Anti-cancer treatment with radioactive PARP inhibitors

Abstract

The present invention relates to a novel anti-cancer radiopharmaceutical treatment that is particularly suitable for targeting PARP enzyme expressing cancers. The novel treatment comprises a compound of formula (I): 1 , A 2 and A 3 and m is as defined herein, or a pharma- ceutical acceptable salt or solvate thereof. The present invention also relates to methods for the preparation of these compounds, pharmaceutical compositions containing them, and their use in the treatment of cancer. [Case 1] JPEG2025506693000017.jpg75170

Description

[Technical field]

[0001] preface The present invention relates to novel anti-cancer radiopharmaceutical treatments. The anti-cancer treatments are particularly suitable for targeting PARP enzyme expressing cancers. The novel treatments include Auger-emitting radiopharmaceutical compounds that target PARP, and the present invention relates to these compounds, methods for making these compounds, their therapeutic use and / or their use as imaging agents. [Background technology]

[0002] Cancer is caused by altered cell proliferation and is characterized by increased genomic instability. The expression of DNA damage repair enzyme PARP and closely related enzymes is increased in tumor tissues compared to surrounding normal tissues. PARP enzymes bind to DNA with single-strand breaks and use the substrate NAD+ to catalytically convert the latter to adenosyl ribose adducts on target proteins. Compounds that bind to PARP enzymes and prevent the binding of NAD+ can inhibit PARP and trap the enzyme in DNA.

[0003] There remains a need for new and effective treatments for cancer, particularly for difficult-to-treat cancers such as pancreatic ductal adenocarcinoma and glioblastoma, a significant proportion of which are PARP enzyme-expressing cancers.

[0004] It is with the above in mind that the present invention has been made. Summary of the Invention

[0005] According to a first aspect of the present invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0006] According to a further aspect of the invention there is provided a pharmaceutical composition comprising a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, in admixture with a pharma- ceutically acceptable diluent or carrier.

[0007] According to a further aspect of the present invention there is provided a method of treating cancer in a patient in need of such treatment, said method comprising the step of administering to said patient a therapeutically effective amount of a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein.

[0008] According to a further aspect of the present invention there is provided a method of treating cancer in which PARP enzyme expression is associated with a patient in need of such treatment, said method comprising the step of administering to said patient a therapeutically effective amount of a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein.

[0009] According to a further aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use in therapy.

[0010] According to a further aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use as a medicament.

[0011] According to a further aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use in the treatment of cancer.

[0012] According to a further aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use in the treatment of a cancer associated with PARP enzyme expression.

[0013] According to a further aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, in the manufacture of a medicament for the treatment of cancer.

[0014] According to a further aspect of the invention there is provided the use of a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, in the manufacture of a medicament for the treatment of a cancer associated with PARP enzyme expression.

[0015] According to a further aspect of the present invention there is provided a process for the preparation of a compound defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0016] According to a further aspect of the present invention there is provided a compound obtainable by, obtained by or directly obtained by a process for preparing a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0017] According to a further aspect of the present invention there is provided a novel intermediate, as defined herein, suitable for use in any one of the synthetic methods described herein.

[0018] Features, including optional, suitable and preferred features, in relation to one aspect of the invention may also be features, including optional, suitable and preferred features, in relation to any other aspect of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] definition Unless otherwise stated, the following terms used in the specification and claims have the following meanings, as set forth below.

[0020] References to "treating" or "treatment" shall be understood to include prevention as well as the alleviation of established symptoms of a condition. Thus, "treating" a condition, disorder, or condition or "treatment" of a condition, disorder, or condition includes (1) preventing or delaying the appearance of clinical symptoms of the condition, disorder, or condition as they occur in a person who may be afflicted by or predisposed to the condition, disorder, or condition, but who has not yet experienced or exhibited clinical or subclinical symptoms of the condition, disorder, or condition, (2) inhibiting the condition, disorder, or condition, i.e., (in the case of maintenance treatment) arresting, reducing, or delaying the onset of the disease or its recurrence or at least one of its clinical or subclinical symptoms, or (3) relieving or attenuating the disease, i.e., causing regression of the condition, disorder, or condition or at least one of its clinical or subclinical symptoms.

[0021] References herein to "PARP" or the "PARP enzyme" refer to all isoforms of the PARP enzyme, including isoforms PARP1 through PARP17.

[0022] "Therapeutically effective amount" means an amount of a compound that, when administered to a mammal to treat a disease, is sufficient to affect such treatment for the disease. "Therapeutically effective amount" varies depending on the compound, its molar activity, the disease and its severity of the mammal to be treated, as well as the age, weight, etc. It should be understood that, for example, in humans or other mammals, a therapeutically effective amount can be experimentally determined in a laboratory or clinical setting, or the therapeutically effective amount can be the amount required by the guidelines of the U.S. Food and Drug Administration (FDA) or corresponding foreign regulatory agencies for a certain disease and subject to be treated. It should be understood that the determination of the appropriate dosage form, dosage, and route of administration is within the level of ordinary skill in the pharmaceutical and medical fields.

[0023] As used herein, alone or in conjunction with one or more other terms, "subject(s)" and "patient(s)" refer to animals (e.g., mammals), particularly humans. Suitably, "subject(s)" and "patient(s)" may be non-human animals (e.g., livestock or household pets) or humans.

[0024] As used herein alone or in conjunction with one or more other terms, "pharmacologically acceptable" generally refers to a material that is chemically and / or physically compatible with other ingredients (e.g., as with respect to a formulation), and / or generally physiologically compatible with its recipient (e.g., a subject, etc.).

[0025] As used herein, the term "alkyl" includes straight and branched chain alkyl groups. Reference to a particular alkyl group, such as "propyl," is specific for the straight chain version only, and reference to a particular branched chain alkyl group, such as "isopropyl," is specific for the branched chain version only. For example, "(1-6C)alkyl" includes (1-4C)alkyl, (1-3C)alkyl, propyl, isopropyl, and t-butyl.

[0026] The term "(m-nC)" or "(m-nC) group" used alone or as a prefix, refers to any group having from m to n carbon atoms.

[0027] An "alkylene" group is an alkyl group that serves to position and connect two other chemical groups. Thus, "(1-6C)alkylene" means a straight chain, saturated, divalent hydrocarbon group of 1 to 6 carbon atoms or a branched, saturated, divalent hydrocarbon group of 3 to 6 carbon atoms, such as methylene (-CH2-), the ethylene isomers (-CH(CH3)- and -CH2CH2-), the propylene isomers (-CH(CH3)CH2-, -CH(CH2CH3)-, -C(CH3)2-, and -CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), and the like.

[0028] The term "alkyenyl" refers to straight and branched chain alkyl groups containing two or more carbon atoms and at least one carbon-carbon double bond is present within the group. Examples of alkenyl groups include ethenyl, propenyl, and but-2,3-enyl, including all possible geometric (E / Z) isomers.

[0029] The term "alkynyl" refers to straight and branched chain alkyl groups containing two or more carbon atoms and at least one carbon-carbon triple bond is present within the group. Examples of alkynyl groups include acetylenyl and propynyl.

[0030] "(m-nC)cycloalkyl" means a saturated hydrocarbon ring system containing m to n carbon atoms. Exemplary cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and bicyclo[2.2.1]heptyl.

[0031] The term "alkoxy" refers to O-linked straight and branched chain alkyl groups. Examples of alkoxy groups include methoxy, ethoxy and t-butoxy.

[0032] The term "haloalkyl" as used herein refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen (e.g., fluorine) atoms. Examples of haloalkyl groups include -CHF, -CHF, and -CF.

[0033] The terms "halo" or "halogeno" refer to fluoro, chloro, bromo, iodo and astatine, suitably fluoro, chloro, bromo and iodo, more suitably fluoro, chloro and iodo.

[0034] The term "carbocyclyl", "carbocyclic" or "carbocycle" means a non-aromatic saturated or partially saturated monocyclic, fused, bridged, or spiro bicyclic carbon-containing ring system(s). Monocyclic carbocyclic rings contain about 3 to 12 (suitably 3 to 7) ring atoms. Bicyclic carbocyclic rings contain 6 to 17 member atoms, suitably 7 to 12 member atoms in the ring. Bicyclic carbocyclic ring(s) may be fused, spiro, or bridged ring systems. Examples of carbocyclic groups include cyclopropyl, cyclobutyl, cyclohexyl, cyclohexenyl, and spiro[3.3]heptanyl.

[0035] The term "heterocyclyl", "heterocyclic" or "heterocycle" refers to a non-aromatic saturated or partially saturated monocyclic, fused, bridged, or spiro bicyclic heterocyclic ring system(s). A monocyclic heterocyclic ring contains about 3-12 (suitably 3-7) ring atoms, with 1-5 (suitably 1, 2 or 3) heteroatoms selected from nitrogen, oxygen, or sulfur in the ring. A bicyclic heterocyclic ring contains 7-17 member atoms, suitably 7-12 member atoms in the ring. The bicyclic heterocyclic ring(s) may be a fused, spiro, or bridged ring system. Examples of heterocyclic groups include cyclic ethers, such as oxiranyl, oxetanyl, tetrahydrofuranyl, dioxanyl, and substituted cyclic ethers. Heterocycles containing nitrogen include, for example, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrotriazinyl, tetrahydropyrazolyl, etc. Exemplary heterocycles containing sulfur include tetrahydrothienyl, dihydro-1,3-dithiol, tetrahydro-2H-thiopyran, and hexahydrothiepin. Other heterocycles include dihydro-oxathiolyl, tetrahydro-oxazolyl, tetrahydro-oxadiazolyl, tetrahydrodioxazolyl, tetrahydro-oxathiazolyl, hexahydrotriazinyl, tetrahydro-oxazinyl, morpholinyl, thiomorpholinyl, tetrahydropyrimidinyl, dioxolinyl, octahydrobenzofuranyl, octahydrobenzimidazolyl, and octahydrobenzothiazolyl. In the case of heterocycles containing sulfur, sulfur oxide heterocycles containing SO or SO2 groups are also included. Examples include the sulfoxide and sulfone forms of tetrahydrothienyl and thiomorpholinyl, such as tetrahydrothienyl 1,1-dioxide and thiomorpholinyl 1,1-dioxide, etc. The heterocycle can contain one or two oxo (=O) or thioxo (=S) substituents.Suitable values ​​for a heterocyclyl group having one or two oxo (=O) or thioxo (=S) substituents are, for example, 2-oxopyrrolidinyl, 2-thioxopyrrolidinyl, 2-oxoimidazolidinyl, 2-thioxoimidazolidinyl, 2-oxopiperidinyl, 2,5-dioxopyrrolidinyl, 2,5-dioxoimidazolidinyl or 2,6-dioxopiperidinyl. Particular heterocyclyl groups are saturated monocyclic 3-7 membered heterocyclyls containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulfur, such as azetidinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, tetrahydrothienyl, tetrahydrothienyl 1,1-dioxide, thiomorpholinyl, thiomorpholinyl 1,1-dioxide, piperidinyl, homopiperidinyl, piperazinyl or homopiperazinyl. As would be understood by one skilled in the art, any heterocycle can be linked to another group by any suitable atom, for example, by a carbon or nitrogen atom. However, references herein to piperidino or morpholino refer to piperidin-1-yl or morpholin-4-yl rings linked by the ring nitrogen.

[0036] "Bridged ring system" means a ring system in which two rings share two or more atoms, e.g., Advanced Organic Chemistry, by Jerry March, 4 th Edition, Wiley Interscience, pages 131-133, 1992. Examples of bridged heterocyclyl ring systems include aza-bicyclo[2.2.1]heptane, 2-oxa-5-azabicyclo[2.2.1]heptane, aza-bicyclo[2.2.2]octane, aza-bicyclo[3.2.1]octane, and quinuclidine.

[0037] By "spiro bicyclic ring system", we mean that two ring systems share one common spiro carbon atom, i.e., the heterocyclic ring is linked to a further carbocyclic or heterocyclic ring by a single common spiro carbon atom. Examples of spiro ring systems include 6-azaspiro[3.4]octane, 2-oxa-6-azaspiro[3.4]octane, 2-azaspiro[3.3]heptane, 2-oxa-6-azaspiro[3.3]heptane, 7-oxa-2-azaspiro[3.5]nonane, 6-oxa-2-azaspiro[3.4]octane, 2-oxa-7-azaspiro[3.5]nonane and 2-oxa-6-azaspiro[3.5]nonane.

[0038] As used herein alone or in conjunction with another term or terms, "aromatic" refers to monocyclic and polycyclic ring systems containing 4n+2 pi electrons, where n is an integer. Aromatic should be understood to refer to and include ring systems containing only carbon atoms (i.e., "aryl") as well as ring systems containing at least one heteroatom selected from N, O, or S (i.e., "heteroaromatic" or "heteroaryl"). Aromatic ring systems may be substituted or unsubstituted.

[0039] As used herein alone or in conjunction with another term or terms, "non-aromatic" refers to a monocyclic or polycyclic ring system having at least one double bond that is not part of a wide conjugated π system. As used herein, non-aromatic refers to and includes ring systems that contain only carbon atoms as well as ring systems that contain at least one heteroatom selected from N, O, or S. Non-aromatic ring systems may be substituted or unsubstituted.

