HK1166076B - Isoquinolin-1 (2h) - one derivatives as parp-1 inhibitors - Google Patents

Description

isoquinolin-1 (2H) -one derivatives as PARP-1 inhibitors

The present invention relates to substituted isoquinolin-1 (2H) -one derivatives that selectively inhibit the activity of poly (ADP-ribose) polymerase PARP-1 relative to poly (ADP-ribose) polymerase PARP-2. The compounds of the invention are therefore suitable for the treatment of diseases such as cancer, cardiovascular diseases, central nervous system injury and different forms of inflammation. The invention also provides methods of preparing these compounds, pharmaceutical compositions comprising these compounds, and methods of treating diseases using pharmaceutical compositions comprising these compounds.

Poly (ADP-ribose) polymerases belong to the 17-member family that catalyzes the addition of ADP-ribose units to DNA or to different receptor proteins, which affect a variety of cellular processes: replication, transcription, differentiation, gene regulation, protein degradation and spindle maintenance. PARP-1 and PARP-2 are the only enzymes in PARPs that are activated by DNA damage and are involved in DNA repair.

PARP-1 is a nuclear protein consisting of 3 domains: an N-terminal DNA-binding domain containing two zinc fingers, an auto-modifying domain, and a C-terminal catalytic domain. PARP-1 binds to DNA fragmentation Single Strands (SSBs) through the zinc-finger domain, cleaves NAD +, and attaches multiple ADP-ribose units to target proteins such as histones and various DNA repair enzymes. This results in a highly negatively charged target, which in turn leads to the melting and repair of damaged DNA through base excision repair pathways. In a mouse model where specific genes were artificially disrupted, deletion of PARP-1 impairs DNA repair, but it is not embryo lethal. Whereas double knockout PARP-1 and PARP-2 mice die early in embryonic development, it suggests that the two enzymes do not show fully overlapping functions. Enhanced PARP-1 expression and/or activity has been shown in different tumor cell lines, including malignant lymphoma, hepatocellular carcinoma, cervical cancer, colorectal cancer, leukemia. This may allow tumor cells to resist genotoxic stress and increase their resistance to DNA-damaging agents. Thus, it has been shown that inhibition of PARP-1 by small molecules sensitizes tumor cells to cytotoxic therapies (e.g., temozolomide, platinum, topoisomerase inhibitors, and radiation). A significant window (window) appears to exist between the ability of PARP inhibitors to achieve therapeutic benefit and undesirable side effects. However, the therapeutic use of combinations of PARP inhibitors with DNA damaging agents is not new and the use of these agents as monotherapy, especially in the case of a lack of background in tumor genetics in homologous recombinant DNA repair, represents a new approach. Individuals with heterozygous germline mutations in BRCA-1 or BRCA-2 homologous recombination repair genes exhibit a high risk of developing breast and other cancers when surviving. Tumors that appear in mutant vectors usually lose the wild-type allele and do not express functional BRCA-1 and BRCA-2 proteins.

Thus, the loss of these two proteins results in tumor-specific dysfunction in the repair of double-strand breaks caused by homologous recombination. It is known that when PARP-1 is inhibited, base excision repair is reduced and single strand breaks produced during the normal cell cycle persist. It has also been determined that the occurrence of an unrepaired break in a replication fork can form a double-stranded break, which is typically repaired by homologous recombination. Tumor cells that are deficient in homologous recombination repair, e.g., BRCA-1 and BRCA-2 mutants, are therefore highly sensitive to PARP inhibition compared to wild-type cells. This is consistent with the concept of synthetic lethality, where the two pathways are individually deficient to be harmless, but their combined deficiency becomes lethal: PARP inhibitors may be more effective in tumor patients with specific DNA repair defects, while the inhibitors do not affect normal heterozygous tissues. In addition to BRCA mutants representing the majority of inherited breast and ovarian cancers, the putative patient population also includes a significant proportion of sporadic cancers that have defects in homologous recombination repair, a phenomenon known as "brcataness". For example, the promoter of the methylated BRCA-1 or FANCF gene and the amplified EMSY gene, which encodes a BRCA-2 interacting protein. By reasonable inference of synthetic lethality of PARP and BRCA-1 and BRCA-2, it is possible that any gene deficiency not redundant in double strand break repair should be sensitive to PARP inhibition. For example, defects in ATM found in patients with T-cell prolymphocytic leukemia and B-cell chronic lymphocytic leukemia and breast cancer, as well as mutations in the CHK2 germ line identified in sarcomas, breast cancer, ovarian cancer and brain tumors, have also been shown to be synthetic lethal in combination with PARP defects and deficiencies in other known HR pathway proteins (including RAD51, DSS1, RAD54, RPA1, NBS1, ATR, CHK1, CHK2, FANCD2, FANCA and FANCC).

FANCC and FANCG mutations have been found in pancreatic cancer. FANCF promoters have been found that are methylated in ovarian, breast, cervical, and lung cancers. The first clinical evidence that BRCA-mutated cancers may be susceptible to PARP inhibitor monotherapy comes from preliminary phase I trial data of the oral small molecule PARP inhibitor AZD 2281. In the enriched BRCA mutant vector phase I population, a partial response was observed in 4 of 10 ovarian cancer patients in whom BRCA-1 mutation was confirmed. It is currently known that other PARP inhibitors, such as AG014699 and BSI-201, are combined with DNA damaging agents and used as single agents for BRCA deficient tumors in phase II clinical trials. Early indications were that these treatments showed low toxicity. In any event, compounds with high selectivity for PARP-1 are expected to be even less toxic in terms of chronic treatment schedules.

PARP-1 has also been implicated in angiogenesis. In particular, PARP-1 inhibition appears to result in a reduction in the accumulation of important regulators of transcriptional hypoxia-inducible factor 1 tumor cells adaptive to hypoxia.

The pro-inflammatory stimulus triggers the release of pro-inflammatory mediators that lead to the production of peroxynitrate and hydroxyl residues, which in turn produce DNA single strand breaks and subsequent activation of PARP-1. Over-activation of PARP-1 consumes NAD + and energy storage, ultimately leading to cell dysfunction and necrosis. This cellular suicide principle has been implicated in the pathological mechanism of the following disorders: stroke, myocardial ischemia, diabetes-related cardiovascular dysfunction, shock, traumatic central nervous system injury, arthritis, colitis, allergic encephalomyelitis and various other forms of inflammation. Of particular interest is that PARP-1 enhances nuclear factor kB-mediated transcription, which plays an important role in the expression of inflammatory cytokines, chemokines and inflammatory mediators.

The present invention relates to substituted isoquinolin-1 (2H) -one derivatives that selectively inhibit the activity of poly (ADP-ribose) polymerase PARP-1 relative to poly (ADP-ribose) polymerase PARP-2. The compounds described herein are therefore useful in the treatment of proliferative diseases such as cancer and cardiovascular diseases, nervous system injury and inflammation.

In the Journal of the Chemical Society, Perkin Transactions 1, (1977), (9), 959-65 on nitrogen heterocycles, 3-phenyl-1 (2H) -isoquinolinone is described. Isoquinoline-1 (2H) -ones with pharmacological activity are described in Science of Synthesis (2005), 15, 839-906. Certain patent applications describe isoquinoline derivatives for the treatment of glaucoma, EP389995, and for the treatment of arteriosclerosis and hyperlipoproteinemia, EP 591937. WO2002090334 in the name of KUDOS PHARM describes isoquinolone derivatives for inhibiting PARP activity. WO2008092292 describes methods of treating pathological conditions related to melatonin receptors using 2-substituted (2H) -isoquinolinones.

The present invention provides novel 1(2H) -isoquinolinones endowed with selective inhibitory activity against PARP-1, relative to PARP-2, and therefore useful in the treatment of cancer, cardiovascular diseases, nervous system injury and inflammation.

Accordingly, a first object of the present invention is to provide a compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein L is an optionally substituted straight or branched chain C₂-C₆Alkyl radical, C₃-C₇Cycloalkyl radicals, or heterocyclic groups C optionally substituted by containing the nitrogen atom to which it is bonded₁-C₆An alkyl group;

r and R₁Independently a hydrogen atom, an optionally substituted linear or branched C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl, heterocyclyl, aryl, heteroaryl or COR₅A group, or together with the nitrogen atom to which they are bonded, forms an optionally substituted heterocyclyl or heteroaryl group;

R₂and R₃Independently is a hydrogen or halogen atom; cyano, nitro, NHCOR₅、COR₅、NR₆R₇、NR₆COR₅、OR₈、SR₈、SOR₁₁、SO₂R₁₁、NHSOR₁₁、NHSO₂R₁₁、R₉R₁₀N-C₁-C₆Alkyl radical, R₉O-C₁-C₆Alkyl radicals, or optionally substituted, linear or branched C₁-C₆Alkyl radical、C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₇A cycloalkyl, heterocyclyl, aryl, or heteroaryl group;

R₄is optionally substituted straight or branched C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, aryl C₁-C₆Alkyl, aryl C₃-C₇Cycloalkyl, aryl C₂-C₆Alkenyl, aryl C₂-C₆Alkynyl, heterocyclic radical C₁-C₆Alkyl, heterocyclic radical C₃-C₇Cycloalkyl, heterocyclyl C₂-C₆Alkenyl, heterocyclyl C₂-C₆Alkynyl, heteroaryl C₁-C₆Alkyl, heteroaryl C₃-C₇Cycloalkyl, heteroaryl C₂-C₆Alkenyl, heteroaryl C₂-C₆An alkynyl group;

R₅is a hydrogen atom or NR₆R₈、OR₈、SR₈、R₉R₁₀N-C₁-C₆Alkyl radical, R₉O-C₁-C₆Alkyl, or optionally substituted straight or branched C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₇Cycloalkyl, heterocyclyl, aryl or heteroaryl groups;

R₆and R₇Independently a hydrogen atom, an optionally substituted linear or branched C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₇Cycloalkyl radical, R₉R₁₀N-C₂-C₆Alkyl radical, R₉O-C₂-C₆An alkyl, heterocyclyl, aryl or heteroaryl group, or R₆And R₇May form, together with the nitrogen atom to which they are bonded, an optionally substituted heterocyclyl group;

R₈is hydrogen, COR₆、SOR₁₁、SO₂R₁₁、R₉R₁₀N-C₂-C₆Alkyl or R₉O-C₂-C₆Alkyl radicals, or optionally substituted, linear or branched C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₇Cycloalkyl, heterocyclyl, aryl or heteroaryl group, wherein R₆As defined above;

R₉and R₁₀Independently of hydrogen, COR₅Or optionally substituted straight or branched C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₇Cycloalkyl, heterocyclyl, aryl or heteroaryl group, or R₉And R₁₀Together with the nitrogen atom to which they are bonded may form an optionally substituted heterocyclyl group, wherein R₅As defined above;

R₁₁is a hydrogen atom, NR₆R₇、OR₈、R₉R₁₀N-C₁-C₆Alkyl radical, R₉O-C₁-C₆Alkyl, or optionally substituted straight or branched C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₃-C₇Cycloalkyl, heterocyclyl, aryl or heteroaryl group, wherein R₆、R₇、R₈、R₉And R₁₀As defined above.

A second object of the present invention relates to a screening method for identifying compounds capable of binding several PARP proteins, as well as probes for use in such methods.

Accordingly, a screening method comprising the steps of:

a) providing a reaction mixture comprising:

the PARP protein isoforms studied,

A compound of formula (II):

wherein R is₁₃Is a hydrogen atom or a methyl group, B is (CH)₂)_n-NH group, wherein n is 2 to 6; m is 0 or 1 and X^-Is a counterion, and

serial dilutions of test compounds;

b) comparing the polarization signal produced in the absence of the test compound with the polarization signal produced in the presence of different concentrations of the test compound, and

c) the ability of the test compound to displace the compound of formula (II) as defined above is evaluated as indicated from the reduced level of fluorescence polarization.

The compounds of formula (II) as defined above are also an object of the present invention for use as probes in screening methods, and methods for preparing them.

The present invention also provides a method for the synthesis of isoquinolin-1 (2H) -one derivatives of formula (I) as defined above, by a process consisting of standard synthetic transformations.

As mentioned above, the inventors have found that the compounds of formula (I) as defined above are potent and selective PARP-1 inhibitors relative to PARP-2 and are therefore useful in the treatment of cancer, cardiovascular diseases, nervous system injuries and in anti-inflammatory therapy. Accordingly, the present invention also provides methods for treating diseases mediated by the PARP-1 protein.

Preferred methods of the invention are for treating diseases mediated by the PARP-1 protein selected from the group consisting of cancer, cardiovascular disease, nervous system injury and inflammation.

Another preferred method of the invention is for treating a specific type of cancer, including but not limited to: cancers such as bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer including small cell lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, and skin cancer including squamous cell carcinoma; hematopoietic cancers of the lymphoid lineage (lymphoblastic carcinomas) including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma; hematopoietic cancers of the myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndromes, and promyelocytic leukemia; carcinomas of mesenchymal origin including fibrosarcoma and rhabdomyosarcoma; central and peripheral nervous system cancers including astrocytomas, neuroblastoma, glioma, and schwannoma; other carcinomas including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma (keratoxanthoma), thyroid follicular carcinoma and kaposi's sarcoma.

Another preferred method of the invention is for the treatment of certain types of cardiovascular disease, including but not limited to: myocardial reperfusion injury, cardiomyopathy, and diabetic cardiovascular dysfunction.

Another preferred method of the invention is for treating certain types of central nervous system injuries, including but not limited to stroke, brain injury, and neurodegenerative disorders.

The invention further provides methods of treatment comprising the compounds of formula (I) for simultaneous, separate or sequential use in anticancer therapy in conjunction with radiation therapy or chemotherapeutic regimens.

In addition, the present invention provides a method for selectively inhibiting the activity of PARP-1 protein, said method comprising contacting said protein with an effective amount of a compound of formula (I).

The present invention also provides a pharmaceutical composition comprising 1 or more compounds of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, carrier or diluent.

The invention also provides pharmaceutical compositions comprising a compound of formula (I) in combination with known cytostatic or cytotoxic agents, antibiotic-type agents, alkylating agents, antimetabolite agents, hormonal agents, immunological agents, interferon-type agents, cyclooxygenase inhibitors (e.g., COX-2 inhibitors), matrix metalloproteinase inhibitors, telomerase inhibitors, tyrosine kinase inhibitors, anti-growth factor receptor agents, anti-HER agents, anti-EGFR agents, anti-angiogenesis agents (e.g., angiogenesis inhibitors), farnesyl transferase inhibitors, ras-raf signal transduction pathway inhibitors, cell cycle inhibitors, other cdks inhibitors, tubulin binding agents, topoisomerase I inhibitors, topoisomerase II inhibitors, and the like.

Furthermore, the present invention provides a product or kit comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof as defined above and 1 or more chemotherapeutic agents, in the form of a combined preparation, for simultaneous, separate or sequential use in anticancer therapy.

In yet another aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined above, for use as a medicament.

In addition, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined above for the manufacture of a medicament for the treatment of a disease mediated by the PARP-1 protein.

Finally, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined above for use in a method of treating a disease mediated by PARP-1 protein, preferably cancer, cardiovascular disease, nervous system injury and inflammation.

The invention also provides synthetic methods for preparing substituted derivatives of formula (I) by processes that include standard synthetic transformations.

If a chiral center or another form of an isomeric center is present in a compound of the present invention, all forms of the isomer, including enantiomers and diastereomers, are intended to be encompassed herein. Compounds containing chiral centers can be used as racemic mixtures, enantiomerically enriched mixtures or racemic mixtures can be separated by well-known techniques, and the individual enantiomers can be used alone. In the case of compounds containing unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of the invention.

The term "pharmaceutically acceptable salts" of the compounds of formula (I) means those salts which retain the biological effectiveness and properties of the parent compound. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid and the like, or with organic acids such as acetic acid, ascorbic acid, trifluoroacetic acid, propionic acid, glycolic acid, (D) or (L) lactic acid, (D) or (L) malic acid, oxalic acid, fumaric acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, benzoic acid, p-toluenesulfonic acid, salicylic acid, cinnamic acid, mandelic acid, tartaric acid, citric acid, succinic acid, isethionic acid and malonic acid.

Pharmaceutically acceptable salts of the compounds of formula (I) also include salts with inorganic or organic bases such as hydroxides, carbonates or bicarbonates of alkali or alkaline earth metals, especially sodium, potassium, calcium, ammonium or magnesium, acyclic or cyclic amines, preferably methylamine, ethylamine, diethylamine, triethylamine, piperidine and the like.

In a preferred embodiment of the screening method representing the second object of the present invention, the PARP protein and the compound of formula (II) as defined above are pre-mixed.

In another preferred embodiment of the screening method, the PARP protein and the test compound are premixed. In another preferred embodiment of the screening method, the PARP protein is PARP-1, PARP-2 and PARP-3. The term "PARP protein" includes full-length native proteins as well as fragments thereof.

Unless otherwise indicated, the present invention includes all isomers, tautomers, hydrates, solvates, complexes, carriers, N-oxides, and pharmaceutically acceptable salts of the compounds of the present invention when referring to the compounds of formula (I) per se as well as any pharmaceutical compositions thereof or any therapeutic treatment comprising the same.

In cases where a compound may exist in tautomeric forms, such as keto-enol tautomers, each tautomeric form is contemplated as being encompassed within the invention, whether it exists in equilibrium or predominantly in one form.

By the term halogen atom we mean a fluorine, chlorine, bromine or iodine atom.

For the term "straight or branched C₁-C₆By alkyl "we mean any group such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, n-hexyl and the like.

For the term "C₂-C₆By alkenyl ", we mean aliphatic C which contains at least one carbon-carbon double bond and which may be straight or branched₂-C₆A hydrocarbon chain. Representative examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-or 2-butenyl, and the like.

For the term "C₂-C₆By alkynyl "we mean aliphatic C containing at least one carbon-carbon triple bond and which may be straight or branched₂-C₆A hydrocarbon chain. Representative examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-or 2-butynyl, and the like.

For the term "C₃-C₇Cycloalkyl ", unless otherwise specified, we mean a 3-to 7-membered all-carbon (all-carbon) monocyclic ring which may contain 1 or more double bonds, but which does not contain a completely conjugated pi-an electronic system. Non-limiting examples of cycloalkyl groups are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene and cyclohexadiene.

By the term "heterocyclyl" we mean a 3-to 8-membered saturated or partially unsaturated carbocyclic ring in which 1 or more carbon atoms are replaced by heteroatoms selected from nitrogen, oxygen or sulfur. Non-limiting examples of heterocyclyl groups are, for example, pyran, pyrrolidine, pyrroline, imidazoline, imidazolidine, pyrazolidine, pyrazoline, thiazoline, thiazolidine, dihydrofuran, tetrahydrofuran, 1, 3-dioxolane, piperidine, piperazine, morpholine, and the like.

The term "aryl" means a mono-, di-or poly-carbocyclic hydrocarbon having from 1 to 4 ring systems, which are optionally further fused to each other or connected by single bonds, wherein at least one carbocyclic ring is "aromatic", wherein the term "aromatic" means a completely conjugated pi-electron bond system. Non-limiting examples of such aryl groups are phenyl, alpha-or beta-naphthyl or biphenyl groups.

The term "heteroaryl" as used herein means an aromatic heterocyclic ring, typically a 5-to 8-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O or S; heteroaryl rings may optionally be further fused or linked to aromatic and non-aromatic carbocyclic and heterocyclic rings. Non-limiting examples of such heteroaryl groups are, for example, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, imidazolyl, thiazolyl, isothiazolyl, pyrrolyl, phenyl-pyrrolyl, furanyl, phenyl-furanyl, oxazolyl, isoxazolyl, pyrazolyl, thienyl, benzothienyl, isoindolyl, benzimidazolyl, indazolyl, quinolinyl, isoquinolinyl, 1,2, 3-triazolyl, 1-phenyl-1, 2, 3-triazolyl, 2, 3-dihydroindolyl, 2, 3-dihydrobenzofuranyl, 2, 3-dihydrobenzothienyl; benzopyranyl, 2, 3-dihydrobenzoxazinyl, 2, 3-dihydroquinoxalinyl, and the like.

According to the invention, any of the above L, R-R unless otherwise indicated₁₁The groups may optionally be substituted at any free position by 1 or more groups, for example 1 to 6Substituted with groups independently selected from: halogen, nitro, oxo (═ O), cyano, C₁-C₆Alkyl, polyfluoroalkyl, polyfluoroalkoxy, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, hydroxyalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, C₃-C₇Cycloalkyl, hydroxy, alkoxy, aryloxy, heterocyclyloxy, methylenedioxy, alkylcarbonyloxy, arylcarbonyloxy, cycloalkenyloxy, heterocyclylcarbonyloxy, alkyleneaminooxy, carboxy, alkoxycarbonyl, aryloxycarbonyl, cycloalkyloxycarbonyl, heterocyclylalkyloxycarbonyl, amino, ureido, alkylamino, dialkylamino, arylamino, diarylamino, heterocyclylamino, formylamino, alkylcarbonylamino, arylcarbonylamino, heterocyclylcarbonylamino, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, arylaminocarbonyl, heterocyclylaminocarbonyl, alkoxycarbonylamino, hydroxyaminocarbonyl, alkoxyimino, alkylsulfonylamino, arylsulfonylamino, heterocyclylsulfonylamino, formyl, alkylcarbonyl, arylcarbonyl, cycloalkylcarbonyl, heterocyclylcarbonyloxy, heterocyclylcarbonylamino, carboxyl, alkylcarbonylamino, alkoxycarbonyl, carbonyl halide, carbonyl, Heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, alkylaminosulfonyl, dialkylaminosulfonyl, arylaminosulfonyl, heterocyclylaminosulfonyl, arylthio, alkylthio, phosphonate, phosphonic acid, phosphonate, and alkylphosphonate. Conversely, each of the above substituents may be further substituted, where appropriate, with 1 or more of the foregoing groups, as described above "independently selected from: "substituted by the groups listed thereafter.

