Heterocyclic compounds for use in cancer treatment

Abstract

The present invention relates to heterocyclic derivatives and their use in the treatment and prevention of cancer, as well as compositions containing said derivatives and processes for their preparation.

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present invention relates to heterocyclic derivatives and their use in the treatment and prevention of cancer, as well as compositions containing said derivatives and methods for their preparation. [Background technology]

[0002] BACKGROUND OF THEINVENTION Robust repair of DNA double-strand breaks (DSBs) is essential for maintaining genome stability and cell viability. DSBs can be repaired by one of three major pathways: homologous recombination (HR), non-homologous end joining (NHEJ), and alternative NHEJ (alt-NHEJ). Microhomology-mediated end joining (MMEJ) is the best-characterized alt-NHEJ mechanism. HR-mediated repair is a high-fidelity mechanism essential for accurate error-free repair and impedes the stability of cancer-prone genomes. Conversely, NHEJ and MMEJ are error-prone pathways that may leave mutational scars at the repair site. MMEJ can function in parallel in both the HR and NHEJ pathways (Truong et al., PNAS 2013, 110(19), 7720-7725).

[0003] Unlike normal cells, the survival of cancer cells often depends on misregulation of DNA damage response (DDR) pathways, for example, increased reliance on one pathway (often mutagenic) to cope with either inactivation of another pathway or increased replication stress due to increased proliferation. Aberrant DDR can also sensitize cancer cells to specific types of DNA damage, so that defective DDR can be exploited to develop targeted cancer therapies. Crucially, cancer cells with impaired or inactivated HR and NHEJ become highly dependent on MMEJ-mediated DNA repair. Genetic, cell biological, and biochemical data have identified Polθ (UniProtKB-O75417(DPOLQ_HUMAN) as a key protein in MMEJ (Kent et al., Nature Structural & Molecular Biology((2015), 22(3), 230-237; Mateos-Gomez et al., Nature(2015), 518(7538), 254-257). Polθ is a multifunctional enzyme that contains an N-terminal helicase domain (SF2 HEL308 type) and a C-terminal low-fidelity DNA polymerase domain (A type) (Wood and Doublie, DNA Repair(2016), 44, 22-32). Both domains have been shown to have coordinated mechanistic functions in MMEJ: the helicase domain mediates the removal of RPA proteins from the ssDNA ends and stimulates annealing, while the polymerase domain extends the ssDNA ends and fills in the remaining gap.

[0004] Thus, therapeutic inactivation of Polθ would abolish the ability of cells to perform MMEJ and provide a novel targeting strategy in various defined tumor contexts. First, Polθ has been shown to be essential for the survival of HR-deficient (HRD) cells (e.g., synthetic lethality due to FA / BRCA deficiency) and is upregulated in HRD tumor cell lines (Ceccaldi et al., Nature (2015), 518(7538), 258-262). In vivo studies have also shown that Polθ is significantly overexpressed in a subset of HRD ovarian, uterine, and breast cancers and is associated with poor prognosis (Higgins et al., Oncotarget (2010), 1, 175-184, Lemee et al., PNAS (2010), 107(30), 13390-13395, Ceccaldi et al., (2015), supra). Importantly, Polθ is largely suppressed in normal tissues, but upregulated in matched cancer samples, and thus its increased expression has been shown to be disease-associated (Kawamura et al., International Journal of Cancer (2004), 109(1), 9-16). Secondly, suppression or inhibition of Polθ confers radiosensitivity to tumor cells. Finally, Polθ inhibition could prevent MMEJ-dependent functional restoration of BRCA2 mutations, which underlies the emergence of cisplatin- and PARP inhibitor (PARPi) resistance in tumors.

[0005] In view of the above, there is a demand to provide Pol θ inhibitors that are effective in cancer treatment. Summary of the Invention

[0006] (Summary of the invention) In a first aspect of the present invention there is provided a compound of formula (I) or a tautomeric or stereochemically isomeric form, a pharma- ceutically acceptable salt, or solvate thereof: [ka] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] Detailed Description of the Invention In one embodiment, the present invention provides a compound of formula (I), which is the free base of the compound of formula (I), (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (E1).

[0008] Reference to compounds of formula (I) and subgroups thereof also includes ionic forms, salts, solvates, isomers (including geometric and stereochemical isomers), tautomers, N-oxides, esters, prodrugs, isotopes and protected forms thereof, such as those described below; preferably, salts or tautomers or isomers or N-oxides or solvates thereof; more preferably, salts or tautomers or N-oxides or solvates thereof, and even more preferably, salts or tautomers or solvates thereof. Hereinafter, the compounds as defined in any aspect of the invention (except intermediate compounds in a chemical process), as well as ionic forms, salts, solvates, isomers (including geometric and stereochemical isomers), tautomers, N-oxides, esters, prodrugs, isotopes and protected forms thereof, are referred to as "compounds of the invention".

[0009] (salt) Certain compounds of formula (I) may exist in the form of salts, such as acid addition salts or, in certain cases, salts of organic and inorganic bases, such as carboxylates, sulfonates, phosphates, etc. All such salts are within the scope of the present invention, and a reference to a compound of formula (I) includes the salt forms of the compound.

[0010] The salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods such as those described in "Pharmaceutical Salts: Properties, Selection, and Use", P. Heinrich Stahl (ed.), Camille G. Wermuth (ed.), ISBN: 3-90639-026-8, Hardcover, page 388, August 2002. Generally, such salts can be prepared by reacting the free acid or free base forms of these compounds with the appropriate base or acid in water or in an organic solvent, or in a mixture of both; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used.

[0011] Acid addition salts (mono- or di-salts) can be formed with a wide variety of acids, both inorganic and organic. Examples of acid addition salts include acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, butanoic acid, (+)camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, and acetic acid. carboxylic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid, glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxoglutaric acid, glycolic acid, hippuric acid, hydrohalic acids (e.g., hydrobromic acid, hydrochloric acid, acid, hydroiodic acid), isethionic acid, lactic acid (e.g., (+)-L-lactic acid, (±)-DL-lactic acid), lactobionic acid, maleic acid, malic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, The mono- or di-salts include those formed with acids selected from the group consisting of carboxylic acid, ...

[0012] A group of particular salts of 1 consists of the salts formed from acetic acid, hydrochloric acid, hydroiodic acid, phosphoric acid, nitric acid, sulfuric acid, citric acid, lactic acid, succinic acid, maleic acid, malic acid, isethionic acid, fumaric acid, benzenesulfonic acid, toluenesulfonic acid, methanesulfonic acid (mesylic acid), ethanesulfonic acid, naphthalenesulfonic acid, valeric acid, acetic acid, propanoic acid, butanoic acid, malonic acid, glucuronic acid, and lactobionic acid. A particular salt of 1 is the hydrochloride salt.

[0013] When the compounds of formula (I) contain an amine group, they may be formed into quaternary ammonium salts, for example, by reaction with alkylating agents in a manner well known to those skilled in the art. Such quaternary ammonium compounds are within the scope of formula (I).

[0014] The compounds of the present invention have a pK value similar to that of the acid from which the salt is formed. a Depending on the compound, it may exist as a mono- or di-salt.

[0015] It will be appreciated that for pharmaceutical use, the salts of the compounds of formula (I) should be pharma- ceutically acceptable. Suitable pharma-ceutically acceptable salts will be apparent to one skilled in the art. Pharmaceutically acceptable salts include those reported by Berge, Bighley, and Monkhouse, J. Pharm. Sci. 1977, 66, pp. 1-19. Such pharma-ceutically acceptable salts include acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, or phosphoric acid, and organic acids, such as succinic acid, maleic acid, acetic acid, fumaric acid, citric acid, tartaric acid, benzoic acid, p-toluenesulfonic acid, methanesulfonic acid, or naphthalenesulfonic acid. Other salts, such as oxalates or formates, may be used, for example, in the isolation of the compounds of formula (I), and are included within the scope of the present invention. However, pharma-ceutically unacceptable salts may also be prepared as intermediate forms which can be subsequently converted to pharma-ceutically acceptable salts. Such non-pharmacologically acceptable salts forms, which may be useful, for example, in the purification or separation of the compounds of the invention, also form part of the invention.

[0016] Certain compounds of formula (I) may form acid addition salts with one or more equivalents of the acid. The present invention includes within its scope all possible stoichiometric and non-stoichiometric forms.

[0017] (Solvate) Those skilled in the art of organic chemistry will recognize that many organic compounds can form complexes with the solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates." For example, complexes with water are known as "hydrates." Pharmaceutically acceptable solvates of the compounds of the present invention are within the scope of the present invention. In one embodiment, the pharma- ceutically acceptable solvates of the compounds of the present invention include hydrates thereof.

[0018] In one embodiment, the crystalline form of the compound of formula (I) is a co-crystal or coformer. Such co-crystals or coformers may be prepared with water-soluble molecules such as saccharin, caffeine, nicotinamide, or carboxylic acids. Coformers may be prepared as described in Emami S et al., (2018) BioImpacts 8(4), 305-320, the teachings of which are incorporated herein by reference.

[0019] It will be understood that the present invention includes pharma- ceutically acceptable derivatives of compounds of formula (I) and these are included within the scope of the present invention.

[0020] As used herein, "pharmaceutical acceptable derivative" includes any pharma-ceutically acceptable ester of a compound of formula (I), or a salt of such an ester, which, upon administration to a recipient, is capable of providing (directly or indirectly) a compound of formula (I) or an active metabolite or residue thereof.

[0021] (N-oxide) Compounds of formula (I) that contain an amine group may also form N-oxides and a reference herein to a compound of formula (I) that contains an amine group also includes the N-oxide.

[0022] If the compound contains several amine groups, one or more of the nitrogen atoms may be oxidized to form an N-oxide. Particular examples of N-oxides are the N-oxides of tertiary amines or of a nitrogen atom of a nitrogen-containing heterocycle.

[0023] N-oxides can be formed by treating the corresponding amine with an oxidizing agent such as hydrogen peroxide or a peracid (e.g., peroxycarboxylic acid), see, for example, Advanced Organic Chemistry, Jerry March, 4th Edition, Wiley Interscience. More specifically, N-oxides can be prepared by the procedure of LW Deady (Syn. Commun. 1977, 7, 509-514), in which the amine compound is reacted with m-chloroperbenzoic acid (mCPBA) in an inert solvent, such as dichloromethane.

[0024] (Prodrug) Those skilled in the art will recognize that certain protected derivatives of the compounds of formula (I), which are prepared before the final deprotection step, may not have pharmacological activity in themselves, but in some instances may be metabolized in the body after oral or parenteral administration to form the pharmacologically active compounds of the invention. Thus, such derivatives may be described as "prodrugs". All such prodrugs of the compounds of the invention are included within the scope of the invention. Examples of functionally suitable prodrugs of the compounds of the invention are described in Drugs of Today, 19, 9, 1983, 499-538, and Topics in Chemistry, Chapter 31, pp. 306-316, and Design of Prodrugs, H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which are incorporated herein by reference). Certain moieties known to those of skill in the art as "promoieties" are described, for example, in H. Bundgaard's "Design of Prodrugs," the disclosure of which is incorporated herein by reference, and it will be further recognized by those of skill in the art that suitable functional groups, if any, may be disposed on such functional groups in the compounds of the invention.

[0025] Also included within the scope of the compounds and various salts of the present invention are polymorphs thereof.

[0026] (Enantiomers) When chiral centers are present in the compounds of formula (I), the present invention includes within its scope all possible enantiomers and diastereoisomers, including mixtures thereof. The different isomeric forms may be separated or resolved one from the other by conventional methods, or any given isomer may be obtained by conventional synthetic methods or by stereospecific or asymmetric syntheses. The invention also extends to any tautomers or mixtures thereof.

[0027] (Isotopes) The present invention also includes all pharma- ceutically acceptable isotopically labeled compounds which are identical to those recited in formula (I) except for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature.

[0028] Examples of isotopes suitable for inclusion in the compounds of the invention include: 2 H(D) and 3 Hydrogen isotopes such as H(T), 11 C. 13 C, and 14 Carbon isotopes such as C, 36 Chlorine isotopes such as Cl, 18 Fluorine isotopes such as F, 123 I, 125 I, and 131 Iodine isotopes such as I, 13 N and 15 Nitrogen isotopes such as N 15 O. 17 O, and 18 Oxygen isotopes such as O 32 Phosphorus isotopes such as P, 35 Sulfur isotopes such as S are included.

