C. for 18 hours. The resulting solution was concentrated under vacuum and taken up on a 1 ml silica cartridge. The material eluted with pure ethyl acetate. A pure amide product was obtained with an Rf of 0.65. To obtain a single peak in LC-MS (8 mg, 100% purity), no further purification was necessary; m / z (LC-MS, ESP), RT = 2.84 min (M + H) 424;

h) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo Imidazolidin-1-yl} -N-methyl-acetamide (40b) and 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazine- 1-ylmethyl) -phenyl] -5-methyl-2,4-dioxo-imidazolidin-1-yl} -N, N-dimethyl-acetamide (40c)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-2,4- in DCM (0.5 mL) HBTU (15 mg, 0.04 mmol), Hunig base (0.85 mmol) and the appropriate amine (0.06 mmol) in a suspension of dioxo-imidazolidin-1-yl} -acetic acid (36A) (14 mg, 0.032 mmol) Was added. The reaction mixture was stirred for 18 hours and then submitted to preparative HPLC purification.

Long method (see above)

i) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4- Dioxo-imidazolidin-1-yl} -acetamide (40d)

(i) {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4-di Oxo-imidazolidin-1-yl} -acetic acid methyl ester

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2 in methanol (2 mL), To a solution of 4-dioxo-imidazolidin-1-yl} -acetic acid (36B) (0.044 g, 0.10 mmol) was added concentrated sulfuric acid (0.25 mL), heated at 70 ° C. for 90 minutes and then at room temperature. Cooled. The reaction mixture was then concentrated in vacuo, diluted with water (5 mL) and then extracted with ethyl acetate (2 × 10 mL). The organic layer was then dried over magnesium sulfate and concentrated in vacuo to give a colorless oil. Obtaining a single peak in LC-MS (42 mg, 91% purity), no further purification was needed; m / z (LC-MS, ESP), RT = 3.31 min (M + H) 453;

(ii) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4 -Dioxo-imidazolidin-1-yl} -acetamide (40d)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4-dioxo-imi Dazolidin-1-yl} -acetic acid methyl ester (0.025 g, 0.055 mmol) was dissolved in a solution of 7N ammonia in methanol (3 mL, 21 mmol) and heated in a sealed tube for 18 hours. The resulting solution was concentrated under vacuum and submitted to preparative HPLC purification.

Long method (see above)

j) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4- Dioxo-imidazolidin-1-yl} -N-methyl-acetamide (40e) and 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthala Jin-1-ylmethyl) -phenyl] -5,5-dimethyl-2,4-dioxo-imidazolidin-1-yl} -N, N-dimethyl-acetamide (40f)

{3- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5,5-dimethyl-2 in DCM (1 mL), In a suspension of 4-dioxo-imidazolidin-1-yl} -acetic acid (36B) (20 mg, 0.045 mmol) HBTU (19 mg, 0.06 mmol), Hunig base (0.85 mmol) and the appropriate amine (0.06 mmol) was added. The reaction mixture was stirred for 18 hours and then submitted to preparative HPLC purification.

Long method (see above)

Example  7

(a) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -ureido} succinic acid (42)

4- (3-isocyanato-benzyl) -2H-phthalazin-1-one (2) (1.4 g, 4.75 in anhydrous DCM (30 mL) in the presence of activated 4 ′ molecular sieve (about 2 g) DL-aspartic acid (0.60 g, 5.0 mmol) was added to the suspension, followed by triethylamine (l0.0 mmol). The reaction was stirred at rt for 18 h. The reaction mixture was then diluted with bicarbonate solution (20 mL) and the mixture was filtered while removing unwanted urea byproducts. The aqueous layer contained the title compound and was washed with DCM (2 × 10 mL). The aqueous layer was then concentrated in vacuo to yield a beige solid, and HPLC suggested that this was a mixture of components (41 & 42).

(ii) The mixture of components 41 & 42 was diluted with water (100 mL) and HCl (about 1 mL, 1 N), heated in an open beaker and evaporated to dryness. This resulted in a brown solid, LC-MS showing greater than 90% purity for the title compound. Obtaining a single peak in LC-MS (1.8 g, 90% purity), no further purification was needed; m / z (LC-MS, ESP), RT = 3.54 min (M + H) 411.