Preferably, such substituents are selected from halogen, cyano, nitro, NHCOR₅、COR₅、NR₆R₇、NR₆COR₅、OR₈、SR₈、SOR₁₁、SO₂R₁₁、NHSOR₁₁、NHSO₂R₁₁、R₉R₁₀N-C₁-C₆Alkyl radical, R₉O-C₁-C₆Alkyl, optionally furtherStep (b) substituted straight or branched chain C₁-C₆Alkyl radical, C₃-C₇Cycloalkyl, heterocyclyl, aryl and heteroaryl groups, wherein R₅、R₆、R₇、R₈、R₉、R₁₀And R₁₁As defined above.

By the term polyfluoroalkyl or polyfluoroalkoxy we mean any of the above straight or branched C₁-C₆Alkyl or alkoxy substituted with more than one fluorine atom, e.g., trifluoromethyl, trifluoroethyl, 1, 1, 1, 3, 3, 3-hexafluoropropyl, trifluoromethoxy, and the like.

By the term hydroxyalkyl we mean any of the above C's containing a hydroxyl group₁-C₆Alkyl groups such as hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl and the like.

From all the above descriptions, it is clear to the skilled person that any group named compound name shall conventionally mean consisting of a moiety derived therefrom, e.g. arylamino is an amino group further substituted by aryl, wherein aryl is as defined above.

Also, any term, for example, alkylthio, alkylamino, dialkylamino, alkoxycarbonyl, alkoxycarbonylamino, heterocyclylcarbonyl, heterocyclylcarbonylamino, cycloalkyloxycarbonyl and the like includes wherein alkyl, alkoxy, aryl, C₃-C₇Cycloalkyl and heterocyclyl moieties are groups as defined above.

Preferably, the present invention provides a compound of formula (I) as defined above or a pharmaceutically acceptable salt thereof, said compound or salt being characterized in that L represents an optionally substituted linear or branched C₂-C₆Alkyl, or heterocyclic group C optionally substituted by containing the nitrogen atom to which it is bonded₁-C₆An alkyl group;

r and R₁Independently a hydrogen atom, an optionally substituted linear or branched C₁-C₆Alkyl or COR₅A group, or together with the nitrogen atom to which they are bonded, forms an optionally substituted heterocyclyl or heteroaryl group;

R₂and R₃Independently is a hydrogen or halogen atom; nitro, amino, hydroxy, COR₅Or optionally substituted straight or branched C₁-C₆An alkyl group;

R₄is an optionally substituted aryl or heteroaryl group;

R₅is optionally substituted straight or branched C₁-C₆An alkyl group.

More preferably, there is provided a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, said compound being characterized in that L is optionally substituted straight or branched C₂-C₄An alkyl group, or a heterocyclic group optionally substituted by containing a nitrogen atom to which it is bonded;

r and R₁Independently a hydrogen atom, an optionally substituted linear or branched C₁-C₄Alkyl or COR₅A group, or together with the nitrogen atom to which they are bonded, form an optionally substituted piperidinyl, pyrrolidinyl, piperazinyl, or pyrrolyl group;

R₂and R₃Independently is a hydrogen, chlorine or fluorine atom; or nitro, amino, hydroxy, COR₅Or optionally substituted straight or branched C₁-C₄An alkyl group;

R₄is an optionally substituted phenyl or thienyl group;

R₅is optionally substituted straight or branched C₁-C₄An alkyl group.

Particularly preferred compounds according to the invention are listed below:

4- (2-amino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

n- [2- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -acetamide,

3.4- (3-amino-propoxy) -3-phenyl-2H-isoquinolin-1-one,

n- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -acetamide,

3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

6.3-phenyl-4- (2-piperazin-1-yl-ethoxy) -2H-isoquinolin-1-one,

7.4- [2- (4-acetyl-piperazin-1-yl) -ethoxy ] -3-phenyl-2H-isoquinolin-1-one,

n- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -benzamide,

9.3- (3-methoxy-phenyl) -4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

11.4- (2-amino-ethoxy) -3- (3-methoxy-phenyl) -2H-isoquinolin-1-one,

12.4- (3-methylamino-propoxy) -3-phenyl-2H-isoquinolin-1-one,

13.3-phenyl-4- (2-pyrrol-1-yl-ethoxy) -2H-isoquinolin-1-one,

14.3-phenyl-4- (3-piperazin-1-yl-propoxy) -2H-isoquinolin-1-one,

15.3-phenyl-4- (3-pyrrol-1-yl-propoxy) -2H-isoquinolin-1-one,

16.4- (2-methylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

17.4- (2-dimethylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

18.4- (3-diethylamino-propoxy) -3-phenyl-2H-isoquinolin-1-one,

19.4- [2- (4-methyl-piperazin-1-yl) -ethoxy ] -3-phenyl-2H-isoquinolin-1-one,

20.4- [3- (4-methyl-piperazin-1-yl) -propoxy ] -3-phenyl-2H-isoquinolin-1-one,

21.3-phenyl-4- (3-pyrrolidin-1-yl-propoxy) -2H-isoquinolin-1-one,

22.4- (4-amino-butoxy) -3-phenyl-2H-isoquinolin-1-one,

3- (4-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

24.4- (2-amino-ethoxy) -3- (4-methoxy-phenyl) -2H-isoquinolin-1-one,

25.4- (2-amino-ethoxy) -7-fluoro-3- (4-methoxy-phenyl) -2H-isoquinolin-1-one,

26.4- (2-amino-ethoxy) -3- (4-hydroxy-phenyl) -2H-isoquinolin-1-one,

27.3- (4-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

28.4- (2-amino-ethoxy) -7-fluoro-3- (4-hydroxy-phenyl) -2H-isoquinolin-1-one,

29.4- (2-amino-ethoxy) -3-thiophen-3-yl-2H-isoquinolin-1-one,

30.7-acetyl-3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

31.6-Nitro-3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

32.4- (2-amino-ethoxy) -3- (3-hydroxy-phenyl) -2H-isoquinolin-1-one,

33.3- (3-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

34.6-Nitro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

35.7-Nitro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

36.7-amino-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

37.7-chloro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

38.7-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

39.4- (2-amino-ethoxy) -7-fluoro-3-phenyl-2H-isoquinolin-1-one,

40.6-amino-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

41.6-chloro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

42.6-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

43.7-fluoro-3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

44.3- (3-chloro-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

45.7-fluoro-3- (3-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

46.4- (2-amino-ethoxy) -7-fluoro-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one,

47.4- (2-amino-ethoxy) -7-fluoro-3- (3-hydroxy-phenyl) -2H-isoquinolin-1-one,

48.4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -2H-isoquinolin-1-one,

49.4- (2-amino-ethoxy) -5-methyl-3-phenyl-2H-isoquinolin-1-one,

50.4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -7-fluoro-2H-isoquinolin-1-one,

51.3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

52.4- (2-diethylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

53.3-phenyl-4- (2-pyrrolidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

54.4- (2-morpholin-4-yl-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

55.5-methyl-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

56.4- (2-amino-ethoxy) -7-fluoro-5-methyl-3-phenyl-2H-isoquinolin-1-one

57.7-fluoro-5-methyl-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

58.4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -5-methyl-2H-isoquinolin-1-one,

59.3- (3-chloro-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

60.4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -7-fluoro-5-methyl-2H-isoquinolin-1-one,

61.3- (3-chloro-phenyl) -7-fluoro-5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

62.4- (2-amino-ethoxy) -7-fluoro-3- (4-methoxy-phenyl) -5-methyl-2H-isoquinolin-1-one,

63.7-fluoro-3- (4-methoxy-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

64.4- (2-amino-ethoxy) -7-fluoro-3- (3-methoxy-phenyl) -5-methyl-2H-isoquinolin-1-one,

65.7-fluoro-3- (3-methoxy-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

66.4- (2-amino-ethoxy) -3- (4-chloro-3-methoxy-phenyl) -7-fluoro-5-methyl-2H-isoquinolin-1-one,

67.3- (4-chloro-3-methoxy-phenyl) -7-fluoro-5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

68.4- (2-amino-ethoxy) -3- (3-chloro-4-methoxy-phenyl) -7-fluoro-5-methyl-2H-isoquinolin-1-one,

69.3- (3-chloro-4-methoxy-phenyl) -7-fluoro-5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

4- (2-amino-ethoxy) -3- (3-chloro-4-methoxy-phenyl) -5-methyl-2H-isoquinolin-1-one,

71.3- (3-chloro-4-methoxy-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

72.4- (2-amino-ethoxy) -3- (4-chloro-3-methoxy-phenyl) -5-methyl-2H-isoquinolin-1-one,

73.3- (4-chloro-3-methoxy-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

74.4- (2-amino-ethoxy) -3- (3-methoxy-phenyl) -5-methyl-2H-isoquinolin-1-one,

3- (3-methoxy-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

76.4- (2-amino-ethoxy) -3- (4-methoxy-phenyl) -5-methyl-2H-isoquinolin-1-one,

77.3- (4-methoxy-phenyl) -5-methyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

78.4- (2-amino-ethoxy) -7-fluoro-3- (4-phenoxy-phenyl) -2H-isoquinolin-1-one,

79.7-fluoro-3- (4-phenoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

80.4- (2-amino-ethoxy) -3-benzyl-7-fluoro-2H-isoquinolin-1-one,

81.3-benzyl-7-fluoro-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

82.4- (2-amino-ethoxy) -8-fluoro-3-phenyl-2H-isoquinolin-1-one,

83.8-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

84.4- [2- (dimethylamino) ethoxy ] -7-fluoro-3-phenylisoquinolin-1 (2H) -one,

85.7-fluoro-3-phenyl-4- (piperidin-4-yloxy) isoquinolin-1 (2H) -one,

86.4- (3-aminopropoxy) -7-fluoro-3-phenylisoquinoline-1 (2H) -one hydrochloride,

87.4- [3- (benzylamino) propoxy ] -7-fluoro-3-phenylisoquinolin-1 (2H) -one,

88.4- [2- (diethylamino) ethoxy ] -7-fluoro-3-phenylisoquinolin-1 (2H) -one,

89.7-fluoro-4- [2- (4-methylpiperazin-1-yl) ethoxy ] -3-phenylisoquinolin-1 (2H) -one and

7-fluoro-3-phenyl-4- [2- (phenylamino) ethoxy ] isoquinolin-1 (2H) -one.

The present invention also provides a process for the preparation of a compound of formula (I) as defined above.

Thus, the method of the invention comprises the following steps:

step 1) reacting a compound of formula (III):

wherein R is₂And R₃Alkylation with a compound of formula (IV), as defined above:

wherein R is₄As defined above and Lg represents a suitable leaving group;

step 2) cyclodehydration of the resulting compound of formula (V):

wherein R is₂，R₃And R₄As defined above;

step 3) rearranging the resulting compound of formula (VI):

wherein R is₂，R₃And R₄As defined above, to give a compound of formula (VII):

wherein R is₂，R₃And R₄As defined above;

or

Step 4) reacting a compound of formula (VIII) with a compound of formula (IX):

wherein R is₄As defined above and R₁₂Is C₁-C₆Alkyl or aryl radicals C₁-C₆An alkyl group;

step 3a) rearranging the resulting compound of formula (X):

wherein R is₄And R₁₂As defined above, to give a compound of formula (VII) as defined above, wherein R is₂And R₃Are each a hydrogen atom;

and

step 5) alkylating a compound of formula (VII) as defined above with a compound of formula (XI):

r, R therein₁L and Lg are as defined above to give a compound of formula (I) as defined above;

or by alkylation with a compound of formula (XII) X' -L-Lg, wherein L is optionally substituted, linear or branched C₂-C₆Alkyl or C₃-C₇A cycloalkyl group, Lg being as defined above and X' represents a suitable leaving group or a group which can be converted to a suitable leaving (living) group;

step 6) the resulting compound of formula (XIII):

wherein R is₂、R₃、R₄And X' is as defined above, L is optionally substituted straight or branched C₂-C₆Alkyl or C₃-C₇Cycloalkyl groups, with formula (XIV): R-NH-R₁Wherein R and R are₁As defined above, to give a compound of formula (I) as defined above.

If necessary or desired, the method comprises converting a compound of formula (I) to a different compound of formula (I) by known chemical reactions; and/or, if necessary, converting a compound of formula (I) into a pharmaceutically acceptable salt thereof or converting a salt into a free compound of formula (I).

For possible conversions of a compound of formula (I) or formula (XIII) into a different compound of formula (I) or formula (XIII), known chemical reactions are for example:

transformation A) Compounds of formula (I) as defined above, when R and/or R₁When protecting groups are present, deprotection is carried out, if necessary, to give the corresponding deprotected compound of formula (I) as hereinbefore defined.

Transformation B) A compound of formula (I) as defined above, wherein R₄is-OR-ed at any empty position of the ring₁₂Substituted aryl, aryl C₁-C₆Alkyl, heteroaryl, or heteroaryl C₁-C₆Alkyl radical, wherein R₁₂As defined above, into a compound of formula (I) as defined above, wherein R is₄Is aryl, aryl C substituted in any vacant position by-OH₁-C₆Alkyl, heteroaryl or heteroaryl C₁-C₆An alkyl group.

Conversion C) of a compound of formula (I) as defined above, in which R is₂Or R₃Is a nitro group, into a compound of formula (I) as defined above, wherein R is₂Or R₃Is an amino group.

Transformation D) reacting a compound of formula (I) as defined above, wherein R is₂Or R₃Is an amino group, is converted by a corresponding diazo derivative into a compound of formula (I) as defined above, wherein R is₂Or R₃Is a halogen atom such as Cl, Br, F or I.

Transformation E) Compounds of formula (I) as defined above, wherein R and/or R₁Is a hydrogen atom, is converted intoA compound of formula (I) wherein R and R₁As defined above, but not COR₅Or R and R₁Are not hydrogen atoms.

Conversion F) Compounds of formula (I) as defined above, in which R and/or R₁Is a hydrogen atom, into a compound of formula (I) as defined above, wherein R or R₁Is COR₅Wherein R is₅As defined above.

Transformation G) a compound of formula (XIII) as defined above is converted into another compound of formula (XIII) by the method described in transformation a to D or by converting group X' into a suitable leaving group.

All the above methods are similar methods, which can be performed according to well known methods and under suitable conditions known in the art.

Scheme 1 below illustrates the sequence of steps of the method according to the invention.

Scheme 1

According to step 1 of the process, a compound of formula (III) as defined above is reacted with a compound of formula (IV) wherein Lg is a leaving group such as e.g. halogen like bromine, chlorine or iodine, p-toluenesulfonate, methanesulfonate or trifluoromethanesulfonate in a suitable solvent such as N, N-dimethylformamide, N-dimethylacetamide, acetonitrile, acetone, ethyl acetate, tetrahydrofuran, dioxane, dichloromethane etc. in the presence of a base such as sodium carbonate, potassium carbonate or cesium carbonate, sodium hydroxide or potassium hydroxide, triethylamine, diisopropylethylamine etc. at a temperature range of 0 ℃ to reflux to give a compound of formula (V) as defined above.

According to step 2 of the process, the compound of formula (V) as defined above is refluxed in a solvent such as ethyl acetate, benzene, toluene, xylene and the like in the presence of a catalytic amount of an acid such as p-toluenesulfonic acid, camphorsulfonic acid and the like in a vessel suitable for the continuous removal of water to provide the compound of formula (VI) as defined above. Cyclodehydration can be carried out as described in Wang, S. et al, J.Bioorg.Med chem.Lett.2002, 12, 2367-.

According to step 3 of the process, the compound of formula (VI) as defined above is heated in a suitable solvent such as N, N-dimethylformamide, N-dimethylacetamide, tetrahydrofuran, dioxane and the like in the presence of a base such as sodium hydride, sodium amide and the like under an inert atmosphere to give the compound of formula (VII) as defined above. May be prepared as described in SchenkeR, k.helv.chim.acta 1968, 51, 413-21; or Wang, S. et al, J.Bioorg.Medchem.Lett.2002, 12, 2367-.

According to step 4 of the process, a compound of formula (VIII) as defined above, used in the form of a racemic mixture, an enantiomerically enriched mixture or an individual enantiomer, as described in Peukert, S. et al Synthesis 2005, 9, 1550-.

According to step 3a of the process, the compound (X) obtained as racemic mixture, enantiomerically enriched mixture or individual enantiomer obtained in step 4 is treated with a base such as sodium methoxide, sodium ethoxide etc. in a solvent such as methanol or ethanol at reflux temperature to form the compound (VII) as defined above. Rearrangements can be made as described in Peukert, S. et al Synthesis 2005, 9, 1550-.

According to step 5 of the process, a compound of formula (VII) as defined above is reacted with a compound of formula (XI) as defined above in a suitable solvent such as N, N-dimethylformamide, N-dimethylacetamide, acetonitrile, acetone, methanol, ethanol, tetrahydrofuran, dioxane, dichloromethane, etc., in the presence of a suitable base such as sodium carbonate, potassium carbonate or cesium carbonate, sodium or potassium bicarbonate, triethylamine, diisopropylethylamine, pyridine, sodium or potassium hydride, etc., at a temperature range of 0 ℃ to reflux to give a compound of formula (I) as defined above. When Lg is bromine, the reaction is preferably carried out at room temperature using N, N-dimethylacetamide as a solvent and cesium carbonate as a base. When Lg is chlorine, potassium iodide is added, and the reaction is preferably carried out under microwave irradiation using potassium carbonate as a base and methanol as a solvent.

The compound of formula (VII) as defined above is also reacted with the compound of formula (XII) as defined above, wherein Lg and X' are as defined above, in a solvent such as N, N-dimethylformamide, N-dimethylacetamide, acetonitrile, acetone, methanol, ethanol, tetrahydrofuran, dioxane, dichloromethane and the like, at a temperature range of 0 ℃ to reflux, in the presence of a suitable base such as sodium carbonate, potassium carbonate or cesium carbonate, sodium or potassium bicarbonate, triethylamine, diisopropylethylamine, pyridine, sodium or potassium hydride and the like, to give the compound of formula (XIII) as defined above.

According to step 6 of the process, the compound of formula (XIII) as defined above is then reacted with the compound of formula (XIV) as defined above, or alternatively using microwave conditions, in the presence of a base such as sodium, potassium or cesium carbonate, sodium or potassium bicarbonate, triethylamine, diisopropylethylamine, pyridine, etc., in a suitable solvent such as acetonitrile, dioxane, methanol, ethanol or N, N-dimethylformamide, at a temperature range of 0 ℃ to reflux to give the compound of formula (I) as defined above.

Transformation A according to this process when R or R in the compound of formula (I)₁Nitrogen protecting groups, containing protecting groups such as tert-butoxycarbonyl, 4-methoxybenzyl, 2, 4-dimethoxybenzyl and trityl groups, may be protected by treatment under acidic conditions, preferably in the presence of an inorganic or organic acid such as hydrochloric acid, trifluoroacetic acid or methanesulfonic acid, boron tribromide or aluminium trichloride, in the presence of an inorganic or organic acid such as dichloromethaneThese protecting groups are removed in a suitable solvent of an alkane, dichloroethane, dioxane, lower alcohol such as methanol or ethanol at a temperature ranging from room temperature to reflux, to give the different compounds of formula (I) as defined above.

R or R in the compound of formula (I)₁When nitrogen protecting groups such as benzyloxycarbonyl and the like are present, different compounds of formula (I) as defined above may be obtained by removing these protecting groups under reducing conditions, for example in the presence of hydrogen and a hydrogenation catalyst in a suitable solvent such as ethanol, methanol, ethyl acetate or mixtures thereof. The catalyst is typically a metal, most often a palladium derivative, such as palladium on carbon, palladium hydroxide or palladium black.

R or R in the compound of formula (I)₁When nitrogen protecting groups such as methoxycarbonyl, ethoxycarbonyl, 9-fluorenylmethoxycarbonyl and the like are contained, different compounds of formula (I) as defined above can be obtained by removing these protecting groups in a suitable solvent such as methanol, ethanol, water, N-dimethylformamide, N-dimethylacetamide and the like under basic conditions such as sodium carbonate, potassium carbonate or cesium carbonate, sodium hydroxide, potassium hydroxide or barium hydroxide, hydrazine, piperidine, morpholine and the like at a temperature ranging from room temperature to reflux.

According to the process of transformation B, a compound of formula (I) as defined above, wherein R is a compound of formula (I) wherein R is as defined above, in a suitable solvent such as acetonitrile, dichloromethane, dichloroethane, acetic acid, acetic anhydride, etc., by using an acid such as hydrobromic acid, boron tribromide, aluminium chloride, etc., in a temperature range of from 0 ℃ to reflux₄is-OR-ed at any empty position of the ring₁₂Substituted aryl, aryl C₁-C₆Alkyl, heteroaryl or heteroaryl C₁-C₆Alkyl radical, wherein R₁₂As defined above, may be converted into different compounds of formula (I) wherein R₄Is aryl, aryl C substituted in any vacant position by-OH₁-C₆Alkyl, heteroaryl or heteroaryl C₁-C₆An alkyl group.

According to transformation C of the process, a compound of formula (I) as defined above, wherein R is R, wherein R is palladium on carbon, palladium hydroxide, palladium black, raney nickel, etc., in a suitable solvent such as methanol, ethanol, dioxane, etc., in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, palladium black, raney nickel, etc., at a temperature ranging from room temperature to reflux, is prepared₂Or R₃Is a nitro group which can be converted into different compounds of the formula (I) in which R is the same as R, by reaction with a suitable reducing agent, for example molecular hydrogen, cyclohexene, cyclohexadiene, formic acid, ammonium formate and the like₂Or R₃Is an amino group. Alternatively, the conversion may be accomplished with metals such as tin, iron, zinc, and the like, optionally in a suitable solvent such as methanol, ethanol, and the like, in the presence of a protic acid such as hydrochloric acid, acetic acid, and the like, at a temperature ranging from room temperature to reflux.

Transformation D according to this process, a compound of formula (I) as defined above, wherein R is₂Or R₃Is an amino group which can be converted into different compounds of the formula (I) by reaction with alkyl nitrites and copper (II) chloride or bromide in suitable solvents such as acetonitrile, tetrahydrofuran, etc., at temperatures ranging from 0 ℃ to room temperature₂Or R₃Is a halogen such as a chlorine or bromine atom. Alternatively, the same conversion can be carried out by using sodium nitrite or potassium nitrite and copper (I) chloride or bromide in the presence of hydrochloric acid or hydrobromic acid at a temperature range of 0 ℃ to room temperature. Furthermore, a compound of formula (I) as defined above, wherein R₂Or R₃Is NH₂The radicals, in a solvent such as o-dichlorobenzene, can be converted into different compounds of formula (I) by reaction with nitrosonium tetrafluoroborate at a temperature ranging from 0 ℃ to reflux, wherein R is₂Or R₃Is fluorine. Alternatively, the same conversion can be accomplished with alkyl nitrites and boron trifluoride in a solvent such as o-dichlorobenzene, or with silicon tetrafluoride in methylene chloride, or with sodium nitrite or potassium nitrite and hydrofluoric acid pyridine complex at a temperature range of 0 ℃ to reflux.