[0029] Certain isotopically labeled compounds of formula (I), for example those incorporating a radioisotope, are useful in drug and / or substrate tissue distribution studies. Compounds of formula (I) may also have valuable diagnostic properties in that they can be used to detect or identify the formation of complexes between the labeled compounds and other molecules, peptides, proteins, enzymes, or receptors. Detection or identification methods can use compounds labeled with labeling agents such as radioisotopes, enzymes, fluorescent substances, luminescent substances (e.g., luminol, luminol derivatives, luciferin, aequorin, and luciferase). Radioisotopes such as tritium, i.e. 3 H(T), and carbon-14, i.e. 14 C are particularly useful for this purpose given their ease of incorporation and ready means of detection.

[0030] Deuterium, i.e. 2 Substitution with heavier isotopes, such as H(D), may be preferable in some circumstances because they may confer certain therapeutic advantages due to greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements.

[0031] 11 C. 18 F, 15 O, and 13 Substitution with positron emitting isotopes, such as N, can be utilized in Positron Emission Topography (PET) studies for examining target occupancy.

[0032] Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the Examples and Preparations below, substituting an appropriate isotopically labeled reagent for the non-labeled reagent conventionally employed.

[0033] (purity) It will be readily appreciated that, because the compounds of formula (I) are intended for use as pharmaceutical compositions, they are each preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure, and preferably at least 85%, especially at least 98% pure (percentages being on a weight to weight basis). Impure preparations of the compounds may be used to prepare the more pure forms used in the pharmaceutical compositions.

[0034] (process) In a further aspect of the present invention, there is provided a process for the preparation of compounds of formula (I) and derivatives thereof. The following schemes are examples of synthetic schemes that may be used to synthesize compounds of the present invention. In the following schemes, reactive groups can be protected and deprotected with protecting groups according to well-established techniques.

[0035] A further aspect of the present invention relates to a process for the preparation of a compound of formula (I) as defined herein, comprising: (a) a compound of formula (II): [ka] Interconversion to compounds of formula (I); and (b) the optional formation of a pharma- ceutically acceptable salt of a compound of formula (I). The method of manufacturing is provided, comprising:

[0036] Process (a) typically involves an interconversion reaction to prepare a compound of formula (I), for example as described in step (xvii) of Example 1.

[0037] The compound of formula (II) can be prepared according to the procedures described herein.For example, the compound of formula (II) can be prepared according to the experimental procedures described in Example 1, particularly in step (xvi) of Example 1.

[0038] Where appropriate, the reactions described herein may be followed or preceded by one or more reactions known to those skilled in the art, carried out in an appropriate order to achieve the necessary substitutions for each of the variables defined herein, to give other compounds of formula (I). Non-limiting examples of such reactions, whose reaction conditions can be found in the literature, include: Protection of reactive groups, Deprotection of reactive groups, Halogenation, Dehalogenation, Dealkylation, Alkylation of amines, anilines, alcohols, and phenols; Mitsunobu reaction for hydroxyl groups, Cycloaddition reactions to suitable groups, Reduction of nitro, ester, cyano, and aldehydes, Transition metal catalyzed coupling reactions, Acylation, Sulfonylation / introduction of sulfonyl groups, Saponification / hydrolysis of ester groups, amidation or transesterification of ester groups; Esterification or amidation of carboxyl groups, Halogen exchange, Nucleophilic substitution with amines, thiols, or alcohols; Reductive amination, Oxime formation of carbonyl and hydroxylamine groups, S-oxidation, N-oxidation, chloride Includes:

[0039] It will be appreciated that the order of the reactions involving the aryl coupling and reduction may be varied. It will also be appreciated that a wide range of palladium-based catalysts are suitable for carrying out the aryl coupling reaction.

[0040] It will also be appreciated that isomer separations may be performed at appropriate stages in the synthetic sequence. It should be emphasized that such chiral separations form an important aspect of the present invention, and that such separations may be performed according to the methods described herein or according to known methods. It will also be appreciated that temporary formation of protected derivatives of intermediates during the synthesis, such as Boc-protected amines or SEM-protected amides, may be beneficial to facilitate chromatographic separations, chiral resolution, or to achieve improved solubility or increased yields at certain steps.

[0041] In many of the reactions described above, it may be necessary to protect one or more groups to prevent reaction from occurring at undesirable sites on the molecule. Examples of protecting groups and methods for protecting and deprotecting functional groups can be found in "Protective Groups in Organic Synthesis" (T. Green and P. Wuts; 4th Edition; John Wiley and Sons, 2007).

[0042] Hydroxy groups may be protected, for example, as ethers (-OR) or esters (-OC(=O)R), such as: tert-butyl ethers; tetrahydropyranyl (THP) ethers; benzyl, benzhydryl (diphenylmethyl), or trityl (triphenylmethyl) ethers; trimethylsilyl or tert-butyldimethylsilyl ethers; or acetyl esters (-OC(=O)CH3).

[0043] Amine groups can be, for example, as amides (-NRCO-R) or carbamates (-NRCO-OR), for example: as methylamides (-NHCO-CH3); as benzyl carbamates (-NHCO-OCH2C6H5, -NH-Cbz, or NH-Z); as tert-butyl carbamates (-NHCOOC(CH3)3, NH-Boc); as 2-biphenyl-2-propyl carbamates (-NHCO-OC(CH3)2C6H4C6H5, NH- Boc), as 9-fluorenylmethyl carbamate (-NH-Fmoc), as 6-nitroveratryl carbamate (-NH-Nvoc), as 2-trimethylsilylethyl carbamate (-NH-Teoc), as 2,2,2-trichloroethyl carbamate (-NH-Troc), as allyl carbamate (-NH-Alloc), or as 2(-phenylsulfonyl)ethyl carbamate (-NH-Psec).

[0044] Other protecting groups for amines, such as cyclic amines and heterocyclic NH groups, include toluenesulfonyl (tosyl) and methanesulfonyl (mesyl) groups, benzyl groups, such as para-methoxybenzyl (PMB), and tetrahydropyranyl (THP) groups.

[0045] Carboxylic acid groups can be used as esters, e.g. 1-7 Alkyl esters (e.g., methyl esters, tert-butyl esters); C 1-7 Haloalkyl esters (e.g., C 1-7 Trihaloalkyl esters); TriC 1-7 Alkylsilyl-C 1-7 Alkyl ester; or C 5-20 Aryl-C 1-7 It may be protected as an alkyl ester (eg, benzyl ester, nitrobenzyl ester, para-methoxybenzyl ester).

[0046] It will be appreciated by those skilled in the art that certain compounds of the invention can be converted to other compounds of the invention by standard chemical methods.

[0047] Pharmaceutically acceptable salts may be prepared conventionally by reaction with the appropriate acid or acid derivative.

[0048] (therapeutic utility) The compounds of the present invention, subgroups and examples thereof, are inhibitors of Pol θ polymerase activity and may be useful for preventing or treating disease states or conditions described herein. Furthermore, the compounds of the present invention and subgroups thereof may be useful for preventing or treating diseases or conditions mediated by Pol θ. Reference to preventing or treating a disease state or condition, such as cancer, includes within its scope alleviating cancer or reducing the incidence of cancer.

[0049] Thus, for example, it is envisaged that the compounds of the invention will be useful in alleviating or reducing the incidence of cancer.

[0050] The compounds of the invention may be useful in the treatment of the adult population.The compounds of the invention may be useful in the treatment of the pediatric population.

[0051] As a result of these inhibitions of Polθ, the compounds may be useful in providing a means to neutralize the ability of cells to perform MMEJ. Thus, the compounds may prove useful in treating or preventing proliferative disorders such as cancer. Furthermore, the compounds of the present invention may be useful in treating diseases in which there is a disorder associated with cell accumulation.

[0052] Without being bound by theory, it is predicted that the Pol θ inhibitors of the invention will demonstrate characteristics that make them particularly useful in the therapeutic treatment of certain cancers. For example, in one embodiment, the Pol θ inhibitors of the invention are preferably lethal in BRCA1 and BRCA2 deficient primary and secondary solid tumors, including breast, ovarian, prostate, and pancreatic solid tumors.

[0053] In further embodiments, the Pol θ inhibitors of the present invention are preferably lethal in a variety of primary and secondary solid tumors that are HRD by mechanisms other than BRCA deficiency, such as those involving promoter hypermethylation. In these tumors where DSB repair pathways may not be completely downregulated, the Pol θ inhibitors (Pol θi) can also be administered together with another DDR modulator, such as a PARP inhibitor, a DNA-PK inhibitor, an ATR inhibitor, an ATM inhibitor, a wee1 inhibitor, or a CHK1 inhibitor.

[0054] In a further embodiment, the Pol θ inhibitors of the present invention are preferably lethal in primary and secondary breast, ovarian, prostate, and pancreatic tumors that retain BRCA1 deficiency but are resistant to PARPi treatment with or without exposure to PARPi drug therapy.

[0055] In further embodiments, the Pol θ inhibitors of the invention, preferably when given with a PARPi therapeutic program, will increase ORR, including CRR, will delay the onset of PARPi resistance, will prolong time to recurrence and DFS, and will prolong OS in HRD (BRCA1 / 2-deficient and other HRD mechanisms) primary and secondary tumors (breast, ovarian, prostate, and pancreatic).

[0056] In a further embodiment, the Pol θ inhibitors of the present invention preferably inhibit ATM activity (ATM), particularly in the context of WT p53. - / - ) in various tumors with loss of the DDR. The tumor types would include about 10% of all solid tumors including gastric, lung, breast, and CRC, along with CLL. Co-drug treatment with another DDR modifier, such as a DNA-PK inhibitor, a PARP inhibitor, or an ATR inhibitor, could further enhance such activity. Polθ inhibitors would resensitize CLL to classical chemotherapy and immunochemotherapy where drug resistance has developed. Thus, in a further embodiment, the pharmaceutical composition of the present invention further comprises a DNA-PK inhibitor, a PARP inhibitor, or an ATR inhibitor.

[0057] In a further embodiment, the Polθ inhibitors of the present invention suitably exhibit synthetic disease and / or synthetic lethality in various tumors defective in the DNA double strand break repair process of non-homologous end joining (NHEJ-D). would comprise approximately 2-10% of all solid tumors, including prostate, pancreas, cervix, breast, lung, bladder, and esophagus. Co-drug treatment with another DDR modifier, such as a PARP inhibitor, ATM inhibitor, wee1 inhibitor, CHK inhibitor, or ATR inhibitor, may further enhance such activity. Polθ inhibitors would further sensitize NHEJD cancer cells to DNA DSB-inducing chemotherapy and ionizing radiation-based therapy. Thus, in a further embodiment, the pharmaceutical composition of the present invention further comprises a PARP inhibitor, ATM inhibitor, wee1 inhibitor, CHK inhibitor, or ATR inhibitor.

[0058] In a further embodiment, the Pol θ inhibitors of the present invention preferably reduce the DNA replication stress response during chemotherapy in HR proficient tumors, such as ovarian, NSCL, and breast tumors, that overexpress Pol θ. This will increase the ORR to treatment and increase OS. Such effects are particularly likely to occur with cytarabine (Ara-C) and hydroxyurea, which are used in a wide variety of leukemias, including CML, and in the management of squamous cell carcinomas.

[0059] In a further embodiment, the Pol θ inhibitors of the present invention preferably selectively sensitize solid tumors with little or no sensitization of normal tissues in radiation therapy, including EBRT and brachytherapy. In a segmented targeted therapy setting, this may improve locoregional control resulting in increased survival. This may be particularly evident in the management of NSCLC, SCCH&N, rectal cancer, prostate cancer, and pancreatic cancer.

[0060] In a further embodiment, the Pol θ inhibitors of the present invention, preferably with or without co-pharmaceutical therapy with a PARPi, exhibit synthetic disease and / or synthetic lethality in PTEN-deficient tumors, such as CaP, and furthermore, such tumors will be exquisitely sensitive to radiation therapy due to both PTEN deficiency and Pol θ inhibitor-induced radiosensitivity.

[0061] In further embodiments, the Pol θ inhibitors of the present invention preferably inhibit TLS polymerase activity, sensitizing primary and secondary solid tumors (e.g., breast, lung, ovarian, CRC) to drugs (e.g., cisplatin, mitomycin, and cyclophosphamide) and reducing the acquisition of drug-induced mutations associated with tumor resistance, resulting in prolonged remission and increased TTR.