(b) Synthesis of Library Compounds

{1- [2-Fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -2,5-dioxo-imid in DCM (5 mL) To a suspension of dazolidin-4-yl} -acetic acid (42) (145 mg, 0.34 mmol) was added HBTU (0.32 g, 0.85 mmol), Hunig base (0.85 mmol) and the appropriate amine (0.85 mmol). The reaction mixture was stirred at rt for 18 h and then purified by preparative HPLC purification.

Example  8

(a) 2- {3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -urido} -propionic acid (45)

D-aspartic acid (0.455 g, in a suspension of 4- (3-isocyanato-benzyl) -2H-phthalazin-1-one (2) (1.4 g, 4.7 mmol) in anhydrous DCM (40 mL), 5.0 mmol), then triethylamine (1.4 mL, l0.0 mmol) was added. The reaction was stirred for 4 days at room temperature. The reaction mixture was then filtered. The filtrate was then diluted with water (20 mL) and washed twice with DCM (2 × 20 mL). The combined DCM layer was dried over sodium sulfate and then concentrated in vacuo to afford a crude oil, which was subjected to flash chromatography treatment with pure ethyl acetate as eluent to remove impurities and then eluent to remove desired components. Flash chromatography was performed with ethyl acetate: methanol (1: 1) as Rf 0.2 in 1: 1 ethyl acetate: methanol. A single peak was obtained in the LC-MS analysis (0.79 g), so no further purification was necessary; m / z (LC-MS, ESP), RT = 2.70 (M + H) 384.

(b) 3- [2-fluoro-5- (4-oxo-3,4-dihydro-phthalazin-1-ylmethyl) -phenyl] -5-methyl-imidazolidine-2,4 Dion (5b ')

To a premixed solution of trimethyl acetyl chloride (240 mg, 2.0 mmol) and triethylamine (0.35 mL, 2.5 mmol), 2- {3- [2-fluoro-5- (4-oxo-3,4-di Hydro-phthalazin-1-ylmethyl) -phenyl] -urido} -propionic acid (45) (0.76 g, 2.0 mmol) was added. The reaction proceeded slowly to the desired product with stirring at room temperature for 4 days. The reaction mixture was then concentrated in vacuo and the crude material submitted to preparative HPLC [40 mg isolated, 95% purity, RT (min): 4.35].

Example  9

To assess the inhibitory action of the compounds, IC ₅₀ values were determined using the following assays [Dillon, et al., JBS . , 8 (3), 347-352 (2003)].

By varying the concentration of inhibitor added in 96 well FlashPlates ™ (NEN, UK), mammalian PARP isolated from HeLa cell nuclear extracts was treated with Z-buffer [25 mM Hepes (Sigma); 12.5 mM MgCl ₂ (Sigma); 50 mM KCl (Sigma); 1 mM DTT (Sigma); 10% glycerol (sigma); 0.001% NP-40 (Sigma); pH 7.4]. All compounds were diluted in DMSO to get a final assay concentration of 10 to 0.01 μM, with DMSO having a final concentration of 1% per well. The total assay volume per well was 40 μl.

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

After incubation, the reaction was quenched by adding 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. Recorded values are counts per minute (cpm) after counting each well for 30 seconds.

Enzyme activity (%) for each compound was then calculated using the following formula:

IC ₅₀ values (concentrations where 50% of enzyme activity was inhibited) were calculated, which were usually measured over different concentration ranges from 10 μM to 0.001 μM. These IC ₅₀ values were used as comparisons to confirm increased compound potency.

All compounds tested had an IC ₅₀ of less than 0.1 μM. The following compounds had an IC ₅₀ of less than 0.1 μM: 5e-5j, 9a, 9c, 13a-d, 35B, 37Aa-Ac, 37Ba, 37Bb, 37Be-Bg, 37Bi, 37Bl, 39, 40a, 43a, 43c, 43d , 43f, 43 g, 5b '.