Transformation according to the methodE, compounds of formula (I) wherein R and/or R are R, R is H, by reacting the starting materials with the appropriate aldehyde or ketone in the presence of a reducing agent such as sodium triacetoxyborohydride, tetramethylammonium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, zinc, optionally in the presence of a protic acid such as hydrochloric acid, acetic acid, trifluoroacetic acid, formic acid, or in the presence of a Lewis acid such as zinc chloride, zinc bromide, tin (IV) chloride, titanium (IV) chloride, boron trifluoride, in a suitable solvent such as methanol, ethanol, dichloromethane, acetic acid, N-dimethylformamide, and the like, at a temperature in the range of 0 ℃ to room temperature₁Is a hydrogen atom, can be converted into different compounds of formula (I) in which R and/or R₁Not COR₅Or both are hydrogen atoms. Alternatively, by using the appropriate R or R, e.g., corresponding iodide, bromide, chloride, mesylate, triflate, etc₁Derivatives, the conversion can be accomplished in the temperature range of 0 ℃ to reflux in the presence of a suitable base such as sodium, potassium or cesium carbonate, sodium or potassium bicarbonate, triethylamine, diisopropylethylamine, pyridine, and the like.

Conversion F according to this process, in the presence of a suitable base such as triethylamine, diisopropylethylamine, pyridine and the like, in a solvent such as dichloromethane, tetrahydrofuran and the like, at a temperature ranging from 0 ℃ to reflux, a compound of formula (I) as defined above, wherein R and/or R₁Is a hydrogen atom, by reaction with R₅COCl, which can be converted into different compounds of formula (I) wherein R and/or R₁Is COR₅Wherein R is₅As defined above. Alternatively, in the presence of a suitable coupling agent, e.g. an alkyl chloroformate, a suitable carbodiimide such as dicyclohexylcarbodiimide, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide and the like, optionally in the presence of HOBt (1-hydroxybenzotriazole), or e.g. carbonyldiimidazole, BOP (benzotriazol-1-yloxy) tris (dimethylamino) phosphonium hexafluorophosphate), TBTU (2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate), HATU (O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluoro-luoridePhosphate) and the like, by reacting with a compound of formula R in the presence of a base such as triethylamine, diisopropylethylamine, pyridine, 4-N, N-dimethylaminopyridine, sodium carbonate or potassium carbonate in a solvent such as dichloromethane, N-dimethylformamide, tetrahydrofuran, acetonitrile and the like at a temperature ranging from 0 ℃ to reflux₅Compound of COOH, wherein R₅As defined above, the transformation may be accomplished.

According to this method of converting G, a compound of formula (XIII) as defined above, wherein X 'is OH, may be converted into a different compound of formula (XIII), wherein X' is a different leaving group. When for example I₂/Ph₃P or CBr₄The conversion can be achieved in a suitable solvent such as dichloromethane, N-dimethylformamide, tetrahydrofuran, acetonitrile, etc., in the presence of a suitable halogenation system with imidazole, at a temperature range of 0 ℃ to reflux. All the above methods are analogous methods which may be carried out according to well known methods and under suitable conditions known in the art, for example according to the methods described in: smith, Michael-March's Advanced Organic Chemistry: interactions mechanisms and structure-6^thEdition, Michael B.Smith and Jerry March, John Wiley&Sons Inc, New York (NY), 2007.

Any of the intermediates of the above-described processes can be converted to a different intermediate by operating in a similar manner to any of the above-described conversion reactions.

From all of the above, it is clear to the skilled person that any compound of formula (I) containing a functional group which can be further converted into another functional group by working according to methods well known in the art, thus yielding another compound of formula (I), is intended to be included in the scope of the present invention. The starting materials and any other reactants are known or can be readily prepared according to known methods, according to any variant of the process for preparing the compounds of formula (I). The compounds of formulae (III), (IV), (VIII), (IX), (XI) and (XII) are commercially available or can be prepared according to known methods.

From all the above, it is clear to the skilled person that when preparing the compounds of formula (I) according to variants of any of the aforementioned methods, the optional functional groups of the starting materials or intermediates thereof, which may cause undesired side reactions, need to be suitably protected according to conventional techniques. Likewise, the conversion of these latter compounds into deprotected compounds can be carried out according to known methods, for example: methods described by Greene, Theodora W. and Wuts, Peter G.M. -Protective Groups in Organic Synthesis, Third Edition, John Wiley & Sons Inc, New York (NY), 1999.

It will be readily understood that if the resulting compounds of formula (I) prepared according to the above-described process are mixtures of isomers, it is within the scope of the present invention to separate them into the individual isomers of formula (I) using conventional techniques.

Conventional techniques for racemate resolution include, for example, fractional crystallization of diastereomeric salt derivatives or preparative chiral HPLC, etc. General methods for isolating compounds containing 1 or more asymmetric centers have been reported, for example: jacques, Jean; collet, Andre; wilen, Samuel H, -Enantiomers Racemates, and solutions, John Wiley & Sons Inc, New York (NY), 1981.

Furthermore, the compounds of formula (I) according to the invention can also be prepared according to combinatorial chemistry techniques well known in the art, for example by carrying out the reactions between the aforementioned intermediates in a parallel and/or serial manner and by operating under Solid Phase Synthesis (SPS) conditions.

As mentioned above, the probe used in the screening method is a compound of formula (II) as defined above. Preferably, the present invention provides a probe of formula (II) as defined above, said probe being characterized in that: is represented by X^-The counter ion of (A) is perchlorate, trifluoroacetate or the like, R₁₃Is a hydrogen atom or a methyl group, m is 0 or 1; when m is 1, n is preferably 6; more preferably X^-Is trifluoroacetate.

The compounds of formula (II) as defined above are very efficient probes binding to PARP proteins, including full-length native proteins and fragments thereof.

The polarization signal may be measured, for example, by a plate reader such as Saphire2 (Tecan). The displacement capacity of the test compound correlates with the affinity of the compound for the NAD pocket of the enzyme. Affinity binding constant (KD) and DC of test Compounds₅₀s can be determined as explained in the examples section.

The test of the invention is based on the use of a compound of formula (II) as defined above as a probe, which provides a fluorescent signal.

Preferred compounds of the invention of formula (II) as defined above are:

p19-dimethylamino-11, 11-dimethyl-1- (3- { methyl- [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -carbamoyl } -propyl) -2, 3, 4, 11-tetrahydro-naphtho [2, 3-g ] quinolinium trifluoroacetate;

p29-dimethylamino-11, 11-dimethyl-1- [3- (3- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -propylcarbamoyl) -propyl ] -2, 3, 4, 11-tetrahydro-naphtho [2, 3-g ] quinolinium trifluoroacetate or

P39-dimethylamino-11, 11-dimethyl-1- [3- (6- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -hexylcarbamoyl) -propyl ] -2, 3, 4, 11-tetrahydro-naphtho [2, 3-g ] quinolinium trifluoroacetate.

The present invention also provides a process for the preparation of a compound of formula (II) as defined above, said process comprising:

step i) reacting a compound of formula (XV):

wherein Lg is as aboveAs defined in (1) and formula (XVI) R₁₃-NH₂Wherein R is₁₃And B is as defined hereinbefore, to give a compound of formula (XVII):

wherein R is₁₃And B is as defined hereinbefore and m is 0;

or step i_a) Reacting a compound of formula (XV) as defined above with a compound of formula (XVIa):

wherein R is₁₃And B is as defined hereinbefore, m is 1 and R₁₄Hydrogen atom, methoxycarbonyl group, ethoxycarbonyl group, tert-butoxycarbonyl group, benzyloxycarbonyl group, 9-fluorenylmethoxycarbonyl group or the like; and

step i_b) If necessary, the resulting compound of formula (XVIIa):

wherein R is₁₃And B is as defined above, m is 1 and R₁₄Is a methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl or 9-fluorenylmethoxycarbonyl group or the like, to a compound of formula (XVII) wherein R is₁₄Is a hydrogen atom;

step ii) coupling the resulting compounds of formula (XVII) and (XVIII) as defined above:

wherein X^-As hereinbefore defined, to obtain the final compound of formula (II) as hereinbefore defined; and, if necessary, converting the compound of formula (II) into another compound of formula (II), wherein X^-Different.

The starting compounds of formula (XV) may be prepared as broadly described, see, for example

WO 2001042219; compounds of formula (XVIII) are described in Cha, J.H. et al, J.Med.chem.2005, 48, 7513-E7516, the fluorescent moiety of ATTO 610 activated as its NHS ester is commercially available (ATTO-TEC GmbH, Siegen, Germany).

Scheme 2 below illustrates the preparation of compounds of formula (II) wherein R₁₃、R₁₄B and m are as defined above.

Scheme 2

According to steps i and i of the method_aReacting a compound of formula (XV) as defined above with a compound of formula (XVI) as defined above in a suitable solvent such as acetonitrile, dioxane, methanol, ethanol or N, N-dimethylformamide at a temperature ranging from 0 ℃ to reflux in the presence of a base such as sodium or potassium hydroxide, sodium or potassium carbonate or cesium carbonate, sodium or potassium bicarbonate, triethylamine, diisopropylethylamine, pyridine, etc., starting from compound (XVI) as defined above to give a compound of formula (XVII) as defined above or starting from compound (XVIa) as defined above to give a compound of formula (XVIIa) as defined above;

according to step i of the method_b(xviiia) compounds of formula (XVIIa) wherein m is 1, R, by removal of the corresponding nitrogen protecting group₁₄Is methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl group or the like, toA compound of formula (XVI) as hereinbefore defined. Specifically, the protecting group such as t-butoxycarbonyl group may be removed under acidic conditions, preferably in a suitable solvent such as dichloromethane, dichloroethane, dioxane, a lower alcohol such as methanol or ethanol, in the temperature range of room temperature to reflux, in the presence of an inorganic or organic acid such as hydrochloric acid, trifluoroacetic acid or methanesulfonic acid. The protecting group such as benzyloxycarbonyl can be removed with a suitable reducing agent such as molecular hydrogen, cyclohexene, cyclohexadiene, formic acid, ammonium formate and the like in a suitable solvent such as methanol, ethanol, dioxane and the like at a temperature ranging from room temperature to reflux in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, palladium black, Raney nickel (Ni Raney) and the like.

The protecting groups such as methoxycarbonyl, ethoxycarbonyl, 9-fluorenylmethoxycarbonyl and the like can be removed in a suitable solvent such as methanol, ethanol, water, N-dimethylformamide, N-dimethylacetamide and the like under basic conditions such as sodium carbonate, potassium carbonate or cesium carbonate, sodium hydroxide, potassium hydroxide or barium hydroxide, hydrazine, piperidine, morpholine and the like at a temperature ranging from room temperature to reflux.

According to step II of the process, a compound of formula (XVII), as defined above, is reacted with a compound of formula (XVIII) in the presence of a suitable base, such as triethylamine, diisopropylethylamine, pyridine, etc., in a solvent, such as N, N-dimethylformamide, N-dimethylacetamide, etc., at a temperature ranging from 0 ℃ to room temperature, to give the desired compound of formula (II).

Pharmacology of

Compound potency was assessed by measuring inhibition of poly (ADP-ribose) (PAR) chain formation as a marker of PARP-1 inhibition.

PARP-1 is a DNA damage-inducing polymerase that catalyzes the cleavage of NAD + into nicotinamide and ADP-ribose, which are then used to synthesize the branched nucleic-acid-like polymer poly (ADP-ribose). The most abundant poly (ADP-ribosylated) protein in vivo is PARP-1 itself, followed by histones. PARP-1 is responsible for 90% of this DNA damage-inducing activity, while the remaining 10% is responsible for PARP-2.

The abbreviations and abbreviations used herein have the following meanings:

PAR (poly (ADP-ribose))

MEM (minimum essential medium)

FCS (fetal calf serum)

FBS (fetal bovine serum)

PBS (phosphate buffered saline)

LC-MS (liquid chromatography-mass spectrometry)

HPLC (high performance liquid chromatography)

DC₅₀(half maximum replacement concentration)

IC₅₀(half maximal inhibitory concentration)

STD (standard deviation)

Testing of cells

The cellular activity of PARP-1 inhibitors was assessed by measuring inhibition of PAR formation by hydrogen peroxide in HeLa cells (ECACC). PAR levels of cells were measured using immunocytochemistry and quantified using an ArrayScan vTi instrument (Cellomics thermosentific).

The study was carried out as follows: 6000 cells/well were seeded in 96 well plates (Perkin Elmer) in MEM/10% FCS and incubated at 37 ℃ with 5% carbon dioxide for 24 hours. The desired concentration of test compound is then added over a period of 30'. DNA damage was then induced by addition of 0.1mM hydrogen peroxide over a 15min period. Concentration curves were prepared in MEM/10% FCS using compound stocks in DMSO, with a final DMSO concentration of 0.002% (v/v). Duplicate wells were prepared at each concentration point using a typical maximum compound concentration of 20 μ M and serial dilutions 1: 3. Plates were dried and fixed with cold methanol/acetone (70: 30) solution at room temperature for 15min, the fixing solution was aspirated, the wells were air dried for 5min, and then in PBSAnd (4) dehydrating. Non-specific binding sites were blocked by incubating the wells for 30min in PBS containing 5% (w/v) FBS 0.05% tween 20. The wells were then incubated at room temperature for 1 hour in PBS containing Anti-PAR Mouse monoclonal antibody (Anti-PAR, Mouse mAb 10H, Tulip Cat N.degree.1020) at a dilution of 1: 200 in blocking solution. After 3 washes in PBS, wells were incubated in PBS (w/v) 5% FBS 0.05% Tween20 containing 2. mu.g/ml Cy 2-conjugated secondary goat anti-mouse antibody (Amersham Pharmacia Biotech cat.N ° PA 42002) (maximum absorption 489nm maximum fluorescence 506nm) and 1. mu.g/ml DAPI (maximum absorption 359nm maximum fluorescence 461nm) (4', 6-diamidino-2-phenylindole hydrogen lactate) (Sigma cat.N ° D9564), a highly sensitive nucleotide staining dye. After 3 additional washes in PBS, cells were evaluated for PAR immunoreactivity as follows: ArrayScan vTi instrument equipped with Zeiss 10X 0.5N.A. eyepiece was used, and Cytoxicity.V3 algorithm (Cellomics/Thermo Fisher) equipped with XF100 filter was used. Each well reads at least 10 fields, corresponding to at least 900 cells. IC (integrated circuit)₅₀Values represent compound concentrations at which the PAR signal of cells is reduced by 50% compared to untreated controls.

The following formula was used:

IC₅₀bottom + (top-bottom)/(1 +10^ ((LogEC 50-X))); x is the logarithm of the concentration. IC (integrated circuit)₅₀Is the reaction value; IC (integrated circuit)₅₀Starting from the bottom and going to the top in an S-shape.

In the above assay, the compounds of formula (I) inhibit PAR formation, the IC thereof₅₀Values below 10. mu.M (Table 3).

Biochemical testing

The affinities of the test compounds and their selectivities to the different PARP isoforms of interest are quantified in displacement assays.

The assay is based on the use of probes of formula (II) bound to the NAD binding pocket and utilises the significant change in polarisation signal observed when the probe binds to PARP-1, -2 and-3.

In titration experiments, probes (II) were tested for their ability to bind FL PARP-1, -2 and-3. The test performance (Z' factor) was then evaluated as well as the displacement of the probe through its backbone and known commercially available PARP inhibitors. In all experiments, the polarization signal was measured using a Saphire2 plate reader (Tecan). Data analysis was performed using Dynafit software. In particular, titration data were fit to the following balance: enzyme + probe<＝＝>Complex enzyme-probe, and displacement data were fitted to the following balance: enzyme + probe<＝＝>Complex enzyme-probe, enzyme + compound<＝＝>Complex enzyme-compound, it follows that the binding of probe and compound to the enzyme is mutually exclusive (purely competitive mechanism). The displacement data was also fitted to a 4-parameter logistic model (4PL), or a Hill-Slope model, using Excel speedsheet (Microsoft Inc. Seattle, USA) to calculate DC₅₀Wherein DC₅₀Values represent the compound concentration at which the polarized signal is reduced by 50% compared to the untreated control.

Titration experiments were performed as follows: 50nM probe (compound P1), FL PARP-1, 2 and 3 at concentrations from 5. mu.M to 0, dilution step 1: 1.5 at 50mM TrisHCl, pH 7.8, 150mM NaCl, 10mM MgCl₂0.001% Triton X100, 1% DMSO (buffer 1). Compound P3 was titrated in a similar manner.

The results obtained (shown in table 1 below) indicate that the probe (compound P1) is able to bind PARP of all the isoforms tested. Only compound P3 was reported to be able to bind PARP-1 KD. The Z 'factor (Z' ═ 1- (3 × (SD probe + protein + SD probe)/(mean probe + protein-mean probe))) was determined as follows: 50nM probe (Compound P1), 250nM PARP-1 and 2,200nM PARP-3. When compound P3 was used as a probe, the PARP-1 concentration was equal to 100 nM. In all cases, the value of Z' was higher than 0.7, indicating that the test was stable (Table 1).

TABLE 1

In the displacement test, the test was validated using 3-aminobenzamide (3-AB) and PJ-34, which was performed as follows: serial dilutions of test compounds were first prepared in 100% DMSO and further diluted in test buffer 1 to give 1% final DMSO concentration. 3-AB was tested at 100. mu.M as the highest concentration, while 10. mu.M was the highest concentration of PJ-34. The final concentration of enzyme used for PARP-1 (100 nM when compound P3 was used as a probe) and PARP-2 was 250nM, while 200nM was used for PARP-3. The final concentration of probe (compound P1 or compound P3) was 50 nM. The enzyme and probe (compound P1 or compound P3) mixture was added to the previously diluted compound. The results (Table 2) indicate that the probe (compound P1 or compound P3) can be completely replaced by 3-AB and PJ-34 from all the PARP isoforms tested, indicating that probe (compound P1 or compound P3) binding is specific. Consistently, the affinity binding constant (KD) was determined by fitting a purely competitive mechanism. KD values are the average of 3 independent experiments.

As expected, 3-AB is not selective for the PARP isoform and shows lower affinity for PJ-34.

TABLE 2

^＊The test sensitivity limit based on fitting error is less than 50%

Taken together, these results show that the displacement assay is specific. Furthermore, it enables quantitative potency evaluation of standard PARP inhibitors tested, thus enabling in-test selectivity evaluation.

The same test was used to evaluate certain representative compounds of formula (I) by using compounds P1 or P3, as reported in table 3.

TABLE 3

^＊The test was carried out using compound P3 as a probe. In all other cases, compound P1 was used as a probe.

From the above, it is clear to the skilled person that the compounds of formula (I) are potent and selective PARP-1 inhibitors in biochemical and cellular assays.

Pharmacokinetics

Mice (Balb, Nu/Nu, Harlan, Italy) were used in a dedicated (ad hoc) pharmacokinetic study to investigate the pharmacokinetic properties and oral bioavailability of the compounds. For intravenous bolus (bolus) administration, compounds were formulated in 10% tween 80/glucose, while oral administration was carried out using compounds formulated in 0.5% methylcellulose. A single administration of 10mg/kg dose was given, using 3 males for each route. All blood samples were taken from the saphenous vein 5min, 30min, 1h, 6h, 24h after intravenous administration and 15min, 30min, 1h, 6h, 24h after oral administration. Plasma samples were prepared by adding 200l of methanol to 10l of plasma protein precipitate of plasma in 96-well plates. After capping and vortex mixing, the plates were centrifuged at 3700rpm for 15min at 6 ℃. The supernatant was considered the final extract and was injected into a LC-MS-MS system (HPLC system: Hewlett Packard 1100series, using an Atlantis HILIC Silica 50X 2.1mm 5.0M analytical column; MS instrument: PerkinElmer SCIEX API 2000, ionizing cationically using Turbo ion spray). The lower limit of quantitation was 5.0ng/ml, and the upper limit of quantitation was 10000 ng/ml. The analysis was performed using Watsonpackage (version 6.4.0.04) and Excel spaadsheet (Microsoft inc. seattle, USA). Non-compartmental methods (linear trapezoidal rule and linear regression analysis of natural log-transformed plasma concentration vs. time data) were used. After intravenous administration, C is administered₀Is provided withIs equal to C_0.083. Absolute bioavailability (F) was calculated as the ratio of mean oral dose-normalized plasma AUC (area under curve) value to IV (intravenous dose-normalized plasma AUC (area under curve) value.

The abbreviations used herein have the following meanings:

AUC (area under plasma concentration vs. time curve up to last concentration tested)

Cl (plasma clearance rate)

Cmax (maximum plasma concentration)

T1/2 (terminal half-life)

Vdss (Steady distribution volume)

Certain representative compounds of formula (I) were evaluated for pharmacokinetic parameters, which are reported in table 4 below as mean values.

TABLE 4

From the above, it is clear to the skilled person that the compounds of formula (I) have good to excellent pharmacokinetic properties and oral bioavailability.

In vivo efficacy studies

Balb, athomic Nu/Nu male mice, from Harlan (Italy), were housed in cages with sterile filter paper lids, food, bedding and acidified water, according to the European Community Commission guide No. 86/609/EEC, directed to the protection of animals for experimental or other scientific purposes. A fragment of the Capan-1 human pancreatic cancer tumor was implanted subcutaneously. Selection of the tumor size with palpability (100-³) And randomly assigned to control and treatment groups. Each group included 7 animals. Treatment was started one day after randomization. In the form of a methylcellulose (methocel) suspension at a dose of 100mg/kg per doseThe compound of formula (I) is administered by the oral route once daily or twice daily for the indicated time. Tumor size was routinely measured by caliper during the experiment, and tumor mass was calculated as described in Simeoni m. et al, Cancer Res 64, 1094-. Tumor growth inhibition (TGI,%) was calculated according to the following equation: % TGI is 100 ═ 100 (mean tumor weight in the treated group/mean tumor weight in the control group) × 100.