[0062] In a further embodiment, the Pol θ inhibitors of the present invention preferably resensitize BCR-ABL positive CML that have developed resistance to imatinib, as well as other solid tumors that exhibit elevated levels of ligase IIIα, decreased levels of ligase IV, and increased dependency on altEJ DSB repair.

[0063] In a further embodiment, the Pol θ inhibitors of the present invention suitably inhibit aromatase inhibitor-resistant ER tumors that also exhibit elevated levels of ligase IIIα, decreased levels of ligase IV, and increased dependency on altEJ DSB repair. - Synthetic disease and / or synthetic lethality in primary and secondary breast cancer.

[0064] In a further aspect of the invention there is provided a compound of formula (I) as defined herein for use in the treatment of a tumour characterised by homologous recombination deficiency (HRD).

[0065] It will be appreciated that reference herein to "homologous recombination deficiency (HRD)" refers to any genetic alteration that results in the deficiency or loss of function of a homologous recombination gene. Examples of such genetic alterations include mutations (e.g., point mutations), substitutions, deletions, single nucleotide polymorphisms (SNPs), haplotypes, chromosomal abnormalities, copy number variations (CNVs), epigenetics, DNA inversions, reduced expression, and mislocalization.

[0066] In one embodiment, the homologous recombination gene is selected from ATM, ATR, BRCA1, BRCA2, BARD1, RAD51C, RAD50, CHEK1, CHEK2, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, PALB2 (FANCN), FANCP (BTBD12), ERCC4 (FANCQ), PTEN, CDK12, MRE11, NBS1, NBN, CLASPIN, BLM, WRN, SMARCA2, SMARCA4, LIG1, RPA1, RPA2, BRIP1, and PTEN.

[0067] It will be appreciated that reference herein to "non-homologous end joining deficiency (NHEJD)" refers to any genetic mutation that results in the deficiency or loss of function of a homologous recombination gene. Examples of such genetic mutations include mutations (e.g., point mutations), substitutions, deletions, single nucleotide polymorphisms (SNPs), haplotypes, chromosomal abnormalities, copy number variations (CNVs), epigenetics, DNA inversions, reduced expression, and mislocalization.

[0068] In one embodiment, the non-homologous end joining gene is selected from any one or more of LIG4, NHEJ1, POLL, POLM, PRKDC, XRCC4, XRCC5, XRCC6, and DCLRE1C.

[0069] In a further aspect of the invention there is provided a compound of formula (I) as defined herein for use in the treatment of tumours overexpressing Pol theta.

[0070] In a further aspect of the invention there is provided a compound of formula (I) as defined herein for use in the treatment of a tumour which has elevated levels of ligase IIIα, decreased levels of ligase IV and increased dependency on altEJ DSB repair.

[0071] Examples of cancers (and their benign counterparts) that may be treated (or inhibited) include, but are not limited to, tumors of epithelial origin (adenomas and various types of carcinomas, including adenocarcinoma, squamous cell carcinoma, transitional cell carcinoma, and other carcinomas), such as tumors of the bladder and urinary tract, breast, gastrointestinal tract (including esophagus, stomach (gastric), small intestine, colon, rectum, and anus), liver (hepatocellular carcinoma), gallbladder and biliary system, exocrine pancreas, kidney, lung (e.g., adenocarcinoma, small cell lung carcinoma, non-small cell lung carcinoma, bronchoalveolar carcinoma, and mesothelioma), head and neck (e.g., tongue, buccal cavity, larynx, pharynx, nasal pharynx, tonsils, and the like), carcinomas of the ovary, fallopian tubes, peritoneum, vagina, vulva, penis, cervix, myometrium, endometrium, thyroid (e.g., follicular thyroid carcinoma), adrenal gland, prostate, skin, and adnexa (e.g., melanoma, basal cell carcinoma, squamous cell carcinoma, keratoacanthoma, dysplastic nevi); hematological malignancies (i.e., leukemia, lymphoma) and premalignant hematological disorders and disorders of borderline malignancies, including lymphatic hematological malignancies and related lesions (e.g., acute lymphocytic leukemia [ALL], chronic lymphocytic leukemia [CLL], B-cell lymphomas, such as diffuse large intestine, rectal cavity, and sinuses); and hematological malignancies and related disorders of myeloid lineage (e.g., acute myeloid leukemia [AML], chronic myeloid leukemia [CML], chronic myelomonocytic leukemia [CMML], hypereosinophilic syndromes, myeloproliferative disorders, e.g., myeloproliferative disorders, etc.), including B-cell lymphoma [DLBCL], follicular lymphoma, Burkitt lymphoma, mantle cell lymphoma, MALT lymphoma, T-cell lymphoma and leukemia, natural killer [NK] cell lymphoma, Hodgkin lymphoma, hairy cell leukemia, monoclonal gammopathy of undetermined significance, plasmacytoma, multiple myeloma, and post-transplant lymphoproliferative disorders, etc.). tumors of mesenchymal origin, including sarcomas of the soft tissue, bone, or cartilage, such as osteosarcoma, fibrosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, angiosarcoma, Kaposi's sarcoma, Ewing's sarcoma, synovial sarcoma, epithelioid sarcoma, gastrointestinal stromal tumors, benign and malignant histiocytoma, and dermatofibrosarcoma protuberans; tumors of the central or peripheral nervous system (e.g., astrocytoma, glioma and glioblastoma, meningioma, ependymoma, pineal tumor, and schwannoma);These include endocrine tumors (e.g., pituitary tumors, adrenal tumors, pancreatic islet cell tumors, parathyroid tumors, carcinoid tumors, and medullary carcinoma of the thyroid); ocular and adnexal tumors (e.g., retinoblastoma); germ cell and trophoblastic tumors (e.g., teratomas, seminomas, dysgerminomas, hydatidiform moles, and choriocarcinomas); and pediatric and fetal tumors (e.g., medulloblastomas, neuroblastomas, Wilms' tumors, and primitive neuroectodermal tumors); or congenital or non-congenital syndromes that predispose a patient to malignancies (e.g., xeroderma pigmentosum);

[0072] Many diseases are characterized by persistent uncontrolled angiogenesis. Chronic proliferative diseases are often accompanied by significant angiogenesis, which can contribute to or maintain inflammatory and / or proliferative conditions, or lead to tissue destruction through the infiltrative proliferation of blood vessels. Tumor growth and metastasis have been found to be angiogenesis-dependent. Compounds of the present invention may therefore be useful in preventing and disrupting the initiation of tumor angiogenesis. In particular, compounds of the present invention may be useful in metastasis and metastatic cancer treatment.

[0073] Metastasis or metastatic disease is the spread of disease from one organ or part to another non-adjacent organ or part. Cancers treatable by the compounds of the present invention include primary tumors (i.e., cancer cells at the site of origin), local invasion (cancer cells that invade and invade surrounding normal tissues in that local area), and metastatic (or secondary) tumors, i.e., tumors that arise from malignant cells that have circulated through the bloodstream (hematogenous spread), or via the lymphatic system, or across body cavities (transcatheterial spread) to other sites and tissues in the body.

[0074] Particular cancers include hepatocellular carcinoma, melanoma, esophageal, renal, colon, colorectal, lung cancer (eg, mesothelioma or lung adenocarcinoma), breast, bladder, gastrointestinal, ovarian, and prostate cancer.

[0075] In a further aspect, there is provided the use of a compound as described herein for the manufacture of a medicament for the treatment of a disease or condition, particularly cancer.

[0076] The compounds may also be useful in the treatment of tumor growth, pathology, resistance to chemotherapy and radiotherapy caused by sensitizing cells to chemotherapy, and may be useful as anti-metastatic agents.

[0077] The efficacy of the compounds of the invention as inhibitors of Pol θ can be measured using the biological and biophysical assays described in the Examples herein, and the level of affinity exhibited by a given compound can be measured using the IC 50 Certain compounds of the invention have an IC value of less than 1 μM, more particularly less than 0.1 μM. 50 It is a compound having a value.

[0078] The role of the loss of Polθ in enhancing the efficacy of CRISPR-mediated gene editing is described in WO2017 / 062754. Thus, Polθ inhibitory compounds may be useful for enhancing the efficiency of CRISPR-based editing methods and / or CRISPR-based editing treatments. Furthermore, compound-mediated Polθ inhibition may reduce the frequency of random integration events, thus providing a way to improve any safety concerns of CRISPR-mediated techniques. Thus, in a further aspect of the present invention, the use of the compound of formula (I) as defined herein in CRISPR-based editing methods and / or CRISPR-based editing treatments, for example, enhancing the efficiency of CRISPR-based editing methods and / or CRISPR-based editing treatments, is provided.

[0079] Pharmaceutical Composition While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (eg, formulation), which in one embodiment is a sterile pharmaceutical composition.

[0080] Accordingly, the present invention further provides pharmaceutical compositions as identified above, as well as methods of making (e.g. mixing) pharmaceutical compositions comprising at least one compound of formula (I) (and subgroups thereof as defined herein), together with one or more pharma- ceutically acceptable excipients and optionally other therapeutic or prophylactic agents, as described herein.

[0081] The pharma- ceutically acceptable excipient(s) can be selected, for example, from carriers (such as solid, liquid, or semi-solid carriers), adjuvants, diluents, fillers or extenders, granulating agents, coating agents, release-controlling agents, binders, disintegrants, lubricants, preservatives, antioxidants, buffers, suspending agents, thickening agents, flavoring agents, sweeteners, taste-masking agents, stabilizers, or any other excipients conventionally used in pharmaceutical compositions. Examples of excipients for various types of pharmaceutical compositions are described in more detail below.

[0082] As used herein, the term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of prudent medical judgment, suitable for use in contact with the tissues of a subject (e.g., a human) without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0083] Pharmaceutical compositions containing compounds of formula (I) can be formulated according to known techniques, see, for example, "Remington's Pharmaceutical Sciences", Mack Publishing Company, Easton, PA, USA.

[0084] The pharmaceutical composition may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic, intraaural, rectal, intravaginal, or transdermal administration. If the composition is intended for parenteral administration, it may be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration, or for direct delivery into a target organ or tissue by injection, infusion, or other delivery means. Delivery may be by bolus injection, short-term infusion, or long-term infusion, by passive delivery, or by utilizing a suitable infusion pump or syringe-driven device.

[0085] Pharmaceutical formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, cosolvents, surfactants, organic solvent mixtures, cyclodextrin complexing agents, emulsifiers (to form and stabilize emulsion formulations), liposomal components to form liposomes, gellable polymers to form polymeric gels, lyoprotectants, and combination agents, among others, to stabilize the active ingredient in soluble form and to render the formulation isotonic with the blood of the intended recipient. Pharmaceutical formulations for parenteral administration may also take the form of aqueous and non-aqueous sterile suspensions, which may contain suspending agents and thickening agents (RG Strickly, Solubilizing Excipients in oral and injectable formulations, Pharmaceutical Research, Vol 21(2)2004, pp. 201-230).

[0086] The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampoules, vials, and pre-filled syringes and may be stored in a lyophilised (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injection, immediately prior to use. In one embodiment, the formulation is provided as the active pharmaceutical ingredient in a bottle for subsequent reconstitution with a suitable diluent.

[0087] Pharmaceutical formulations can be prepared by lyophilisation of a compound of formula (I) or any subgroup thereof. Lyophilisation refers to the procedure of freeze-drying a composition. Thus, freeze-drying and lyophilisation are used synonymously herein.

[0088] Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0089] Pharmaceutical compositions of the invention for parenteral injection may also include pharma- ceutically acceptable sterile, aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions immediately prior to use.

[0090] Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or vehicles include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (sunflower oil, safflower oil, corn oil, olive oil, etc.), and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of thickening or coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0091] The compositions of the present invention may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial action may be ensured by including various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include agents for adjusting tonicity, such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0092] In a particular embodiment of the invention, the pharmaceutical composition is in a form suitable for intravenous (iv) administration, for example by injection or infusion. Solutions for intravenous administration can be administered as is or can be injected into an infusion bag (containing a pharma- ceutical acceptable excipient, such as 0.9% saline or 5% glucose, etc.) before administration.