Synergistic factor (PF ₅₀ ) for the compound was calculated as the ratio of IC ₅₀ of control cell growth divided by the PA ₅₀ inhibitor + IC ₅₀ of cell growth. Growth inhibition curves for both control and compound treated cells are shown in the presence of alkyl methanesulfonate (MMS). Test compounds were used at fixed concentrations of 0.2 or 0.5 micromolar. The concentration of MMS was in the range of 0-10 μg / ml. Cell growth was assessed using the sulfohodamine 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 a 100 μl volume into each well of a 96 well microtiter plate with a flat bottom and incubated at 37 ° C. for 6 hours. Cells were replaced only with media at a final concentration of 0.5, 1 or 5 μM or with PARP inhibitor containing media. After the cells were grown for an additional hour, various concentration ranges (typically 0, 1, 2, 3, 5, 7 and 10 μg / ml) of MMS were added to untreated cells or PARP inhibitor treated cells. Only cells treated with PPAR inhibitors were used for the evaluation of growth inhibition by PARP inhibitors.

After leaving the cells for an additional 16 hours, the medium was replaced and the cells were grown at 37 ° C. for an additional 72 hours. The medium was then removed and the cells fixed with 100 μl of ice cold 10% (w / v) trichloroacetic acid. The plates were incubated at 4 ° C. for 20 minutes and then washed four times with water. Each cell well was then contaminated with 0.4% (w / v) SRB in 100 μl of 1% acetic acid for 20 minutes, followed by 4 washes with 1% acetic acid. The plate was then dried at room temperature for 2 hours. 100 μl of 10 mM Tris base was added to each well to lyse the dye from the contaminated cells. The plate was gently shaken and left at room temperature for 30 minutes before the optical concentration was measured at 564 nM on a Microquant microtiter plate reader.

Most of the compounds tested had at least one PF ₅₀ at 200 nM.

Solubility Assay

Typical assays that can be used to evaluate the solubility of the compounds of the present invention are as follows. The solubility of the compound was assessed in water or phosphate buffered saline (pbs) at pH 7.4. The samples were all allowed to equilibrate in solvent (with shaking) at room temperature for 20 hours. After this period, the sample was visually inspected to determine the presence / absence of undissolved solids. Samples can be centrifuged or filtered as needed to remove insoluble material, and solutions can be analyzed to determine the solubility of DS while diluting both aqueous and DMSO samples to a concentration similar to DMSO. The peak area obtained by HPLC (using a diode array detector) from the sample can be compared to the peak area obtained from DMSO solution (diluted to the same concentration as the sample) and quantified in consideration of the sample weight taken for initial dissolution. It is assumed that the sample will be completely dissolved in DMSO at the concentration used for the test.

By comparing the ratio of the peak areas, the solubility can be calculated from the concentration of the original sample.

Manufacture of samples

About 1 mg of sample was accurately weighed into a 4 ml glass vial and exactly 1.0 ml of water, aqueous buffer or DMSO was added by pipette. Each vial was sonicated for up to 2 minutes to aid in dissolving the solids. Samples were kept at room temperature for 20 hours with shaking on an orbital shaker. After this period the vials were inspected to determine the presence / absence of undissolved solids. Samples should be centrifuged or filtered through a 0.45 μm filter, if necessary, to remove insoluble material and, if appropriate, all samples should be diluted with DMSO and the filtrate analyzed to determine the compound concentration in solution. 20 μl are injected onto HPCL using the method shown below, with all samples injected twice. The maximum solubility that could be measured using this method was usually 1.0 mg / ml, and the weight was taken by dividing by the volume of solvent used.

Analysis technology

Typically, samples were LC / MS treated using a Waters Micromass ZQ instrument (or equivalent) with the following test parameters.

Waters Micromass ZQ in Cationic Mode

injection at m / z 100 to 800

Mobile phase A-0.1% aqueous formic acid

Mobile Phase B-0.1% Formic Acid in Acetonitrile

Column-Jones Chromatography Genesis 4 μ C18 Column, 4.6 × 50 mm

Flow rate-2.0 ml / min

Injection volume-30 μl injection into 20 μl loop

Gradient-starting at 95% A / 5% B, rising to 95% B after 4 minutes, holding there for 4 minutes, then returning to starting conditions (which can be changed to better isolate the peak if necessary)

PDA detection scans from 210 to 400 nm

Quantification of Sample

Initial testing of sample vials containing an aqueous dilution indicates whether the compound is dissolved in its buffer at that concentration. If it does not dissolve it should be reflected in the concentration obtained in solution by HPLC / MS. If the solution is clear, the concentration in the aqueous solvent should be similar to that in DMSO, and if no decomposition of the compound occurs, this should be shown on the chromatogram.