Certain representative compounds of formula (I), compound (25), compound (39) and compound (11) were evaluated as single agents for antitumor activity against a mouse model of the Capan-1BRCA-2 mutation. Toxicity was assessed on the basis of weight loss and animal survival. The results are reported in table 5.

TABLE 5

Compound (I)	Dosage form	Schedule sheet	Max TGI(％)	Toxicity
					25	100mg/kg	1-20 times per day	18％	0/7
39	100mg/kg	1-10 times per day for 2 times	48％	0/7
					11	150mg/kg	1-14 times per day	31％	0/7

Representative compounds of formula (I), compound (39), in combination with temozolomide, were evaluated for anti-tumor activity in a mouse model of the Capan-1BRCA-2 mutation. Compound (39) was administered by the oral route twice daily at a dose of 100mg/kg for 10 consecutive days (1 to 10 days). Temozolomide was administered by the oral route at a dose of 50mg/kg on days 3, 4, 5, 6 and 7. Tumor growth and body weight were measured every 3 days. Tumor growth was evaluated by caliper. Two diameters were recorded and tumor weights were calculated according to the following formula: length (mm) x width²/2. The efficacy of anti-tumor therapy is assessed as the exponential growth onset delay of the tumor (see literature anticancer drugs 7: 437-60, 1996). This delay (T-C value) is defined as the difference in the time (in days) required for the treated (T) and control (C) tumors to reach a predetermined size (1 g). Toxicity was assessed on the basis of weight loss and animal survival. The results are reported in table 6.

TABLE 6

^＊Oral treatment is carried out twice daily on days 1 to 10

^＊＊Treatment by oral route once daily on days 3, 4, 5, 6 and 7

^＊＊＊Compound (39) treatment is 1 to 10 days, temozolozoleAmine treatment was on days 3, 4, 5, 6, 7.

When compound (39) was combined with temozolomide, the observed T-C was superior to that expected by simple addition of T-C obtained by monotherapy, indicating that there was significant synergy.

From the above, it is clear to the skilled person that the compounds of formula (I) have good tumor growth inhibitory activity as single agents and synergistic tumor growth inhibitory activity in combination with cytotoxic agents.

Examples section

When referring to any particular compound of formula (I) of the present invention, said compound is optionally present in the form of a pharmaceutically acceptable salt, see examples section and claims. The compounds of the present invention are synthesized using the methods described herein or other methods well known in the art with reference to the following examples.

The abbreviations and acronyms used herein have the following meanings:

min (minutes)

mmol (millimole)

DMSO (dimethyl sulfoxide)

ESI (electrospray ionization)

In order to better illustrate the invention without posing any limitation thereto, the following examples are now given.

The symbols used herein and conventions used in the methods, schemes and examples are consistent with those used in scientific literature of the same generation, such as the Journal of the American Chemical Society or Journal of Biological Chemistry.

All materials were obtained from commercial suppliers, of optimal grade, and used without further purification, unless otherwise indicated. Anhydrous solvents such as N, N-dimethylformamide, tetrahydrofuran, dichloromethane and toluene were obtained from Aldrich chemical company. All reactions involving air or moisture sensitive compounds were carried out under nitrogen or argon atmosphere.

General purification and analytical methods

Flash chromatography was performed on silica gel (Merck grade 9395, 60A). HPLC was performed on a Waters X TerrarP 18(4, 6X 50mm, 3.5 μm) column using a Waters 2790HPLC system equipped with a 996Waters PDA detector and a microscale mode (Micromass mod). A ZQ single quadrupole mass spectrometer equipped with an Electrospray (ESI) ion source. Mobile phase a was ammonium acetate 5mM buffer (pH 5.2 with acetic acid-acetonitrile 95: 5) and mobile phase B was water-acetonitrile (5: 95). The gradient was from 10 to 90% B over 8min, maintaining 90% B2 min. UV detection was at 220nm and 254 nm. The flow rate was 1 mL/min. The injection volume was 10. mu.L. Full scan, mass ranging from 100 to 800 atomic mass. The capillary voltage was 2.5 KV; the source temperature was 120 ℃; the taper hole voltage is 10V. The mass is given in m/z ratio.

When necessary, compounds were purified by preparative HPLC 600 on a Waters Symmetry C18(19X 50mm, 5m) column or a Waters X Terra RP 18(30X 150mm, 5 μm) column using Waters equipped with a 996Waters PDA detector and a microscale mode. ZMD Single quadrupole Mass Spectrometry, electrospray ionization, Positive ion mode. Mobile phase a was water-0.01% trifluoroacetic acid and mobile phase B was acetonitrile. The gradient was from 10 to 90% B over 8min, maintaining 90% B2 min. The flow rate was 20 mL/min. Alternatively, mobile phase A is water-0.1% NH₃And the mobile phase B is acetonitrile. The gradient was from 10 to 100% B over 8min, maintaining 100% B2 min. The flow rate was 20 mL/min.

In Mercury VX 400[1H (15N-31P) ID _ PFG Varian equipped with 5mm dual resonance probe]Operating at 400.45MHz, performing¹H NMR spectrum.

Example 1

Step 1

2- [2- (3-methoxy-phenyl) -2-oxo-ethoxy ] -benzamide

8.36g (36.5mmol) of 2-bromo-1- (3-methoxy-phenyl) -ethanone and 7.57g (54.7mmol) of potassium carbonate are added to a suspension of 5g (36.5mmol) of salicylamide in 60ml of N, N-dimethylformamide. The mixture was stirred at room temperature overnight. Water was added with stirring, the precipitate was filtered, washed with water and dried in vacuo to give 9g of the title compound (90%).

¹H NMR(DMSO-d₆)(ppm)：8.41(br.s，1H)，7.99(dd，J＝7.8，1.8Hz，1H)，7.69(br.s，1H)，7.67(ddd，J＝7.7，1.5，0.9Hz，1H)，7.57(dd，J＝2.5，1.5Hz，1H)，7.52(t，J＝8.0Hz，1H)，7.51(ddd，J＝8.4，7.3，2.0Hz，1H)，7.29(ddd，J＝8.3，2.6，0.9Hz，1H)，7.29(dd，J＝8.3，0.9Hz，1H)，7.09(ddd，J＝7.8，7.3，1.0Hz，1H)，5.77(s，2H)，3.85(s，3H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

2- (2-oxo-2-phenyl-ethoxy) -benzamides

¹H NMR(DMSO-d₆)(ppm)：8.41(br.s，1H)，8.08(dd，J＝8.4，1.2Hz，2H)，7.99(dd，J＝7.7，1.8Hz，1H)，7.72(t，J＝7.4Hz，1H)，7.69(br.s，1H)，7.60(t，J＝7.7Hz，2H)，7.51(ddd，J＝8.2，7.4，2.0Hz，1H)，7.28(d，J＝8.2Hz，1H)，7.09(t，J＝7.5Hz，1H)，5.78(s，2H)。

5-acetyl-2- (2-oxo-2-phenyl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.56(d，J＝2.4Hz，1H)，8.35(br.s，1H)，8.08(dd，J＝8.2，1.5Hz，2H)，8.09(dd，J＝8.8，2.4Hz，1H)，7.86(br.s，1H)，7.73(t，J＝7.4Hz，1H)，7.61(t，J＝7.6Hz，2H)，7.40(d，J＝8.8Hz，1H)，5.90(s，2H)，2.57(s，3H)。

2- [2- (4-methoxy-phenyl) -2-oxo-ethoxy ] -benzamide

¹H NMR(DMSO-d₆)(ppm)：3.88(s，3H)5.71(s，2H)7.06-7.09(m，1H)7.09-7.14(m，2H)7.24-7.29(m，1H)7.47-7.53(m，1H)7.66(d，J＝1.46Hz，1H)7.98(dd，J＝7.75，1.77Hz，1H)8.02-8.10(m，2H)8.46(br.s，1H)。

5-chloro-2- (2-oxo-2-phenyl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.39(br.s，1H)，8.07(dd，J＝7.8，1.2Hz，2H)，7.91(d，J＝2.8Hz，1H)，7.87(br.s，1H)，7.72(t，J＝7.4Hz，1H)，7.60(t，J＝8.0Hz，2H)，7.57(dd，J＝8.8，2.9Hz，1H)，7.34(d，J＝8.9Hz，1H)，5.80(s，2H)。

5-fluoro-2- (2-oxo-2-phenyl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.47(br.s，1H)，8.07(dd，J＝8.4，1.3Hz，2H)，7.87(br.s，1H)，7.72(t，J＝7.6Hz，1H)，7.69(dd，J＝9.6，3.2Hz，1H)，7.60(t，J＝7.7Hz，2H)，7.39(ddd，J＝9.0，7.6，3.3Hz，1H)，7.34(dd，J＝9.1，4.5Hz，1H)，5.78(s，2H)。

4-nitro-2- (2-oxo-2-phenyl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.42(br.s，1H)，8.15(d，J＝8.7Hz，1H)，8.13(d，J＝2.1Hz，1H)，8.12(dd，J＝7.7，1.3Hz，2H)，8.02(br.s，1H)，7.94(dd，J＝8.5，2.1Hz，1H)，7.75(tt，J＝7.4，1.3Hz，1H)，7.62(t，J＝7.7Hz，2H)，5.97(s，2H)。

5-Nitro-2- (2-oxo-2-phenyl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.76(d，J＝3.0Hz，1H)，8.38(dd，J＝9.1，3.0Hz，1H)，8.36(br.s，1H)，8.08(dd，J＝8.3，1.1Hz，2H)，8.05(br.s，1H)，7.74(tt，J＝7.4，1.3Hz，1H)，7.61(t，J＝7.7Hz，2H)，7.52(d，J＝9.3Hz，1H)，5.97(s，2H)。

3-methyl-2- (2-oxo-2-phenyl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：7.96(dd，J＝8.4，1.2Hz，2H)，7.93(br.s，1H)，7.68(tt，J＝7.3，1.3Hz，1H)，7.55(t，J＝8.0Hz，2H)，7.51(ddd，J＝7.7，1.7，0.4Hz，1H)，7.48(br.s，1H)，7.35(ddd，J＝7.5，1.7，0.7Hz，1H)，7.13(t，J＝7.6Hz，1H)，5.42(s，2H)，2.31(s，3H)。

2- [2- (3-chloro-phenyl) -2-oxo-ethoxy ] -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.35(br.s，1H)，8.13(t，J＝1.8Hz，1H)，8.02(ddd，J＝7.7，1.5，1.0Hz，1H)，7.98(dd，J＝7.8，1.8Hz，1H)，7.79(ddd，J＝8.1、2.2，1.0Hz，1H)，7.70(br.s，1H)，7.63(t，J＝7.9Hz，1H)，7.51(ddd，J＝8.4，7.3，2.0Hz，1H)，7.31(dd，J＝8.4，0.7Hz，1H)，7.09(ddd，J＝7.8，7.3，1.0Hz，1H)，5.78(s，2H)。

5-fluoro-2- [2- (3-methoxy-phenyl) -2-oxo-ethoxy ] -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.47(br.s，1H)，7.87(br.s，1H)，7.69(dd，J＝9.5，3.1Hz，1H)，7.66(ddd，J＝7.7，1.6，1.0Hz，1H)，7.56(dd，J＝2.4，1.6Hz，1H)，7.51(t，J＝7.9Hz，1H)，7.39(ddd，J＝9.0，7.6，3.2Hz，1H)，7.34(dd，J＝9.1，4.5Hz，1H)，7.29(ddd，J＝8.3，2.7，0.8Hz，1H)，5.76(s，2H)，3.85(s，3H)。

5-fluoro-2- [2- (4-methoxy-phenyl) -2-oxo-ethoxy ] -benzamide

¹H NMR(DMSO-d₆)(ppm)：8.53(br.s，1H)，8.04(d，J＝9.0Hz，2H)，7.85(br.s，1H)，7.68(dd，J＝9.6，3.2Hz，1H)，7.38(ddd，J＝9.1，7.6，3.3Hz，1H)，7.32(dd，J＝9.1，4.4Hz，1H)，7.11(d，J＝8.9Hz，2H)，5.71(s，2H)，3.87(s，3H)。

2- [2- (3-chloro-phenyl) -2-oxo-ethoxy ] -5-fluoro-benzamide

¹H NMR(DMSO-d₆)(ppm)：8.41(br.s，1H)，8.12(t，J＝1.8Hz，1H)，8.01(dt，J＝8.0，1.2Hz，1H)，7.88(br.s，1H)，7.79(ddd，J＝8.0，2.2，1.0Hz，1H)，7.69(ddd，J＝9.6，3.0，0.5Hz，1H)，7.63(t，J＝7.9Hz，1H)，7.39(ddd，J＝9.1，7.4，3.2Hz，1H)，7.36(ddd，J＝9.1，4.6，0.5Hz，1H)，5.77(s，2H)。

2- (2-oxo-2-thiophen-3-yl-ethoxy) -benzamide

¹H NMR(DMSO-d₆)(ppm)：1.31-1.40(m，2H)1.40-1.52(m，4H)1.67(quin，J＝6.59Hz，2H)2.24(br.s，6H)3.51(t，J＝6.23Hz，2H)6.85(d，J＝8.79Hz，2H)7.51(ddd，J＝8.00，7.08，1.28Hz，3H)7.51(d，J＝8.67Hz，2H)7.77(ddd，J＝8.18，7.02，1.28Hz，1H)7.86(d，J＝7.95Hz，1H)8.20(ddd，J＝7.93，1.10，0.49Hz，1H)9.75(s，1H)11.05(s，1H)。

Step 2

3- (3-methoxy-phenyl) -4H-benzo [ f ] [1, 4] oxazepin-5-one

A well stirred suspension of 9g (31.5mmol) of 2- [2- (3-methoxy-phenyl) -2-oxo-ethoxy ] -benzamide and 0.3g (1.58mmol) of p-toluenesulfonic acid in toluene (0.3L) was refluxed with a Dean-Stark apparatus for 2 hours. After concentrating the solvent under reduced pressure, the residue was treated with ethyl acetate and diethyl ether, the resulting solid was filtered, and the mixture was washed with the aforementioned solvent and dried in vacuo to give 7.55g of the title compound (83%).

¹H NMR(DMSO-d₆)(ppm)：9.76(s，1H)，7.78(dd，J＝7.8，1.6Hz，1H)，7.57(ddd，J＝8.1，7.3，1.8Hz，1H)，7.30(t，J＝8.0Hz，1H)，7.29(td，J＝7.5，1.2Hz，1H)，7.14(dd，J＝8.1，0.8Hz，1H)，7.05(ddd，J＝7.7，1.6，0.9Hz，1H)，7.01(dd，J＝2.3，1.8Hz，1H)，6.94(ddd，J＝8.3，2.5，0.8Hz，1H)，6.91(d，J＝0.4Hz，1H)，3.78(s，3H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.77(s，1H)，7.78(dd，J＝7.7，1.6Hz，1H)，7.57(ddd，J＝8.2，7.4，1.8Hz，1H)，7.36-7.50(m，5H)，7.29(td，J＝7.6，0.9Hz，1H)，7.14(dd，J＝8.2，0.7Hz，1H)，6.87(s，1H)。

7-acetyl-3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.97(s，1H)，8.36(d，J＝2.2Hz，1H)，8.13(dd，J＝8.5，2.4Hz，1H)，7.38-7.52(m，5H)，7.28(d，J＝8.5Hz，1H)，6.90(s，1H)，2.61(s，3H)。

3- (4-methoxy-phenyl) -4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：3.76(s，3H)6.77(s，1H)6.94(d，J＝8.91Hz，3H)7.13(dd，J＝8.06，0.73Hz，1H)7.28(td，J＝7.54，1.04Hz，1H)7.39(d，J＝8.91Hz，2H)7.56(ddd，J＝8.06，7.32，1.83Hz，1H)7.77(dd，J＝7.81，1.71Hz，1H)9.72(s，1H)。

7-chloro-3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.95(s，1H)，7.75(d，J＝2.7Hz，1H)，7.63(dd，J＝8.7，2.8Hz，1H)，7.37-7.52(m，5H)，7.20(d，J＝8.7Hz，1H)，6.90(s，1H)。

7-fluoro-3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.93(s，1H)，7.51(dd，J＝9.0，3.2Hz，1H)，7.38-7.49(m，6H)，7.20(dd，J＝8.9，4.5Hz，1H)，6.90(s，1H)。

8-Nitro-3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：10.15(s，1H)，8.12(dd，J＝8.5，2.2Hz，1H)，8.04(d，J＝8.5Hz，1H)，7.94(d，J＝2.2Hz，1H)，7.48-7.52(m，2H)，7.38-7.44(m，3H)，6.95(s，1H)。

7-Nitro-3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：10.13(s，1H)，8.55(d，J＝2.8Hz，1H)，8.40(dd，J＝9.0，3.0Hz，1H)，7.48-7.53(m，2H)，7.41(s，4H)，6.92(s，1H)。

9-methyl-3-phenyl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.76(s，1H)，7.58(dd，1H)，7.42-7.50(m，3H)，7.33-7.42(m，3H)，7.16(t，J＝7.6Hz，1H)，6.97(s，1H)，2.31(s，3H)。

3- (3-chloro-phenyl) -4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.82(s，1H)，7.78(dd，J＝7.7，1.7Hz，1H)，7.57(td，1H)，7.53-7.54(m，1H)，7.40-7.47(m，3H)，7.30(td，J＝7.6，1.1Hz，1H)，7.14(dd，J＝8.1，0.8Hz，1H)，6.97(s，1H)。

7-fluoro-3- (3-methoxy-phenyl) -4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.91(s，1H)，7.51(dd，J＝8.9，3.2Hz，1H)，7.43(ddd，J＝8.9，8.1，3.3Hz，1H)，7.30(t，J＝7.9Hz，1H)，7.20(dd，J＝8.9，4.5Hz，1H)，7.04(ddd，J＝7.7，1.6，0.9Hz，1H)，7.01(t，J＝2.0Hz，1H)，6.93(s，1H)，6.94(ddd，J＝8.3，2.6，0.9Hz，1H)，3.78(s，3H)。

7-fluoro-3- (4-methoxy-phenyl) -4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.87(s，1H)，7.50(dd，J＝9.0，3.2Hz，1H)，7.42(ddd，J＝8.9，8.1，3.3Hz，1H)，7.39(d，J＝8.9Hz，2H)，7.19(dd，J＝8.9，4.6Hz，1H)，6.95(d，J＝8.8Hz，2H)，6.79(s，1H)，3.76(s，3H)。

3- (3-chloro-phenyl) -7-fluoro-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：9.97(s，1H)，7.53-7.56(m，1H)，7.50(dd，J＝8.9，3.2Hz，1H)，7.36-7.47(m，4H)，7.21(dd，J＝8.9，4.5Hz，1H)，6.99(s，1H)。

3-thiophen-3-yl-4H-benzo [ f ] [1, 4] oxazepin-5-one

¹H NMR(DMSO-d₆)(ppm)：7.10(s，1H)7.14(dd，J＝8.12，0.92Hz，1H)7.28(td，J＝7.50，1.15Hz，1H)7.31(dd，J＝5.13，1.34Hz，1H)7.56(ddd，J＝8.39，7.11，1.83Hz，1H)7.57(dd，J＝5.13，2.93Hz，1H)7.67(dd，J＝2.87，1.28Hz，1H)7.75(dd，J＝7.75，1.65Hz，1H)9.80(s，1H)。

Step 3

4-hydroxy-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one

0.225g (5.6mmol) of sodium hydride (60% dispersion in mineral oil) was added under a strictly oxygen-free atmosphere to a suspension of 1g (3.74mmol) of 3- (3-methoxy-phenyl) -4H-benzo [ f ] [1, 4] oxazepin-5-one in dioxane (20mL) and the mixture was heated at 100 ℃ for 45 min. After cooling to room temperature, 2N hydrochloric acid was added until pH 1, then water was added until complete precipitation of the product was achieved. The solid was filtered, washed with water and dried in vacuo to provide 0.82g of the title compound (82%).