[0093] In another particular embodiment, the pharmaceutical composition is in a form suitable for subcutaneous (sc) administration.

[0094] Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), caplets, pills, lozenges, syrups, liquids, powders, granules, elixirs, and suspensions, sublingual tablets, wafers, or patches such as an intraoral buccal patch.

[0095] Thus, the tablet composition may contain a unit dose of active compound together with an inert diluent or carrier, such as a sugar or sugar alcohol, specifically lactose, sucrose, sorbitol, or mannitol; and / or a non-sugar derived diluent, such as sodium carbonate, calcium phosphate, calcium carbonate, or a cellulose or its derivatives, such as microcrystalline cellulose (MCC), methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, and starch, such as corn starch. Tablets may also contain standard ingredients, such as binders and granulating agents (such as polyvinylpyrrolidone), disintegrants (e.g., swellable cross-linked polymers, such as cross-linked carboxymethylcellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate or citrate buffers), and effervescent agents (e.g., citrate / bicarbonate mixtures). These excipients are well known and need not be described in detail here.

[0096] Tablets may be designed to release the drug upon contact with gastric fluids (immediate release tablets) or to release in a controlled manner over an extended period of time or in specific areas of the gastrointestinal tract (controlled release tablets).

[0097] Capsule formulations may be of the hard or soft gelatin variety and can contain the active ingredient in solid, semi-solid, or liquid form. Gelatin capsules can be made from animal gelatin or its equivalents of synthetic or vegetable origin.

[0098] Solid dosage forms (e.g., tablets, capsules, etc.) may be coated or uncoated. Coatings may function as protective films (e.g., polymers, waxes, or varnishes) or as mechanisms for controlling drug release or for aesthetic or identification purposes. Coatings (e.g., Eudragit™-type polymers) can be designed to release the active ingredient at a desired site in the gastrointestinal tract. Thus, coatings can be selected to degrade under specific pH conditions in the gastrointestinal tract, thereby selectively releasing the compound in the stomach or in the ileum, duodenum, jejunum, or colon.

[0099] Instead of or in addition to a coating, the drug may be provided in a solid matrix that includes a release control agent, such as a release retarding agent that can be adapted to release the compound in the gastrointestinal tract in a controlled manner. Alternatively, the drug may be provided in a polymer coating, such as a polymethacrylate polymer coating, that can be adapted to selectively release the compound in the gastrointestinal tract under conditions of varying acidity or alkalinity. Alternatively, the matrix material or release retarding coating may be in the form of a readily degradable polymer (e.g., maleic anhydride polymer) that degrades substantially continuously as the dosage form passes through the gastrointestinal tract. In another alternative embodiment, the coating may be designed to degrade by microbial action in the gastrointestinal tract. In a further alternative embodiment, the active compound may be formulated in a delivery system that provides osmotic control of the release of the compound. Osmotic release and other delayed or sustained release formulations (e.g., ion exchange resin-based formulations) may be made according to methods well known to those skilled in the art.

[0100] The compounds of formula (I) may be formulated with carriers and administered in the form of nanoparticles, the increased surface area of ​​which aids in their absorption. Furthermore, nanoparticles offer the possibility of direct penetration into cells. Nanoparticle drug delivery systems are described in "Nanoparticle Technology for Drug Delivery" edited by Ram B Gupta and Uday B. Kompella, Informa Healthcare, ISBN 9781574448573, published March 13, 2006. Nanoparticles for drug delivery are also described in J. Control. Release, 2003, 91(1-2), 167-172 and Sinha et al., Mol. Cancer Ther. August 1, (2006) 5, 1909.

[0101] The pharmaceutical composition usually comprises from about 1% (w / w) to about 95% (w / w) of the active ingredient and from 99% (w / w) to 5% (w / w) of a pharma- ceutical acceptable excipient or combination of excipients. In particular, the composition comprises from about 20% (w / w) to about 90%, % (w / w) of the active ingredient and from 80% (w / w) to 10% of a pharma- ceutical acceptable excipient or combination of excipients. The pharmaceutical composition comprises from about 1% to about 95%, in particular from about 20% to about 90%, of the active ingredient. The pharmaceutical composition according to the invention may be, for example, in unit dose form, for example in the form of ampoules, vials, suppositories, pre-filled syringes, dragees, tablets, or capsules.

[0102] The pharma- ceutically acceptable excipient(s) can be selected according to the desired physical form of the formulation and can be selected, for example, from diluents (e.g., solid diluents such as fillers or bulking agents; and liquid diluents such as solvents and cosolvents), disintegrants, buffers, lubricants, flow aids, release-controlling agents (e.g., release-retarding or release-extending polymers or waxes), binders, granulating agents, dyes, plasticizers, antioxidants, preservatives, flavorings, taste-masking agents, tonicity modifiers, and coating agents.

[0103] A person skilled in the art would have the expertise to select the appropriate amounts of ingredients for formulation use. For example, tablets and capsules typically contain 0-20% disintegrant, 0-5% lubricant, 0-5% flow aid, and / or 0-99% (w / w) filler / or extender (depending on drug dose). They may also contain 0-10% (w / w) polymer binder, 0-5% (w / w) antioxidant, 0-5% (w / w) dye. Extended release tablets may further contain 0-99% (w / w) release-controlling (e.g. retarding) polymer (depending on dose). Film coatings for tablets or capsules typically contain 0-10% (w / w) polymer, 0-3% (w / w) dye, and / or 0-2% (w / w) plasticizer.

[0104] Parenteral formulations typically contain 0-20% (w / w) buffer, 0-50% (w / w) co-solvent, and / or 0-99% (w / w) water for injection (WFI) (depending on dose, in the case of lyophilisates). Intramuscular depot formulations may also contain 0-99% (w / w) oils.

[0105] Pharmaceutical compositions for oral administration can be obtained by combining the active ingredient with a solid carrier, optionally granulating the resulting mixture, and processing the mixture into tablets, dragee cores, or capsules, if necessary or necessary after the addition of suitable excipients. These can also be incorporated into polymer or wax matrices that allow the active ingredient to diffuse or be released in measured amounts.

[0106] The compounds of the present invention can also be formulated as solid dispersions. A solid dispersion is a homogeneous and extremely finely dispersed phase of two or more solids. A type of solid dispersion, solid solution (molecular dispersion system), is well known for use in pharmaceutical technology (see Chiou and Riegelman, J. Pharm. Sci., 60, 1281-1300 (1971)), and is useful for increasing the dissolution rate and bioavailability of poorly water-soluble drugs.

[0107] The present invention also provides solid dosage forms comprising the above-mentioned solid solutions. Solid dosage forms include tablets, capsules, chewable tablets, and dispersible or effervescent tablets. Known excipients can be blended with the solid solutions to obtain the desired dosage forms. For example, capsules can comprise the solid solutions blended with (a) a disintegrant and a lubricant, or (b) a disintegrant, a lubricant, and a surfactant. Additionally, capsules can comprise a filler, such as lactose or microcrystalline cellulose. Tablets can comprise the solid solutions blended with at least one disintegrant, a lubricant, a surfactant, a filler, and a lubricant. Chewable tablets can comprise the solid solutions blended with a filler, a lubricant, and optionally additional sweeteners (e.g., artificial sweeteners), and suitable flavors. Solid solutions can also be formed by spraying a solution of the drug and a suitable polymer onto the surface of an inert carrier, such as sugar beads ("non-pareils"). These beads can then be filled into capsules or compressed into tablets.

[0108] The pharmaceutical formulations may be provided to patients in "patient packs" that contain the entire source of treatment in a single package, usually a blister pack. Patient packs have the advantage over traditional prescriptions, where bulk supplies of pharmaceuticals are distributed by pharmacists as patient supplies, that patients always have access to the package insert, which is not included in typical patient prescriptions but is included in patient packs. The inclusion of the package insert has been shown to improve patient compliance with physician instructions.

[0109] Compositions for topical and nasal delivery include ointments, creams, sprays, patches, gels, liquid drops, and inserts (e.g., intraocular inserts). Such compositions can be formulated according to known methods.

[0110] Examples of formulations for rectal or vaginal administration include pessaries and suppositories which may be formed, for example, from a moldable or waxy material containing the active compound. Solutions of the active compound may also be used for rectal administration.

[0111] Compositions for inhalation administration may be in the form of inhalable powder compositions or liquid or powder sprays, and can be administered in standard forms using powder inhalation devices or aerosol dispensing devices. Such devices are well known. For inhalation administration, powdered formulations usually contain the active compound together with an inert solid powdered diluent, such as lactose.

[0112] The compounds of formula (I) are generally provided in unit dosage form and therefore usually contain sufficient compound to provide a desired level of biological activity. For example, a formulation may contain 1 nanogram to 2 grams of active ingredient, e.g., 1 nanogram to 2 milligrams of active ingredient. Within these ranges, specific subranges of the compound are 0.1 milligrams to 2 grams of active ingredient (more generally 10 milligrams to 1 gram, e.g., 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (e.g., 1 microgram to 10 milligrams, e.g., 0.1 milligram to 2 milligrams of active ingredient).

[0113] For oral compositions, a unit dosage form may contain from 1 milligram to 2 grams, more typically from 10 milligrams to 1 gram, for example, 50 milligrams to 1 gram, for example, 100 milligrams to 1 gram of active compound.

[0114] The active compounds will be administered to a patient in need thereof (eg, a human or animal patient) in an amount sufficient to achieve the desired therapeutic effect.

[0115] (Treatment method) The compounds of formula (I) and subgroups as defined herein may be useful in the prevention or treatment of various disease states or conditions mediated by Pol θ. Thus, in a further aspect of the present invention, there is provided a method of treating a disease state or condition mediated by Pol θ (e.g., cancer), comprising administering to a subject in need thereof a compound of formula (I) as described herein. Examples of such disease states and conditions are set out above and include, in particular, cancer.

[0116] The compounds are generally administered to a subject in need of such administration, such as a human or animal patient, particularly a human.

[0117] The compound will usually be administered in an amount that is therapeutically or prophylactically useful and generally non-toxic. However, in certain circumstances (e.g., in the case of severe disease), the benefits of administering the compound of formula (I) may outweigh the disadvantages of any toxic or side effects, in which case it may be considered desirable to administer the compound in an amount associated with some toxicity.

[0118] The compounds may be administered over an extended period of time to maintain beneficial therapeutic effects, or for only a short period of time, or alternatively, the compounds may be administered in a continuous or intermittent manner (e.g., in a pulsatile manner) to provide dosing.

[0119] Typical daily dosages of a compound of formula (I) can range from 100 picograms to 100 milligrams per kilogram of body weight, more typically from 5 nanograms to 25 milligrams per kilogram of body weight, more usually from 10 nanograms to 15 milligrams per kilogram of body weight (e.g., 10 nanograms to 10 milligrams, more typically from 1 microgram per kilogram to 20 milligrams per kilogram, e.g., 1 microgram to 10 milligrams per kilogram), although higher or lower doses can be administered if necessary. Compounds of formula (I) can be administered daily or repeatedly, for example, every 2, or 3, or 4, or 5, or 6, or 7, or 10, or 14, or 21, or 28 days.

[0120] The compounds of the present invention may be administered orally in a dosage range of, for example, 1-1500 mg, 2-800 mg, or 5-500 mg, e.g., 2-200 mg or 10-1000 mg, with specific examples of dosages including 10, 20, 50, and 80 mg. The compounds may be administered once a day or more than once a day. The compounds may be administered continuously (i.e., taken every day without interruption for the entire duration of the treatment regimen). Alternatively, the compounds may be administered intermittently (i.e., taken continuously for a given period of time, such as a week, then discontinued for a period of time, such as a week, then taken continuously for another period of time, such as a week, and so on throughout the duration of the treatment regimen). Examples of treatment regimens involving intermittent administration include regimens in which administration is repeated in cycles of 1 week on, 1 week off; or 2 weeks on, 1 week off; or 3 weeks on, 1 week off; or 2 weeks on, 2 weeks off; or 4 weeks on, 2 weeks off; or 1 week on, 3 weeks off, for one or more cycles, e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more cycles.

[0121] In one particular dosing schedule, the patient would receive an infusion of a compound of formula (I) for one hour daily for up to 10 days, particularly up to 5 days, for one week, with the treatment repeated at desired intervals such as 2-4 weeks, particularly every 3 weeks.