[0040] The term "heteroaryl" or "aromatic heterocycle" refers to an aromatic monocyclic, bicyclic, or polycyclic ring incorporating one or more (e.g., 1-4, particularly 1, 2, or 3) heteroatoms selected from nitrogen, oxygen, or sulfur. The term heteroaryl includes monovalent and divalent species. Examples of heteroaryl groups are monocyclic and bicyclic groups containing 5-12 ring members, more usually 5-10 ring members. Heteroaryl groups can be, for example, 5- or 6-membered monocyclic rings or 9- or 10-membered bicyclic rings, e.g., 5- and 6-membered fused rings or a bicyclic structure formed from two 6-membered fused rings. Each ring can contain up to about 4 heteroatoms, typically selected from nitrogen, sulfur, and oxygen. Typically, heteroaryl rings contain up to 3 heteroatoms, more usually up to 2, e.g., a single heteroatom. In one embodiment, a heteroaryl ring contains at least one ring nitrogen atom. The nitrogen atoms in the heteroaryl ring may be basic, as in the case of an imidazole or pyridine, or essentially non-basic, as in the case of an indole or pyrrole nitrogen. Generally, the number of basic nitrogen atoms present in a heteroaryl group, including any amino group substituents on the ring, will be fewer than five.

[0041] Examples of heteroaryl include furyl, pyrrolyl, thienyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazenyl, benzofuranyl, indolyl, isoindolyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, benzothiazolyl, indazolyl, purinyl, benzofurazanyl, quinolyl, isoquinolyl, quinazolyl, and aryl. These include aryl, quinoxalinyl, cinnolinyl, pteridinyl, naphthyridinyl, carbazolyl, phenazinyl, benzisoquinolinyl, pyridopyrazinyl, thieno[2,3-b]furanyl, 2H-furo[3,2-b]-pyranyl, 5H-pyrido[2,3-d]-o-oxazinyl, 1H-pyrazolo[4,3-d]-oxazolyl, 4H-imidazo[4,5-d]thiazolyl, pyrazino[2,3-d]pyridazinyl, imidazo[2,1-b]thiazolyl, and imidazo[1,2-b][1,2,4]triazinyl. "Heteroaryl" also encompasses partially aromatic bicyclic or polycyclic ring systems in which at least one ring is aromatic and one or more of the other ring(s) is non-aromatic, saturated or partially saturated, provided that at least one ring contains one or more heteroatoms selected from nitrogen, oxygen or sulfur. Examples of partially aromatic heteroaryl groups include, for example, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 2-oxo-1,2,3,4-tetrahydroquinolinyl, dihydrobenzthienyl, dihydrobenzfuranyl, 2,3-dihydro-benzo[1,4]dioxinyl, benzo[1,3]dioxolyl, 2,2-dioxo-1,3-dihydro-2-benzothienyl, 4,5,6,7-tetrahydrobenzofuranyl, indolinyl, 1,2,3,4-tetrahydro-1,8-naphthyridinyl, 1,2,3,4-tetrahydropyrido[2,3-b]pyrazinyl, and 3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazinyl.

[0042] Examples of 5-membered heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, furazanyl, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl and tetrazolyl groups.

[0043] Examples of 6-membered heteroaryl groups include, but are not limited to, pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl, and triazinyl.

[0044] Bicyclic heteroaryl groups include, for example: a benzene ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; a pyridine ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; a pyrimidine ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; a pyrrole ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; a pyrazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; a pyrazine ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; an imidazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; an oxazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; an isoxazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; a thiazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; an isothiazole ring fused to a 5- or 6-membered ring containing 1 or 2 ring heteroatoms; a thiophene ring fused to a 5- or 6-membered ring containing 1, 2, or 3 ring heteroatoms; a furan ring fused to a 5- or 6-membered ring containing 1, 2 or 3 ring heteroatoms; a cyclohexyl ring fused to a 5- or 6-membered aromatic heterocycle containing 1, 2, or 3 ring heteroatoms; and The radical may be a group selected from a cyclopentyl ring fused to a 5- or 6-membered aromatic heterocycle containing 1, 2 or 3 ring heteroatoms.

[0045] Particular examples of bicyclic heteroaryl groups containing a 6-membered ring fused to a 5-membered ring include, but are not limited to, benzfuranyl, benzthiophenyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzthiazolyl, benzisothiazolyl, isobenzofuranyl, indolyl, isoindolyl, indolizinyl, indolinyl, isoindolinyl, purinyl (e.g., adeninyl, guaninyl), indazolyl, benzodioxolyl and pyrazolopyridinyl groups.

[0046] Particular examples of bicyclic heteroaryl groups containing two fused six-membered rings include, but are not limited to, quinolinyl, isoquinolinyl, chromanyl, thiochromanyl, chromenyl, isochromenyl, chromanyl, isochromanyl, benzodioxanyl, quinolidinyl, benzoxazinyl, benzodiazinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl and pteridinyl groups.

[0047] The term "aryl" refers to a cyclic or polycyclic aromatic ring having 5 to 12 carbon atoms. The term aryl includes monovalent and divalent species. Examples of aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl, and the like. In a particular embodiment, the aryl is phenyl.

[0048] The present specification also uses some compound terms to describe groups that contain two or more functional groups. Such terms are understood by those of skill in the art. For example, (3-6C)cycloalkyl(m-nC)alkyl includes (m-nC)alkyl substituted with (3-6C)cycloalkyl.

[0049] The term "optionally substituted" refers to groups, structures, or molecules that are substituted and to groups, structures, or molecules that are not substituted. 1 "One / any CH, CH, CH group or heteroatom (i.e., NH) in the group is optionally substituted" means that R 1 Suitably it means that (any) one of the hydrogen radicals of the group is replaced by the associated specified group.

[0050] When an optional substituent is selected from "one or more" groups, it is to be understood that this definition includes all substituents selected from one of the specified groups or substituents selected from two or more of the specified groups. In some embodiments, one or more refers to one, two, or three. In other embodiments, one or more refers to one or two. In particular embodiments, one or more refers to one.

[0051] The phrase "compounds of the invention" refers generally and specifically to the compounds disclosed herein.

[0052] As used herein in connection with a measurable value, such as an amount or duration, "about" is meant to encompass a reasonable variation of the value, for example, allowing for experimental error in determining said value.

[0053] compound In one aspect, the present invention provides a compound having structural formula (I), as shown below: [ka] [In the formula, A1 and A2 are each selected from CR1 and CH, CR1 and CF, CR1 and N, CH and CR1, CF and CR1, or NH and CR1; R1 is a radioactive isotope of iodine (I) or astatine (At); A3 is selected from N-X1-R3 or C(R4)OR5; X1 is absent, -C(O)-, -[CH2] 1~3 -, -C(O)NH-, -C(O)O-, or -S(O) 0~2 - Selected from; R3 is H, (1-10C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl or heteroaryl, each alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl group being selected from one or more R A optionally substituted by substituents; R4 is H or C 1~4 alkyl; R5 is selected from (1-7C)alkyl, heterocyclyl and aryl, which groups are each independently selected from one or more R A optionally substituted by substituents; or R4 and R5, together with the carbon and oxygen atoms attached thereto, can be one or more R A spiro-C optionally substituted with substituents or fused to a phenyl or 5- or 6-membered heteroaryl ring 5~7 Forming an oxygen-containing heterocyclyl; Each R A The substituents are fluoro, cyano, (1-4C) alkyl, (1-4C) haloalkyl, (1-4C) hydroxyalkyl, phenyl, 4- to 6-membered heterocyclyl, 5- to 6-membered heteroaryl, -OR 1a , -NR 1a R 1b , -C(O)R 1a , -C(O)N(R 1b )R 1a , N(R 1b )C(O)R 1a , -S(O) p R 1a (p is 0, 1 or 2), -SON(R 1b )R 1a , -N(R 1b )SO2R 1a (In the formula, R 1a and R 1bare each independently selected from hydrogen, (1-4C)alkyl or (3-6C)alkyl), or a pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0054] Certain compounds of the present invention include, for example, compounds of structural formulae Ia-Ij shown below: [ka] [ka] [ka] [wherein, where appropriate, R1, R3 and R5 are each as defined herein], or a pharma- ceutically acceptable salt, hydrate and / or solvate thereof.

[0055] Certain compounds of the present invention include, for example, compounds of formula (I) [including subformulae Ia-Ii], or pharma- ceutically acceptable salts, hydrates and / or solvates thereof, and unless otherwise indicated, A1, A2, A3, R1, X1, R3, R4, R5 and R A Each of has any of the meanings defined above or in any of paragraphs (1) to (34) below: (1) A1 and A2 are each selected from CR1 and CH, CR1 and CF, CH and CR1, or CF and CR1; (2) A1 and A2 are each selected from CR1 and CH, CR1 and N, CH and CR1, or NH and CR1; (3) A1 and A2 are selected from CR1 and CH, CH and CR1, or CF and CR1; (4) A1 is CR1 and A2 is CH; (5) R1 is a radioactive isotope of iodine (I); (6) R1 is a radioactive isotope of astatine (At); (7) R1 is 123 I, 131 I, 127 I, 124 I or 211 At, or 123 I, 131 I, 124 I or 211 Selected from At (8) R1 is 123 I, 131 I or 211 Selected from At; (9) R1 is 123 I or 131 I (10) R1 is 123 I am; (11) A3 is N-X1-R3; (12) A3 is C(R4)OR5; (13) X1 is absent, -C(O)-, -[CH2] 1~3 - or -C(O)NH-; (14) X1 is -C(O)-; (15) R3 is H, (1-6C)alkyl, (3-8C)cycloalkyl, phenyl, 4- to 10-membered heterocyclyl, or 5- or 6-membered heteroaryl, each alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl group being selected from one or more R A Optionally substituted by substituents; (16) R3 is H, (1-6C)alkyl, (3-8C)cycloalkyl, phenyl, 4- to 8-membered heterocyclyl, or 5- or 6-membered heteroaryl, each alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl group being selected from one or more R A Optionally substituted by substituents; (17) R3 is (1-6C)alkyl or (3-8C)cycloalkyl, each of which may be one or more R A Optionally substituted by substituents; (18) R3 is one or more R A (3-8C)cycloalkyl optionally substituted by substituents; (19) R3 is one or more R A (3-6C)cycloalkyl optionally substituted by substituents; (20) R3 is cyclopropyl; (21) R4 is H or C 1~4 selected from alkyl; (22) R4 is selected from H or methyl; (23) R4 is H; (24) R5 is selected from (1-7C)alkyl, (3-8C)cycloalkyl, 4- to 10-membered heterocyclyl, and phenyl, which groups are each independently selected from one or more R A Optionally substituted by substituents; (25) R5 is selected from (1-7C)alkyl, 4- to 10-membered heterocyclyl, and phenyl, which groups are selected from one or more R A Optionally substituted by substituents; (26) R5 is selected from (1-7C)alkyl, (3-6C)cycloalkyl, 4- to 10-membered heterocyclyl, and phenyl, which groups are each independently selected from one or more R A Optionally substituted by substituents; (27) R5 is one or more R A is (1-6C)alkyl optionally substituted by substituents; (28) R5 is methyl; (29) R4 and R5, together with the carbon and oxygen atoms attached thereto, represent one or more R A spiro-C optionally substituted with substituents or fused to a phenyl or 5- or 6-membered heteroaryl ring 5~6 Forming an oxygen-containing heterocyclyl; (30) R4 and R5, together with the carbon and oxygen atoms attached thereto, form a spiro-C 5~6 Forming an oxygen-containing heterocyclyl; (31)Each R A The substituents are fluoro, cyano, (1-4C) alkyl, (1-4C) haloalkyl, (1-4C) hydroxyalkyl, -OR 1a , -NR 1a R 1b , -C(O)R 1a , -C(O)N(R 1b )R 1a , N(R 1b )C(O)R 1a , -S(O) p R 1a (p is 0, 1 or 2), -SON(R 1b )R 1a , -N(R 1b )SO2R 1a [In the formula, R 1a and R 1b are each independently selected from hydrogen or (1-4C)alkyl; (32)Each R A The substituents are fluoro, cyano, (1-4C) alkyl, (1-4C) haloalkyl, (1-4C) hydroxyalkyl, -OR 1a , -NR 1a R 1b , -C(O)R 1a , -C(O)N(R 1b )R 1a , N(R 1b )C(O)R 1a , -S(O) p R 1a (p is 0, 1 or 2) 1a and R 1b are each independently selected from hydrogen or (1-2C)alkyl; (33)Each R A The substituents are fluoro, cyano, (1-4C) alkyl, (1-4C) haloalkyl, (1-4C) hydroxyalkyl, or -OR. 1a [In the formula, R 1a are independently selected from hydrogen or (1-2C)alkyl; (34)Each R A The substituent is fluoro.

[0056] Suitably, A1 and A2 are as defined in any one of numbered paragraphs (1) to (4) above. More suitably, A1 and A2 are as defined in numbered paragraphs (3) or (4) above. Most suitably, A1 and A2 are as defined in paragraph (4) above.

[0057] Suitably, R1 is as defined in any one of numbered paragraphs (5) to (10) above. More suitably, R1 is as defined in any one of numbered paragraphs (7) to (10) above. Most suitably, R1 is as defined in paragraph (10) above.