¹H NMR(DMSO-d₆)(ppm)：10.93(br.s，1H)，8.28(s，1H)，8.22(ddd，J＝8.0，1.4，0.6Hz，1H)，7.96(d，J＝8.0Hz，1H)，7.79(ddd，J＝8.3，7.0，1.5Hz，1H)，7.54(ddd，J＝8.0，7.0，1.2Hz，1H)，7.37(t，J＝7.9Hz，1H)，7.28(dt，J＝7.7，1.2Hz，1H)，7.24(dd，J＝2.6，1.6Hz，1H)，6.96(ddd，J＝8.2，2.5，1.0Hz，1H)，3.82(s，3H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

4-hydroxy-3-phenyl-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：10.96(s，1H)，8.26(s，1H)，8.22(dd，J＝8.0，0.8Hz，1H)，7.96(d，J＝7.8Hz，1H)，7.79(td，J＝7.6，1.3Hz，1H)，7.66-7.71(m，2H)，7.54(ddd，J＝8.0，7.1，1.1Hz，1H)，7.43-7.49(m，2H)，7.36-7.41(m，1H)。

7-acetyl-4-hydroxy-3-phenyl-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.27(s，1H)，8.77(d，J＝1.6Hz，1H)，8.44(s，1H)，8.28(dd，J＝8.5，2.0Hz，1H)，8.04(d，J＝8.5Hz，1H)，7.71(d，J＝7.7Hz，2H)，7.49(t，J＝7.3Hz，2H)，7.42(t，J＝7.2Hz，1H)，2.69(s，3H)。

4-hydroxy-3- (4-methoxy-phenyl) -2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：3.81(s，3H)7.02(d，J＝8.91Hz，2H)7.51(ddd，J＝8.03，7.05，1.16Hz，1H)7.63(d，J＝8.79Hz，2H)7.77(ddd，J＝8.24，7.02，1.34Hz，1H)7.93(ddd，J＝8.06，0.98，0.61Hz，1H)8.14(s，1H)8.20(ddd，J＝8.06，1.34，0.60Hz，1H)10.90(s，1H)。

7-fluoro-4-hydroxy-3-phenyl-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.13(s，1H)，8.38(s，1H)，8.02(dd，J＝9.1，5.4Hz，1H)，7.87(dd，J＝9.5，2.8Hz，1H)，7.66-7.71(m，1H)，7.65-7.68(m，2H)，7.44-7.49(m，2H)，7.37-7.41(m，1H)。

4-hydroxy-6-nitro-3-phenyl-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.45(s，1H)，8.75(d，J＝2.2Hz，1H)，8.72(s，1H)，8.44(d，J＝8.7Hz，1H)，8.25(dd，J＝8.8，2.3Hz，1H)，7.70(d，J＝7.8Hz，2H)，7.50(t，J＝7.3Hz，2H)，7.44(t，J＝7.3Hz，1H)。

4-hydroxy-7-nitro-3-phenyl-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.55(s，1H)，8.93(d，J＝2.3Hz，1H)，8.64(s，1H)，8.54(dd，J＝9.0，2.5Hz，1H)，8.14(d，J＝8.9Hz，1H)，7.71(dd，J＝7.3，1.4Hz，2H)，7.43-7.54(m，3H)。

3- (3-chloro-phenyl) -4-hydroxy-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.05(s，1H)，8.46(s，1H)，8.22(ddd，J＝8.1，1.3，0.5Hz，1H)，7.97(d，J＝8.1Hz，1H)，7.80(ddd，J＝8.2，7.0，1.3Hz，1H)，7.74(t，J＝1.6Hz，1H)，7.64(dt，J＝7.4，1.4Hz，1H)，7.56(ddd，J＝8.0，7.0，1.2Hz，1H)，7.49(td，J＝7.8，0.4Hz，1H)，7.45(ddd，J＝7.9，2.0，1.3Hz，1H)。

4-hydroxy-5-methyl-3-phenyl-2H-isoquinolin-1-one

MS calculated: 251.0946, respectively; MS found: 251.0950

ESI(+)MS：m/z 252(MH⁺)。

7-fluoro-4-hydroxy-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.10(s，1H)，8.41(s，1H)，8.02(dd，J＝9.1，5.3Hz，1H)，7.87(dd，J＝9.5，2.7Hz，1H)，7.68(td，J＝8.8，2.8Hz，1H)，7.37(t，J＝7.9Hz，1H)，7.27(dt，J＝7.8，1.2Hz，1H)，7.23(dd，J＝2.4，1.6Hz，1H)，6.96(ddd，J＝8.2，2.6，0.9Hz，1H)，3.82(s，3H)。

7-fluoro-4-hydroxy-3- (4-methoxy-phenyl) -2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.06(s，1H)，8.27(s，1H)，7.99(dd，J＝9.1，5.2Hz，1H)，7.85(dd，J＝9.5，2.7Hz，1H)，7.67(td，J＝8.8，2.8Hz，1H)，7.62(d，J＝8.9Hz，2H)，7.02(d，J＝8.9Hz，2H)，3.81(s，3H)。

3- (3-chloro-phenyl) -7-fluoro-4-hydroxy-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：11.20(s，1H)，8.58(s，1H)，8.03(dd，J＝8.9，5.2Hz，1H)，7.88(dd，J＝9.4，2.7Hz，1H)，7.73(t，J＝1.5Hz，1H)，7.70(td，J＝8.8，2.8Hz，1H)，7.63(dt，J＝7.5，1.3Hz，1H)，7.42-7.52(m，2H)。

4-hydroxy-3-thiophen-3-yl-2H-isoquinolin-1-one

¹H NMR(DMSO-d₆)(ppm)：7.52(ddd，J＝7.99，7.08，1.16Hz，1H)7.63(dd，J＝5.07，2.99Hz，1H)7.78(ddd，J＝8.24，7.02，1.34Hz，1H)7.80(dd，J＝5.07，1.28Hz，1H)7.97(ddd，J＝8.18，0.98，0.60Hz，1H)8.11(dd，J＝2.99，1.28Hz，1H)8.20(ddd，J＝8.00，1.34，0.49Hz，1H)8.55(s，1H)10.85(s，1H)。

Step 5

{2- [3- (3-methoxy-phenyl) -1-oxo-1, 2-dihydro-isoquinolin-4-yloxy ] -ethyl } -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₂＝R₃＝H，R₁Tert-butoxycarbonyl and R₄3-methoxyphenyl).

3.18g (14.19mmol) of tert-butyl (2-bromo-ethyl) -carbamate and 5.09g (15.6mmol) of cesium carbonate are added to a solution of 3.79g (14.19mmol) of 4-hydroxy-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one in N, N-dimethylacetamide (90mL), and the resulting mixture is stirred at room temperature for 2H under argon. Water was added to the reaction mixture, which was extracted 2 times with ethyl acetate. The combined organic layers were washed with water at least 3 times and then evaporated to dryness. The crude product was purified by flash chromatography (eluent: ethyl acetate/hexane 1/1) to provide 3.2g of the title compound (55%).

¹H NMR(DMSO-d₆)(ppm)：11.18(s，1H)，8.23(ddd，J＝7.9，1.2，0.6Hz，1H)，7.89(d，J＝8.0Hz，1H)，7.78(ddd，J＝8.1，7.0，1.3Hz，1H)，7.57(ddd，J＝8.0，7.0，1.2Hz，1H)，7.40(t，J＝7.9Hz，1H)，7.30(dt，J＝8.2，1.2Hz，1H)，7.24(dd，J＝2.4，1.5Hz，1H)，7.01(ddd，J＝8.2，2.6，0.9Hz，1H)，6.84(t，J＝5.4Hz，1H)，3.83(s，3H)，3.49(t，J＝5.8Hz，2H)，3.08(q，J＝6.0Hz，2H)，1.36(s，9H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

[2- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl]-carbamic acid tert-butyl ester (I, L ═ CH)₂-CH₂，R＝R₂＝R₃＝H，R₁Tert-butoxycarbonyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.36(s，9H)3.06(q，J＝5.61Hz，2H)3.46(t，J＝5.79Hz，2H)6.82(t，J＝5.55Hz，1H)7.56(ddd，J＝8.08，7.04，1.22Hz，1H)7.70(dd，J＝8.11，1.52Hz，2H)7.78(ddd，J＝8.17，7.07，1.34Hz，1H)7.88(d，J＝7.92Hz，1H)8.24(ddd，J＝7.92，1.34，0.61Hz，1H)11.21(s，1H)。

[3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂-CH₂，R＝R₂＝R₃＝H，R₁Tert-butoxycarbonyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.35(s，9H)1.61(quin，J＝6.70Hz，2H)2.91(q，J＝6.40Hz，2H)3.49(t，J＝6.22Hz，2H)6.65(t，J＝5.97Hz，1H)7.56(ddd，J＝8.01，6.73，1.46Hz，1H)7.67(dd，J＝8.11，1.52Hz，2H)7.80(ddd，J＝8.05，6.83，1.22Hz，1H)7.84(dd，J＝8.15，1.40Hz，1H)8.24(ddd，J＝8.05，1.22，0.70Hz，1H)11.22(s，1H)。

4- [2- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -piperazine-1-carboxylic acid tert-butyl ester

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁co-N-tert-butoxycarbonyl-piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.39(s，9H)2.20(t，J＝5.00Hz，4H)2.34(t，J＝5.00Hz，2H)2.46(t，J＝5.73Hz，2H)3.42(t，J＝5.79Hz，2H)3.56(t，J＝5.42Hz，2H)7.56(ddd，J＝8.05，7.07，1.10Hz，1H)7.68(dd，J＝8.05，1.46Hz，2H)7.82(ddd，J＝8.17，7.07，1.34Hz，1H)7.98(d，J＝8.17Hz，1H)8.23(dd，J＝7.98，0.79Hz，1H)11.21(s，1H)。

{2- [3- (4-methoxy-phenyl) -1-oxo-1, 2-dihydro-isoquinolin-4-yloxy ] -ethyl } -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₂＝R₃＝H，R₁Tert-butoxycarbonyl and R₄4-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：1.37(s，9H)3.08(q，J＝5.25Hz，2H)3.46(t，J＝5.74Hz，2H)3.83(s，3H)6.83(t，J＝5.61Hz，1H)7.04(d，J＝8.91Hz，2H)7.54(ddd，J＝8.03，7.05，1.16Hz，1H)7.66(d，J＝8.91Hz，2H)7.77(ddd，J＝8.12，7.02，1.34Hz，1H)7.86(d，J＝7.93Hz，1H)8.22(ddd，J＝7.93，1.34，0.50Hz，1H)11.14(s，1H)。

[2- (7-fluoro-1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₃＝H，R₂＝7-F，R₁Tert-butoxycarbonyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.37(s，1H)，7.95(dd，J＝8.8，5.3Hz，1H)，7.90(dd，J＝9.4，2.7Hz，1H)，7.69(dd，J＝7.9，1.5Hz，2H)，7.67(td，J＝8.9，2.8Hz，1H)，7.44-7.53(m，3H)，6.81(t，J＝5.3Hz，1H)，3.45(t，J＝5.8Hz，2H)，3.05(q，J＝5.4Hz，2H)，1.36(s，9H)。

{2- [3- (3-chloro-phenyl) -1-oxo-1, 2-dihydro-isoquinolin-4-yloxy ] -ethyl } -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₂＝R₃＝H，R₁Tert-butoxycarbonyl and R₄3-chlorophenyl).

¹H NMR(DMSO-d₆)(ppm)：11.29(s，1H)，8.24(ddd，J＝7.9，1.1，0.6Hz，1H)，7.89(d，J＝8.1Hz，1H)，7.80(ddd，J＝8.1，7.0，1.3Hz，1H)，7.71-7.75(m，1H)，7.66-7.70(m，1H)，7.59(ddd，1H)，7.50-7.54(m，2H)，6.84(t，J＝5.4Hz，1H)，3.49(t，J＝5.7Hz，2H)，3.08(q，J＝5.4Hz，2H)，1.36(s，9H)。

[2- (5-methyl-1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₂＝H、R₃Not being 5-methyl, R₁Tert-butoxycarbonyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.23(s，1H)，8.14(ddd，J＝8.1，1.5，0.5Hz，1H)，7.70(dd，J＝8.0，1.5Hz，2H)，7.53-7.58(m，J＝7.4，1.4，0.5，0.5，0.5，0.5Hz，1H)，7.44-7.52(m，3H)，7.41(t，J＝7.6Hz，1H)，6.55(t，J＝5.6Hz，1H)，3.28-3.31(m，2H)，2.92(q，J＝5.9Hz，2H)，2.75(s，3H)，1.34(s，9H)。

{2- [ 7-fluoro-3- (3-methoxy-phenyl) -1-oxo-1, 2-dihydro-isoquinolin-4-yloxy]-ethyl } -carbamic acid tert-butyl ester (I, L ═ CH)₂-CH₂，R＝R₃＝H，R₂＝7-F，R₁Tert-butoxycarbonyl and R₄3-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.34(s，1H)，7.96(dd，J＝8.8，5.2Hz，1H)，7.90(dd，J＝9.4，2.7Hz，1H)，7.67(td，J＝8.8，2.9Hz，1H)，7.40(t，J＝7.9Hz，1H)，7.29(d，J＝7.9Hz，1H)，7.21-7.25(m，1H)，7.01(ddd，J＝8.2，2.6，0.8Hz，1H)，6.77-6.87(m，1H)，3.83(s，3H)，3.48(t，J＝5.7Hz，2H)，3.08(q，J＝5.6Hz，2H)，1.36(s，9H)。

{2- [ 7-fluoro-3- (4-methoxy-phenyl) -1-oxo-1, 2-dihydro-isoquinolin-4-yloxy]-ethyl } -carbamic acid tert-butyl ester (I, L ═ CH)₂-CH₂，R＝R₃＝H，R₂＝7-F，R₁Tert-butoxycarbonyl and R₄4-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.29(s，1H)，7.93(dd，J＝9.0，5.3Hz，1H)，7.88(dd，J＝9.3，2.7Hz，1H)，7.65(d，J＝8.8Hz，2H)，7.66(ddd，J＝9.0，8.0，2.8Hz，1H)，7.04(d，J＝8.9Hz，2H)，6.82(t，J＝5.2Hz，1H)，3.82(s，3H)，3.45(t，J＝5.6Hz，2H)，3.08(q，J＝5.7Hz，2H)，1.36(s，9H)。

{2- [3- (3-chloro-phenyl) -7-fluoro-1-oxo-1, 2-dihydro-isoquinolin-4-yloxy ] -ethyl } -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₃＝H，R₂＝7-F，R₁Tert-butoxycarbonyl and R₄3-chlorophenyl).

¹H NMR(DMSO-d₆)(ppm)：11.43(s，1H)，7.96(dd，J＝8.8，5.3Hz，1H)，7.90(dd，J＝9.3，2.7Hz，1H)，7.70-7.74(m，1H)，7.64-7.71(m，2H)，7.50-7.53(m，2H)，6.83(t，J＝5.6Hz，1H)，3.49(t，J＝5.7Hz，2H)，3.07(q，J＝5.4Hz，2H)，1.36(s，9H)。

[2- (1-oxo-3-thiophen-3-yl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂，R＝R₂＝R₃＝H，R₁Tert-butoxycarbonyl and R₄Thien-3-yl).

¹H NMR(DMSO-d₆)(ppm)：1.40(s，6H)3.21-3.28(m，2H)3.60(t，J＝5.55Hz，2H)7.08(br.s，1H)7.55(td，J＝7.54，1.16Hz，1H)7.67(dd，J＝5.07，2.99Hz，1H)7.74(dd，J＝5.13，1.34Hz，1H)7.78(ddd，J＝8.15，6.99，1.34Hz，1H)7.84-7.88(m，1H)8.17(dd，J＝2.81，1.22Hz，1H)8.22(dd，J＝7.99，0.67Hz，1H)11.09(s，1H)。

7-acetyl-3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one (Compound 30)

(I，L＝CH₂-CH₂，R₂7-acetyl, R₃H, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.49(br.s，1H)，8.77(d，J＝1.5Hz，1H)，8.27(dd，J＝8.5，1.9Hz，1H)，8.13(d，J＝8.5Hz，1H)，7.71(dd，J＝8.0，1.5Hz，2H)，7.44-7.57(m，3H)，3.54(br.s，2H)，2.69(s，3H)，2.40(br.s，2H)，2.21(br.s，4H)，1.43(br.s，4H)，1.34(br.s，2H)。

6-Nitro-3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one (Compound 31)

(I，L＝CH₂-CH₂，R₂6-nitro, R₃H, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.67(br.s，1H)，8.80(br.s，1H)，8.45(d，J＝8.8Hz，1H)，8.27(dd，J＝8.7，2.1Hz，1H)，7.69-7.74(m，2H)，7.48-7.59(m，3H)，3.55(br.s，2H)，2.39(br.s，2H)，2.23(br.s，4H)，1.28-1.45(m，6H)。

4- [2- (dimethylamino) ethoxy ] -7-fluoro-3-phenylisoquinolin-1 (2H) -one (Compound 84)

(I，L＝CH₂CH₂，R₂＝F，R₃＝H，R＝R₁Methyl, R₄Arbityl phenyl

¹H NMR(DMSO-d₆)(ppm)：11.37(s，1H)，8.02(dd，J＝5.25，8.91Hz，1H)，7.89(dd，J＝2.69，9.40Hz，1H)，7.64-7.76(m，4H)，7.34-7.55(m，4H)，3.53(t，J＝5.74Hz，2H)，2.35-2.45(m，2H)，2.07(s，6H)。

Tert-butyl 4- [ (7-fluoro-1-oxo-3-phenyl-1, 2-dihydroisoquinolin-4-yl) oxy ] piperidine-1-carboxylate

(I, L taken together with the nitrogen to which it is bonded is piperidin-4-yl, R₂＝F，R₃H, R is tert-butoxycarbonyl, R₄Arbityl phenyl

¹H NMR(DMSO-d₆)(ppm)：11.39(s，1H)，7.87-7.96(m，1H)，7.63-7.74(m，1H)，7.43-7.53(m，1H)，4.82(dt，J＝4.26，8.21Hz，1H)，4.14(dd，J＝3.30，5.74Hz，1H)，3.56-3.64(m，1H)，3.14-3.22(m，1H)。

3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 10)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ 1-piperidinyl and R₄3-methoxyphenyl).

1.45g (5.73mmol) of 1- (3-chloro-propyl) -piperidine hydrochloride, 1.58g (11.5mmol) of potassium carbonate and 47mg (0.286mmol) of potassium iodide were added to a suspension of 766mg (2.86mmol) of 4-hydroxy-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one in methanol (29mL), and the mixture was placed in a microwave at 85 ℃ while cooling for 15 min. The solvent was concentrated under vacuum and the mixture was treated with a saturated solution of dichloromethane and sodium bicarbonate. The organic layer was washed with brine and then evaporated to dryness. The crude product was purified by flash chromatography (eluent: dichloromethane/methanol 96/4) to provide 456mg of the title compound (41%).

¹H NMR(DMSO-d₆)(ppm)：11.19(s，1H)，8.23(dd，J＝7.9，0.6Hz，1H)，7.90(d，J＝7.9Hz，1H)，7.80(ddd，J＝8.2，7.0，1.3Hz，1H)，7.56(ddd，J＝8.0，7.0，1.1Hz，1H)，7.40(t，J＝7.9Hz，1H)，7.27(dt，J＝7.9，1.1Hz，1H)，7.23(dd，J＝2.5，1.5Hz，1H)，7.02(ddd，J＝8.2，2.6，0.7Hz，1H)，3.83(s，3H)，3.55(t，J＝6.2Hz，2H)，2.22(br.s，6H)，1.68(br.s，2H)，1.45(br.s，4H)，1.37(br.s，2H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

3- (3-methoxy-phenyl) -4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one (Compound 9)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁Are combined together1-piperidinyl and R₄3-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：1.29-1.39(m，2H)1.39-1.50(m，4H)2.24(br.s，4H)2.44(t，J＝4.82Hz，2H)3.57(t，J＝5.42Hz，2H)3.84(s，3H)7.03(ddd，J＝8.29，2.56，0.85Hz，1H)7.25(dd，J＝2.38，1.52Hz，1H)7.29(ddd，J＝7.68，1.34，0.98Hz，1H)7.41(t，J＝7.92Hz，1H)7.56(td，J＝7.56，1.10Hz，1H)7.81(ddd，J＝8.17，7.07，1.34Hz，1H)7.56(ddd，J＝7.98，7.13，1.10Hz，1H)8.23(ddd，J＝8.05，1.22，0.49Hz，1H)11.17(s，1H)。

3- (4-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 23)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ 1-piperidinyl and R₄4-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：1.40(br.s，2H)1.51(br.s，4H)1.76(br.s，2H)2.35(s，6H)3.53(t，J＝6.10Hz，2H)3.83(s，3H)7.06(d，J＝8.79Hz，2H)7.54(ddd，J＝8.15，6.93，1.04Hz，1H)7.63(d，J＝8.79Hz，2H)7.79(ddd，J＝8.06，7.05，1.10Hz，1H)7.85(d，J＝8.06Hz，1H)8.22(d，J＝7.81Hz，1H)11.17(br.s，1H)。

3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one trifluoroacetate salt (Compound 5)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(400MHz，DMSO-d₆)(ppm)：1.36(qt，J＝12.80，3.29Hz，1H)1.80(d，J＝14.14Hz，2H)1.89(dq，J＝10.80，5.80Hz，2H)2.72-2.85(m，2H)2.87-2.98(m，2H)3.55(t，J＝5.91Hz，2H)7.56-7.62(m，1H)7.83(d，J＝3.66Hz，2H)8.26(d，J＝7.92Hz，1H)8.92(b r.s，1H)11.29(s，1H)。

4- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -piperazine-1-carboxylic acid tert-butyl ester

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁co-N-tert-butoxycarbonyl-piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.39(s，9H)1.65(quin，J＝6.43Hz，2H)2.18(t，J＝4.20Hz，4H)2.23(t，J＝6.89Hz，2H)3.23(t，J＝4.50Hz，4H)3.53(t，J＝6.16Hz，2H)7.56(ddd，J＝8.17，7.19，1.22Hz，1H)7.65-7.70(m，2H)7.81(ddd，J＝7.80，7.30，1.22Hz，1H)7.88(ddd，J＝8.00，1.10，0.60Hz，1H)8.23(ddd，J＝8.05，1.22，0.50Hz，1H)11.21(s，1H)。

6-Nitro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 34)

(I，L＝CH₂-CH₂-CH₂，R₂＝H，R₃6-Nitro, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.69(br.s，1H)，8.52(d，J＝2.2Hz，1H)，8.46(d，J＝8.8Hz，1H)，8.27(dd，J＝8.7，2.3Hz，1H)，7.70(dd，J＝7.9，1.6Hz，2H)，7.46-7.58(m，3H)，3.56(t，J＝6.2Hz，2H)，2.25(br.s，6H)，1.69(br.s，2H)，1.45(br.s，4H)，1.36(br.s，2H)。

7-Nitro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 35)

(I，L＝CH₂-CH₂-CH₂，R₂＝H，R₃7-nitro, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.77(br.s，1H)8.91(d, J ═ 2.3Hz, 1H), 8.49(dd, J ═ 9.0, 2.5Hz, 1H), 8.06(d, J ═ 9.0Hz, 1H), 7.62 to 7.71(m, 2H), 7.48 to 7.56(m, 3H), 3.51(t, J ═ 6.2Hz, 2H), 2.04 to 2.91(m, 6H), 1.32 to 1.74(m, 8H) 7-chloro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 37)

(I，L＝CH₂-CH₂-CH₂，R₂＝7-Cl，R₃H, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.42(s，1H)，8.17(d，J＝2.0Hz，1H)，7.93(d，J＝8.5Hz，1H)，7.82(dd，J＝8.7，2.3Hz，1H)，7.67(dd，J＝8.1，1.6Hz，2H)，7.43-7.55(m，3H)，3.51(t，J＝6.3Hz，2H)，2.18(br.s，6H)，1.62(quin，J＝6.7Hz，2H)，1.42(br.s，4H)，1.28-1.38(m，2H)。

7-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 38)

(I，L＝CH₂-CH₂-CH₂，R₂＝7-F，R₃H, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.37(s，1H)，7.97(dd，J＝8.9，5.2Hz，1H)，7.89(dd，J＝9.4，2.8Hz，1H)，7.69(td，J＝8.7，2.8Hz，1H)，7.67(dd，J＝8.1，1.5Hz，2H)，7.42-7.53(m，3H)，3.51(t，J＝6.2Hz，2H)，2.22(br.s，6H)，1.64(quin，J＝6.4Hz，2H)，1.43(br.s，4H)，1.35(br.s，2H)。

3- (3-chloro-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 44)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ 1-piperidinyl and R₄3-chlorophenyl).