[0122] More particularly, a patient receives an infusion of a compound of formula (I) for one hour each day for five days, which treatment may be repeated every three weeks.

[0123] In another particular dosing schedule, the patient receives an infusion over 30 minutes to 1 hour, followed by a continuous infusion of variable duration, for example 1 to 5 hours, for example 3 hours.

[0124] In a more particular dosing schedule, the patient receives a continuous infusion over a period of 12 hours to 5 days, particularly a continuous infusion over a period of 24 hours to 72 hours.

[0125] In another particular dosing schedule, the patient is given the compound orally once a week.

[0126] In another particular dosing schedule, the patient is given the compound orally once daily for 7 to 28 days, for example, 7, 14, or 28 days.

[0127] In another particular dosing schedule, patients are given the compound orally once daily for 1, 2, 3, 5, or 1 week, followed by the necessary number of days off drug to complete a 1 or 2 week cycle.

[0128] In another particular dosing schedule, patients are given the compound orally once daily for two weeks, followed by a two-week rest period.

[0129] In another particular dosing schedule, patients are given the compound orally once daily for two weeks, followed by one week of rest.

[0130] In another particular dosing schedule, patients are given the compound orally once daily for one week, followed by one week of rest.

[0131] Ultimately, however, the amount of compound administered and the type of composition used will correspond to the nature of the disease or physiological condition being treated and will be at the discretion of the physician.

[0132] It will be appreciated that Pol theta inhibitors can be used as single agents or in combination with other anticancer agents. Combination studies can be performed, for example, as described in Chou TC, Talalay P., "Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors," Adv Enzyme Regulat 1984;22:27-55.

[0133] The compounds defined herein can be administered as a sole therapeutic agent for the treatment of a particular disease state, e.g., a neoplastic disease such as cancer as defined herein above, or they can be administered in combination therapy with one or more other compounds (or therapies). For the treatment of the above conditions, the compounds of the invention may be advantageously employed in combination with one or more other pharmaceutical agents, more particularly with other anti-cancer agents or adjuvants in cancer therapy. Examples of other therapeutic agents or treatments that may be administered together with the compounds of formula (I) (whether simultaneously overlapping or at different time intervals) include, but are not limited to: Topoisomerase I inhibitors; ·Antimetabolites; ·Tubulin targeting agents; ·DNA binders and topoisomerase II inhibitors; · Alkylating agents; · Monoclonal antibodies; ·Anti-hormonal drugs; Signal transduction inhibitors; · Proteasome inhibitors; DNA methyltransferase inhibitors; · Cytokines and retinoids; · Chromatin-targeted therapies; Radiation therapy; and Other therapeutic or prophylactic drugs Includes:

[0134] Specific examples of anti-cancer agents or adjuvants (or salts thereof) include, but are not limited to, any of the agents selected from groups (i) to (xlvi), and optionally group (xlvii) below: (i) platinum compounds, such as cisplatin (optionally in combination with amifostine), carboplatin, or oxaliplatin; (ii) a taxane compound, such as paclitaxel, paclitaxel protein-bound particles (Abraxane™), docetaxel, cabazitaxel, or larotaxel; (iii) a topoisomerase I inhibitor, for example, a camptothecin compound, such as camptothecin, irinotecan (CPT11), SN-38, or topotecan; (iv) topoisomerase II inhibitors, such as antitumor epipodophyllotoxins or podophyllotoxin derivatives, such as etoposide or teniposide; (v) Vinca alkaloids, such as vinblastine, vincristine, liposomal formulations of vincristine (Onco-TCS), vinorelbine, vindesine, vinflunine, or vinvesir; (vi) nucleoside derivatives, such as 5-fluorouracil (5-FU, optionally in combination with leucovorin), gemcitabine, capecitabine, tegafur, UFT, S1, cladribine, cytarabine (Ara-C, cytosine arabinoside), fludarabine, clofarabine, or nelarabine; (vii) antimetabolites, such as clofarabine, aminopterin, or methotrexate, azacitidine, cytarabine, floxuridine, pentostatin, thioguanine, thiopurine, 6-mercaptopurine, or hydroxyurea (hydroxycarbamide); (viii) alkylating agents, such as nitrogen mustards or nitrosoureas, e.g., cyclophosphamide, chlorambucil, carmustine (BCNU), bendamustine, thiotepa, melphalan, treosulfan, lomustine (CCNU), altretamine, busulfan, dacarbazine, estramustine, fotemustine, ifosfamide (optionally in combination with mesna), pipobroman, procarbazine, streptozocin, temozolomide, uracil, mechlorethamine, methylcyclohexylchloroethylnitrosurea, or nimustine (ACNU); (ix) anthracyclines, anthracenediones, and related drugs, such as daunorubicin, doxorubicin (optionally in combination with dexrazoxane), liposomal formulations of doxorubicin (e.g., Caelyx™, Myocet™, Doxil™), idarubicin, mitoxantrone, epirubicin, amsacrine, or valrubicin; (x) Epothilones, such as ixabepilone, patupilone, BMS-310705, KOS-862 and ZK-EPO, epothilone A, epothilone B, desoxyepothilone B (also known as epothilone D or KOS-862), azaepothilone B (also known as BMS-247550), aulimalide, isolaulimalide, or luetherobin;

[0135] (xi) DNA methyltransferase inhibitors, such as temozolomide, azacitidine or decitabine, or SGI-110; (xii) antifolates, such as methotrexate, pemetrexed disodium, or raltitrexed; (xiii) cytotoxic antibiotics, such as antinomycin D, bleomycin, mitomycin C, dactinomycin, carminomycin, daunomycin, levamisole, plicamycin, or mithramycin; (xiv) tubulin binding agents, such as combrestatin, colchicine, or nocodazole; (xv) Signal transduction inhibitors, for example, kinase inhibitors (e.g., EGFR (epidermal growth factor receptor) inhibitors, VEGFR (vascular endothelial growth factor receptor) inhibitors, PDGFR (platelet-derived growth factor receptor) inhibitors, MTKI (multi-targeted kinase inhibitors), Raf inhibitors, mTOR inhibitors, for example, imatinib mesylate, erlotinib, gefitinib, dasatinib, lapatinib, dovotinib, axitinib, nilotinib, vandetanib, vatalinib, pazopanib, sorafenib, sunitinib, temsirolimus, everolimus (RAD 001), vemurafenib (PLX4032 / RG7204), dabrafenib, encorafenib, or IκB kinase inhibitors such as SAR-113945, bardoxolone, BMS-066, BMS-345541, IMD-0354, IMD-2560, or IMD-1041, or MEK inhibitors such as selumetinib (AZD6244) and trametinib (GSK121120212); (xvi) Aurora kinase inhibitors, such as AT9283, barasertib (AZD1152), TAK-901, MK0457 (VX680), cenisertib (R-763), danusertib (PHA-739358), alisertib (MLN-8237), or MP-470; (xvii) CDK inhibitors, such as AT7519, roscovitine, seliciclib, alvocidib (flavopiridol), dinaciclib (SCH-727965), 7-hydroxy-staurosporine (UCN-01), JNJ-7706621, BMS-387032 (also known as SNS-032), PHA533533, PD332991, ZK-304709, or AZD-5438; (xviii) PKA / B inhibitors and PKB (akt) pathway inhibitors, for example, AKT inhibitors, for example KRX-0401 (Perifosine / NSC 639966), ipatasertib (GDC-0068; RG-7440), afuresertib (GSK-2110183; 2110183), MK-2206, MK-8156, AT13148, AZD-5363, triciribine phosphate (VQD-002; triciribine phosphate monohydrate (API-2; TCN-P; TCN-PM; VD-0002), RX-0201, NL-71-101, SR-13668, PX-316, AT13148, AZ-5363, semaphore, SF1126, or enzastaurin hydrochloride (LY317615), or an MTOR inhibitor, such as a rapamycin analogue, for example, RAD 001 (everolimus), CCI 779 (temsirolemus), AP23573 and ridaforolimus, sirolimus (originally known as rapamycin), AP23841 and AP23573, calmodulin inhibitors such as CBP-501 (forkhead translocation inhibitor), enzastaurin hydrochloride (LY317615), or PI3K inhibitors such as dactolisib (BEZ235) , buparlisib (BKM-120; NVP-BKM-120), BYL719, copanlisib (BAY-80-6946), ZSTK-474, CUDC-907, apitolisib (GDC-0980; RG-7422), pictilisib (pictorelisib, GDC-0941, RG-7321), GDC-0032, GDC-0068, GSK-2636771, idelalisib (formerly CAL-101, GS 1101, GS-1101), MLN1117 (INK1117), MLN0128 (INK128), IPI-145 (INK1197), LY-3023414, ipatasertib, afuresertib, MK-2206, MK-8156, LY-3023414, LY294002, SF1126 or PI-103, or sonolisib (PX-866); (xix) Hsp90 inhibitors, such as AT13387, herbimycin, geldanamycin (GA), 17-allylamino-17-desmethoxygeldanamycin (17-AAG), such as NSC-330507, Kos-953, and CNF-1010, 17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride (17-DMAG), such as NSC-707545 and Kos-1022, NVP-AUY922 (VER-52296), NVP-BEP800, CNF-2024 (BIIB-021, oral purine), ganetespib (STA-9090), SNX-5422 (SC-102112), or IPI-504; (xx) Monoclonal antibodies (unconjugated or conjugated to radioisotopes, toxins, or other drugs), antibody derivatives and related drugs, such as anti-CD, anti-VEGFR, anti-HER2, anti-CTLA4, anti-PD-1, or anti-EGFR antibodies, such as rituximab (CD20), ofatumumab (CD20), ibritumomab tiuxetan (CD20), GA101 (CD20), tositumomab (CD20), epratuzumab (CD22). , lintuzumab (CD33), gemtuzumab ozogamicin (CD33), alemtuzumab (CD52), galiximab (CD80), trastuzumab (HER2 antibody), pertuzumab (HER2), trastuzumab-DM1 (HER2), ertumaxomab (HER2 and CD3), cetuximab (EGFR), panitumumab (EGFR), necitumumab (EGFR), nimotuzumab (EGFR), bevacizumab (VEGF), catumaxumab ( mab) (EpCAM and CD3), abagovomab (CA125), farletuzumab (folate receptor), elotuzumab (CS1), denosumab (RANK ligand), figitumumab (IGF1R), CP751,871 (IGF1R), mapatumumab (TRAIL receptor), metMAB (met), mitumomab (GD3 ganglioside), naptumomab estafenatox (5T4), siltuximab (IL6), or immunomodulatory agents, e.g., CTLA-4 blockade Antibodies and / or antibodies against PD-1 or PD-L1 and / or PD-L2, such as ipilimumab (CTLA4), MK-3475 (pembrolizumab, formerly lambrolizumab, anti-PD-1), nivolumab (anti-PD-1), BMS-936559 (anti-PD-L1), MPDL320A, AMP-514 or MEDI4736 (anti-PD-L1), or tremelimumab (formerly ticilimumab, CP-675,206, anti-CTLA-4);

[0136] (xxi) estrogen receptor antagonists or selective estrogen receptor modulators (SERMs) or estrogen synthesis inhibitors, such as tamoxifen, fulvestrant, toremifene, droloxifene, faslodex, or raloxifene; (xxii) Aromatase inhibitors and related drugs, such as exemestane, anastrozole, letrazole, testolactone aminoglutethimide, mitotane, or vorozole; (xxiii) antiandrogens (i.e., androgen receptor antagonists) and related drugs, such as bicalutamide, nilutamide, flutamide, cyproterone, or ketoconazole; (xxiv) Hormones and their analogues, such as medroxyprogesterone, diethylstilbestrol (also known as diethylstilboestrol) or octreotide; (xxv) steroids, such as dromostanolone propionate, megestrol acetate, nandrolone (decanoate, phenpropionate), fluoxymestrone, or gossypol, (xxvi) steroidal cytochrome P450 17α-hydroxylase-17,20-lyase inhibitors (CYP17), such as abiraterone; (xxvii) gonadotropin-releasing hormone agonists or antagonists (GnRAs), such as abarelix, goserelin acetate, histrelin acetate, leuprolide acetate, triptorelin, buserelin, or deslorelin; (xxviii) Glucocorticoids, such as prednisone, prednisolone, dexamethasone; (xxix) differentiation inducers, such as retinoids, rexinoids, vitamin D, or retinoic acid, and retinoic acid metabolism blockers (RAMBAs), such as accutane, alitretinoin, bexarotene, or tretinoin; (xxx) Farnesyltransferase inhibitors, such as tipifarnib;