[0058] Suitably, A3 is as defined in paragraph (11) above.

[0059] Suitably, X1 is as defined above in paragraph (13) or (14). Most suitably, X1 is as defined above in paragraph (14).

[0060] Suitably, R3 is as defined in any one of numbered paragraphs (15) to (20) above. More suitably, R3 is as defined in any one of numbered paragraphs (18) to (20) above. Most suitably, R3 is as defined in numbered paragraph (20) above.

[0061] Suitably, R4 is as defined in any one of numbered paragraphs (21) to (23) above. More suitably, R4 is as defined in numbered paragraphs (22) or (23). Most suitably, R4 is as defined in numbered paragraph (23) above.

[0062] Suitably, R5 is as defined in any one of numbered paragraphs (24) to (28) above. More suitably, R5 is as defined in any one of numbered paragraphs (26) to (28) above. Most suitably, R5 is as defined in numbered paragraph (28) above.

[0063] Suitably, when R4 and R5 are both linked, they are as defined in paragraph (29) or (30) above.

[0064] Appropriately, R A is as defined in any one of numbered paragraphs (31) to (34) above. More suitably, R A is as defined in numbered paragraph (33) or (34) above. Most suitably, R A is as defined in paragraph (34) above.

[0065] Suitably, R5 is as defined in any one of numbered paragraphs (28) to (30) above. More suitably, R5 is as defined in numbered paragraphs (29) or (30) above. Most suitably, R5 is as defined in numbered paragraph (30) above.

[0066] Suitably, y is as defined in numbered paragraph (36) above.

[0067] Suitably, x is as defined in numbered paragraph (40) above.

[0068] As noted above, certain compounds of the present invention include compounds of formula (I) as defined herein, for example having one of the structural formulae Ia-Ij shown above, where R1, R3 and R5, as appropriate, each have any one of the definitions set forth herein, or a pharma- ceutically acceptable salt, hydrate and / or solvate thereof.

[0069] Certain compounds of the present invention have the structural formulas Ij and Ik shown below: [ka] (also referred to herein as CC1(4-[(3-[(4-cyclopropylcarbonyl)piperazin-1-yl]carbonyl-4-iodophenyl]methyl(2H)phthalazin-1-one) [ka] or pharma-ceutically acceptable salts, hydrates and / or solvates thereof.

[0070] Certain compounds of the present invention have the structural formula Ij above, or a pharma- ceutically acceptable salt, hydrate and / or solvate thereof.

[0071] The various functional groups and substituents which make up the compounds of formula (I) or sub-formulae (Ia) to (Ie) are typically selected such that the molecular weight of the compounds of formula (I) does not exceed 1000.

[0072] Suitable pharma- ceutically acceptable salts of the compounds of the invention are, for example, sufficiently basic, such as acid-addition salts of the compounds of the invention, for example with inorganic or organic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, trifluoroacetic acid, formic acid, citric acid, methane sulfonate or maleic acid.Furthermore, suitable pharma- ceutically acceptable salts of the compounds of the invention that are sufficiently acidic are alkali metal salts, for example sodium or potassium salts, alkaline earth metal salts, for example calcium or magnesium salts, ammonium salts or salts with organic bases that generate pharma- ceutically acceptable cations, for example salts with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.

[0073] Compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or in the arrangement of their atoms in space are called "isomers". Isomers that differ in the arrangement of their atoms in space are called "stereoisomers". Stereoisomers that are not mirror images of one another are called "diastereomers" and those that are non-superimposable mirror images of one another are called "enantiomers". When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. Enantiomers can be characterized by the absolute configuration of their asymmetric center and described by the R- and S-sequencing rules of Cahn and Prelog, or by the way the molecule rotates a plane of polarized light and can be named as dextrorotatory or levorotatory (i.e., (+) or (-)-isomer, respectively). Chiral compounds can exist as either enantiomer or as a mixture thereof. A mixture containing equal proportions of enantiomers is called a "racemic mixture".

[0074] The compounds of the present invention may have one or more asymmetric centers; therefore, such compounds can be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless otherwise indicated, the description or naming of a particular compound in this specification and claims is intended to include both individual enantiomers and their mixtures, racemic or otherwise. Methods for determining stereochemistry and separating stereoisomers are well known in the art, for example, by synthesis from optically active starting materials or by resolution of racemates (see those discussed in Chapter 4 of "Advanced Organic Chemistry", 4th edition J. March, John Wiley and Sons, New York, 2001). Some of the compounds of the present invention may have geometric isomeric centers (E- and Z-isomers).

[0075] It is to be understood that the invention encompasses all optical, diastereomeric and geometric isomers and mixtures thereof that have anti-cancer activity or are suitable as imaging agents.

[0076] The present invention also includes compounds of the invention as defined herein that contain one or more isotopic substitutions. For example, H is 1 H, 2 H(D), and 3 H(T) can be in any isotopic form; C is 12 C, 13C, and 14 It can be in any isotopic form, including C; O is 16 O and 18 It may be in any isotopic form, including O, etc.

[0077] It is also to be understood that some of the compounds of formula (I) or subformulas Ia to Ik can exist in solvated as well as unsolvated forms, such as, for example, hydrated forms, etc. It is to be understood that the present invention encompasses all such solvated forms that possess activity.

[0078] It is also to be understood that some compounds of formula (I) or subformulas Ia to Ik may exhibit polymorphism, and that the invention encompasses all such forms that possess activity.

[0079] The compounds of formula (I) or sub-formulae Ia to Ik can exist in many different tautomeric forms, and reference to a compound of formula (I) or sub-formulae Ia to Ik includes all such forms. For the avoidance of doubt, a compound may exist in one of several tautomeric forms, only one of which is described or shown in detail, while all others are encompassed by formula (I) or sub-formulae (Ia) to (Ie). Examples of tautomeric forms include, for example, the keto, enol, and enolate forms for the following tautomeric pairs: keto / enol (illustrated below), imine / enamine, amide / iminoalcohol, amidine / amidine, nitroso / oxime, thioketone / enethiol, and nitro / aci-nitro.

[0080] [ka]

[0081] Compounds of formula (I) or sub-formulas Ia to Ik that contain amine functional groups can also form N-oxides. References herein to compounds of formula (I) or sub-formulas Ia to Ik that contain amine functional groups also include N-oxides. When a compound contains several amine functional groups, one or more nitrogen atoms can be oxidized to form N-oxides. Particular examples of N-oxides are the N-oxides of tertiary amines or the nitrogen atoms of nitrogen-containing heterocycles. N-oxides can be formed by treatment of the corresponding amines with oxidizing agents, such as hydrogen peroxide or peracids (e.g. peroxycarboxylic acids), see, for example, Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience, pages. More specifically, the N-oxides can be prepared, for example, by the procedure of LW Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compounds are reacted with m-chloroperoxybenzoic acid (mCPBA) in an inert solvent such as dichloromethane.

[0082] The compounds of formula (I) or sub-formulae Ia to Ik can be administered in the form of prodrugs that are broken down in the human or animal body to release the compounds of the invention. Prodrugs can be used to modify the physical and / or pharmacokinetic properties of the compounds of the invention. Prodrugs can be formed when the compounds of the invention contain a suitable group or substituent to which a property-modifying group can be attached. Examples of prodrugs include in vivo cleavable ester derivatives that can be formed at the carboxy or hydroxy groups in the compounds of formula (I) or sub-formulae Ia to Ik, and in vivo cleavable amide derivatives that can be formed at the carboxy or amino groups in the compounds of formula (I) or sub-formulae Ia to Ik.

[0083] The present invention therefore includes compounds of formula (I) or sub-formulas Ia to Ik as defined above when available by organic synthesis and when available in the human or animal body as cleavage of their prodrugs. The present invention therefore includes compounds of formula (I) or sub-formulas Ia to Ik produced by organic synthetic means and also produced in the human or animal body as metabolism of precursor compounds, which compounds of formula (I) or sub-formulas Ia to Ik, may be synthetically produced compounds or metabolically produced compounds.

[0084] Suitable pharma- ceutically acceptable prodrugs of the compounds of formula (I) or subformulas Ia through Ik are those based on sound medical judgment, such that they are suitable for administration to the human or animal body without undesirable pharmacological activity and without undue toxicity.

[0085] Various forms of prodrugs are described, for example, in the following documents: a)Methods in Enzymology, Vol.42, p.309-396, edited by K.Widder, et al.(Academic Press, 1985); b)Design of Pro-drugs, edited by H.Bundgaard,(Elsevier, 1985); c)A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H.Bundgaard, Chapter 5 “Design and Application of Pro-drugs”, by H.Bundgaard p.113-191(1991); d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992); e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285(1988); f) N.Kakeya, et al., Chem.Pharm.Bull., 32, 692(1984); g) T. Higuchi and V. Stella, “Pro-Drugs as Novel Delivery Systems”, ACSSymposium Series, Volume 14; and h) E. Roche (editor), “Bioreversible Carriers in Drug Design”, Pergamon Press, 1987.

[0086] Suitable pharma- ceutically acceptable prodrugs of the compounds of formula (I) or sub-formulae Ia to Ik that contain a carboxy group are, for example, in vivo cleavable esters thereof. In vivo cleavable esters of the compounds of formula (I) or sub-formulae Ia to Ik that contain a carboxy group are, for example, pharma- ceutically acceptable esters that are cleaved in the human or animal body to produce the parent acid or alcohol. Suitable pharma- ceutically acceptable esters for carboxy include (1-6C)alkyl esters such as methyl, ethyl and tert-butyl, (1-6C)alkoxymethyl esters such as methoxymethyl ester, (1-6C)alkanoyloxymethyl esters such as pivaloyloxymethyl ester, 3-phthalidyl ester, (3-8C)cycloalkylcarbonyloxy-(1-6C)alkyl esters such as cyclopentylcarbonyloxymethyl and 1-cyclohexylcarbonyloxyethyl ester, 2-oxo-1,3-dioxolenylmethyl esters such as 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl ester and (1-6C)alkoxycarbonyloxy-(1-6C)alkyl esters such as methoxycarbonyloxymethyl and 1-methoxycarbonyloxyethyl ester.

[0087] Suitable pharma- ceutically acceptable prodrugs of the compounds of formula (I) or sub-formulae Ia to Ik having a hydroxy group are, for example, in vivo cleavable esters or ethers thereof. The in vivo cleavable esters or ethers of the compounds of formula (I) or sub-formulae Ia to Ik containing a hydroxy group are, for example, pharma- ceutically acceptable esters or ethers which are cleaved in the human or animal body to produce the parent hydroxy compound. Suitable pharma- ceutically acceptable esters forming groups for hydroxy groups include inorganic esters, for example, phosphate esters (including phosphoramidate cyclic esters), and the like. Further suitable pharma- ceutically acceptable esters forming groups for hydroxy groups include (1-10C)alkanoyl groups, for example, acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, (1-10C)alkoxycarbonyl groups, for example, ethoxycarbonyl, N,N-(1-6C)2carbamoyl, 2-dialkylaminoacetyl and 2-carboxyacetyl groups, and the like. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-1-ylmethyl and 4-(1-4C)alkylpiperazin-1-ylmethyl.Suitable pharma-ceutically acceptable ether forming groups for hydroxy groups include α-acyloxyalkyl groups, such as acetoxymethyl and pivaloyloxymethyl groups.

[0088] Suitable pharma- ceutically acceptable prodrugs of the compounds of formula (I) or sub-formulae Ia to Ik bearing a carboxy group are, for example, in vivo cleavable amides thereof, such as amides formed with amines, such as ammonia, (1-4C)alkylamines, such as methylamine, [(1-4C)alkyl]2amines, such as dimethylamine, N-ethyl-N-methylamine or diethylamine, (1-4C)alkoxy-(2-4C)alkylamines, such as 2-methoxyethylamine, phenyl-(1-4C)alkylamines, such as benzylamine and amino acids, such as glycine or its esters.

[0089] Suitable pharma- ceutically acceptable prodrugs of the compounds of formula (I) or sub-formulas Ia to Ik bearing an amino group are, for example, their amide derivatives which are cleavable in vivo. Suitable pharma- ceutically acceptable amides derived from an amino group include, for example, amides formed with (1-10C)alkanoyl groups, such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups. Examples of ring substituents on phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-1-ylmethyl and 4-(1-4C)alkyl)piperazin-1-ylmethyl.

[0090] The in vivo effects of the compounds of formula (I) or sub-formulae Ia to Ik may be exerted in part by one or more metabolic products formed in the human or animal body following administration of the compounds of formula (I) or sub-formulae Ia to Ik. As described below, the in vivo effects of the compounds of formula (I) or sub-formulae Ia to Ik may also be exerted as a result of the metabolism of precursor compounds (prodrugs).

[0091] While the invention may relate to any compound or particular group of compounds defined herein as an optional, preferred or suitable feature or otherwise in relation to certain particular embodiments, the invention may relate to any compound or particular group of compounds that specifically excludes said optional, preferred or suitable feature or certain particular embodiment.

[0092] Suitably, the present invention excludes any individual compound that does not have biological activity as defined herein.

[0093] synthesis The compounds of the present invention can be prepared by any suitable technique known in the art. Certain methods for the preparation of these compounds are further described in the accompanying examples.