¹H NMR(DMSO-d₆)(ppm)：11.29(s，1H)，8.24(ddd，J＝7.9，1.2，0.5Hz，1H)，7.91(ddd，J＝8.1，1.1，0.6Hz，1H)，7.81(ddd，J＝8.2，7.0，1.3Hz，1H)，7.73(td，J＝1.6，0.9Hz，1H)，7.64(tt，J＝3.7，1.7Hz，1H)，7.58(ddd，J＝8.0，7.0，1.2Hz，1H)，7.51-7.54(m，2H)，3.55(t，J＝6.2Hz，2H)，2.19(t，J＝7.0Hz，6H)，1.65(quin，J＝6.6Hz，2H)，1.43(quin，J＝5.4Hz，4H)，1.30-1.39(m，2H)。

Methyl- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -carbamic acid tert-butyl ester

(I，L＝CH₂-CH₂-CH₂R is methyl, R₁Tert-butoxycarbonyl, R₂＝R₃＝H，R₄Phenyl).

MS calculated: 408.2049, respectively; MS found: 408.2046

ESI(+)MS：m/z 409(MH⁺)。

4- (3-diethylamino-propoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 18)

(I，L＝CH₂-CH₂-CH₂，R＝R₁Ethyl, R₂＝R₃H, and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.09(br.s，6H)1.80(br.s，2H)3.57(t，J＝5.85Hz，2H)7.47-7.55(m，3H)7.58(ddd，J＝8.02，6.55，1.52Hz，1H)7.68(dd，J＝7.99，1.52Hz，2H)7.82(td，J＝8.05，1.10Hz，1H)8.25(d，J＝7.93Hz，1H)8.91(br.s，1H)11.27(br.s，1H)。

3-phenyl-4- (2-pyrrol-1-yl-ethoxy) -2H-isoquinolin-1-one (Compound 13)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ pyrrole and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：3.65(t，J＝5.12Hz，2H)4.04(t，J＝5.12Hz，2H)6.06(t，J＝2.13Hz，2H)6.75(t，J＝2.07Hz，2H)7.23(d，J＝8.05Hz，1H)7.44-7.49(m，2H)7.49-7.54(m，1H)7.59-7.65(m，1H)7.65-7.70(m，2H)8.20(dd，J＝7.93，0.73Hz，1H)11.21(s，1H)。

3-phenyl-4- (3-pyrrol-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 15)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ pyrrole and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.90(quin，J＝6.60Hz，2H)3.50(t，J＝5.97Hz，2H)3.80(t，J＝7.13Hz，2H)5.91(t，2H)6.55(t，J＝2.07Hz，2H)7.56(dd，J＝8.05，4.15Hz，1H)7.65-7.71(m，2H)7.80(d，J＝3.54Hz，2H)8.24(dt，J＝7.93，0.91Hz，1H)11.23(s，1H)。

3-phenyl-4- (3-pyrrolidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 21)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ pyrrolidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.74(br.s，6H)2.63(br.s，6H)3.55(t，J＝6.10Hz，2H)7.57(ddd，J＝8.05，6.89，1.28Hz，1H)7.68(dd，J＝8.05，1.46Hz，1H)7.81(ddd，J＝8.17，6.95，1.22Hz，1H)7.87(dd，J＝8.20，1.20Hz，1H)8.24(d，J＝7.56Hz，1H)11.24(s，1H)。

7-fluoro-3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one hydrochloride (Compound 43)

(I，L＝CH₂-CH₂-CH₂，R₂＝H，R₃7-F, R and R₁Together ═ 1-piperidinyl and R₄3-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.41(s，1H)，9.43(br.s，1H)，7.91(dd，J＝9.2，4.9Hz，1H)，7.91(dd，J＝9.5，2.8Hz，1H)，7.72(td，J＝8.7，2.8Hz，1H)，7.44(t，J＝8.0Hz，1H)，7.26(dt，J＝7.9，1.0Hz，1H)，7.21(dd，J＝2.4，1.6Hz，1H)，7.06(ddd，J＝8.3，2.6，0.9Hz，1H)，3.84(s，3H)，3.57(t，J＝5.9Hz，2H)，3.24-3.39(m，2H)，2.92(td，J＝8.1，5.1Hz，2H)，2.70-2.83(m，2H)，1.94(dq，J＝11.0，5.8Hz，2H)，1.57-1.83(m，4H)，1.26-1.70(m，2H)。

4- (2-chloro-ethoxy) -3-phenyl-2H-isoquinolin-1-one

315mg (2.28mmol) of potassium carbonate and 190L (2.28mmol) of 1-bromo-2-chloroethane are added to a suspension of 180mg (0.76mmol) of 4-hydroxy-3-phenyl-2H-isoquinolin-1-one in methanol (6mL), and the mixture is placed in a microwave at 120 ℃ for 10 min. The solvent was concentrated under reduced pressure, and the resulting crude product was purified by flash chromatography (eluent: ethyl acetate/hexane 1/2) to give 100mg of the title compound (44%).

¹H NMR(DMSO-d₆)(ppm)：11.21(br.s，1H)，8.23(dd，J＝7.9，0.7Hz，1H)，7.94(d，J＝7.7Hz，1H)，7.81(ddd，J＝8.2，7.0，1.4Hz，1H)，7.66-7.72(m，2H)，7.55(ddd，J＝8.0，7.1，1.2Hz，1H)，7.42-7.52(m，3H)，3.55(t，J＝5.9Hz，2H)，2.55(t，J＝5.9Hz，2H)，2.25-2.35(m，4H)，1.62(dt，4H)

Tert-butyl {3- [ (7-fluoro-1-oxo-3-phenyl-1, 2-dihydroisoquinolin-4-yl) oxy ] propyl } carbamate

(I，L＝CH₂CH₂CH₂，R₂＝F，R₃＝R＝H，R₁Tert-butoxycarbonyl, R₄Arbityl phenyl

0.38g (1.58mmol) of tert-butyl (3-bromo-propyl) -carbamate and 0.084g (3.56mmol) of sodium hydride are added to a solution of 0.45g (1.76mmol) of 7-fluoro-4-hydroxy-3-phenyl-2H-isoquinolin-1-one in N, N-dimethylformamide (formammide) (18mL), and the resulting mixture is stirred under argon at room temperature for 2 hours. Water was added to the reaction mixture, which was extracted 2 times with ethyl acetate. The combined organic layers were washed with water at least 3 times and then evaporated to dryness. The crude product was purified by flash chromatography (eluent: ethyl acetate/hexane 4/6) to afford 0.4g of the title compound (55%).

1H NMR(DMSO-d6)(ppm)：11.37(s，1H)，7.85-7.95(m，2H)，7.61-7.72(m，3H)，7.41-7.54(m，3H)，6.65(br.s，1H)，3.48(t，J＝6.16Hz，2H)，2.86-2.95(m，2H)，1.60(t，J＝6.77Hz，2H)，1.34-1.37(m，8H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

7-fluoro-3-phenyl-4- [2- (tetrahydro-2H-pyran-2-yloxy) ethoxy ] isoquinolin-1 (2H) -one

¹H NMR(DMSO-d₆)(ppm)：11.36(s，1H)，8.08(dd，J＝5.25，8.91Hz，1H)，7.89(dd，J＝2.75，9.34Hz，1H)，7.66-7.74(m，3H)，

7.41-7.54(m，3H)，4.47(t，J＝2.50Hz，1H)。tep 6

4- (2-Morpholin-4-yl-ethoxy) -3-phenyl-2H-isoquinolin-1-one (Compound 54)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁Co ═ morpholino and R₄Phenyl).

11mg (0.08mmol) of potassium carbonate and 7L (0.08mmol) of morpholine are added to a suspension of 20mg (0.07mmol) of 4- (2-chloro-ethoxy) -3-phenyl-2H-isoquinolin-1-one in acetonitrile (0.35mL) and the mixture is left for 1H in a microwave at 120 ℃. The solvent was concentrated under reduced pressure and the resulting crude product was purified by preparative HPLC to provide 7mg of the title compound (28%).

¹H NMR(DMSO-d₆)(ppm)：11.22(br.s，1H)，8.24(d，J＝7.7Hz，1H)，8.00(d，J＝7.8Hz，1H)，7.80-7.86(m，1H)，7.67-7.71(m，2H)，7.57(t，J＝7.6Hz，1H)，7.43-7.53(m，3H)，3.54-3.60(m，2H)，3.51(br.s，4H)，2.42-2.48(m，2H)，2.25(br.s，4H)

Following the same methodology, but employing appropriately substituted derivatives, the following compounds were prepared:

3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one (Compound 51)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ 1-piperidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.19(br.s，1H)，8.23(ddd，J＝7.9，1.3，0.7Hz，1H)，8.01(ddd，J＝8.1，1.0，0.4Hz，1H)，7.80(ddd，J＝8.2，7.0，1.3Hz，1H)，7.66-7.70(m，2H)，7.55(ddd，J＝8.0，7.1，1.1Hz，1H)，7.43-7.52(m，3H)，3.53(t，J＝5.5Hz，2H)，2.40(t，J＝5.5Hz，2H)，2.21(t，J＝4.9Hz，4H)，1.42(quin，J＝5.4Hz，4H)，1.33(q，J＝5.6Hz，2H)。

4- (2-diethylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one (Compound 52)

(I，L＝CH₂-CH₂，R＝R₁Ethyl, R₂＝R₃H and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.20(br.s，1H)，8.23(ddd，J＝8.1，1.3，0.5Hz，1H)，7.98(ddd，J＝8.2，1.1，0.6Hz，1H)，7.81(ddd，J＝8.2，7.0，1.3Hz，1H)，7.66-7.71(m，2H)，7.56(ddd，J＝8.1，7.0，1.2Hz，1H)，7.43-7.53(m，3H)，3.50(t，J＝6.3Hz，2H)，2.51-2.54(m，2H)，2.36(q，J＝7.1Hz，4H)，0.85(t，J＝7.1Hz，6H)。

3-phenyl-4- (2-pyrrolidin-1-yl-ethoxy) -2H-isoquinolin-1-one (Compound 53)

(I，L＝CH₂-CH₂-，R₂＝R₃H, R and R₁Together ═ pyrrolidinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.21(br.s，1H)，8.23(dd，J＝7.9，0.7Hz，1H)，7.94(d，J＝7.7Hz，1H)，7.81(ddd，J＝8.2，7.0，1.4Hz，1H)，7.66-7.72(m，2H)，7.55(ddd，J＝8.0，7.1，1.2Hz，1H)，7.42-7.52(m，3H)，3.55(t，J＝5.9Hz，2H)，2.55(t，J＝5.9Hz，2H)，2.25-2.35(m，4H)，1.62(dt，4H)。

4- [3- (benzylamino) propoxy ] -7-fluoro-3-phenylisoquinoline-1 (2H) -one (Compound 87)

(I，L＝CH₂CH₂CH₂，R₂＝F，R₃＝R＝H，R₁Is benzyl, R₄Arbityl phenyl

23mg (0.28mmol) of sodium acetate, 10.5mg (1.12mmol) of zinc chloride and 21l (0.21mmol) of benzaldehyde are added to a suspension of 50mg (0.14mmol) of 4- (3-aminopropoxy) -7-fluoro-3-phenylisoquinoline-1 (2H) -one hydrochloride in methanol (1mL) and the resulting mixture is stirred at room temperature for 18 hours. Then, 10.5mg (0.28mmol) of sodium cyanoborohydride was added, and the reaction was stirred for 4 hours. The solvent was removed under reduced pressure and the resulting solid residue was diluted with ethyl acetate and washed 2 times with water. The combined organic layers were evaporated to dryness and the crude product was purified by flash chromatography (eluent: dichloromethane/methanol 9/1) to afford 32mg of the title compound (57%).

¹H NMR(DMSO-d₆)(ppm)：11.17(s，1H)，7.83-7.95(m，2H)，7.61-7.70(m，3H)，7.39-7.49(m，3H)，7.26-7.34(m，5H)，7.18-7.26(m，1H)，3.64(s，2H)，3.55(t，J＝6.16Hz，2H)，1.66(quin，J＝6.16Hz，2H)。

4- [2- (diethylamino) ethoxy ] -7-fluoro-3-phenylisoquinoline-1 (2H) -one trifluoroacetate salt (Compound 88)

(I，L＝CH₂CH₂，R₂＝F，R₃＝H，R＝R₁Ethyl, R₄Arbityl phenyl

13.5mg (0.097mmol) of potassium carbonate and 0.01mL (0.097mmol) of diethylamine were added to a solution of 20mg (0.048mmol) of 7-fluoro-4- (2-iodoethoxy) -3-phenylisoquinoline-1 (2H) -one in N, N-dimethylformamide (1 mL). The mixture was heated at 100 ℃ for 3 hours. The solvent was removed under reduced pressure, the residue diluted with dichloromethane and washed 2 times with water. The combined organic layers were evaporated to dryness and the crude product was purified by reverse phase chromatography to afford 2mg of the title compound (3%).

¹H NMR(DMSO-d₆) (ppm): 0.85(t, J ═ 7.1Hz, 6H), 2.32-2.40(m, 4H), 2.52-2.55(m, 2H), 3.48(t, J ═ 6.1Hz, 2H), 7.44-7.52(m, 3H), 7.65-7.70(m, 2H), 7.71(ddd, J ═ 8.8, 8.8, 2.7Hz, 1H), 7.89(dd, J ═ 9.3, 2.7Hz, 1H), 8.07(dd, J ═ 8.8, 5.2Hz, 1H), 8, 32 (wide signal. 1H).

Following the same methodology, but employing appropriately substituted derivatives, the following compounds were prepared:

7-fluoro-4- [2- (4-methylpiperazin-1-yl) ethoxy ] -3-phenylisoquinoline-1 (2H) -one trifluoroacetate salt (Compound 89)

(I，L＝CH₂CH₂，R₂＝F，R₃H, R and R₁Together ═ 4-methylpiperazin-1-yl, R₄Arbityl phenyl

¹H NMR(DMSO-d₆)(ppm)：11.40(s，1H)，8.05(dd，J＝5.31，8.97Hz，1H)，7.91(dd，J＝2.75，9.34Hz，1H)，7.75(td，J＝2.81，8.79Hz，1H)，7.64-7.71(m，2H)，7.43-7.56(m，3H)。

7-fluoro-3-phenyl-4- [2- (phenylamino) ethoxy ] isoquinolin-1 (2H) -one trifluoroacetate salt (Compound 90)

(I，L＝CH₂CH₂，R₂＝F，R₃＝R＝H，R₁＝R₄Arbityl phenyl

MS +1 calculated: 375.1504, respectively; MS +1 found: 375.1504

ESI(+)MS：m/z 375(MH⁺)。

¹H NMR(DMSO-d₆)(ppm)：11.39(s，1H)，7.98(dd，J＝5.13，9.03Hz，1H)，7.88(dd，J＝2.75，9.34Hz，1H)，7.67-7.73(m，2H)，7.61(td，J＝2.81，8.73Hz，1H)，7.44-7.53(m，3H)，6.99-7.06(m，2H)，6.48-6.54(m，1H)，6.45(dd，J＝1.04，8.61Hz，2H)，5.43-5.50(m，1H)，3.59(t，J＝5.92Hz，2H)，3.13-3.22(m，2H)。

Example 2

4- (3-Aminopropoxy) -7-fluoro-3-phenylisoquinoline-1 (2H) -one hydrochloride Thing 86)

(I，L＝CH₂CH₂CH₂，R₂＝F，R₃＝R＝R₁＝H，R₄Arbityl phenyl

A solution of 620mg (1.5mmol) of tert-butyl {3- [ (7-fluoro-1-oxo-3-phenyl-1, 2-dihydroisoquinolin-4-yl) oxy ] propyl } carbamate in 4M dioxane solution of hydrochloric acid (7mL) was stirred at room temperature for 4 hours. The solvent was evaporated to dryness and the solid residue was treated with diethyl ether until precipitation of the hydrochloride salt occurred. The product was collected by filtration and washed with diethyl ether to provide the title compound (95%) as a white solid, 490 mg.

¹H NMR(DMSO-d₆)(ppm)：11.42(s，1H)，7.87-7.94(m，2H)，7.72(td，J＝2.81，8.73Hz，4H)，7.64-7.68(m，2H)，7.44-7.55(m，3H)，3.52-3.60(m，2H)，2.67-2.75(m，2H)，1.72-1.83(m，2H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

7-fluoro-3-phenyl-4- (piperidin-4-yloxy) isoquinolin-1 (2H) -one hydrochloride (Compound 85)

(I, L together with the nitrogen to which it is bonded is piperidin-4-yl, R₂＝F，R₃＝R＝H，R₄Arbityl phenyl

¹H NMR(DMSO-d₆)(ppm)：11.43(s，1H)，8.22-8.55(m，2H)，7.92(dd，J＝2.81，9.28Hz，1H)，7.84-7.89(m，1H)，7.72(td，J＝2.81，8.73Hz，1H)，7.61-7.68(m，2H)，7.44-7.55(m，4H)，3.80-3.92(m，1H)，2.86(br.s，2H)，2.67(dt，J＝1.85，3.75Hz，2H)，1.79(br.s，2H)，1.52(br.s，2H)。

Example 3

Transformation A

4- (2-amino-ethoxy) -3- (3-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride (Compound 11)

(I，L＝CH₂-CH₂-，R＝R₁＝R₂＝R₃H and R₄3-methoxyphenyl).

4M hydrochloric acid in dioxane (10mL) was added to a solution of 3.2g (7.8mmol) of {2- [3- (3-methoxy-phenyl) -1-oxo-1, 2-dihydro-isoquinolin-4-yloxy ] -ethyl } -carbamic acid tert-butyl ester in methanol (10mL) and the mixture was stirred at room temperature under an argon atmosphere for 4 h. The solvent was evaporated to dryness and the solid residue was treated with methanol and then ether until precipitation of the hydrochloride salt occurred. The product was collected by filtration and washed with diethyl ether to provide 2.62g of the title compound as a white solid (97%).

¹H NMR(DMSO-d₆)(ppm)：11.27(s，1H)，8.25(dd，J＝8.0，0.8Hz，1H)，8.11(br.s，3H)，7.98(d，J＝7.8Hz，1H)，7.83(ddd，J＝8.1，7.1，1.3Hz，1H)，7.59(ddd，J＝8.0，7.1，1.1Hz，1H)，7.43(t，J＝7.9Hz，1H)，7.33(dt，J＝7.8，1.0Hz，1H)，7.24(dd，J＝2.3，1.8Hz，1H)，7.05(ddd，1H)，3.84(s，3H)，3.72(t，J＝5.5Hz，2H)，2.96(sxt，J＝5.5Hz，2H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

4- (2-amino-ethoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 1)

(I，L＝CH₂-CH₂，R＝R₁＝R₂＝R₃H, and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：2.95(sxt，J＝5.36Hz，2H)3.68(t，J＝5.42Hz，2H)7.59(ddd，J＝8.02，7.04，1.16Hz，1H)7.83(ddd，J＝8.17，7.07，1.34Hz，1H)7.96(d，J＝8.17Hz，1H)7.98(br.s，3H)8.26(ddd，J＝8.05，1.22，0.50Hz，1H)11.30(s，1H)。

4- (3-amino-propoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 3)

(I，L＝CH₂-CH₂-CH₂，R＝R₁＝R₂＝R₃H, and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.74-1.85(m，2H)2.67-2.77(m，2H)3.57(t，J＝6.10Hz，2H)7.58(ddd，J＝8.11，4.88，3.35Hz，1H)7.67(dd，J＝7.92，1.58Hz，2H)7.70(br.s，3H)8.25(d，J＝7.92Hz，1H)11.27(s，1H)。

4- (3-methylamino-propoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 12)

(I，L＝CH₂-CH₂-CH₂R is methyl, R₁＝R₂＝R₃H, and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.77-1.87(m，2H)2.49(t，J＝5.49Hz，3H)2.73-2.83(m，2H)3.57(t，J＝6.04Hz，2H)7.58(dt，J＝8.05，4.15Hz，1H)7.67(dd，J＝7.99，1.52Hz，2H)7.83(d，J＝3.66Hz，2H)8.25(d，J＝8.05Hz，1H)8.48(br.s，2H)11.28(s，1H)。

4- (4-amino-butoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 22)

(I，L＝CH₂-CH₂-CH₂-CH₂，R＝R₁＝R₂＝R₃H, and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.57(br.s，4H)2.69(br.s，2H)3.49(br.s，2H)7.58(dt，J＝7.99，4.18Hz，1H)7.73(br.s，3H)7.69(dd，J＝8.05，1.34Hz，2H)7.83(d，J＝3.66Hz，2H)8.26(d，J＝7.93Hz，1H)8.73(br.s，1H)11.26(s，1H)。

3-phenyl-4- (2-piperazin-1-yl-ethoxy) -2H-isoquinolin-1-one di-hydrochloride (Compound 6)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：3.66-3.73(m，2H)3.77(br.s，2H)7.59(ddd，J＝8.05，7.07，1.10Hz，1H)7.66-7.72(m，2H)7.83(ddd，J＝8.11，7.13，1.34Hz，1H)7.96(d，J＝8.05Hz，1H)8.25(dd，J＝7.92，0.73Hz，1H)9.43(br.s，3H)11.30(s，1H)。

4- (2-amino-ethoxy) -3- (4-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride (Compound 24)

(I，L＝CH₂-CH₂，R＝R₁＝R₂＝R₃H and R₄4-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：2.97(sxt，J＝5.54Hz，2H)3.68(t，J＝5.43Hz，2H)3.84(s，3H)7.02-7.09(m，2H)7.52-7.60(m，1H)7.66-7.72(m，2H)7.81(td，J＝7.63，1.34Hz，1H)7.95(d，J＝7.81Hz，1H)8.06-8.14(m，3H)8.24(dd，J＝7.93，0.73Hz，1H)11.22(s，1H)。

3-phenyl-4- (3-piperazin-1-yl-propoxy) -2H-isoquinolin-1-one di-hydrochloride (Compound 14)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.94(br.s，2H)3.57(t，J＝5.79Hz，2H)7.58(ddd，J＝8.02，6.49，1.71Hz，1H)7.65-7.72(m，2H)7.82(ddd，J＝8.05，7.02，1.22Hz，1H)7.86(dd，J＝8.20，1.22Hz，0H)8.25(d，J＝7.93Hz，1H)9.37(br.s，2H)11.28(s，1H)11.44(br.s，1H)。

4- (2-amino-ethoxy) -7-fluoro-3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 39)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂7-F and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.46(br.s，1H)，8.07(dd，J＝9.1，5.2Hz，1H)，8.07(br.s，3H)，7.92(dd，J＝9.3，2.7Hz，1H)，7.66-7.77(m，3H)，7.40-7.57(m，3H)，3.67(t，J＝5.2Hz，2H)，2.94(t，J＝5.4Hz，2H)。

4- (2-amino-ethoxy) -7-fluoro-3- (4-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride (Compound 25)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂7-F and R₄4-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.38(s，1H)，8.02(dd，J＝8.9，5.2Hz，1H)，7.98(br.s，3H)，7.90(dd，J＝9.3，2.8Hz，1H)，7.71(td，J＝8.7，2.8Hz，1H)，7.67(d，J＝8.8Hz，2H)，7.06(d，J＝8.9Hz，2H)，3.83(s，3H)，3.65(t，J＝5.4Hz，2H)，2.98(sxt，J＝5.4Hz，2H)。

4- (2-amino-ethoxy) -7-fluoro-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride (Compound 46)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂7-F and R₄3-methoxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.42(s，1H)，8.05(dd，J＝8.8，5.2Hz，1H)，7.99(br.s，3H)，7.92(dd，J＝9.3，2.7Hz，1H)，7.73(td，J＝8.7，2.8Hz，1H)，7.43(t，J＝8.0Hz，1H)，7.29-7.33(m，1H)，7.21-7.25(m，1H)，7.05(ddd，J＝8.3，2.6，0.9Hz，1H)，3.84(s，3H)，3.69(t，J＝5.4Hz，2H)，2.92-3.04(m，2H)。

4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -2H-isoquinolin-1-one hydrochloride (Compound 48)

(I，L＝CH₂-CH₂，R＝R₁＝R₂＝R₃H and R₄3-chlorophenyl).