[0137] (xxxi) chromatin targeted therapy, e.g., histone deacetylase (HDAC) inhibitors, e.g., panobinostat, resminostat, abexinostat, vorinostat, romidepsin, belinostat, entinostat, xinostat, pracinostat, tefinostat, mocetinostat, gibinostat, CUDC-907, CUDC-101, ACY-1215, MGCD-290, EVP-0334, RG-2833, 4SC-202, romidepsin, AR-42 (Ohio State University), CG-200745, valproic acid, CKD-581, sodium butyrate, suberoylanilide hydroxamic acid (SAHA), depsipeptide (FR 901228), dacinostat (NVP-LAQ824), R306465 / JNJ-16241199, JNJ-26481585, trichostatin A, chlamydocin, A-173, JNJ-MGCD-0103, PXD-101, or apicidin; (xxxii) proteasome inhibitors, such as bortezomib, carfilzomib, delanzomib (CEP-18770), ixazomib (MLN-9708), oprozomib (ONX-0912), or marizomib; (xxxiii) photodynamic therapeutic agents, such as porfimer sodium or temoporfin; (xxxiv) Anticancer drugs derived from marine organisms, such as trabectidin, etc.; (xxxv) Radiolabeled drugs for radioimmunotherapy, e.g., those containing beta particle emitting isotopes (e.g., Iodine-131, Yttrium-90) or alpha particle emitting isotopes (e.g., Bismuth-213 or Actinium-225), such as Ibritumomab or Iodo-tositumomab; (xxxvi) telomerase inhibitors, such as telomestatin; (xxxvii) matrix metalloproteinase inhibitors, such as batimastat, marimastat, prinostat, or metastat; (xxxviii) recombinant interferons (such as interferon-gamma and interferon-alpha) and interleukins (such as interleukin 2), such as aldesleukin, denileukin diftitox, interferon alpha 2a, interferon alpha 2b, or pegylated interferon alpha 2b; (xxxix) selective immune response modifiers, such as thalidomide or lenalidomide; (xl) therapeutic vaccines, such as sipuleucel-T (Provenge) or OncoVex;

[0138] (xli) a cytokine activator, picibanil, romurtide, sizofiran, virulidin, or thymosin; (xlii) arsenic trioxide; (xliii) G protein-coupled receptor (GPCR) inhibitors, such as atrasentan; (xliv) an enzyme, such as L-asparaginase, pegaspargase, rasburicase, or pegademase; (xlv) DNA repair inhibitors, such as PARP inhibitors, for example, olaparib, velaparib, iniparib, rucaparib (AG-014699 or PF-01367338), talazoparib, or AG-014699; (xlvi) DNA damage response inhibitors, such as ATM inhibitors AZD0156 MS3541, ATR inhibitors AZD6738, M4344, M6620, wee1 inhibitors AZD1775; (xlvii) agonists of death receptors (e.g., TNF-related apoptosis-inducing ligand (TRAIL) receptors), such as mapatuzumab (formerly HGS-ETR1), conatumumab (formerly AMG 655), PRO95780, lexatumumab, dulanermin, CS-1008, apomab, or recombinant TRAIL ligands, such as recombinant human TRAIL / Apo2 ligand;

[0139] (xlviii) prophylactic (adjuvant) agents; i.e., agents that reduce or ameliorate some of the side effects associated with chemotherapeutic agents, e.g. -Antiemetics, - agents that prevent or shorten the duration of chemotherapy-associated neutropenia and prevent complications resulting from decreased levels of platelets, red blood cells, or white blood cells, such as interleukin-11 (e.g., oprelvekin), erythropoietin (EPO) and its analogs (e.g., darbepoetin alfa), colony-stimulating factor analogs, such as granulocyte-macrophage colony-stimulating factor (GM-CSF) (e.g., sargramostim), and granulocyte colony-stimulating factor (G-CSF) and its analogs (e.g., filgrastim, pegfilgrastim); - bone resorption inhibitors, such as denosumab or bisphosphonates, such as zoledronate, zoledronic acid, pamidronate, and ibandronate; - inflammatory response suppressants, such as dexamethasone, prednisone, and prednisolone; - Medications used to lower blood levels of growth hormone and IGF-I (and other hormones) in patients with acromegaly or other rare hormone-producing tumors, such as synthetic forms of the hormone somatostatin, e.g. octreotide acetate; -Antidote to drugs that lower folate levels, e.g. leucovorin or folinic acid, - pain medications, such as opiates, e.g. morphine, diamorphine, and fentanyl; nonsteroidal anti-inflammatory drugs (NSAIDs), such as COX-2 inhibitors, for example celecoxib, etoricoxib, and lumiracoxib; - Medications for mucositis, e.g. Palifermin, - Medications for treating side effects including anorexia, cachexia, edema, or thromoembolic events, such as megestrol acetate.

[0140] In one embodiment, the anticancer drugs include recombinant interferons (e.g., interferon-gamma and interferon-alpha), and interleukins (e.g., interleukin 2), such as aldesleukin, denileukin diftitox, interferon-alpha 2a, interferon-alpha 2b, or pegylated interferon-alpha 2b; interferon-alpha 2 (500 μg / ml), particularly interferon-beta; and signal transduction inhibitors, such as kinase inhibitors (e.g., E GFR (epidermal growth factor receptor) inhibitors, VEGFR (vascular endothelial growth factor receptor) inhibitors, PDGFR (platelet-derived growth factor receptor) inhibitors, MTKI (multi-target kinase inhibitors), Raf inhibitors, mTOR inhibitors, such as imatinib mesylate, erlotinib, gefitinib, dasatinib, lapatinib, dovotinib, axitinib, nilotinib, vandetanib, vatalinib, pazopanib, sorafenib, sunitinib, temsirolimus, everolimus (RAD 001), vemurafenib (PLX4032 / RG7204), dabrafenib, encorafenib, or IκB kinase inhibitors such as SAR-113945, bardoxolone, BMS-066, BMS-345541, IMD-0354, IMD-2560, or IMD-1041, or MEK inhibitors such as selumetinib (AZD6244) and trametinib (GSK121120212), in particular Raf inhibitors (e.g. vemurafenib) or MEK inhibitors (e.g. trametinib).

[0141] Each compound present in the combination of the present invention can be administered individually by different routes with various dosage schedules.Thus, the dosage of each of the two or more drugs can be different, and each of the two or more drugs can be administered simultaneously or at different times.Those skilled in the art will know by their general knowledge the dosage regimen and combination therapy to be used.For example, the compound of the present invention can be used in combination with one or more other drugs that are administered according to their existing combination regimen.An example of a standard combination regimen is shown below.

[0142] The taxane compound is advantageously administered at a dose of 50 to 400 mg per square meter of body surface area (mg / m 2 ), e.g. 75-250 mg / m 2 In particular, paclitaxel is at a dosage of about 175-250 mg / m 2 and docetaxel at about 75-150 mg / m 2 is administered at a dosage of

[0143] The camptothecin compound is advantageously administered at a dose of 0.1 to 400 mg per square meter of body surface area (mg / m 2 ), e.g. 1-300 mg / m 2 In particular, irinotecan is administered at a dose of approximately 100-350 mg / m 2 , and topotecan at about 1-2 mg / m 2 is administered at a dosage of

[0144] The antitumor podophyllotoxin derivative is advantageously administered at a dose of 30 to 300 mg per square meter of body surface (mg / m 2 ), e.g. 50-250 mg / m 2 In particular, etoposide is administered at a dose of about 35-100 mg / m 2 , and teniposide at about 50-250 mg / m 2 is administered at a dosage of

[0145] The antitumor vinca alkaloids are advantageously administered at a dose of 2 to 30 mg per square meter of body surface area (mg / m 2 ) at a dosage of about 3–12 mg / m 2 At a dosage of 1–2 mg / m 2 and vinorelbine at a dosage of about 10-30 mg / m 2 is administered at a dosage of

[0146] The antitumor nucleoside derivative is advantageously administered at a dose of 200 to 2500 mg per square meter of body surface area (mg / m 2 ), e.g. 700-1500 mg / m 2The dosage of 5-FU is 200-500 mg / m 2 At a dose of 100 mg / m, gemcitabine is approximately 800–1200 mg / m 2 and capecitabine at a dosage of about 1000-2500 mg / m 2 is administered at a dosage of

[0147] Alkylating agents such as nitrogen mustards or nitrosoureas are advantageously administered at a dose of 100 to 500 mg per square meter of body surface (mg / m) per course of treatment. 2 ), e.g., 120-200 mg / m 2 In particular, cyclophosphamide is administered at a dose of about 100-500 mg / m 2 Chlorambucil is administered at a dose of about 0.1-0.2 mg / kg, and carmustine is administered at a dose of about 150-200 mg / m 2 and lomustine at a dosage of about 100-150 mg / m 2 is administered at a dosage of

[0148] The antitumor anthracycline derivative is advantageously administered at a dose of 10 to 75 mg per square meter of body surface (mg / m 2 ), e.g., 15-60 mg / m 2 In particular, doxorubicin is administered at a dose of approximately 40-75 mg / m 2 At a dosage of 25-45 mg / m2, daunorubicin 2 and idarubicin at a dosage of about 10-15 mg / m 2 is administered at a dosage of

[0149] The antiestrogens are advantageously administered in a dosage of about 1-100 mg per day depending on the particular agent and the condition being treated. Tamoxifen is advantageously administered orally in a dosage of 5-50 mg, especially 10-20 mg, twice a day, the therapy being continued for a sufficient time to achieve and maintain a therapeutic effect. Toremifene is advantageously administered orally in a dosage of about 60 mg once a day, the therapy being continued for a sufficient time to achieve and maintain a therapeutic effect. Anastrozole is advantageously administered orally in a dosage of about 1 mg once a day. Droloxifene is advantageously administered orally in a dosage of about 20-100 mg once a day. Raloxifene is advantageously administered orally in a dosage of about 60 mg once a day. Exemestane is advantageously administered orally in a dosage of about 25 mg once a day.

[0150] The antibody is advantageously administered at a dose of about 1 to 5 mg per square meter of body surface area (mg / m 2 Trastuzumab is advantageously administered at a dosage of 1 to 5 mg per square meter of body surface area (mg / m) per course of treatment. 2 ), especially 2-4 mg / m 2 is administered at a dosage of

[0151] When the compound of formula (I) is administered in combination therapy with one, two, three, four or more other therapeutic agents (particularly one or two, more particularly one), the compounds may be administered simultaneously or sequentially. In the latter case, the two or more compounds may be administered for a period and in an amount and manner sufficient to ensure the achievement of a beneficial or synergistic effect. When administered sequentially, they may be administered at short intervals (e.g., between 5-10 minutes) or at longer intervals (e.g., 1, 2, 3, 4 hours or more apart, or even longer if necessary), with the exact dosing regimen being commensurate with the properties of the therapeutic agent(s). These doses may be administered, for example, once, twice or more times per course of treatment, which may be repeated, for example, every 7, 14, 21 or 28 days.

[0152] In one embodiment, there is provided a compound of formula (I) for the manufacture of a medicament for use in therapy, wherein the compound is used in combination with one, two, three, or four other therapeutic agents. In another embodiment, there is provided a medicament for the treatment of cancer comprising a compound of formula (I), wherein the medicament is used in combination with one, two, three, or four other therapeutic agents. The invention further provides the use of a compound of formula (I) for the manufacture of a medicament for enhancing or potentiating the response rate in a patient suffering from cancer, wherein the patient is being treated with one, two, three, or four other therapeutic agents.

[0153] It will be appreciated that the particular method and sequence of administration of each component of the combination, and their respective dosage amounts, and treatment regimens may depend on the particular other medicinal agents and compounds of the invention being administered, their routes of administration, the particular tumor being treated, and the particular host being treated. Optimal method and sequence of administration, and dosage amounts, and treatment regimens can be readily determined by one of skill in the art using conventional methods and in view of the information presented herein.