Since it is assumed that the sample is completely dissolved in DMSO, the peak size obtained from that sample will reflect 100% solubility. Assuming the dilution of all samples is the same, the solubility expressed in mg / ml is (area from pbs solution / area from DMSO solution) × (original weight / dilution in DMSO solution).

Stability test

Typical assays that can be used to assess the stability of the compounds of the present invention are as follows. The stability of the compounds was evaluated in various aqueous solutions and phosphate buffered saline (pbs). Samples will be tested at nominal pH 2, 7.4 (pbs) and 9. This value was chosen to reflect the conditions encountered in the intestine (about pH 2 to about pH 9) and in plasma (nominal pH 7.4) during digestion. Samples were dissolved in methanol / DMSO to prepare mother liquor. The mother liquor was then diluted to give an aqueous solution of nominal pH 2, 7.4 and 9. Samples were analyzed immediately to obtain initial values for purity and possible related compounds. Samples were then kept (typically) at room temperature and reanalyzed after 2 hours, 6 hours, 24 hours and 2 days (nominal).

The chromatogram of the initial sample and the chromatogram of the sample in aqueous buffer after a predetermined time can be compared to assess the stability of the compound in this aqueous buffer over the test period.

Sample Preparation and Analysis

About 5-6 mg of sample was accurately measured in a 4 ml glass vial and about 2 ml of methanol was added thereto. If the solution was not completely dissolved in this organic solvent, an additional 0.5 to 1.0 mL of DMSO was added and the final solution concentration should be about 2.0 mg / mL. This 2 mg / ml organic solution is then (a) with water for use as a 'initial' sample, (b) very dilute HCl at about pH 2, (c) pbs at pH 7.4, and (d) Dilute to 1 + 3 with very dilute NaOH at about pH 9. The pH of each dilution was then checked and recorded and, if not close to a predetermined value, the pH can be adjusted, if appropriate, with dilute acid or alkali. These dilutions were prepared at predetermined intervals after the 'initial' sample to be delayed by HPLC analysis. All samples must be diluted 50/50 with DMSO prior to injection on HPLC.

After the sample was initially kept at room temperature for 2 hours, the subsample as described above was diluted 50/50 with DMSO prior to injection. 20 μl was injected on HPLC using the method shown below, with all samples injected twice. The procedure was repeated after 6 hours, 24 hours and 2 days (nominal time interval).

Analysis technology

Typically, samples were LC / MS treated using a Waters Micromass ZQ instrument (or equivalent) with the following test parameters.

Waters Micromass ZQ in Cationic Mode

injection at m / z 150 to 900

Mobile phase A-0.1% aqueous formic acid

Mobile Phase B-0.1% Formic Acid in Acetonitrile

Column-Jones Chromatography Genesis 4 μ C18 Column, 4.6 × 50 mm

Flow rate-2.0 ml / min

Injection volume-30 μl injection into 20 μl loop

Gradient-starting at 95% A / 5% B, rising to 95% B after 5 minutes, holding there for 4 minutes, then returning to starting conditions (which can be changed to better isolate the peak if necessary)

PDA detection scans from 210 to 400 nm

Stability evaluation

After any given time interval the chromatogram peak areas of the samples at various pHs were compared with the areas from the initial analysis at 0 hours. DS peaks should be quantified as% of initial sample and values should be tabulated.