[0094] In the descriptions of synthetic methods described herein and in any referenced synthetic methods used to prepare starting materials, it is to be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment and work-up procedures, can be selected by one of ordinary skill in the art.

[0095] It is understood by one skilled in the art of organic synthesis that the functionality present on various portions of the molecule must be compatible with the reagents and reaction conditions employed.

[0096] It is understood that during the synthesis of the compounds of the invention in the methods defined herein, or during the synthesis of some of the starting materials, it may be desirable to protect some of the substituents to prevent their undesired reactions. The skilled chemist will understand when such protection is required and how such protecting groups can be introduced and subsequently removed.

[0097] For examples of protecting groups, see one of the many general texts on the subject, such as 'Protective Groups in Organic Synthesis' by Theodora Green, published by John Wiley & Sons, Inc. Protecting groups can be removed by any convenient method described in the literature or known to the skilled chemist as appropriate for the removal of the protecting group in question, such method being chosen to effect removal of the protecting group with minimal disturbance of other groups in the molecule.

[0098] Thus, if the reactants contain groups such as amino, carboxy or hydroxy, it may be desirable to protect the group in some of the reactions mentioned herein.

[0099] For example, suitable protecting groups for amino or alkylamino groups are, for example, acyl groups, such as alkanoyl groups, for example acetyl, alkoxycarbonyl groups, for example methoxycarbonyl, ethoxycarbonyl or t-butoxycarbonyl groups, arylmethoxycarbonyl groups, for example benzyloxycarbonyl, or aroyl groups, for example benzoyl. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, acyl groups, such as alkanoyl or alkoxycarbonyl groups or aroyl groups, can be removed, for example, by hydrolysis with a suitable base, for example an alkali metal hydroxide, for example lithium hydroxide or sodium hydroxide. Alternatively, acyl groups, such as tert-butoxycarbonyl groups, may be removed by treatment with a suitable acid, for example hydrochloric acid, sulfuric acid or phosphoric acid or trifluoroacetic acid, and arylmethoxycarbonyl groups, such as benzyloxycarbonyl groups, may be removed, for example, by hydrogenation over a catalyst, for example palladium on carbon, or by treatment with a Lewis acid, for example boron tris(trifluoroacetate).A suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or with hydrazine.

[0100] Suitable protecting groups for hydroxyl groups are, for example, acyl groups, such as alkanoyl groups, for example acetyl, aroyl groups, for example benzoyl, or arylmethyl groups, for example benzyl. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, acyl groups, such as alkanoyl or aroyl groups, can be removed, for example, by hydrolysis with a suitable base, for example an alkali metal hydroxide, for example lithium, sodium hydroxide or ammonia. Alternatively, arylmethyl groups, such as benzyl groups, can be removed, for example, by hydrogenation over a catalyst, for example palladium on carbon.

[0101] A suitable protecting group for a carboxy group is, for example, an esterifying group, a methyl or ethyl group which may be removed, for example, by hydrolysis with a base, such as, for example, sodium hydroxide, or, for example, a t-butyl group which may be removed, for example, by treatment with an acid, for example an organic acid, such as, for example, trifluoroacetic acid, or a benzyl group which may be removed, for example, by hydrogenation, for example, over a catalyst, such as, for example, palladium on carbon.

[0102] Resins can also be used as protecting groups.

[0103] The methodology used to synthesize compounds of formula (I), or subformulas Ia-Ik, will vary depending on the nature of the variable A1, A2 or A3 groups.

[0104] In certain methods of the present invention, the method of synthesizing a compound of formula I comprises the steps of: [ka] [In the formula, A 1a Or A 2a is a CB group, B is a boronic acid or boronate ester (e.g., a boronate pinacol ester); A1 and A2 are each as defined herein; formula: M[R1] wherein M is a metal (e.g., Na) and R1 is as defined above; The method includes reacting in the presence of a copper catalyst (e.g., Cu(OCOCF3)) and a suitable ligand (e.g., 1,10 phenanthroline).

[0105] In one embodiment, the reaction comprises reacting a compound of formula A in a manner as defined above.

[0106] Suitably B is a boronate ester (eg a boronate pinacol ester).

[0107] Suitably, M is Na, the copper catalyst is Cu(OCOCF3) and the ligand is 1,10 phenanthroline.

[0108] Suitably, R1 is 123 I.

[0109] Suitably the reaction is carried out in a suitable solvent, for example a mixture of methanol and water.

[0110] Suitably the reaction is carried out at an elevated temperature of 50 to 85° C. (eg 80° C.) for a period of 10 to 120 minutes (eg 25 minutes).

[0111] Compounds of formula Ij as defined herein are particularly difficult to synthesize. In certain methods of the invention, a method for synthesizing a compound of formula Ij, or a pharma- ceutically acceptable salt thereof, comprises reacting a compound of formula Aj, as shown below: [ka] wherein B is a boronic acid or a boronate ester (e.g., a boronate pinacol ester); and A1 and A2 are each as defined herein; formula: M[ 123 I] wherein M is a metal (e.g., Na); The method includes reacting in the presence of a copper catalyst (e.g., Cu(OCOCF3)) and a suitable ligand (e.g., 1,10 phenanthroline).

[0112] Suitably B is a boronate ester (eg a boronate pinacol ester).

[0113] Suitably, M is Na, the copper catalyst is Cu(OCOCF3) and the ligand is 1,10 phenanthroline.

[0114] Suitably the reaction is carried out in a suitable solvent, for example a mixture of methanol and water.

[0115] Suitably the reaction is carried out at an elevated temperature of 50 to 85° C. (eg 80° C.) for a period of 10 to 120 minutes (eg 25 minutes).

[0116] After a compound of formula (I), or of sub-formulas Ia to Ik, has been synthesised by any one of the processes defined herein, the processes then include the following further steps: (i) removing any protecting groups present; (ii) converting a compound of formula (I) to another compound of formula (I); (iii) forming a pharma- ceutically acceptable salt, hydrate or solvate thereof; and / or (iv) forming a prodrug thereof It may further include:

[0117] An example of (ii) above is where a compound of formula (I) is synthesised and then one or more groups can be further reacted to alter the nature of that group to provide an alternative compound of formula (I).

[0118] The resulting compounds of formula (I), or subformulas Ia to Ik, can be isolated and purified using techniques well known in the art.

[0119] Compounds of formula (I), or subformulas Ia-Ik, may be synthesised according to the synthetic routes set out in the Examples section below.

[0120] Pharmaceutical Compositions According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of the present invention as hereinbefore defined, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, in association with a pharma- ceutically acceptable diluent or carrier.

[0121] The compositions of the invention may be in a form suitable for oral use (e.g., as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), topical use (e.g., as creams, ointments, gels, or aqueous or oily solutions or suspensions), administration by inhalation (e.g., as a finely divided powder or liquid aerosol), administration by insufflation (e.g., as a finely divided powder) or parenteral administration (e.g., as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, intraperitoneal or intramuscular administration or as a suppository for rectal administration).

[0122] The compositions of the present invention can be obtained by conventional procedures using conventional pharmaceutical excipients well known in the art. Thus, compositions intended for oral use can contain, for example, one or more coloring agents, sweeteners, flavoring agents and / or preservatives.

[0123] An effective amount of a compound of the invention for use in therapy is an amount sufficient to treat or prevent, slow the progression of and / or alleviate the symptoms associated with the proliferative conditions referred to herein.

[0124] The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending on the individual treated and the particular route of administration.

[0125] use R1 is 123 I, 124 I, 131 I, 127 I, or 211 The compounds of the invention selected from At are suitable for the therapeutic uses and applications described below. In particular, 123 The compounds of the invention, I, are exceptionally potent and effective therapeutic agents. In particular, compounds of formula Ij are shown to be exceptionally active in the examples described herein.

[0126] R1 is 124The compounds of the present invention, which are I, are suitable for PET imaging applications. PET imaging is well known in the art and is 124 The compounds of the invention, I, are suitable agents for PET imaging of cancer, particularly cancers expressing the PARP enzyme.

[0127] Such compounds can be administered to the body and accumulate in the cancer. The composition for administering such compounds is the same as the pharmaceutical composition defined herein. Similarly, the route of delivery of such compounds is the same as that of the therapeutic compounds defined herein.

[0128] As described herein, R 123 Compounds that are I can be used for therapeutic applications (as described above) and imaging applications. For example, such compounds can be used for dosimetry studies, patient selection and / or treatment follow-up.

[0129] Therapeutic Uses and Applications The present invention provides compounds that are radiopharmaceuticals and capable of delivering a lethal dose of radiation to tumor cells. Suitably, the compounds of the present invention that are suitable for therapeutic applications are those in which R1 is 123 I, 131 I or 211 At, especially 123 It is I.

[0130] As a result, the compounds of the present invention are suitable for the treatment of cancer.

[0131] The compounds of the present invention are particularly suitable for targeting PARP enzyme-expressing cancers.

[0132] According to one aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use in therapy.

[0133] According to a further aspect of the invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use as a medicament.

[0134] According to one aspect of the invention there is provided a method of treating cancer in a patient in need of such treatment, said method comprising the step of administering to said patient a therapeutically effective amount of a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein.

[0135] According to a further aspect of the present invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein, for use in the treatment of cancer.

[0136] According to a further aspect of the invention there is provided the use of a compound defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, in the manufacture of a medicament for the treatment of cancer.

[0137] According to certain aspects of the invention, there is provided a method of treating a PARP enzyme-expressing cancer in a patient in need of such treatment, said method comprising the step of administering to said patient a therapeutically effective amount of a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein.

[0138] According to a further particular aspect of the present invention there is provided a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein, for use in the treatment of a PARP enzyme-expressing cancer.

[0139] According to a further particular aspect of the present invention there is provided the use of a compound as defined herein, or a pharma- ceutically acceptable salt, hydrate or solvate thereof, in the manufacture of a medicament for the treatment of a PARP enzyme-expressing cancer.

[0140] The compounds of the present invention can be used to treat any suitable cancer amenable to treatment.Cancers that may be targeted include (e.g., adenoid cystic carcinoma, adrenal tumors, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxia telangiectasia, Beckwith-Wiedemann syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dubé syndrome, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, Carney complex, central nervous system tumors, cervical cancer, colorectal cancer, Cowden disease, craniopharyngioma, desmoplastic infantile ganglioglioma, ependymoma, esophageal cancer, Ewing's sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial melanoma, familial Non-VHL clear cell renal cell carcinoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell carcinoma, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal cell carcinoma, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumors, laryngeal and hypopharyngeal cancer, leukemia (acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), B-cell prolymphocytic leukemia, hairy cell leukemia, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia ( CML), chronic T-cell lymphocytic leukemia, eosinophilic leukemia), Li-Fraumeni syndrome, liver cancer, lung cancer (non-small cell lung cancer, small cell lung cancer), lymphoma (Hodgkin, non-Hodgkin), Lynch syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia type 1 and type 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndrome (MDS), nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, neuroendocrine tumors (e.g., of the gastrointestinal tract, lung, or pancreas), neurofibromatosis type 1 and type 2, nevoid basal cell carcinoma syndrome, oral or oropharyngeal cancer, osteosarcoma, ovarian / fallopian tube / peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers syndrome, pheochromocytoma, paraganglioma, pituitary tumors, pleuropulmonary blastoma, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma (e.g., Kaposi or soft tissue), skin cancer, small intestine cancer, gastric cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis, uterine cancer, vaginal cancer, von Hippel-Lindau syndrome, vulvar cancer, Waldenström's macroglobulinemia, Werner syndrome, Wilms' tumor, and xeroderma pigmentosum).Particular cancers of interest include hematological cancers, such as lymphomas (including diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkitt's lymphoma (BL) and angioimmunoblastic T-cell lymphoma (AITL)), leukemias (including acute lymphoblastic leukemia (ALL) and chronic myeloid leukemia (CML)), multiple myeloma, breast cancer, non-small cell lung cancer (NSCLC), colorectal cancer, endometrial cancer, gastro-esophageal cancer, neuroendocrine cancer, osteosarcoma, prostate cancer, pancreatic cancer, small intestine cancer, bladder cancer, rectal cancer, cholangiocarcinoma, CNS cancer, thyroid cancer, head and neck cancer, esophageal cancer, and ovarian cancer.

[0141] The compound of the present invention is particularly suitable for treating PARP enzyme expressing cancer.The detailed examples of PARP enzyme expressing cancer include glioma, glioblastoma, thyroid cancer, lung cancer (e.g. NSCLC), esophageal cancer, head and neck cancer, tongue cancer, gastric cancer, liver cancer (e.g. HCC), neuroendocrine cancer, pancreatic cancer (e.g. PDAC), colon cancer, renal cancer, prostate cancer, breast cancer (e.g. TNBC, ductal and invasive subtypes), endometrial cancer, cervical cancer, ovarian cancer, melanoma / skin cancer and lymphoma.

[0142] dose The compounds of the present invention suitable for therapeutic use can be administered in any suitable dosage. For example, the compounds of the present invention can be administered at a dose of 0.1 MBq to 10 GBq per cycle over 1 to 10 separate treatment cycles. A treatment cycle typically includes a period of treatment (e.g., 1 to 7 days) followed by a calling off period (e.g., 1 to 21 days) during which no treatment is administered.