[0154] When administered in combination, the weight ratio of the compound of the present invention and one or more other anticancer drug(s) can be determined by one of skill in the art. The ratio and the exact dosage and frequency of administration will depend on the specific compound of the present invention and other anticancer drug(s) used, the specific condition being treated, the severity of the condition being treated, the age, weight, sex, diet, time of administration, and general health of the particular patient, the mode of administration, and other therapeutic agents that the individual patient may be taking, as will be well known to those of skill in the art. Furthermore, it will be apparent that the effective daily amount may be decreased or increased depending on the response of the subject being treated and / or depending on the judgment of the physician prescribing the compound of the present invention. The specific weight ratio of the compound of formula (I) and another anticancer drug may range from 1 / 10 to 10 / 1, more particularly from 1 / 5 to 5 / 1, and even more particularly from 1 / 3 to 3 / 1.

[0155] The compounds of the invention may also be administered in conjunction with non-chemotherapeutic treatments, such as radiation therapy, photodynamic therapy, gene therapy; surgery and dietary restrictions.

[0156] The compounds of the invention also have therapeutic applications in sensitizing tumor cells to radiation and chemotherapy. Thus, the compounds of the invention can be used as "radiosensitizers" and / or "chemosensitizers" or can be administered in combination with other "radiosensitizers" and / or "chemosensitizers". In one embodiment, the compounds of the invention are used as chemosensitizers.

[0157] The term "radiosensitizer" is defined as a molecule that is administered to a patient in a therapeutically effective amount to increase the sensitivity of cells to ionizing radiation and / or to facilitate the treatment of a disease treatable with ionizing radiation.

[0158] The term "chemosensitizer" is defined as a molecule that is administered to a patient in a therapeutically effective amount to increase the sensitivity of cells to chemotherapy and / or to facilitate the treatment of a chemotherapy-treatable disease.

[0159] In one embodiment, the compounds of the invention are administered in combination with a "radiosensitizer" and / or a "chemosensitizer." In one embodiment, the compounds of the invention are administered in combination with an "immunosensitizer."

[0160] The term "immunosensitizer" is defined as a molecule that is administered to a patient in a therapeutically effective amount to increase the sensitivity of cells to Pol theta inhibitors.

[0161] Many cancer treatment protocols currently employ radiosensitizers in conjunction with X-ray irradiation. Examples of X-ray activated radiosensitizers include, but are not limited to: metronidazole, misonidazole, desmethylmisonidazole, pimonidazole, etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, EO9, RB 6145, nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cisplatin, and their therapeutically effective analogs and derivatives.

[0162] Photodynamic therapy (PDT) of cancer employs visible light as the radiation activator of a sensitizer. Examples of photodynamic radiosensitizers include, but are not limited to: hematoporphyrin derivatives, photofrin, benzoporphyrin derivatives, tin etioporphyrin, pheoborbide-a, bacteriochlorophyll-a, naphthalocyanines, phthalocyanines, zinc phthalocyanines, and their therapeutically effective analogs and derivatives.

[0163] The radiosensitizer may be administered with a therapeutically effective amount of one or more other compounds, including, but not limited to: a compound of the invention; a compound that enhances uptake of the radiosensitizer into target cells; a compound that controls the flux of therapeutic substances, nutrients, and / or oxygen to target cells; a chemotherapeutic agent that acts against the tumor with or without additional radiation; or another compound that is therapeutically effective for treating cancer or other diseases.

[0164] The chemotherapy sensitizer may be administered with a therapeutically effective amount of one or more other compounds, including, but not limited to, the compounds of the present invention; compounds that promote the incorporation of the chemotherapy sensitizer into target cells; compounds that control the flow of therapeutic substances, nutrients, and / or oxygen to target cells; chemotherapy agents acting against tumors, or other compounds therapeutically effective for treating cancer or other diseases. Calcium antagonists, such as verapamil, have been found to be useful in combination with anti-neoplastic agents to establish chemotherapy sensitivity in tumor cells resistant to accepted chemotherapy agents and to enhance the effectiveness of the compounds in drug-sensitive malignancies.

[0165] Examples of immune sensitizers include, but are not limited to: immunomodulators, such as monoclonal antibodies, such as immune checkpoint antibodies (e.g., CTLA-4 blocking antibodies and / or antibodies against PD-1 and PD-L1 and / or PD-L2, such as ipilimumab (CTLA4), MK-3475 (pembrolizumab, formerly lambrolizumab, anti-PD-1), nivolumab (anti-PD-1), BMS-936559 (anti-PD-L1), MPDL320A, AMP-514, or MEDI4736 (anti-PD-L1), or tremelimumab (formerly ticilimumab, CP-675,206, anti-CTLA-4)]; or signal transduction inhibitors; or cytokines (e.g., recombinant interferons, etc.); or oncolytic viruses; or immune adjuvants (e.g., BCG).

[0166] The immunosensitizer may be administered with a therapeutically effective amount of one or more other compounds, including, but not limited to: a compound of the invention; a compound that promotes incorporation of the immunosensitizer into target cells; a compound that controls the flow of therapeutic substances, nutrients, and / or oxygen to target cells; a therapeutic agent that acts on a tumor, or other compound that is therapeutically effective for treating cancer or other diseases.

[0167] For use in combination therapy with another chemotherapeutic agent, the compound of formula (I) and one, two, three, four or more other therapeutic agents can be formulated together, for example, in a dosage form containing two, three, four or more therapeutic agents, i.e., in a unitary pharmaceutical composition containing all agents. In another embodiment, the individual therapeutic agents may be formulated separately and provided together in the form of a kit, optionally including instructions for their use.

[0168] In one embodiment, there is provided a combination of a compound of Formula (I) with one or more (e.g., one or two) other therapeutic agents (e.g., anticancer agents as described above). In a further embodiment, there is provided a combination of a Pol θ inhibitor as described herein and a PI3K / AKT pathway inhibitor selected from: apitolisib, buparlisib, copanlisib, pictilisib, ZSTK-474, CUDC-907, GSK-2636771, LY-3023414, ipatasertib, afuresertib, MK-2206, MK-8156, idelalisib, BEZ235 (dactolisib), BYL719, GDC-0980, GDC-0941, GDC-0032, and GDC-0068.

[0169] In another embodiment, there is provided a compound of Formula (I) in combination with one or more (e.g., one or two) other therapeutic agents (e.g., anti-cancer agents) for use in therapy, e.g., the prevention or treatment of cancer.

[0170] In one embodiment, a pharmaceutical composition comprises a compound of formula (I) together with a pharma- ceutically acceptable carrier and, optionally, one or more therapeutic agent(s).

[0171] In another embodiment the invention relates to the use of a COMBINATION OF THE PRESENT EMBODIMENT in the manufacture of a pharmaceutical composition for inhibiting the growth of tumor cells.

[0172] In a further embodiment, the present invention relates to a product comprising a compound of formula (I) and one or more anti-cancer agents as a combined preparation for simultaneous, separate or sequential use in the treatment of patients suffering from cancer. EXAMPLES

[0173] (Example) The invention will now be described with reference to specific embodiments described in the following examples, but the invention is not limited thereto.

[0174] (abbreviation) [Table 1]

[0175] (Typical Preparative HPLC Method) For purification of intermediates by HPLC, the following columns were typically used: SunFire C18, Xtimate C18, Phenomenex Gemini, Phenomenex Synergi C18, Phenomenex Luna, Waters Xbridge C18, Boston Prime C18, and Shim-pack C18. The mobile phase used was typically water and MeCN with either acidic or basic additives, e.g. formic acid (0.1% (v / v)) or ammonium hydroxide (0.05% (v / v)). A typical procedure started with 95% water:5% MeCN and reduced the polarity ratio of water and MeCN over 5-12 minutes, typically at a flow rate of 25 mL / min. The mass spectrometer typically used for mass detection HPLC was a Waters 3100, detecting masses from 100 to 700 g / mol.

[0176] Example 1 (3aR,11aS)-6,10-Dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-2,11(3H)-dione [ka] TIFF2024542949000005.tif229170

[0177] (step i.) To a mixture of 2-chloro-6-nitroaniline (20.0 g, 116 mmol), triethylamine (23.4 g, 231 mmol), and 4-dimethylaminopyridine (1.42 g, 11.5 mmol) in DCM (200 mL) was added di-tert-butyl dicarbonate (55.6 g, 254 mmol). The reaction mixture was stirred at 25 °C for 16 h. After completion, the mixture was diluted with water (300 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with saturated aqueous citric acid (100 mL), brine (3 x 300 mL), dried over Na2SO4, and evaporated to give tert-butyl (tert-butoxycarbonyl) (2-chloro-6-nitrophenyl)carbamate (43.0 g, 99% yield) as a yellow solid, which was used without purification. [ka]

[0178] (Step ii.) To a solution of tert-butyl (tert-butoxycarbonyl) (2-chloro-6-nitrophenyl)carbamate (53.0 g, 142 mmol) in THF (500 mL) was added 5% Pt-V / C (5 g). The reaction mixture was stirred at 20 °C for 20 h under H2 atmosphere (15 psi). Upon completion, the reaction was filtered and the filtrate was evaporated to give tert-butyl (2-amino-6-chlorophenyl) (tert-butoxycarbonyl)carbamate (48.3 g, 99% yield) as a white solid, which was used without further purification. [ka]

[0179] (step iii.) A mixture of tert-butyl (2-amino-6-chlorophenyl)(tert-butoxycarbonyl)carbamate (46.5 g, 135 mmol) and K2CO3 (37.4 g, 271 mmol) in MeOH (500 mL) was stirred at 60 °C for 3 h. After completion, the mixture was evaporated to remove MeOH, diluted with water (300 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over Na2SO4 and evaporated to give tert-butyl (2-amino-6-chlorophenyl)carbamate (32.3 g, 98% yield) as a yellow solid, which was used without further purification. [ka]

[0180] (step iv.) To a solution of sodium methoxide (33.3 g, 618 mmol) in MeOH (150 mL) was added tert-butyl (2-amino-6-chlorophenyl)carbamate (15.0 g, 61.8 mmol) and paraformaldehyde (2.78 g, 92.7 mmol). The mixture was stirred at 0 °C for 12 h, after which NaBH4 (11.6 g, 309 mmol) was added and the reaction mixture was stirred at 20 °C for an additional 2 h. After completion, the mixture was quenched with water (20 mL) and evaporated to remove MeOH. The aqueous mixture was diluted with water (200 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3×200 mL), dried over Na2SO4, and purified by column chromatography (PE / EtOAc=5 / 1) to give tert-butyl (2-chloro-6-(methylamino)phenyl)carbamate (7.8 g, 49% yield) as a white solid. [ka]

[0181] (step v.) To a solution of tert-butyl 2-((diphenylmethylene)amino)acetate (CAS: 81477-94-3; 100 g, 339 mmol) and dimethyl fumarate (CAS: 624-49-7; 73.2 g, 508 mmol) in EtOAc (1 L) was added (S)-2-((2,3-bis(dicyclohexylamino)cycloprop-2-en-1-ylidene)amino)propan-1-ol (CAS: 1808186-23-3, prepared as described in JS Bander et al., Chem. Sci. 2015, 6, 1537; 18.5 g, 33.9 mmol). The reaction mixture was stirred at room temperature for 48 h. After completion, the reaction mixture was evaporated and the crude product was purified by column chromatography (hexane / EtOAc=4 / 1) to give 1-(tert-butyl) 2,3-dimethyl(1S,2S)-1-((diphenylmethylene)amino)propane-1,2,3-tricarboxylate as a colorless oil (140 g, 94% yield).