¹H NMR(DMSO-d₆)(ppm)：11.34(s，1H)，8.23(ddd，J＝7.9，1.2，0.5Hz，1H)，7.97(br.s，3H)，7.94(d，J＝8.1Hz，1H)，7.82(ddd，J＝8.1，7.1，1.3Hz，1H)，7.72(t，J＝1.8Hz，1H)，7.67(dt，J＝6.4，2.0Hz，1H)，7.59(ddd，J＝8.0，7.1，1.0Hz，1H)，7.48-7.55(m，2H)，3.68(t，J＝5.4Hz，2H)，2.89-3.02(m，2H)。

4- (2-amino-ethoxy) -5-methyl-3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 49)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂Is 5-methyl andR₄phenyl).

¹H NMR(DMSO-d₆)(ppm)：11.33(br.s，1H)，8.16(ddd，J＝7.9，1.4，0.5Hz，1H)，7.74(br.s，3H)，7.70(dd，J＝7.8，1.8Hz，2H)，7.59(ddd，J＝7.3，1.4，0.7Hz，1H)，7.47-7.56(m，3H)，7.44(t，J＝7.6Hz，1H)，3.54(t，J＝6.0Hz，2H)，2.76(s，3H)，2.68(t，J＝6.0Hz，2H)。

4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -7-fluoro-2H-isoquinolin-1-one hydrochloride (Compound 50)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂7-F and R₄3-chlorophenyl).

¹H NMR(DMSO-d₆)(ppm)：11.51(s，1H)，8.05(dd，J＝9.0，5.2Hz，1H)，7.98(br.s，3H)，7.93(dd，J＝9.3，2.7Hz，1H)，7.74(t，J＝1.7Hz，1H)，7.74(td，J＝8.8，2.8Hz，2H)，7.69(dt，J＝6.7，1.8Hz，1H)，7.57(dt，J＝7.9，1.9Hz，1H)，7.54(t，J＝8.1Hz，1H)，3.69(t，J＝5.3Hz，2H)，2.98(br.s，1H)。

4- (2-amino-ethoxy) -3-thiophen-3-yl-2H-isoquinolin-1-one hydrochloride (Compound 29)

(I，L＝CH₂-CH₂，R＝R₁＝R₂＝R₃H and R₄Phenylthio).

¹H NMR(DMSO-d₆)(ppm)：3.13-3.20(m，2H)3.79(t，J＝5.25Hz，2H)7.54-7.61(m，2H)7.69-7.74(m，2H)7.82(td，J＝7.63，1.34Hz，2H)7.95(d，J＝8.18Hz，2H)8.13(br.s，3H)8.18(t，J＝2.07Hz，1H)8.24(dd，J＝7.93，0.73Hz，1H)11.18(s，1H)。

Example 4

Transformation B

7-fluoro-3- (3-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one hydrochloride (Compound 45)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₃＝H，R₂7-F and R₄3-hydroxyphenyl).

Boron tribromide (1M in dichloromethane) (1mL, 1mmol) was added to a solution of 68mg (0.166mmol) of 7-fluoro-3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one hydrochloride (compound 43) in anhydrous dichloromethane (1mL) and the mixture was stirred at room temperature under an argon atmosphere overnight. The solvent was concentrated under reduced pressure and the crude product was purified by flash chromatography (eluent: starting from dichloromethane/methanol/acetone/7N ammonia in methanol: 93/3/3/1, to 88/6/5/1) to afford the free base, which was then converted to the hydrochloride salt according to usual chemical methods (58mg of the title compound, 81%).

¹H NMR(DMSO-d₆)(ppm)：11.35(s，1H)，9.73(s，1H)，9.49(br.s，1H)，7.88-7.92(m，2H)，7.71(td，J＝8.8，2.7Hz，1H)，7.32(t，J＝8.1Hz，1H)，7.05-7.08(m，2H)，6.89(ddd，J＝8.1、2.3，1.0Hz，1H)，3.56(t，J＝5.8Hz，2H)，2.90(dt，J＝10.6，5.3Hz，2H)，2.70-2.82(m，J＝12.3，12.3，9.2、3.5Hz，2H)，1.87-2.01(m，2H)，1.60-1.81(m，6H)，1.29-1.42(m，2H)

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

4- (2-amino-ethoxy) -7-fluoro-3- (3-hydroxy-phenyl) -2H-isoquinolin-1-one (Compound 47)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂7-F and R₄3-hydroxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.35(s，1H)，9.70(s，1H)，8.03(dd，J＝8.9，5.2Hz，1H)，7.99(br.s，3H)，7.90(dd，J＝9.3，2.7Hz，1H)，7.72(td，J＝8.7，2.8Hz，1H)，7.31(t，J＝7.8Hz，1H)，7.12(dt，J＝7.9，1.4Hz，1H)，7.09(t，J＝2.0Hz，1H)，6.88(ddd，J＝8.1、2.4，0.7Hz，1H)，3.69(t，J＝5.4Hz，2H)，2.98(sxt，J＝5.4Hz，2H)。

4- (2-amino-ethoxy) -7-fluoro-3- (4-hydroxy-phenyl) -2H-isoquinolin-1-one hydrobromide (Compound 28)

(I，L＝CH₂-CH₂，R＝R₁＝R₃＝H，R₂7-F and R₄4-hydroxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：3.00(br.s，2H)3.65(t，J＝5.37Hz，2H)6.88(d，J＝8.67Hz，2H)7.55(d，J＝8.67Hz，2H)7.70(td，J＝8.73，2.81Hz，1H)7.90(b r.s，3H)7.89(dd，J＝9.28，2.81Hz，2H)8.00(dd，J＝8.97，5.31Hz，1H)9.85(s，1H)11.30(s，1H)。

3- (3-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 33)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₂＝R₃H and R₄3-hydroxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.10(s，1H)，9.57(s，1H)，8.22(dd，J＝8.0，0.8Hz，1H)，7.89(d，J＝7.9Hz，1H)，7.79(ddd，J＝8.1，7.0，1.3Hz，1H)，7.54(ddd，J＝8.0，7.0，1.1Hz，1H)，7.28(t，J＝8.1Hz，1H)，7.08(t，J＝1.7Hz，1H)，7.07(dt，J＝6.6，1.2Hz，1H)，6.84(ddd，J＝8.1、2.4，0.9Hz，1H)，3.53(t，J＝6.2Hz，2H)，2.22(t，J＝7.0Hz，6H)，1.66(quin，J＝6.7Hz，2H)，1.43(quin，J＝5.1Hz，4H)，1.28-1.39(m，J＝5.3，5.3，5.3，5.3、4.8Hz，2H)。

3- (4-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 27)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₂＝R₃H and R₄4-hydroxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：1.31-1.40(m，2H)1.40-1.52(m，4H)1.67(quin，J＝6.59Hz，2H)2.24(br.s，6H)3.51(t，J＝6.23Hz，2H)6.85(d，J＝8.79Hz，2H)7.51(ddd，J＝8.00，7.08，1.28Hz，3H)7.51(d，J＝8.67Hz，2H)7.77(ddd，J＝8.18，7.02，1.28Hz，1H)7.86(d，J＝7.95Hz，1H)8.20(ddd，J＝7.93，1.10，0.49Hz，1H)9.75(s，1H)11.05(s，1H)。

4- (2-amino-ethoxy) -3- (3-hydroxy-phenyl) -2H-isoquinolin-1-one hydrochloride (Compound 32)

(I，L＝CH₂-CH₂，R＝R₁＝R₂＝R₃H and R₄3-hydroxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：11.19(s，1H)，9.72(s，1H)，8.24(dd，J＝8.1，0.7Hz，1H)，8.08(br.s，3H)，7.96(d，J＝8.1Hz，1H)，7.82(td，J＝7.6，1.3Hz，1H)，7.58(dd，J＝15.1，1.1Hz，1H)，7.31(t，J＝7.9Hz，1H)，7.09-7.15(m，2H)，6.88(ddd，J＝8.1、2.2，1.0Hz，1H)，3.72(t，J＝5.4Hz，2H)，2.91-3.05(m，2H)。

4- (2-amino-ethoxy) -3- (4-hydroxy-phenyl) -2H-isoquinolin-1-one (Compound 26)

(I，L＝CH₂-CH₂，R＝R₁＝R₂＝R₃H and R₄4-hydroxyphenyl).

¹H NMR(DMSO-d₆)(ppm)：2.64(t，J＝5.74Hz，2H)3.46(t，J＝5.74Hz，2H)6.81-6.89(m，2H)7.48-7.56(m，3H)7.78(td，J＝7.60，1.28Hz，1H)7.90(d，J＝7.93Hz，1H)8.20(dd，J＝8.12，0.67Hz，1H)。

Example 5

Transformation C

7-amino-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 36)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₂7-amino, R₃H and R₄Phenyl).

Cyclohexene (5mL, 49.4mmol) and 250mg palladium on carbon (10% by weight loading) were added to a suspension of 575mg (1.41mmol) of 7-nitro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 35) in dioxane (15mL) and the mixture was heated at 100 ℃ under an argon atmosphere for 3H. The mixture was cooled to room temperature, filtered through a pad of celite, and then washed with dioxane. The solvent was concentrated under reduced pressure and the resulting crude product was purified by flash chromatography (eluent: dichloromethane/methanol/acetone/7N ammonia in methanol: 90/5/5/1) to provide 210mg of the title compound (39%).

¹H NMR(DMSO-d₆)(ppm)：10.75(s，1H)，7.64(dd，J＝7.3，1.3Hz，2H)，7.61(d，J＝8.5Hz，1H)，7.45(t，J＝7.4Hz，2H)，7.38(t，J＝7.3Hz，1H)，7.35(d，J＝2.4Hz，1H)，7.07(dd，J＝8.5，2.4Hz，1H)，5.63(s，2H)，3.49(t，J＝6.2Hz，2H)，2.24(br.s，6H)，1.64(br.s，2H)，1.45(br.s，4H)，1.36(br.s，2H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

6-amino-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 40)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₂6-amino, R₃H and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：10.54(br.s，1H)，7.88(d，J＝8.7Hz，1H)，7.63(d，J＝7.7Hz，2H)，7.46(t，J＝7.4Hz，2H)，7.41(t，J＝7.3Hz，1H)，6.79(d，J＝2.2Hz，1H)，6.74(dd，J＝8.7，2.2Hz，1H)，5.97(s，2H)，3.45(t，J＝6.5Hz，2H)，2.16(br.s，4H)，2.12(t，J＝7.4Hz，2H)，1.61(quin，J＝6.9Hz，2H)，1.39(quin，J＝5.3Hz，4H)，1.26-1.36(m，2H)。

Example 6

Transformation D

6-chloro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 41)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₂6-chloro, R₃H and R₄Phenyl).

37L (0.316mmol) of tert-butyl nitrite are added dropwise to a suspension of 34mg (0.25mmol) of copper (II) chloride in acetonitrile (1 mL). The mixture was cooled in an ice bath and 80mg (0.21mmol) of a suspension of 6-amino-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 40) in acetonitrile (1.5mL) was added. The reaction mixture was stirred at room temperature under an argon atmosphere overnight and then treated with water and dichloromethane. The separated organic layer was washed once with a saturated solution of sodium bicarbonate, then concentrated under reduced pressure and the resulting crude product was purified by flash chromatography (eluent: dichloromethane/methanol/acetone/7N ammonia in methanol: 90/5/5/1) to yield 32mg of the title compound (38%).

¹H NMR(DMSO-d₆)(ppm)：11.37(s，1H)，8.22(d，J＝8.7Hz，1H)，7.84(d，J＝2.0Hz，1H)，7.67(dd，J＝8.0，1.5Hz，2H)，7.57(dd，J＝8.5，2.1Hz，1H)，7.43-7.54(m，3H)，3.51(t，J＝6.2Hz，2H)，2.20(t，J＝6.8Hz，6H)，1.61(quin，J＝6.6Hz，2H)，1.44(quin，J＝5.4Hz，4H)，1.28-1.38(m，2H)

6-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (Compound 42)

(I，L＝CH₂-CH₂-CH₂R and R₁Together as 1-piperidinyl, R₂6-fluoro, R₃H and R₄Phenyl).

A suspension of 145mg (0.38mmol) of 6-amino-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one (compound 40) in dichloromethane (1mL) was added with stirring at 0 ℃ under an argon atmosphere to a suspension of 67mg (0.57mmol) of nitrosonium tetrafluoroborate in dichloromethane (1 mL). The reaction mixture was then stirred at room temperature overnight. 1, 2-dichlorobenzene (1mL) was added and the mixture was heated at 170 ℃ for 30 min. The solvent was concentrated under reduced pressure and the reaction was treated with water and dichloromethane. The separated organic layer was concentrated under reduced pressure and the resulting crude product was purified by flash chromatography (eluent: dichloromethane/methanol/acetone/7N ammonia in methanol: 95/2/2/1) to yield 25mg of the title compound (17%).

¹H NMR(DMSO-d₆)(ppm)：11.30(s，1H)，8.29(dd，J＝8.8，5.9Hz，1H)，7.67(dd，J＝8.0，1.5Hz，2H)，7.62(dd，J＝10.2，2.5Hz，1H)，7.44-7.54(m，3H)，7.39(td，J＝8.8，2.6Hz，1H)，3.51(d，J＝12.2Hz，2H)，2.22(br.s，6H)，1.63(quin，J＝6.6Hz，2H)，1.45(quin，J＝5.2Hz，4H)，1.29-1.40(m，2H)。

Example 7

Transformation of E

4- (2-methylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one (Compound 16)

(I，L＝CH₂-CH₂R is methyl, R₁＝R₂＝R₃H and R₄Phenyl).

mu.L (0.08mmol) of formaldehyde (37% in water) was added to a solution of 25mg (0.079mmol) of 4- (2-amino-ethoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride (Compound 1) in formic acid (0.1mL) and the mixture was refluxed for 1H. The solvent was concentrated under reduced pressure, and the resulting crude product was purified by preparative HPLC to provide 12mg of the title compound (51%).

¹H NMR(DMSO-d₆)(ppm)：2.18(s，3H)2.63(t，2H)3.58(t，J＝5.43Hz，2H)7.57(ddd，1H)7.70(dd，J＝8.05，1.46Hz，2H)7.82(ddd，J＝8.14，7.04，1.28Hz，1H)7.92(d，J＝7.80Hz，1H)8.24(ddd，J＝8.05，1.22，0.40Hz，1H)

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

4- (2-dimethylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one (Compound 17)

(I，L＝CH₂-CH₂，R＝R₁Methyl, R₂＝R₃H and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：2.08(s，6H)2.43(br.s，2H)3.54(t，J＝5.73Hz，2H)7.56(ddd，J＝8.02，7.04，1.16Hz，1H)7.69(dd，J＝8.05，1.46Hz，2H)7.81(ddd，J＝8.14，7.04，1.28Hz，1H)7.93(d，J＝7.80Hz，1H)8.23(ddd，J＝8.05，1.10，0.50Hz，1H)11.22(s，1H)

4- [2- (4-methyl-piperazin-1-yl) -ethoxy ] -3-phenyl-2H-isoquinolin-1-one dihydrochloride (Compound 19)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁co-N-methyl-piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：2.78(br.s，3H)7.49(t，J＝7.40Hz，1H)7.53(t，J＝7.40Hz，2H)7.59(ddd，J＝8.05，7.13，0.79Hz，1H)7.69(d，J＝7.90Hz，2H)7.84(ddd，J＝8.11，7.13，1.10Hz，1H)7.96(d，J＝8.05Hz，1H)8.25(d，J＝7.44Hz，1H)11.29(br.s，1H)。

4- [3- (4-methyl-piperazin-1-yl) -propoxy ] -3-phenyl-2H-isoquinolin-1-one dihydrochloride (Compound 20)

(I，L＝CH₂-CH₂-CH₂，R₂＝R₃H, R and R₁co-N-methyl-piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.89(br.s，2H)2.79(br.s，3H)3.56(t，J＝5.85Hz，2H)7.48(t，J＝7.40Hz，1H)7.53(t，J＝7.50Hz，2H)7.58(ddd，J＝8.02，6.43，1.89Hz，1H)7.69(d，J＝7.70Hz，2H)7.82(td，J＝8.05，0.98Hz，1H)7.85(dd，J＝8.05，1.50Hz，1H)8.25(d，J＝7.93Hz，1H)11.27(br.s，1H)。

Example 8

Transformation F

N- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -acetamide (Compound 4)

(I，L＝CH₂-CH₂-CH₂R ═ acetyl, R₁＝R₂＝R₃H and R₄Phenyl).

Triethylamine (190L, 1.36mmol) and acetyl chloride (39.3L, 0.55mmol) were added to a solution of 150mg (0.45mmol) of 4- (2-amino-ethoxy) -3-phenyl-2H-isoquinolin-1-one (compound 1) in dichloromethane (6mL) at 0 ℃ under an argon atmosphere and the mixture was stirred at room temperature overnight. The reaction mixture was then washed with water, the layers were separated and the aqueous layer was extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, concentrated under reduced pressure, and the resulting crude product was purified by flash chromatography (dichloromethane: methanol 97: 3) to provide 64mg of the title compound (42%).

¹H NMR(DMSO-d₆)(ppm)：2.18(s，3H)2.63(t，2H)3.58(t，J＝5.43Hz，2H)7.57(ddd，1H)7.70(dd，J＝8.05，1.46Hz，2H)7.82(ddd，J＝8.14，7.04，1.28Hz，1H)7.92(d，J＝7.80Hz，1H)8.24(ddd，J＝8.05，1.22，0.40Hz，1H)。

The following compounds were prepared according to the same methodology, but using appropriately substituted starting materials:

n- [2- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -acetamide (Compound 2)

(I，L＝CH₂-CH₂R ═ acetyl, R₁＝R₂＝R₃H and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.78(s，3H)3.19(q，J＝5.70Hz，2H)3.46(t，J＝5.61Hz，2H)7.56(ddd，1H)7.81(ddd，J＝8.29，7.02，1.30Hz，1H)7.88(ddd，J＝8.29，0.91，0.51Hz，1H)7.94(t，J＝5.67Hz，1H)8.24(ddd，J＝8.05，1.22，0.49Hz，1H)11.22(s，1H)。

4- [2- (4-acetyl-piperazin-1-yl) -ethoxy ] -3-phenyl-2H-isoquinolin-1-one (Compound 7)

(I，L＝CH₂-CH₂，R₂＝R₃H, R and R₁Together ═ N-acetyl-piperazinyl and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.98(s，3H)2.20(t，J＝4.69Hz，2H)2.27(t，J＝4.40Hz，2H)2.49(t，J＝5.25Hz，2H)3.58(t，J＝5.30Hz，1H)7.57(ddd，J＝7.95，7.10，0.67Hz，1H)7.69(dd，J＝8.11，1.40Hz，2H)7.83(ddd，J＝8.14，7.10，1.22Hz，1H)8.00(d，J＝8.05Hz，1H)8.24(d，J＝7.56Hz，1H)11.22(s，1H)。

N- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -benzamide (Compound 8)

(I，L＝CH₂-CH₂-CH₂R ═ benzoyl, R₁＝R₂＝R₃H and R₄Phenyl).

¹H NMR(DMSO-d₆)(ppm)：1.78(quin，J＝6.64Hz，2H)3.26(q，J＝6.50Hz，2H)3.56(t，J＝6.28Hz，2H)7.55(ddd，J＝8.01，7.04，1.04Hz，1H)7.69(d，J＝7.80Hz，2H)7.86(d，J＝7.90Hz，1H)8.23(ddd，J＝7.98，1.20，0.60Hz，1H)8.32(t，J＝5.42Hz，1H)11.22(s，1H)。

Preparation of 7-fluoro-4- (2-hydroxyethoxy) -3-phenylisoquinoline-1 (2H) -one

88mg of p-toluenesulfonic acid (0.46mmol) were added to a solution of 590mg (1.54mmol) of 7-fluoro-3-phenyl-4- [2- (tetrahydro-2H-pyran-2-yloxy) ethoxy ] isoquinolin-1 (2H) -one in methanol (12mL), and the resulting mixture was heated at 55 ℃ for 2 hours. The solvent was removed under reduced pressure, the residue diluted with dichloromethane and washed 2 times with saturated aqueous sodium bicarbonate solution. The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure to provide 438mg of the title compound (95%).