[0143] In a particular embodiment, the compound of formula Ij is administered at a dose of 0.1 MBq to 10 GBq per cycle for 1 to 10 treatment cycles.

[0144] Route of administration The compounds of the invention, or pharmaceutical compositions containing these compounds, can be administered to a subject by any convenient route of administration, whether systemic, peripheral, or local (i.e., at the desired site of action).

[0145] Routes of administration include, but are not limited to, oral (e.g., by ingestion); buccal; sublingual; transdermal (including, e.g., patches, plasters, etc.); transmucosally (including, e.g., patches, plasters, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eye drops); pulmonary (e.g., by inhalation or insufflation therapy, e.g., via aerosol, e.g., through the mouth or nose, using); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); by injection, including, e.g., intratumor, subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intraarticular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal; parenterally, e.g., subcutaneously or intramuscularly, by placement of a depot or reservoir.

[0146] Combination therapy The compounds of the present invention can be administered as a monotherapy or can include, in addition to the compounds of the present invention, conventional surgery, additional conventional radiation therapy or chemotherapy. Such chemotherapy can include one or more of the following categories: (i) As used in medical oncology, antiproliferative / antineoplastic agents and combinations thereof, including, but not limited to, alkylating agents (e.g., cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, temozolamide, and nitrosoureas); antimetabolites (e.g., gemcitabine and antifolates, such as fluoropyrimidines, such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea). antitumor antibiotics (e.g., anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin); mitotic inhibitors (e.g., vinca alkaloids such as vincristine, vinblastine, vindesine, and vinorelbine, and taxoids such as taxol and taxotere, and polo kinase inhibitors); and topoisomerase inhibitors (e.g., epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, and camptothecin); (ii) Cytostatic agents, such as, but not limited to, steroid hormones including antiestrogens (e.g., tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene, and iodoxyfene), antiandrogens (e.g., bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists or agonists (e.g., goserelin, leuprorelin, and buserelin), progestogens (e.g., megestrol acetate), and corticosteroids (e.g., dexamethasone, prednisone, and prednisolone), aromatase inhibitors (such as anastrozole, letrozole, vorazole, and exemestane), and inhibitors of 5α-reductase, such as finasteride; (iii) anti-invasion agents, such as, but not limited to, the c-Src kinase family inhibitor 4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530; International Patent Application Publication No. WO01 / 94341), N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl ) piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole-5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661), bosutinib (SKI-606), and metalloproteinase inhibitors, such as marimastat, inhibitors of urokinase-type plasminogen activator receptor function or antibodies against heparanase; (iv) inhibitors of growth factor function, including, but not limited to, growth factor antibodies and growth factor receptor antibodies (e.g., the anti-erbB2 antibody trastuzumab [Herceptin™], the anti-EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab [Erbitux, C225] and the antibodies described by Stern et al. (Critical reviews in oncology / haematology, 2005, Vol. 54, pp 11-29); such inhibitors also include tyrosine kinase inhibitors, such as inhibitors of the epidermal growth factor family (e.g., EGFR family tyrosine kinase inhibitors, such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, ZD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (CI1033), erbB2 tyrosine kinase inhibitors, such as lapatinib); inhibitors of the growth factor family; inhibitors of the insulin growth factor family; inhibitors of the platelet-derived growth factor family, such as imatinib and / or nilotinib (AMN107); inhibitors of serine / threonine kinases (e.g., Ras / Raf signaling inhibitors, such as farnesyltransferase inhibitors, e.g., sorafenib (BAY43-9006), tipifarnib (R115777) and lonafarnib (SCH66336)), inhibitors of cell signaling via MEK and / or AKT kinases, c-kit inhibitors, abl kinase inhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1R kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; Aurora kinase inhibitors and cyclin-dependent kinase inhibitors, such as CDK2 and / or CDK4 inhibitors; (v) antiangiogenic agents, including but not limited to those that inhibit the effects of vascular endothelial growth factor, [e.g., the anti-vascular endothelial growth factor antibody bevacizumab (Avastin™) and, for example, VEGF receptor tyrosine kinase inhibitors, such as vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU11248), axitinib (AG-013736) and pazopanib (GW786034); (vi) vascular damaging agents, including but not limited to combretastatin A4 and compounds disclosed in International Patent Application Publications WO99 / 02166, WO00 / 40529, WO00 / 41669, WO01 / 92224, WO02 / 04434 and WO02 / 08213; (vii) endothelin receptor antagonists, such as zibotentan (ZD4054) or atrasentan; (viii) Antisense therapies, including but not limited to those directed to the targets set forth above, e.g., ISIS2503, anti-ras antisense, etc.; (ix) Immunotherapeutic approaches, including, for example, cancer vaccines, antibodies, viruses (oncolytic viruses), and small molecule or cell therapy approaches to increase the immunogenicity of the patient's tumor cells and / or facilitate cell-mediated anti-tumor responses. Such therapies may include, but are not limited to, OX40 agonists, cGAS-STING agonists, A2a receptor antagonists, PI3 kinase inhibitors, TLR7 / 8 agonists, IDO inhibitors, arginase inhibitors, BTK inhibitors, and bromodomain inhibitors; transduction of cancer antigens by microbial vectors, direct transduction of cancer antigens into antigen-presenting cells, treatment with immune cells specific for cancer antigens (e.g., CAR-T), treatment with antibodies, antibody fragments, and antibody-drug conjugates that enable the immune system to recognize tumor cells.

[0147] Such conjoint treatment may be accomplished by the simultaneous, sequential or separate administration of the individual components of the treatment. Such combination products employ the compounds of this invention within the dosage ranges described hereinabove and the other pharmacologic-active agent(s) within their approved dosage ranges.

[0148] According to this aspect of the invention there is provided a combination for use in the treatment of cancer (e.g. cancer involving a solid tumour) comprising a compound of the invention as hereinbefore defined, or a pharma- ceutically acceptable salt or solvate thereof, and a further anti-tumour agent.

[0149] According to this aspect of the invention there is provided a combination comprising a compound of the invention as hereinbefore defined, or a pharma- ceutically acceptable salt or solvate thereof, and any one of the anti-tumour agents indicated in the specification above, for use in the treatment of a proliferative condition, such as cancer (e.g. a cancer involving a solid tumour).

[0150] In a further aspect of the present invention, there is provided a compound of the present invention or a pharma- ceutically acceptable salt or solvate thereof for use in the treatment of cancer, in combination with another anti-tumor agent, optionally selected from those set out in the specification above.

[0151] When the term "combination" is used herein, it is understood that it means simultaneous, separate or sequential administration. In one aspect of the present invention, "combination" means simultaneous administration. In another aspect of the present invention, "combination" means separate administration. In a further aspect of the present invention, "combination" means sequential administration. When administration is sequential or separate, the delay in administering the second component should not, for example, cause the beneficial effect of the combination to be lost.

[0152] According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of the present invention, or a pharma- ceutically acceptable salt or solvate thereof, in combination with an anti-tumor agent (optionally selected from those set out in the specification above), in association with a pharma- ceutically acceptable diluent or carrier. [Brief description of the drawings]

[0153] [Figure 1] A shows the chemical structures of PARP inhibitors Olaparib and CC1. B shows the enzyme inhibition of PARP1, 2 and 3 of CC1 and a selection of known PARP inhibitors. C shows a heat map visualizing the enzyme activity screening of CC1 (10 mM) against 13 related enzymes. D shows the radiosynthesis and structure of [123I]CC1. [Figure 1-2] E shows the uptake of [123I]CC1 into AsPC1, PSN1 and U87MG cells. F shows the blockade of [123I]CC1 uptake into AsPC1, PSN1 and U87MG cells by a panel of non-radioactive non-labeled PARP, ALDH2 or IMPDH2 inhibitors. Asterisks indicate significance levels: ns, P>0.05; *, P<0.05; **, P<0.01; ***, P<0.001; and ****, P<0.0001. [Figure 1-3] Supplementary Figure 1 shows catalytic inhibition of PARP1, 2, or 3 by increasing concentrations of CC1, olaparib, or rucaparib. [Diagram 2] A shows flow cytometric measurement of PARP1, 2 and 3, and γ-H2AX expression in AsPC1, PSN1 and U87MG at 0, 1 or 24 hours after 1 hour exposure to [123I]CC1 (50 kBq, 20 GBq / μmol).B shows flow cytometric measurement of PARP1, 2 and 3, and γ-H2AX expression in AsPC1, PSN1 and U87MG at 0, 1 or 24 hours after 1 hour exposure to [123I]CC1 (50 kBq, 20 GBq / μmol). [Figure 2-2]C shows a clonogenic assay comparing exposure of cells to [123I]CC1 or equivalent concentrations of CC1 or olaparib for 1 hour. [Figure 2-3] Supplementary Figure 2 is a flow cytometry frequency histogram examining PARP1, 2 and 3, and IMPDH2 expression in AsPC1, PSN1 and U87MG cells. [Diagram 3] A, Representative dynamic SPECT images after intravenous administration of [123I]CC1 (3MBq, 20GBq / μmol). The mean time window is shown in minutes. Images are shown as maximum intensity projections. B, VOI analysis of dynamic imaging. [Figure 3-2] C shows the biodistribution of [123I]CC1 in selected tissues in PSN1 xenograft-bearing mice 1, 2 or 24 hours after intravenous administration of [123I]CC1 (3MBq, 20GBq / μmol). [Figure 3-3] Supplementary Figure 3(A) shows the time-dependent uptake of [123I]CC1 into AsPC1, PSN1 and U87MG cells. Supplementary Figure 3(B) shows the cell retention assay of [123I]CC1 in AsPC1, PSN1 and U87MG cells. [Figure 4] (A) Biodistribution in selected tissues in PSN1 xenograft-bearing mice 2 hours after intravenous injection of [123I]CC1 (3MBq, Am=20GBq / μmol) (n=3 / group) with or without excess non-radioactive unlabeled rucaparib (0.5 mg). (B) Autoradiography of PSN1 tumor sections showing 123I localization. Asterisks indicate significance levels: ns, P>0.05; *, P<0.05; **, P<0.01; ***, P<0.001; and ****, P<0.0001. [Figure 4-2] Supplementary Figure 4 shows flow cytometry measuring PARP1, 2, and 3 expression in cells after exposure to [123I]CC1. [Diagram 5]A shows tumor growth after a single dose of CC1 or [123I]CC1 (3MBq, 20GBq / μmol). B shows Kaplan-Meier survival plots after administration of CC1 or [123I]CC1. [Figure 5-2] C shows the body weight of animals after administration of CC1 or [123I]CC1. [Figure 5-3] Supplementary Figure 5 shows VOI analysis of dynamic imaging after intravenous administration of [123I]CC1 to mice. [Figure 6] 6 shows the blockade of [123I]CC1 uptake into U87MG cells by CC1 in PSN1, AsPC1, or CAPAN1 cells. Asterisks indicate statistical significance levels: *, P<0.05. [Figure 6-2] Supplementary Figure 6 shows western blots showing PARP1 and PARP2 expression in AsPC1, PSN-1, and U87MG in vitro whole cell lysates. [Figure 7] Figure 7(E) Autoradiography of tumor sections harvested 2 hours after intravenous injection of [123I]CC1 (3MBq) into mice bearing PSN-1, U87MG, or MDA-MB-231 xenografts. Figure 7(F) Tumor uptake 2 hours after intravenous injection of [123I]CC1 (3MBq) into mice bearing PSN-1, U87MG, or MDA-MB-231 xenografts. Asterisks indicate significance levels: ns, P>0.05; *, P<0.05. [Figure 7-2] Supplementary Figure 7 shows (A) time-dependent uptake of [123I]CC1 into AsPC1, PSN1 and U87MG cells and (B) cell-associated retention of [123I]CC1 after 1 hour exposure of AsPC1, PSN1 and U87MG cells. [Figure 8](A,B) Tumor growth and Kaplan-Meier survival plots after a single intravenous administration of [123I]CC1 (3 MBq, 124.2-341.9 GBq / μmol) or an equivalent amount of unlabeled CC1 to mice bearing PSN-1 xenografts. (C) Representative photographs of mice bearing PSN-1 xenografts 15 days after [123I]CC1 or CC1 administration. (D) Animal weights after administration of [123I]CC1 or CC1. (E,F) Tumor growth curves after a single intravenous administration of [123I]CC1 (3 MBq) or an equivalent amount of unlabeled CC1 to mice bearing U87MG or MDA-MB-231 xenografts. (G) Representative H&E staining in selected normal tissues or untreated C57 / Bl6 mice 28 days after administration of [123I]CC1 (3 MBq). [Figure 8-2] Supplementary Figure 8 shows flow cytometry measurements demonstrating increased numbers of cells with higher PARP2 expression 1 or 24 h after 1 h exposure to [I]CC1 (50 kBq, 18 GBq / μmol) in PSN1, AsPC1, and U87MG cells. [Figure 9] Supplementary Figure 9 shows VOI analysis of blood clearance following intravenous bolus administration of [123I]CC1 (3MBq) to PSN1 xenograft-bearing mice. EXAMPLES

[0154] Example – Part A material and method Synthesis and characterization of CC1 Unless otherwise stated, all reagents were purchased from Sigma-Aldrich and used without further purification. Olaparib, rucaparib, veliparib, talazoparib, and niraparib were purchased from Stratech Scientific Ltd (UK). Daidzin and mycophenolate mofetil were purchased from Cambridge Bioscience Ltd (UK) and Insight Biotechnology Ltd, respectively. CC1 (4-[(3-[(4-cyclopropylcarbonyl)piperazin-1-yl]carbonyl-4-iodophenyl]methyl(2H)phthalazin-1-one) (1) was synthesized using a modified literature procedure. See Supplementary Information for a detailed description of the protocol.