Claims (20)

A compound of formula (I) Formula I In the above formula, A and B together represent a fused aromatic ring optionally substituted; D is (i) Wherein Y ¹ is selected from CH and N, Y ² is selected from CH and N, and Y ³ is selected from CH, CF and N; And (ii) Wherein Q is O or S Is selected from; R ^D is [Wherein, R ^N1 is selected from H and optionally substituted C _{1 -10} alkyl; X is selected from a single bond, NR ^N2 , CR ^C3 R ^C4 and C═O; R ^N2 is selected from H and optionally substituted C _{1 -10} alkyl; R ^C3 and R ^C4 are independently H, R, C (= O) OR (wherein, R is an optionally substituted C _{1 -10} alkyl, optionally substituted C _{5 -20} aryl or optionally substituted hetero C _{3 -20} Is a cyclic); Y is selected from NR ^N3 and CR ^C1 R ^C2 ; R ^C1 and R ^C2 are independently selected from H, R, C (= O) OR (wherein, R is an optionally substituted C _{1 -10} alkyl, optionally substituted C _{5 -20} aryl or optionally substituted hetero C _{3 -20} Is a cyclic); R ^C1 and R ^C2 together with the carbon atom attached thereto may form a spiro-fused C _{5 -7} carbocyclic or heterocyclic ring optionally substituted and; If X is a single bond, R ^N1 and R ^C2 together with the N and C atoms attached thereto an optionally substituted C _{5 -7} may form a heterocyclic ring; When X is CR ^C3 R ^C4 , R ^C2 and R ^C4 may together form an additional bond such that a double bond is present between the atoms substituted with R ^C1 and R ^C3 . The compound according to claim 1, wherein the fused aromatic ring represented by -A-B- is benzene. The compound of claim 2, wherein the fused benzene ring is substituted with halo. The compound of claim 1, wherein D is selected from phenylene, fluoro-phenylene, pyridylene, fluoro-pyridylene, furanylene and thiophenylene. The compound of claim 4, wherein D is The compound selected from. The method of claim 5, wherein D is The compound selected from. The compound of claim 6, wherein D is Compound. 8. The compound of claim 1, wherein X is selected from a single bond, NR ^N2 and CR ^C3 R ^C4 , and Y is CR ^C1 R ^C2 . The compound of claim 8, wherein X is a single bond. The compound of claim 9, wherein at least one of R ^N1 , R ^C1 and R ^C2 is not hydrogen. The compound of claim 9 or claim 10 wherein, R ^C1 and R ^C2 is of more than 1 is selected from C _{1 -4} alkyl. Claims 9 to 11 of the method according to any of the preceding, R ^N1 is at the carboxy terminus, an amido group or ester group to the substituted C _{1 -4} alkyl. 13. The method of claim 12, wherein the amino substituent of the amido group is cyclic with the nitrogen atom attached thereto. The compound of claim 8, wherein X is N ^R2 and R ^C1 , R ^C2 and R ^N2 are H. 10. The compound of claim 8, wherein X is CR ^C3 R ^C4 , and R ^C1 , R ^C2 , R ^C3 and R ^C4 are H. 10. A pharmaceutical composition comprising the compound of any one of claims 1 to 15 and a pharmaceutically acceptable carrier or diluent. Use of the compound of claim 1 in a method of treating a human or animal body. Of the compound of any one of claims 1 to 15, (a) prevention of poly (ADP-ribose) chain formation by inhibiting the activity of cellular PARP (PARP-1 and / or PARP-2); (b) vascular disease; Septic shock; Ischemic damage to both the brain and cardiovascular; Reperfusion injury in both the brain and cardiovascular; Neurotoxicity, including acute and chronic treatment for stroke and Parkinson's disease; Hemorrhagic shock; Inflammatory diseases such as arthritis, inflammatory bowel disease, ulcerative colitis and Crohn's disease; Multiple sclerosis; As well as treatment of secondary effects of diabetes, as well as acute treatment of diseases that are ameliorated by cardiovascular surgery following cytotoxicity or inhibition of PARP activity; (c) use as adjuvant in cancer therapy, or as adjuvant to increase drug efficacy on tumor cells for treatment with ionizing radiation or chemotherapy Use in the manufacture of a medicament for Use of a compound of any one of claims 1 to 15 in the manufacture of a medicament for the treatment of a cancer lacking homologous recombination (HR) dependent DNA double strand break (DSB) repair activity. . A method of treating a disease ameliorated by inhibition of PARP, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1 to 15.