[0182] (step vi.) To a solution of 1-(tert-butyl) 2,3-dimethyl(1S,2S)-1-((diphenylmethylene)amino)-propane-1,2,3-tricarboxylate (140 g, 318.5 mmol) in THF (1.4 L) was added 15% (w / w) aqueous citric acid (1.4 L). The reaction mixture was stirred at room temperature for 48 h. After completion, the reaction was quenched with saturated aqueous NaHCO3 and extracted with EtOAc (3×1 L). The combined organic layers were dried over Na2SO4 and evaporated. The crude product was purified by column chromatography (hexane / EtOAc=3 / 7) to give 2-(tert-butyl) 3-methyl(2S,3S)-5-oxopyrrolidine-2,3-dicarboxylate as a white solid (70 g, 90% yield). [ka]

[0183] (Step vii.) A mixture of 2-(tert-butyl)3-methyl(2S,3S)-5-oxopyrrolidine-2,3-dicarboxylate (40.0 g, 164 mmol) and TFA (250 mL) was stirred at 25 °C for 12 h under N2 atmosphere. After completion, the reaction mixture was evaporated. The residue was triturated with (PE / EtOAc = 10 / 1) for 60 min. The suspension was filtered to give (2S,3S)-3-(methoxycarbonyl)-5-oxopyrrolidine-2-carboxylic acid (27.0 g, 87% yield) as a white solid. [ka]

[0184] (Step viii.) To a solution of (2S,3S)-3-(methoxycarbonyl)-5-oxopyrrolidine-2-carboxylic acid (3.64 g, 19.4 mmol) in MeCN (40 mL) was added Ghosez's reagent (CAS: 26189-59-3; 3.12 g, 23.3 mmol) at 0° C. The mixture was stirred at 20° C. for 1 h, after which the mixture was added dropwise to a mixture of tert-butyl (2-chloro-6-(methylamino)phenyl)carbamate (prepared as described in step iv; 5 g, 19.4 mmol) and N,N-dimethylpyridin-2-amine (4.76 g, 38.9 mmol) in MeCN (40 mL) at 0° C. The reaction mixture was stirred at 0° C. for 30 min. After completion, the mixture was diluted with water (20 mL) and evaporated to remove MeCN. The residue was further diluted with water (60 mL) and extracted with EtOAc (3×80 mL). The combined layers were washed with brine (200 mL), dried over Na2SO4, evaporated and purified by column chromatography (EtOAc) to give methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-5-oxopyrrolidine-3-carboxylate (6.5 g, 71% yield) as a yellow solid. m / z ES+ [M+H] + 425.9.

[0185] (step ix.) A mixture of methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-5-oxopyrrolidine-3-carboxylate (11.4 g, 26.8 mmol), 2-bromo-6-methyl-4-(trifluoromethyl)pyridine (CAS: 451459-17-9; 8.35 g, 34.8 mmol), Pd2(dba)3 (2.45 g, 2.7 mmol), Xantphos (3.10 g, 5.4 mmol), and K2CO3 (11.1 g, 80.3 mmol) in 1,4-dioxane (120 mL) was degassed and purged with N2 three times. The reaction mixture was stirred at 100 °C for 3 h under N2 atmosphere. After completion, the mixture was evaporated, diluted with water (300 mL) and extracted with EtOAc (3×350 mL). The combined organic layers were washed with brine (600 mL), dried over Na2SO4, evaporated and purified by column chromatography (PE / EtOAc=3 / 1) to give methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-3-carboxylate (11.0 g, 66% yield) as a yellow solid. [ka]

[0186] (Step x.) To a solution of methyl (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-3-carboxylate (19.3 g, 32.9 mmol) in THF (200 mL) and MeOH (20 mL) was added NaBH4 (2.50 g, 65.9 mmol) in small portions at 0 °C. The reaction mixture was stirred at 25 °C for 1 h. After completion, the mixture was slowly quenched with saturated aqueous NH4Cl (150 mL) and the aqueous mixture was stirred for 30 min. The mixture was evaporated, diluted with water (100 mL) and extracted with EtOAc (3 x 250 mL). The combined organic layers were washed with brine (600 mL), dried over Na2SO4 and evaporated to give tert-butyl (2-chloro-6-((2S,3S)-3-(hydroxymethyl)-N-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)phenyl)carbamate (18.3 g, crude) as a brown solid which was used without further purification. m / z ES+ [M+H] + 557.1.

[0187] (step xi.) To a solution of tert-butyl (2-chloro-6-((2S,3S)-3-(hydroxymethyl)-N-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)phenyl)carbamate (18.3 g, 32.8 mmol) in DCM (200 mL) was added triethylamine (13.3 g, 131 mmol) followed by dropwise addition of methanesulfonyl chloride (6.02 g, 52.5 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. After completion, the mixture was quenched with water (200 mL). The layers were separated and the aqueous layer was further extracted with EtOAc (2 x 300 mL). The combined organic layers were washed with brine (800 mL), dried over Na2SO4 and evaporated to give ((2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidin-3-yl)methyl methanesulfonate (20.5 g, crude) as a brown solid which was used without further purification. m / z ES+ [M+H] + 635.2.

[0188] (step xii.) To a solution of ((2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-oxopyrrolidin-3-yl)methyl methanesulfonate (20.5 g, 32.2 mmol) in N-methyl-2-pyrrolidone (250 mL) was added K3PO4 (20.5 g, 96.8 mmol). The reaction mixture was stirred at 60 °C for 12 h. After completion, the mixture was diluted with water (1200 mL) and extracted with EtOAc (3 x 800 mL). The combined organic layers were washed with water (2×1000 mL), brine (2×1000 mL), dried over Na2SO4, evaporated, and purified by column chromatography (PE / EtOAc=3 / 1) to give tert-butyl (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-5-carboxylate (14.0 g, 76% yield) as a yellow solid. [ka]

[0189] (Step xiii.) To a solution of tert-butyl (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-5-carboxylate (13.5 g, 25.0 mmol) in DCM (120 mL) was added TFA (36.9 g, 324 mmol). The reaction mixture was stirred at 20 °C for 5 h. After completion, the mixture was evaporated, the residue was diluted with DCM (100 mL) and basified to pH 8 with saturated aqueous NaHCO3. The layers were separated and the aqueous layer was further extracted with EtOAc (2 x 150 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, and evaporated to give (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (10.9 g, crude) as a yellow solid, which was used without further purification. [ka]

[0190] (step xiv.) To a mixture of (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-2,11(3H)-dione (11.3 g, 25.7 mmol), Na2CO3 (8.19 g, 77.2 mmol), and TBAB (830 mg, 2.58 mmol) in DMF (110 mL) was added allyl bromide (15.5 g, 128 mmol). The reaction mixture was stirred at 100 °C for 12 h. After completion, the mixture was diluted with water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with water (2×150 mL), brine (2×100 mL), dried over Na2SO4, evaporated, and purified by column chromatography (PE / EtOAc=3 / 1) to give (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (12.0 g, 97% yield) as an off-white solid. [ka]

[0191] (step xv.) To a mixture of (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (4.45 g, 9.29 mmol) and NaIO4 (5.96 g, 27.8 mmol) in THF (50 mL) and water (10 mL) was added OsO4 (236 mg, 0.93 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. After completion, the reaction mixture was quenched with saturated aqueous Na2S2O3 (50 mL) and stirred at 25 °C for an additional 30 min. The mixture was diluted with water (200 mL) and extracted with EtOAc (3×100 mL). The organic layer was washed with saturated aqueous Na2S2O3 (2×100 mL), saturated aqueous NaHCO3 (2×100 mL), and brine (3×100 mL). The organic layer was dried over Na2SO4 and evaporated to give 2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetaldehyde (4.4 g, 98% yield) as a yellow solid. [ka]

[0192] (step xvi.) To a mixture of 2-((3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-5-yl)acetaldehyde (50 mg, 0.10 mmol), 1-methylpiperazine (11 mg, 0.11 mmol), and 4Å molecular sieves (10 mg) in MeOH (2 mL) was added acetic acid (31 mg, 0.52 mmol). The mixture was stirred at 20 °C for 30 min, after which NaBH3CN (13 mg, 0.21 mmol) was added. The reaction mixture was stirred at 20 °C for another 30 min. After completion, the reaction mixture was quenched with water (0.1 mL) and filtered. The filtrate was evaporated and purified by preparative HPLC to give (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (29 mg, 46% yield) as an off-white solid. [ka]

[0193] (step xvii.) A mixture of (3aR,11aS)-6-chloro-10-methyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocin-2,11(3H)-dione (3.5 g, 6.19 mmol), methylboronic acid (11.1 g, 185 mmol), XPhos-Pd-G2 (974 mg, 1.24 mmol), and Cs2CO3 (6.05 g, 18.5 mmol) in toluene (55 mL) was degassed and purged with N2 three times. The reaction mixture was stirred at 110 °C for 12 h under N2 atmosphere. The mixture was filtered, and the filtrate was evaporated. The residue was purified by column chromatography (Al2O3, EtOAc / MeOH=15 / 1) followed by reversed-phase flash chromatography (water (0.1% NH4OH) / MeCN) to give (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (5.0 g, 74% yield) as a white solid. [ka]

[0194] (Biological Data) (Polθ polymerase domain WT K I Assay) Using the PicoGreen assay, we identified compounds that reversibly inhibited in vitro Polθ activity. I The values ​​were measured.

[0195] Human Pol θ polymerase domain (amino acids (aa) 1820-2590) was expressed in E. coli, purified, aliquoted, and stored at -80°C until needed. Pol θ substrate was made from a 1.2:1 ratio mixture of DNA II Short to DNA II Long, with a final concentration of 20 mM substrate in annealing buffer (20 mM Tris pH 7.5, 50 mM NaCl). Substrate was heated in 50 mL aliquots to 95°C in a heating block for 5 minutes, after which the heating block was switched off and the reaction was cooled to room temperature and stored at -20°C until needed. [Table 2]

[0196] Assay measurements were performed in 1x buffer containing 25 mM Tris pH 7.5, 12.5 mM NaCl, 0.5 mM NaCl, 5% (v / v) glycerol, 0.01% (v / v) Triton X-100, 0.1 mg / ml BSA, 1 mM DTT. Test compounds were prepared by dilution in 100% DMSO to give intermediate stocks of 12 μM each (100x final highest concentration). 100 nL of 23x 1:1.5-fold serial dilutions and DMSO only controls were dispensed into a Greiner 384-well black low volume plate (product code 784076) using a Tecan dispenser. DMSO concentration was maintained at 1% of the final assay volume by backfilling with DMSO.

[0197] 2x working stocks of substrates (200 nM DNA substrate and 100 μM dNTPs) and enzyme (0.312 nM Polθ) were made in assay buffer. 5 μL / well of the 2x solutions of both enzyme and substrate were dispensed into the compound pre-dispensed assay plate using a Tempest dispenser (Formulatrix) to give final assay concentrations of 100 nM DNA substrate, 50 μM dNTPs, and 0.156 nM Polθ. To stop the reaction, 5 μL of a solution containing 25 mM Tris-HCl pH 7.5 and 20 mM EDTA was added at six time points (t=0, 15, 30, 60, 90, 120, 150, 180 min) using the Tempest time delay function. The plate was covered during the time course to prevent evaporation. After the assay was completed, 5 μL of detection reagent (25 mM Tris-HCl pH 7.5, and 2.5% (v / v) PicoGreen) was dispensed into the wells using a Tempest liquid handler (Formulatrix), and the plate was then read on a CLARIOstar Plus (BMG Labtech) using default optical settings for fluorescein and autogain / focus settings.

[0198] All data analyses were performed using GraphPad Prism V.8 (GraphPad Software Inc, San Diego, CA). The time course data for each inhibitor concentration were fitted to a linear regression model in GraphPad Prism. All time points where the control (DMSO only) response was not linear were excluded from the analysis. The initial rates (slope) obtained from the linear regression were then plotted against inhibitor concentration and fitted to a slope (4-parameter) model of inhibitor vs. response variables in GraphPad Prism to obtain the K I value was determined.

[0199] The compound of Example 1 was used to treat the Pol θ polymerase domain WT K I The assay was tested and the results are shown in the table below. [Table 3]

Claims

1. Compounds of formula (I): [Formula 1] or a tautomeric or stereochemically isomeric form, a pharma- ceutically acceptable salt, or a solvate thereof.

2. 2. The compound according to claim 1, which is the free base of the compound of formula (I), which is (3aR,11aS)-6,10-dimethyl-1-(6-methyl-4-(trifluoromethyl)pyridin-2-yl)-5-(2-(4-methylpiperazin-1-yl)ethyl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazocine-2,11(3H)-dione (E1).

3. A pharmaceutical composition comprising a compound of formula (I) according to claim 1 or 2.

4. 10. A pharmaceutical composition comprising a compound of formula (I) as defined in claim 1 or 2 in combination with one or more therapeutic agents.

5. 3. A compound according to claim 1 or 2 for use in therapy.

6. 3. A compound according to claim 1 or 2 for use in the prevention or treatment of cancer.

7. A method for preparing a compound of formula (I) according to claim 1, comprising the steps of: (a) a compound of formula (II): [chemical 2] Interconversion to a compound of formula (I); and (b) the optional formation of a pharma- ceutically acceptable salt of a compound of formula (I). The manufacturing method comprising the steps of:

Images