[0155] PARP-1, 2, and 3 catalytic activity was measured in vitro in the presence of various concentrations of established PARP inhibitors or CC1 using a commercially available colorimetric assay (Trevigen, Gaithersburg, MD) according to the manufacturer's instructions. The ability of 10 nM CC1 to inhibit the catalytic activity of various PARP family enzymes was performed by AMS Biotechnology Europe Ltd. according to the BPS assay kit protocol.

[0156] [ 123 Radiosynthesis of [I]CC1 [ in 0.1N NaOH 123 I] sodium iodide (GE, UK). The appropriate boronic pinacol ester precursor was prepared by a procedure modified from (1, 2) 123 [I]CC1 was synthesized (Figure 1). Radiochemical yields of >95% and molar activities of >20 GBq / μmol were routinely obtained. Activity yields were >75% (ndc) for a 2 h synthesis. See Supplementary Information for a detailed description of the synthesis methodology.

[0157] cell culture Human malignant glioma (U87MG) cells were kindly provided by Professor Nicola Sibson in our laboratory and were maintained in high glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mM L-glutamine, 100 units / mL penicillin, and 0.1 mg / mL streptomycin (Gibco). Pancreatic adenocarcinoma (AsPC1 and PSN1) cells were purchased from ATCC (UK) and maintained in Roswell Park Memorial Institute (RPMI) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mM L-glutamine, 100 units / mL penicillin, and 0.1 mg / mL streptomycin (Gibco). Cells were grown in a humidified environment at 37°C and 5% CO2. Cells were harvested and passaged using trypsin-EDTA solution. Cells were used no more than 25 passages after resuscitation from liquid nitrogen storage. All cells were authenticated by STR profiling and routinely tested for the absence of mycoplasma.

[0158] PARP protein expression in vitro The relative expression levels of PARP isoforms in AsPC1, PSN1, and U87MG cells were determined by live cell flow cytometry. AsPC1, PSN1, and U87MG cells were cultured in 96-well plates (1 × 10 5The cells were separately seeded in 1000 x g (1000 x 1000 cells / well). The cells were washed with FACS buffer (PBS, 2% FBS, 1 mM EDTA, 0.1% NaN3) by centrifugation at 500 x g for 5 min. Immunostaining was performed with Foxp3 / transcription factor staining buffer set (eBioscience™, USA). Intracellular staining was performed in the dark in permeabilization buffer for 30 min at 4° C. with the following antibodies separately: AF488-conjugated anti-PARP-1 (1:100; sc-80070), AF594-conjugated anti-PARP-2 (1:100; sc-393310) and AF488-conjugated anti-PARP-3 (1:100; sc-390771) from Santa Cruz Biotechnology Inc. (USA), and AF488-conjugated anti-IMPDH2 (1:500; ab-200770) from Abcam plc. (UK). The fixable viability dye ef780 (1:4000; eBioscience™; 65-0865-14) was used to distinguish between live and dead cells. Fixation of immunostained cells was performed with 10% formaldehyde for 15 min at RT. Flow cytometry was performed using a CytoFLEX flow cytometer (Beckman Coulter, USA) with appropriate lasers and filters, positive and negative controls. Data was analyzed using FlowJo™ (Tree Star Inc., BD Biosciences, USA).

[0159] [ 123 In vitro uptake and specificity of I]CC1 AsPC1(1×10 5 cells / well), PSN1 cells (7.5 × 10 4 cells / well) or U87MG (1 × 10 5 Cells / well) were seeded separately into 24-well plates containing growth medium and allowed to adhere for at least 20 hours. Cells were washed and exposed to non-radioactive, unlabeled PARP inhibitor (100 μM, in a total of 270 μL growth medium) for 45 minutes at 37°C. Then, 123I]CC1 was added and the cells were incubated at 37°C for 45 min. The cell culture medium was removed and the cells were washed with PBS. The cells were lysed with RIPA buffer (950 mM Tris pH 8.0, 1% NP40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, and 150 mM sodium chloride) at room temperature for 15 min, and the lysate 123 The amount of I was measured using an automated gamma counter (PerkinElmer). In a separate experiment, cells were prepared in a similar manner, except that the cells were washed and 123 The cells were exposed to [I]CC1 at different intervals (1–120 min) at 37°C. 123 The amount of I was measured as described above.

[0160] Cell retention assay Cells were seeded separately in 24-well plates as described above and allowed to adhere for at least 20 hours. 123 The cells were exposed to [I]CC1 for 30 min at 37 °C. The cell culture medium was removed, washed with PBS, and fed with fresh medium (300 μL). The cells were then further incubated at 37 °C. The cell culture medium was removed, and the cells were washed with PBS at different time points (0–24 h). The cells were lysed using RIPA buffer (950 mM Tris pH 8.0, 1% NP40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, and 150 mM sodium chloride) at room temperature for 15 min, and the chromatin in the cell lysate was analyzed by HPLC. 123 The amount of I was measured using an automatic gamma counter (PerkinElmer).

[0161] Colony formation assay Aliquots of cells (PSN1 and U87MG, 5000 cells) were incubated with increasing amounts of 123 Cells were exposed to [I]CC1 (0–100 kBq), or non-radioactive unlabeled olaparib or CC1 (0–0.1 mM, in a total of 200 µL growth medium) for 60 min at 37 °C. Separate aliquots of cells were irradiated (0–10 Gy, 137Cs irradiator, 0.8 Gy / min). The cells were then washed and a portion was then transferred to 6-well plates containing 2 mL of growth medium, and colony formation was measured 2 weeks after treatment. The number of colonies was normalized to the number of cells transferred to the plate to obtain the plating efficiency, which was then normalized to the plating efficiency of untreated cells.

[0162] [ 123 [I] Quantification of nuclear protein expression after CC1 treatment AsPC1, PSN1 or U87MG (1 × 10 in 2 mL growth medium) 6 Cells (1000 cells / well) were seeded into 6-well plates and allowed to adhere for at least 20 hours. The cells were washed and 123I]CC1 (30 μL, 50 kBq, 20 GBq / μmol, in 2 mL of growth medium) for 1 h at 37°C. Cells were harvested using trypsin-EDTA solution, washed with FACS buffer (PBS, 2% FBS, 1 mM EDTA, 0.1% NaN3) and centrifuged at 500×g for 5 min. Immunostaining was performed using Foxp3 / transcription factor staining buffer set (eBioscience™, USA). Intracellular staining was performed in the dark for 30 min at 4° C. in permeabilization buffer with the following antibodies separately: AF488-conjugated anti-PARP-1 (1:100; sc-80070), AF594-conjugated anti-PARP-2 (1:100; sc-393310) and AF488-conjugated anti-PARP-3 (1:100; sc-390771) from Santa Cruz Biotechnology Inc. (USA), AF488-conjugated anti-IMPDH2 (1:500; ab-200770), γH2AX (1:1000; JBW301 Merck-Millipore) from Abcam plc. (UK). The fixable viability dye ef780 (1:4000; eBioscience™; 65-0865-14) was used for live / dead discrimination. Fixation of immunostained cells was performed at RT for 15 min. Flow cytometry was performed using a CytoFLEX flow cytometer (Beckman Coulter, USA) with appropriate lasers and filters, positive and negative controls. Data was analyzed using FlowJo™ (Tree Star Inc., BD Biosciences, USA).

[0163] In xenograft-bearing immunodeficient mice, 123 SPECT / CT imaging and biodistribution of I]CC1 Four to six week-old female Balb / c nu / nu (OlaHsd-Foxn1 nu ) Mice were purchased from Envigo (UK). Animals were housed up to six per cage in IVC cages in an artificial day-night cycle facility. Food and water were provided ad libitum.

[0164] PSN1 cells were harvested using trypsin, washed twice with PBS, and reconstituted in PBS:Matrigel® Matrix High Concentration (1:1). Cell suspension (PSN1: 2×10 6 The mice were subcutaneously injected with 100 μL of ... 3 was allowed to grow to a size of

[0165] Animals were given the following injections intravenously via the lateral tail vein: 123 I]CC1 (3 MBq in 100 μL PBS, A m = 20 GBq / μmol). To assess the specificity of tumor uptake, an excess of unlabeled rucaparib (0.5 mg) was co-administered as a blocker. 60 or 120 min after radiolabeled compound injection, animals were euthanized. Selected organs, tissues and blood were harvested and the percentage of injected dose per gram of tissue (%ID / g) was determined using a HIDEX automated gamma counter.

[0166] In PSN1 xenografts, 123 [I]CC1 localization was determined ex vivo using autoradiography performed on frozen tumor sections (10 μm). Uptake into PSN1 tumors was further compared with immunohistochemistry staining for PARP1, 2, and 3, as well as γ-H2AX. Full details of the procedure and protocol are provided in the Supplementary Information.

[0167] Separately, to the animals 123 I]CC1 was administered and dynamic PET images were acquired over 45 min. Images were analyzed using VOI-based analysis with PMOD.

[0168] In vivo therapy studies The average tumor size was 82 mm 3 PSN1 xenograft-bearing mice were randomly grouped into cohorts and [ 123 I]CC1 (3 MBq in 100 μL PBS, A m= 20 GBq / μmol) or an equivalent amount of non-radioactive, unlabeled CC1 was injected intravenously. Mice and tumor size were monitored daily. The study endpoint was a tumor size >1200 mm 3 or based on humane endpoints including weight loss >15%.

[0169] statistical analysis All data were obtained in at least triplicate. All statistical analyses and nonlinear regressions were performed using GraphPad Prism v6 or higher (GraphPad Software, San Diego, CA, USA). Data were tested for normality and analyzed by one-way or two-way analysis (ANOVA) as appropriate. Results are reported as the mean ± 1 standard deviation unless otherwise stated.

[0170] result CC1 is a selective and potent PARP inhibitor CC1 proved to be a potent PARP inhibitor with in-house determined cell-free IC50 values ​​of 2.9 and 0.6 nM for PARP 1 and 2, respectively (Figure 1B, Supplementary Figure 1). This compared favorably with olaparib (2.6 and 0.8 nM in the same assay). Conversely, PARP3 inhibition of CC1 (68 nM) was lower compared with olaparib (18 nM). A single dose of CC1 (10 nM) was able to inhibit PARP1, PARP2, and PARP6 (Figure 1B), but to a lesser extent compared with olaparib, rucaparib, and other established PARP inhibitors. When interpreted as a proxy for PARP binding, CC1 binds to PARP1, PARP2, and PARP3.

[0171] In vitro 123 I]CC1 uptake AsPC1, PSN1, and U87MG cells expressed PARP1, 2, and 3 to various degrees, with PARP1 expression being AsPC1>U87MG>PSN1. 123 I]CC1 was taken up by all three cell lines within minutes and plateaued after 1 hour, whereas [123 I]CC1 was retained in cells for a short time (Supplementary Figure 3). 123 [I]CC1 was selectively taken up into AsPC1, PSN1 and U87MG cells, with uptake of approximately 0.3 pmol / cell (Fig. 1D). Addition of non-radioactive unlabeled PARP inhibitors significantly reduced uptake, demonstrating PARP selectivity (Fig. 1E). 123 I]CC1 uptake was also reduced by the addition of daidzin and m.mofetil. These two drugs were more potent than other NAD + -binding enzymes, ALDH2 and IMPDH2, respectively. 123 Exposure of all three cell types to I]CC1 resulted in increased expression of PARP1 and PARP2 after 24 hours (Figure 4). Furthermore, yH2AX expression, a marker of DNA double-strand breaks, was increased by [ 123 I] was significantly increased 24 h after exposure to CC1.

[0172] [ 123 [I]CC1 reduces viability in colony formation assays The clonogenic survival rate of cells was significantly improved with relatively low doses of 10 Bq for U87MG cells. 123 I] was significantly decreased by exposure to CC1. 123 [I] The IC50 value for CC1 is equal to 1 kBq / mL on PSN1 cells. Equivalent doses of CC1 or olaparib were ineffective.

[0173] [ 123 I]CC1 biodistribution [ 123 The biodistribution of [I]CC1 (3 MBq, 20 GBq / μmol) was investigated in PSN1 xenograft-bearing mice (Figure 5). 18[F]High uptake into the liver, gallbladder, and large / small intestine showing a hepatobiliary clearance pattern similar to olaparib. Blood clearance showed a two-phase decay with fast and slow half-lives of 16.85 and 1.35 min, resulting in a weighted half-life of 1.86 min (CI95 1.64-2.09 min). Tumor uptake into PSN-1 xenografts was 0.9 ± 0.06% ID / g 1 hour after intravenous administration.