¹H NMR(DMSO-d₆)(ppm)：11.36(s，1H)，8.08(dd，J＝5.25，8.91Hz，1H)，7.89(dd，J＝2.75，9.34Hz，1H)，7.66-7.74(m，3H)，7.41-7.54(m，3H)，4.47(t，J＝2.50Hz，1H)。

Preparation of 7-fluoro-4- (2-iodoethoxy) -3-phenylisoquinoline-1 (2H) -one

350mg (1.33mmol) of triphenylphosphine, 90mg (1.33mmol) of imidazole and 251mg (0.99mmol) of iodide were added to a solution of 100mg (0.33mmol) of 7-fluoro-4- (2-hydroxyethoxy) -3-phenylisoquinoline-1 (2H) -one in acetonitrile (10mL), and the mixture was stirred at room temperature under argon for 18 hours. The mixture was filtered through celite and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (hexane/ethyl acetate: 6/4) to give 57mg of the title compound (42%).

¹H NMR(DMSO-d₆)(ppm)：11.41(s，1H)，8.06(dd，J＝5.13，9.03Hz，1H)，7.90(dd，J＝2.62，9.34Hz，1H)，7.64-7.76(m，3H)，7.43-7.56(m，4H)，3.70(t，J＝6.16Hz，2H)，3.22-3.27(m，2H)。

Example 9

Step i

2-methylamino-N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide

To a stirred suspension of 150mg of 2-chloro-N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide (XV) (0.52mmol) in N, N-dimethylformamide (1.4mL) was added 9mL of a 33% solution of methylamine in ethanol (mM). The reaction mixture was stirred at room temperature for 3 hours. The ethanol was then evaporated under reduced pressure, the mixture was diluted with diethyl ether and filtered. The pale yellow solid was then collected, washed with ether, then with cold water and dried. The title compound was obtained in moderate yield (100mg, 68%).

¹H NMR(DMSO-d₆)(ppm)：11.68(s，1H)，10.33(br.s，1H)，8.61(d，J＝2.1Hz，1H)，8.34(dd，J＝7.9，1.1Hz，1H)，8.26(d，J＝8.2Hz，1H)，7.89(ddd，J＝8.2，7.1，1.4Hz，1H)，7.70(dd，J＝8.7，2.2Hz，1H)，7.67(ddd，J＝7.9，7.2，0.7Hz，1H)，7.35(d，J＝8.8Hz，1H)，3.68(s，2H)，2.54(s，3H)。

Step ia

(3- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -propyl) -carbamic acid tert-butyl ester

0.09mL of (3-amino-propyl) -carbamic acid tert-butyl ester (0.51mmol) and 0.036mL of triethylamine (0.26mmol) were added to a stirred suspension of 50mg of 2-chloro-N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide (XV) (0.17mmol) in N, N-dimethylformamide (3 mL). The reaction mixture was stirred at room temperature overnight, evaporated to dryness and then purified by preparative HPLC on a Waters X Terra RP 18(19 × 250mm, 5m) column. Mobile phase a was 0.05% ammonia/acetonitrile: 95/5, mobile phase B is acetonitrile/water: 95/5. The gradient was from 10 to 75% B in 15 min. The fractions containing the desired compound were dried to provide 32mg (44% yield) of the title compound.

¹H NMR(DMSO-d₆)(ppm)：11.65(s，1H)，10.09(br.s，1H)，8.63(d，J＝2.0Hz，1H)，8.33(dd，J＝7.9，1.2Hz，1H)，8.28(d，J＝8.2Hz，1H)，8.12-8.12(m，1H)，7.88(ddd，J＝8.2，7.1，1.4Hz，1H)，7.72(dd，J＝8.8，2.0Hz，1H)，7.66(ddd，J＝8.0，7.1，0.7Hz，1H)，7.33(d，J＝8.8Hz，1H)，6.83(t，J＝5.7Hz，1H)，3.47(br.s，2H)，3.02(q，J＝6.8Hz，2H)，2.68(t，J＝7.3Hz，2H)，1.63(quin，J＝6.9Hz，2H)，1.37(s，9H)。

(6- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -hexyl) -carbamic acid tert-butyl ester

175mg of (6-amino-hexyl) -carbamic acid tert-butyl ester hydrochloride (0.69mmol) and 0.097mL of triethylamine (0.69mmol) were added to a stirred suspension of 65mg of 2-chloro-N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide (XV) (0.23mmol) in N, N-dimethylformamide (2 mL). The reaction mixture was stirred at room temperature overnight, diluted with dichloromethane and the resulting solution was washed first with water, then with brine, dried over sodium sulfate and evaporated to dryness. The crude product was purified by preparative HPLC on a Waters X Terra RP 18(19X250mm, 5m) column. Mobile phase a was 0.05% ammonia/acetonitrile: 95/5, mobile phase B is acetonitrile/water: 95/5. The gradient is from 10 to 75% B in 15 min. The fractions containing the desired compound were dried to provide 50mg (47% yield) of the title compound.

¹H NMR(DMSO-d₆)(ppm)：11.62(br.s，1H)，9.90(br.s，1H)，8.65(d，J＝1.7Hz，1H)，8.33(dd，J＝8.5，1.2Hz，1H)，8.30(d，J＝8.0Hz，1H)，7.88(ddd，J＝8.3，7.1，1.1Hz，1H)，7.74(dd，J＝8.7，1.9Hz，1H)，7.65(t，J＝7.6Hz，1H)，7.31(d，J＝8.7Hz，1H)，6.73(t，J＝5.5Hz，1H)，3.27-3.33(m，2H)，2.89(q，J＝6.5Hz，2H)，2.55(t，J＝7.0Hz，2H)，1.97-2.61(m，1H)，1.17-1.51(m，8H)，1.36(s，9H)。

Step ib

2- (3-amino-propylamino) -N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide hydrochloride

32mg of (3- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -propyl) -carbamic acid tert-butyl ester (0.075mmol) are dissolved in dichloromethane (1mL) and 4N hydrochloric acid in dioxane (1mL) is added. The reaction mixture was stirred at room temperature overnight and then evaporated to afford 32mg (97% yield) of the title compound.

¹H NMR(DMSO-d₆)(ppm)：11.72(s，1H)，10.90(s，1H)，9.29(d，J＝0.6Hz，2H)，8.62(d，J＝2.0Hz，1H)，8.35(dd，J＝7.9，1.2Hz，1H)，8.22(d，J＝8.2Hz，1H)，7.98(br.s，3H)，7.91(ddd，J＝8.2，7.1，1.3Hz，1H)，7.69(dd，J＝8.8，2.2Hz，1H)，7.68(ddd，J＝7.9，7.2，0.9Hz，1H)，7.38(d，J＝8.8Hz，1H)，4.00(t，J＝4.5Hz，2H)，3.11(br.s，2H)，2.92(sxt，J＝6.4Hz，2H)，2.00(quin，J＝7.6Hz，2H)。

2- (6-amino-hexylamino) -N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide trifluoroacetate salt

45mg of (6- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -hexyl) -carbamic acid tert-butyl ester (0.097mmol) were dissolved in dichloromethane (5mL) and trifluoroacetic acid (1.5mL) was added. The reaction mixture was stirred at room temperature overnight, evaporated, treated with ether, filtered and the collected solid dried under vacuum to provide 30mg (65% yield) of the title compound as a pale yellow solid.

¹H NMR(DMSO-d₆)(ppm)：11.72(s，1H)，10.67(s，1H)，8.93(dt，J＝10.7，5.4Hz，2H)，8.58(d，J＝2.1Hz，1H)，8.35(dd，J＝8.1，1.2Hz，1H)，8.22(d，J＝8.2Hz，1H)，7.91(ddd，J＝8.2，7.1，1.4Hz，1H)，7.69(ddd，J＝7.9，7.2，0.7Hz，1H)，7.65(dd，J＝9.0，2.3Hz，1H)，7.67(br.s，3H)，7.38(d，J＝8.7Hz，1H)，3.99(t，J＝5.4Hz，2H)，2.95-3.06(m，2H)，2.72-2.85(m，2H)，1.64(quin，J＝7.1Hz，2H)，1.53(quin，J＝7.0Hz，2H)，1.33(dt，J＝6.9，3.4Hz，4H)。

Step ii

9-dimethylamino-11, 11-dimethyl-1- (3- { methyl- [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -carbamoyl } -propyl) -2, 3, 4, 11-tetrahydro-naphtho [2, 3-g ] quinolinium trifluoroacetate

(Compound P1, X^-＝CF₃CO₂ ^-，R₁₃Methyl, m 0)

0.0067mL of diisopropylethylamine (0.039mmol) and 5mg of ATTO 610NHS ester perchlorate (XVIII) (0.0085mmol) were added under nitrogen to a stirred solution of 3.65mg of 2-methylamino-N- (6-oxo-5, 6-dihydro-phenanthridin-2-yl) -acetamide (0.013mmol) in N, N-dimethylformamide (0.3mL) and the reaction stirred at room temperature for 3 h. The mixture was then evaporated and the resulting crude product was purified by preparative HPLC on a Hypersil (21X 250mm, 5 μm) column. Mobile phase a was 0.1% trifluoroacetic acid/acetonitrile: 95/5, mobile phase B is acetonitrile/water: 95/5. The gradient was from 0 to 70% B in 20 min. The fractions containing the desired compound were dried to provide 1.6mg of the title compound.

MS calculated: 654.3444, respectively; MS found: 654.3447.

ESI(+)MS：m/z 654(M⁺)。

following the same methodology, but using the appropriate starting materials, the following compounds were prepared:

9-dimethylamino-11, 11-dimethyl-1- [3- (3- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -propylcarbamoyl) -propyl ] -2, 3, 4, 11-tetrahydro-naphtho [2, 3-g ] quinolinium trifluoroacetate

(Compound P2, X^-＝CF₃CO₂ ^-，R₁₃＝H，m＝1，B＝(CH₂)₃-NH-)

MS calculated: 697.3866, respectively; MS found: 697.3864.

ESI(+)MS：m/z 697(M⁺)。

9-dimethylamino-11, 11-dimethyl-1- [3- (6- { [ (6-oxo-5, 6-dihydro-phenanthridin-2-ylcarbamoyl) -methyl ] -amino } -hexylcarbamoyl) -propyl ] -2, 3, 4, 11-tetrahydro-naphtho [2, 3-g ] quinolinium trifluoroacetate

(Compound P3, X^-＝CF₃CO₂ ^-，R₁₃＝H，m＝1，B＝(CH₂)₆-NH-)

MS calculated: 739.4337, respectively; MS found: 739.4333.

ESI(+)MS：m/z 739(M⁺)。

Claims (10)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein L is a straight or branched chain C₂-C₆An alkyl group, or a heterocyclic group formed by containing a nitrogen atom to which it is bonded;

r and R₁Independently a hydrogen atom, a linear or branched chain C₁-C₆Alkyl or COR₅A group, or together with the nitrogen atom to which they are bonded, form a heterocyclyl group;

R₂and R₃Independently is a hydrogen or halogen atom; or straight or branched C₁-C₆An alkyl group;

R₄is optionally substituted phenyl or thienyl;

R₅is OR₈Or straight or branched C₁-C₆An alkyl group;

R₈is straight chain or branched C₁-C₆An alkyl group;

wherein the optional substituents are selected from halogen, hydroxy, C₁-C₆Alkoxy and aryloxy groups.

2. A compound of formula (I) according to claim 1, characterized in that R and R are R₁Independently a hydrogen atom, a linear or branched chain C₁-C₆Alkyl or COR₅A group, or together with the nitrogen atom to which they are bonded, form a heterocyclyl group; and

R₅is straight or branched C₁-C₆An alkyl group.

3. A compound of formula (I) according to claim 2, characterized in that L represents a linear or branched C, or a pharmaceutically acceptable salt thereof₂-C₄An alkyl group, or a heterocyclic group formed by containing a nitrogen atom to which it is attached;

r and R₁Independently a hydrogen atom, a linear or branched chain C₁-C₄Alkyl or COR₅A group, or taken together with the nitrogen atom to which they are bonded, forms a piperidinyl, pyrrolidinyl or piperazinyl group;

R₂and R₃Independently is a hydrogen, chlorine or fluorine atom; or straight or branched C₁-C₄An alkyl group;

R₅is straight or branched C₁-C₄An alkyl group.

4. A compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, selected from:

4- (2-amino-ethoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride,

n- [2- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -ethyl ] -acetamide,

4- (3-amino-propoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride,

n- [3- (1-oxo-3-phenyl-1, 2-dihydro-isoquinolin-4-yloxy) -propyl ] -acetamide,

3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one trifluoroacetate salt,

3-phenyl-4- (2-piperazin-1-yl-ethoxy) -2H-isoquinolin-1-one di-hydrochloride,

4- [2- (4-acetyl-piperazin-1-yl) -ethoxy ] -3-phenyl-2H-isoquinolin-1-one,

3- (3-methoxy-phenyl) -4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

4- (2-amino-ethoxy) -3- (3-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride,

4- (3-methylamino-propoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride,

3-phenyl-4- (2-pyrrol-1-yl-ethoxy) -2H-isoquinolin-1-one,

3-phenyl-4- (3-piperazin-1-yl-propoxy) -2H-isoquinolin-1-one di-hydrochloride,

4- (2-methylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

4- (2-dimethylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

4- (3-diethylamino-propoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride,

4- [2- (4-methyl-piperazin-1-yl) -ethoxy ] -3-phenyl-2H-isoquinolin-1-one di-hydrochloride,

4- [3- (4-methyl-piperazin-1-yl) -propoxy ] -3-phenyl-2H-isoquinolin-1-one di-hydrochloride,

3-phenyl-4- (3-pyrrolidin-1-yl-propoxy) -2H-isoquinolin-1-one,

4- (4-amino-butoxy) -3-phenyl-2H-isoquinolin-1-one hydrochloride,

3- (4-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

4- (2-amino-ethoxy) -3- (4-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -7-fluoro-3- (4-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -3- (4-hydroxy-phenyl) -2H-isoquinolin-1-one,

3- (4-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

4- (2-amino-ethoxy) -7-fluoro-3- (4-hydroxy-phenyl) -2H-isoquinolin-1-one hydrobromide,

4- (2-amino-ethoxy) -3-thiophen-3-yl-2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -3- (3-hydroxy-phenyl) -2H-isoquinolin-1-one hydrochloride,

3- (3-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

7-chloro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

7-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

4- (2-amino-ethoxy) -7-fluoro-3-phenyl-2H-isoquinolin-1-one hydrochloride,

6-chloro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

6-fluoro-3-phenyl-4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

7-fluoro-3- (3-methoxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one hydrochloride,

3- (3-chloro-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one,

7-fluoro-3- (3-hydroxy-phenyl) -4- (3-piperidin-1-yl-propoxy) -2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -7-fluoro-3- (3-methoxy-phenyl) -2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -7-fluoro-3- (3-hydroxy-phenyl) -2H-isoquinolin-1-one,

4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -5-methyl-3-phenyl-2H-isoquinolin-1-one hydrochloride,

4- (2-amino-ethoxy) -3- (3-chloro-phenyl) -7-fluoro-2H-isoquinolin-1-one hydrochloride,

3-phenyl-4- (2-piperidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

4- (2-diethylamino-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

3-phenyl-4- (2-pyrrolidin-1-yl-ethoxy) -2H-isoquinolin-1-one,

4- (2-morpholin-4-yl-ethoxy) -3-phenyl-2H-isoquinolin-1-one,

4- [2- (dimethylamino) ethoxy ] -7-fluoro-3-phenylisoquinolin-1 (2H) -one,

7-fluoro-3-phenyl-4- (piperidin-4-yloxy) isoquinolin-1 (2H) -one hydrochloride,

4- (3-aminopropoxy) -7-fluoro-3-phenylisoquinoline-1 (2H) -one hydrochloride,

4- [3- (benzylamino) propoxy ] -7-fluoro-3-phenylisoquinolin-1 (2H) -one,

4- [2- (diethylamino) ethoxy ] -7-fluoro-3-phenylisoquinoline-1 (2H) -one trifluoroacetate salt, and

7-fluoro-4- [2- (4-methylpiperazin-1-yl) ethoxy ] -3-phenylisoquinolin-1 (2H) -one trifluoroacetate salt.

5. A process for the preparation of a compound of formula (I) as defined in claim 1, as shown in scheme 1 below,

scheme 1

The method comprises the following steps:

the following path comprising the steps of:

step 1) reacting a compound of formula (III):

wherein R is₂And R₃Alkylating, as defined in claim 1, with a compound of formula (IV):

wherein R is₄As defined in claim 1 and Lg represents a suitable leaving group;

step 2) cyclodehydration of the resulting compound of formula (V) by refluxing the compound of formula (V) in the presence of a catalytic amount of an acid in a suitable solvent with continuous removal of water:

wherein R is₂，R₃And R₄As defined above;

step 3) rearranging the resulting compound of formula (VI) by heating in a suitable solvent in the presence of a base:

wherein R is₂，R₃And R₄As defined above, to give a compound of formula (VII):

wherein R is₂，R₃And R₄As defined above;

step 5) alkylating a compound of formula (VII) as defined above with a compound of formula (XI) in a suitable solvent, in the presence of a suitable base, and at a temperature in the range of 0 ℃ to reflux:

wherein L is as defined in claim 1 and R, R₁And Lg is as defined above, to give a compound of formula (I) as defined in claim 1;

or alkylating a compound of formula (VII) as defined above with a compound of formula (XII) X' -L-Lg, wherein L is optionally substituted straight or branched chain C, in a suitable solvent, in the presence of a suitable base, and at a temperature in the range of 0 ℃ to reflux₂-C₆Alkyl, Lg is as defined above and X' represents a suitable leaving group;

step 6) reacting the resulting compound of formula (XIII):

wherein R is₂，R₃，R₄And X' is as defined above, L is an optionally substituted straight or branched chain C₂-C₆Alkyl, with a compound of formula (XIV) R-NH-R₁Wherein R and R are₁As defined above, to obtain a compound of formula (I) as defined above;

optionally converting the compound of formula (I) into a different compound of formula (I) by known chemical reactions; and/or, if necessary, converting a compound of formula (I) into a pharmaceutically acceptable salt thereof or converting a salt into a free compound of formula (I).

6. A process for the preparation of a compound of formula (I) as defined in claim 1, as shown in scheme 1 below,

scheme 1

The method comprises the following steps:

the following path comprising the steps of:

step 4) reacting the compound of formula (VIII) only with the compound of formula (IX) under microwave irradiation:

wherein R is₄As defined above and R₁₂Is C₁-C₆Alkyl or aryl radicals C₁-C₆An alkyl group, a carboxyl group,

step 3a) rearranging the resulting compound of formula (X) in a suitable solvent under reflux using a suitable base:

wherein R is₄And R₁₂As defined above, to give a compound of formula (VII) as defined above, wherein R is₂And R₃Are each a hydrogen atom;

and

step 5) alkylating a compound of formula (VII) as defined above with a compound of formula (XI) in a suitable solvent, in the presence of a suitable base, and at a temperature in the range of 0 ℃ to reflux:

wherein L is as defined in claim 1 and R, R₁And Lg is as defined above, to give a compound of formula (I) as defined in claim 1;

or alkylating a compound of formula (VII) as defined above with a compound of formula (XII) X' -L-Lg, wherein L is optionally substituted straight or branched chain C, in a suitable solvent, in the presence of a suitable base, and at a temperature in the range of 0 ℃ to reflux₂-C₆Alkyl, Lg is as defined above and X' represents a suitable leaving group;

step 6) reacting the resulting compound of formula (XIII):

wherein R is₂，R₃，R₄And X' is as defined above, L is an optionally substituted straight or branched chain C₂-C₆Alkyl, with a compound of formula (XIV) R-NH-R₁Wherein R and R are₁As defined above, to obtain a compound of formula (I) as defined above;

optionally converting the compound of formula (I) into a different compound of formula (I) by known chemical reactions; and/or, if necessary, converting a compound of formula (I) into a pharmaceutically acceptable salt thereof or converting a salt into a free compound of formula (I).

7. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) as defined in claim 1, and at least one pharmaceutically acceptable carrier and/or diluent.

8. The pharmaceutical composition according to claim 7, further comprising one or more agents selected from the group consisting of: cytostatic or cytotoxic agents, antibiotic-type agents, alkylating agents, antimetabolite agents, hormonal agents, immunological agents, interferon-type agents, cyclooxygenase inhibitors, matrix metalloproteinase inhibitors, telomerase inhibitors, tyrosine kinase inhibitors, anti-growth factor receptor agents, anti-HER agents, anti-EGFR agents, anti-angiogenesis agents, farnesyl transferase inhibitors, ras-raf signal transduction pathway inhibitors, cell cycle inhibitors, other cdks inhibitors, tubulin binding agents, topoisomerase I inhibitors, and topoisomerase II inhibitors.

9. A product comprising a compound of formula (I) as defined in claim 1 or a pharmaceutical composition thereof as defined in claim 7, and one or more agents selected from: cytostatic or cytotoxic agents, antibiotic-type agents, alkylating agents, antimetabolite agents, hormonal agents, immunological agents, interferon-type agents, cyclooxygenase inhibitors, matrix metalloproteinase inhibitors, telomerase inhibitors, tyrosine kinase inhibitors, anti-growth factor receptor agents, anti-HER agents, anti-EGFR agents, anti-angiogenesis agents, farnesyl transferase inhibitors, ras-raf signal transduction pathway inhibitors, cell cycle inhibitors, other cdks inhibitors, tubulin binding agents, topoisomerase I inhibitors, and topoisomerase II inhibitors, said product being in the form of a combined preparation of a compound of formula (I) as defined in claim 1 or a pharmaceutical composition thereof as defined in claim 7 and 1 or more chemotherapeutic agents for simultaneous, separate or sequential use in anticancer therapy.

10. Use of a compound of formula (I) as defined in claim 1 in the manufacture of a medicament for the treatment of a disease mediated by the PARP-1 protein.