[0174] [ 123 I]Therapeutic efficacy of CC1 Treatment of PARP-expressing tumors 123 The application of I]CC1 was tested using PSN1-xenograft-bearing mice. 123 A single intravenous administration of [I]CC1 significantly inhibited tumor growth compared with animals exposed to non-radioactive, unlabeled CC1, without any signs of significant toxicity, as judged by the lack of mouse weight loss (p=0.04).

[0175] Consideration PARP inhibitors trap the PARP enzyme to the broken DNA. This proximity to DNA makes them good vehicles for carrying Auger electron-emitting radionuclides. These short-range electron-emitting radionuclides are highly radiotoxic to cells when they decay in close proximity to DNA. Here, we present a novel method for the detection of Auger electron emitters that bind to PARP. 123 We have synthesized compounds containing I. This type of compound can be used for therapeutic benefit. Importantly, the very low concentrations that already allow radionuclide therapy will not lead to a total catalytic inhibition of PARP enzyme activity. On the contrary, it is Auger electron emission that causes DNA damage and leads to growth inhibition. We have [ 123 I]CC1 specifically binds to the PARP enzyme and can detect very small amounts of radiation, even at only 10 Bq[ 123 We found that exposure to [I]CC1 for only 1 h significantly reduced the clonogenic survival of PARP-expressing cancer cells. 123Apart from being an Auger electron emitter, I also emits gamma rays, which allows detection by gamma-detectors and SPECT imaging. We have determined that at a certain dose of [ 123 We found that I]CC1 was already effective in inhibiting tumor growth in human tumor xenograft mouse models. However, other radiolabeled compounds, e.g. 18 F]olaparib can be used as a companion diagnostic. 18 [F]Olaparib can be used for PET imaging of PARP and can serve as a companion diagnostic.

[0176] No significant toxicity was observed in vivo: uptake of other radiolabeled PARP inhibitors into the liver, gallbladder, and intestine, as well as into the kidney, appeared to be intraluminal. Due to the short path length of the Auger electrons, little DNA damage was produced by the radiolabeled compounds in those tissues.

[0177] Example – Part B material and method Synthesis and characterization of CC1 and 123 Radiosynthesis of I]CC1 was carried out as described above in part A.

[0178] cell culture Human malignant glioma (U87MG) cells were kindly provided by Professor Nicola Sibson in our laboratory and were maintained in high glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mM L-glutamine, 100 units / mL penicillin, and 0.1 mg / mL streptomycin (Gibco). Pancreatic adenocarcinoma (AsPC1, PSN1, and CAPAN-1) and breast cancer (MDA-MB-231) cells were purchased from ATCC (UK) and maintained in Roswell Park Memorial Institute (RPMI) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mM L-glutamine, 100 units / mL penicillin, and 0.1 mg / mL streptomycin (Gibco). Cells were grown in a humidified environment at 37°C and 5% CO2. Cells were harvested and passaged using trypsin-EDTA solution. Cells were used no more than 20 passages after resuscitation from liquid nitrogen storage. All cells were donor and authenticated by STR profiling and routinely tested for the absence of mycoplasma.

[0179] PARP protein expression in vitro Relative expression levels of PARP1, 2, and 3 were determined by flow cytometry of live cells as described above in part A. In a separate study, DNA double-strand break damage was quantified using anti-γH2AX antibody (JBW301, 1:800) in combination with AF488-conjugated anti-mouse antibody (Santa Cruz Biotechnology Inc.). Fixable viability dye ef780 (1:4000; eBioscience™; 65-0865-14) was used to distinguish live and dead cells. Fixation of immunostained cells was performed with Foxp3 Transcription Factor Staining Buffer Set (eBioscience™, 00-5523-00) for 15 min at RT. Flow cytometry was performed using a CytoFLEX flow cytometer (Beckman Coulter, USA) with appropriate lasers and filters, positive and negative controls. Data were analyzed using FlowJo™ (Tree Star Inc., BD Biosciences, USA).

[0180] [ 123 In vitro uptake and specificity of I]CC1 AsPC1(1×10 5 cells / well), PSN1 (7.5 × 10 4 cells / well) or U87MG (1 × 10 5 CAPAN-1 cells (7.5 × 10 cells / well) were seeded as described above in part A. 4 Cells / well) were seeded according to the same protocol described above in Part A. In separate experiments, cells were cultured in 100 mL of 100 mL of 1000 mM NaCl, 100 mM NaCl, 100 mM MgCl, 100 mM Na ... 123 I]CC1 for 30 min at 37 °C, washed, fed with fresh growth medium, and associated cells after varying intervals. 123 The amount of I was measured.

[0181] Colony formation assay Colony formation assays were performed on aliquots of PSN1, U87MG and MDA-MB-231 cells following the protocol outlined in part A above.

[0182] [ 123 [I] Quantification of nuclear protein expression after CC1 treatment Quantification of AsPC1, PSN1 or U87MG cells was performed according to the protocol outlined above in part A. Relative expression of PARP1 and 2 was measured using flow cytometry. γH2AX expression was assessed in a similar manner as a measure of DNA double-strand break damage.

[0183] In xenograft-bearing immunodeficient mice, 123 SPECT / CT imaging and biodistribution of I]CC1 Following the protocol described in Part A above, PSN1, U87MG, or MDA-MB-231 cells were harvested with trypsin, washed twice with PBS, and reconstituted in PBS:Matrigel® Matrix High Concentration (1:1). The cell suspension was injected subcutaneously into the right posterior flank to allow tumor formation. Tumors were 100-200 mm 3 Once the animals reached the target concentration, they were administered 100 mg of 10 ... 123 I]CC1 (3 MBq in 100 μL PBS, A m =124.3 GBq / μmol). To assess the specificity of tumor uptake, an excess of unlabeled rucaparib (0.5 mg) was co-administered as a blocker to some animals. At 60 or 120 min after injection of the radiolabeled compound, animals were euthanized (n=3 per group). Selected organs, tissues and blood were harvested and the percentage of injected activity per gram of tissue (%IA / g) was determined using a HIDEX automated gamma counter.

[0184] In a selected number of animals (n=3), animals were anesthetized with 2% isoflurane and dynamic SPECT / CT imaging was performed over 1 h using a MILabs VECTor4 camera with an ultra-high resolution rat / mouse collimator (1.8 mm), followed by a cone-beam CT scan (55 kV, 0.19 mA) for anatomical reference and attenuation correction. Anesthesia was maintained with 2.5% isoflurane throughout the entire period of image acquisition. SPECT images were reconstructed using U-SPECT-Rec3.22 software (MILabs, Utrecht, The Netherlands) and a pixel-based algorithm, subsetting expectation maximization (OSEM), was applied with 4 subsets, 4 iterations and a voxel size of 0.6 mm for iodine-123 (energy window settings 141.3–172.7 keV). Reconstructed SPECT and CT images were viewed and analyzed using PMOD v.3.37 (PMOD Technologies, Zurich, Switzerland).

[0185] In PSN1 xenografts, 123[I]CC1 localization was further determined ex vivo using autoradiography performed on frozen tumor sections (10 μm). Uptake into tumor tissue was further compared with immunohistochemistry staining for PARP1 and 2.

[0186] Toxicology testing H&E staining was performed on naïve C57 / Bl6 mice with 3MBq[ 123 Selected tissues (n=3 per time point) were examined 24 h and 28 days after intravenous administration of [I]CC1. Liver, spleen, kidneys, and intestines were examined by an experienced veterinary pathologist and compared with age-matched untreated control animals.

[0187] Tissues were further stained for γH2AX to investigate DNA double-strand break damage, and liver was additionally stained with PAS as a marker of glycogen stores.

[0188] In vivo therapy studies The average tumor size was 82 mm 3 PSN1 xenograft-bearing mice were randomly grouped into cohorts and [ 123 I]CC1 (3 MBq in 100 μL PBS, A m Mice were intravenously injected with CC1 (124–342 GBq / μmol) or an equivalent amount of unlabeled CC1. Mice were monitored daily. Tumor size was determined by caliper. The study endpoint was a tumor size >1000 mm 3 , or based on humane endpoints including weight loss >15%.

[0189] Dosimetry In vitro, radiation absorbed dose was determined using uptake and retention data in U87MG cells using the MIRD Cell package. Cell and cell nucleus dimensions were approximated as concentric circles with sizes based on confocal microscopy (cell diameter 14 μm, nuclear diameter 8 μm).

[0190] statistical analysis All data were obtained in at least triplicate. All statistical analyses and nonlinear regressions were performed using GraphPad Prism v8 or higher (GraphPad Software, San Diego, CA, USA). Data were tested for normality and analyzed by one-way or two-way analysis (ANOVA) as appropriate. Results are reported as the mean ± 1 standard deviation unless otherwise stated.

[0191] result In vitro cell 123 I]CC1 uptake is PARP-selective [ 123 I]CC1 was synthesized with high A in good radiochemical yield. m AsPC1, PSN1, and U87MG cells expressed PARP1 and 2 to various degrees, with PARP1 expression being AsPC1>U87MG>PSN1 (Supplementary Figure 6). 123 I]CC1 was taken up by all three cell lines within minutes and plateaued after 1 h (Supplementary Figure 7A). 123 I]CC1 was retained in cells for a short time (Supplementary Figure 7B). 123 I]CC1 was selectively taken up into AsPC1, PSN1, and U87MG cells, with uptake of approximately 0.3 fmol / cell (Figures 1E, 1F, and 6). Addition of structurally related and unrelated unlabeled PARP inhibitors reduced [ 123 I] significantly reduced the cell-associated amount of CC1 (P<0.0001), 123 I] CC1, suggesting PARP-selective binding (Fig. 6 ).

[0192] [ 123 I]CC1 causes DNA damage and reduces clonogenic survival γH2AX expression, a marker of DNA double-strand breaks, was detected by low levels of 123 I] was significantly increased 24 hours after 1 hour exposure of PSN-1 and U87MG cells to CC1 (50 kBq) (Figure 2B). 123Exposure of all three cell types to [I]CC1 resulted in increased expression of PARP1 and PARP2 24 h after a 1-h exposure (Figure 2A and Supplementary Figure 8). The radiation absorbed dose (over 24 h) was estimated to be 7.89 Gy by in vitro cell dosimetry calculated by MIRDcell based on uptake and retention values ​​(Supplementary Figure 7).

[0193] The clonogenic survival of cells was significantly improved by relatively small doses of 10 Bq (20 MBq / μmol) for U87MG cells. 123 I] was significantly decreased by exposure to CC1. 123 The IC50 value for [I]CC1 was equivalent to 631 ± 35 Bq (in 200 μl) on U87MG cells. Efficacy on PSN1 cells in vitro was much less pronounced. Equivalent doses of CC1 or olaparib had no effect on clonogenic survival (Figure 1E and Figure 1F).

[0194] [ 123 Biodistribution of I]CC1 shows a hepatobiliary clearance pattern [ 123 Dynamic SPECT / CT imaging and biodistribution of [I]CC1 (3 MBq, 20 GBq / μmol) were investigated in PSN1 xenograft-bearing mice (Figure 3, Figure 4, Figure 7 and Supplementary Figure 5 and Supplementary Figure 9). High uptake in the liver and intestine indicated a similar hepatobiliary clearance pattern to other radiolabeled PARP inhibitors (Figure 3A and Figure 3B). Blood clearance showed a two-stage decay with fast and slow half-lives of 16.85 and 1.35 min, resulting in a weighted half-life of 1.86 min (CI95 1.64–2.09 min) (Supplementary Figure 9B). Cut-and-count biodistribution studies showed tumor uptake in PSN-1 xenografts that amounted to 0.9 ± 0.06% IA / g 1 h after intravenous administration, and normal tissue uptake consistent with dynamic SPECT imaging (Figure 3C). After 24 hours, the [ 123I]CC1 remained in the tumors 24 hours after injection. Coadministration of excess rucaparib to block the NAD+ binding pocket of the PARP enzyme significantly reduced tumor uptake (P<0.05) (Fig. 4A), as confirmed by autoradiography (Fig. 7E). Tumor uptake in U87MG and MDA-MB-231 xenografts was significantly lower (0.46±0.01% IA / g for U87MG xenografts and 0.19±0.01% IA / g for MDA-MB-231 xenografts) compared to PSN-1 xenografts (Fig. 7E and Fig. 7F).

[0195] [ 123 I]CC1 exhibits limited normal tissue toxicity For radionuclide therapy of tumors 123 With the intention of using I]CC1, an evaluation was carried out as to whether the radiolabeled compound induces any toxicity in normal tissues. Based on its biodistribution pattern, 123 Radiation-induced damage from exposure to [I]CC1 can be predicted in the liver and intestine. Due to partial renal clearance, the kidneys were also evaluated, and radiolabeled PARP inhibitor [ 18 F] Given the ability of olaparib to specifically bind to splenic tissue, we also considered the spleen ( Figure 8 ).

[0196] [ 123 Intestines of mice administered [I]CC1 (3MBq) intravenously showed minimal proprial infiltration by lymphocytes and plasma cells. Scattered intact eosinophils were present within the lamina propria, but enterocytes from none of the mice showed signs of formation or necrosis, and the apical brush border remained intact. Mitotic figures were regularly present and within normal numbers. Observations were not different 24 hours or 28 days after administration. Kidneys showed no observable changes, but mild to moderate numbers of hemosiderophages were observed in the spleen at 24 hours and 28 days after administration, similar to those observed in the spleen. No signs of necrosis were seen.

[0197] In the liver, hepatocyte nuclei were centrally located and showed no signs of necrosis. Several small foci of extramedullary hematopoiesis were present. Mild anisocytosis and anisocytosis were seen. A few individual scattered hepatocytes (0.1-0.2 per field) showed atrophic shape, hypereosinophilic cytoplasm, and atrophic nuclei with condensed chromatin that was interpreted as pyknosis. The one notable effect in the liver consisted of diffuse cytoplasmic pallor / rarefaction created by optically empty feathery spaces and vacuoles, and some residual flocculent granular cytoplasmic material, often at the periphery.[ 123 In animals 28 days after administration of I]CC1, the effect was slightly more pronounced, although not statistically significantly so.

[0198] In PDAC-xenograft tumor-bearing mice, 123 Potent tumor growth inhibition by I]CC1 A relatively small amount of [ 123 Intravenous administration of I]CC1 (3MBq) showed significant tumor growth retardation in PSN1-xenograft-bearing mice (Figure 8). 123 A single intravenous dose of [I]CC1 significantly inhibited tumor growth compared to animals exposed to non-radioactive unlabeled CC1 (p=0.04) without any signs of significant toxicity as judged by the lack of mouse weight loss (p>0.05). U87MG xenografts responded less than PSN-1 xenografts (p>0.05), whereas MDA-MB-231 xenografts showed a similar amount of [I]CC1 in their growth. 123 I] was not affected by CC1 (p>0.05).

[0199] Consideration [ 123 I]CC1 was shown to selectively bind to PARP, induce DNA double-strand break damage, and reduce clonogenic survival in vitro and tumor growth in vivo. 123 I-CC1 also induced increased expression of PARP1 and PARP2 in tumor tissues, which may form the basis of a possible feedback mechanism for multiple administration. 123This was not investigated, as a single dose of I-CC1 (3 MBq) was therapeutically effective in PSN-1 xenograft-bearing mice.

[0200] Although the in vitro efficacy in PSN1 cells was less pronounced than the in vitro response in U87MG cells, in vivo uptake in PSN1 tumors was higher than that in U87MG tumor xenografts, resulting in better therapeutic efficacy. This resulted in a higher absorbed radiation dose. 123 This could be due to higher uptake of I, differences in radiosensitivity in vitro versus in vivo, and / or differences in proliferation rates and regrowth in vitro versus in vivo.

[0201] 123 I-MAPi, 123 / 125 I-KX1, 125 I-KX-02-019, 211 At-MM4, 125 I-PARP-01 and 77 Compared with Br-GD1, [ 123 I]CC1 was shown to be effective after a single intravenous administration at a relatively low dose. Previous studies have already shown that small changes in the structure of PARP inhibitors produce large changes in their characteristics. Factors that affect the efficacy of radioligand therapy with radiolabeled PARP inhibitors may include binding affinity, binding spectrum, bioavailability, pharmacokinetics, and tumor uptake. In addition, trapping of the PARP enzyme by PARP inhibitors, and thus by radiolabeled PARP inhibitors, may play an important role in their efficacy. It is therefore expected that different radiolabeled PARP inhibitors may have very different therapeutic indices.

[0202] Many approaches to radiolabeling PARP inhibitors incorporate Auger electron emitters, given that trapping of the PARP enzyme brings the PARP inhibitors into close proximity with DNA, providing an excellent match with short-range Auger electron emitters. 125I is a very efficient Auger electron emitter, with approximately 23 low-energy electrons emitted per decay, but considering the half-life 123 It is preferable to use I. 123 I has the advantages of 14 AEs per disintegration, a more manageable half-life, and worldwide availability.

[0203] The effect on normal tissue toxicity was minimal. The absence of normal tissue toxicity was 123 This can be explained by the very short range of the AE of I: despite the high uptake in the gallbladder, intestine and urinary bladder, the radiation dose due to Auger electron emission is minimal due to the luminal uptake of the PARP inhibitor in these organs. In the liver, the uptake of another labeled PARP inhibitor was shown to be in the cytoplasm and not in the nucleus. Given that Auger electron emitters that decay in the cytoplasm are 30 times less cytotoxic compared to those that decay in the nucleus, the [ 123 Cytoplasmic uptake of [I]CC1 would be much less cytotoxic.

[0204] To tumor tissue 18 Uptake of F-labeled olaparib has previously been shown to be highly dependent on administered mass and molar activity. 123 The same may be true for I-CC1 and other PARP inhibitor-based radioligand therapies.

[0205] 123 The gamma emission from I also makes it an imaging agent. Thus, 123 I]CC1 can be considered a true therapeutic diagnostic agent, with lower doses used for SPECT imaging and higher doses for therapy. 18 F-(6), or even 124 Using the I-labeled variants, the relative expression of the target enzyme in tumor tissue can be measured by PET imaging.

[0206] In summary, 123I]CC1 is a promising Auger-electron emitting PARP-inhibitor based therapeutic radiopharmaceutical.

Claims

1. Formula I: 【Chemistry 1】 [In the formula, A １ and A ２ are CR １ and C.H., C.R. １ and C.F., C.R. １ and N, C.H. and C.R. １ , C.F. and C.R. １ or NH and CR １ Selected from: R １ is a radioactive isotope of iodine (I) or astatine (At); A ３ is N-X １ -R ３ or C(R ４ ) OR ５ Selected from: X １ is not present, or is —C(O)—, —[CH ２ ] １～３ -, -C(O)NH-, -C(O)O-, or -S(O) ０～２ - selected from; R ３ is H, (1-10C)alkyl, (3-8C)cycloalkyl, aryl, heterocyclyl or heteroaryl, each alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl group being selected from one or more R Ａ optionally substituted by substituents; R ４ is H or C １～４ alkyl; R ５ is selected from (1-7C)alkyl, (3-8C)cycloalkyl, heterocyclyl and aryl, which groups are each independently selected from one or more R Ａ optionally substituted by substituents; Or R ４ and R ５ together with the carbon and oxygen atoms attached thereto, represent one or more R Ａ spiro-C optionally substituted with substituents or fused to a phenyl or 5- or 6-membered heteroaryl ring ５～７ forming an oxygen-containing heterocyclyl; Each R Ａ Substituents include fluoro, cyano, (1-4C)alkyl, (1-4C)haloalkyl, (1-4C)hydroxyalkyl, phenyl, 4- to 6-membered heterocyclyl, 5- to 6-membered heteroaryl, -OR １ａ , -NR １ａ R １ｂ , -C(O)R １ａ , -C(O)N(R １ｂ ) R １ａ , N(R １ｂ ) C(O)R １ａ , -S(O) ｐ R １ａ (wherein p is 0, 1 or 2), -SO ２ N (R １ｂ ) R １ａ , -N(R １ｂ ) S.O. ２ R １ａ (In the formula, R １ａ and R １ｂ are each independently selected from hydrogen, (1-4C)alkyl or (3-6C)alkyl), or a pharma- ceutically acceptable salt or solvate thereof.

2. (i) A １ But, CR １ And A ２ is selected from CH, CF, or N; or (ii) A １ is selected from CH, CF, and N; A ２ But, CR １ 2. The compound of claim 1, wherein:

3. A １ But, CR １ And A ２ 3. The compound of claim 2, or a pharma- ceutically acceptable salt or solvate thereof, wherein:

4. R １ but, １２３ I, １３１ I, １２７ I or １２４ a radioisotope of iodine (I) selected from I; or ２１１ 4. The compound according to claim 1, wherein At is a radioactive isotope of astatine (At), or a pharma- ceutically acceptable salt or solvate thereof.

5. R １ but, １２３ I, １２４ I or １３１ 3. The compound of claim 1 or claim 2, wherein the compound is a radioactive isotope of iodine (I) selected from I, or a pharma- ceutically acceptable salt or solvate thereof.

6. R １ but, １２３ 4. The compound of claim 1 or claim 3, wherein R is H, or a pharma- ceutically acceptable salt or solvate thereof.

7. A ３ But N-R ３ 7. The compound of any one of claims 1 to 6, wherein:

8. R ３ is (1-6C)alkyl, (3-6C)cycloalkyl, phenyl, 4- to 6-membered heterocyclyl, or 5- to 6-membered heteroaryl, each of which is selected from one or more R Ａ 8. A compound according to any one of claims 1 to 7, or a pharma- ceutically acceptable salt or solvate thereof, optionally substituted by a substituent.

9. R ３ is (1-6C)alkyl or (3-6C)cycloalkyl, each of which is one or more R Ａ 9. A compound according to any one of claims 1 to 8, or a pharma- ceutically acceptable salt or solvate thereof, optionally substituted by a substituent.

10. R ３ 10. The compound according to any one of claims 1 to 9, wherein is (1-6C)alkyl or (3-6C)cycloalkyl, or a pharma- ceutically acceptable salt or solvate thereof.

11. R ３ The compound according to any one of claims 1 to 10, or a pharma- ceutically acceptable salt or solvate thereof, wherein: is (3-6C)cycloalkyl.

12. R ３ 12. The compound of any one of claims 1 to 11, or a pharma- ceutically acceptable salt or solvate thereof, wherein: is cyclopropyl.

13. A ３ But C(R ４ ) OR ５ 7. The compound of any one of claims 1 to 6, wherein:

14. R ４ But, H, C １～４ alkyl; R ５ is selected from (1-7C)alkyl, 4- to 6-membered heterocyclyl, and phenyl, which groups are joined by one or more R Ａ 14. The compound of claim 13, or a pharma- ceutically acceptable salt or solvate thereof, optionally substituted by a substituent.

15. R ４ is selected from H; R ５ is (1-4C)alkyl, or a pharma- ceutically acceptable salt or solvate thereof.

16. R ４ is selected from H; R ５ 14. The compound of claim 13, or a pharma- ceutically acceptable salt or solvate thereof, wherein: is methyl.

17. R Ａ but: (i) fluoro, cyano, (1-4C)alkyl, (1-4C)haloalkyl, (1-4C)hydroxyalkyl, phenyl, 4- to 6-membered heterocyclyl, 5- to 6-membered heteroaryl, -OR １ａ , -NR １ａ R １ｂ , -C(O)R １ａ , -C(O)N(R １ｂ ) R １ａ , N(R １ｂ ) C(O)R １ａ , -S(O) ｐ R １ａ (p is 0, 1 or 2), -SO ２ N (R １ｂ ) R １ａ , -N(R １ｂ ) S.O. ２ R １ａ [In the formula, R １ａ and R １ｂ are each independently selected from hydrogen or (1-4C)alkyl; or (ii) fluoro, cyano, (1-4C) alkyl, (1-4C) haloalkyl, (1-4C) hydroxyalkyl, -OR １ａ , -NR １ａ R １ｂ , -C(O)R １ａ , -C(O)N(R １ｂ ) R １ａ , N(R １ｂ ) C(O)R １ａ , -S(O) ｐ R １ａ (p is 0, 1 or 2), -SO ２ N (R １ｂ ) R １ａ , -N(R １ｂ ) S.O. ２ R １ａ [In the formula, R １ａ and R １ｂ are each independently selected from hydrogen or (1-4C)alkyl.

17. The compound of any one of claims 1 to 16, selected from: or a pharma- ceutically acceptable salt or solvate thereof.

18. The compound of formula Ia, Ib, Ic, Id, Ie, If, Ig, Ih, Ii, Ij or Ik shown below: 【Chemistry 2】 【Chemistry 2-2】 【Chemistry 2-3】 【Chemistry 2-4】 [In the formula, as appropriate: R １ is as defined in any one of claims 1 or 4 to 6; R ３ is as claimed in any one of claims 1 to 17; R ５ or a pharma- ceutically acceptable salt or solvate thereof.

19. formula: 【Chemistry 3】 2. The compound of claim 1, which is a compound of the formula:

20. 20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19, or a pharma- ceutically acceptable salt or solvate thereof, and a pharma- ceutically acceptable excipient.

21. 21. A compound according to any one of claims 1 to 19, or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 20, for use in therapy.

22. 21. A compound according to any one of claims 1 to 19, or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 20, for use in the treatment of cancer.

23. 21. The compound of claim 20, or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition, for use in the treatment of cancer, wherein the cancer is a PARP-expressing cancer.

24. 24. The compound or a pharma- ceutically acceptable salt or solvate thereof, or a pharmaceutical composition for use in the treatment of cancer according to claim 22 or claim 23, wherein the cancer is selected from glioma, glioblastoma, thyroid cancer, lung cancer (e.g. NSCLC), esophageal cancer, head and neck cancer, tongue cancer, gastric cancer, liver cancer (e.g. HCC), neuroendocrine cancer, pancreatic cancer (e.g. PDAC), colon cancer, renal cancer, prostate cancer, breast cancer (e.g. TNBC, ductal and invasive subtypes), endometrial cancer, cervical cancer, ovarian cancer, melanoma / skin cancer and lymphoma.

25. 21. A method of treating cancer comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 19, or a pharma- ceutically acceptable salt or solvate thereof, or administering a pharmaceutical composition according to claim 20.

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