Thiadiazolyl derivatives as DNA polymerase theta inhibitors and their uses

Abstract

Disclosed herein are compounds of Formula (I) that inhibit DNA polymerase theta (Pol θ) activity, particularly by inhibiting the activity of the ATP-dependent helicase domain of Pol θ. Also disclosed are pharmaceutical compositions containing such compounds and methods for treating and / or preventing diseases treatable by inhibiting Pol θ, such as cancers, including homologous recombination (HR)-deficient cancers. [C1] JPEG2025541823000057.jpg36170

Description

[Background technology]

[0001] Targeting DNA repair deficiencies has become a proven and effective strategy in cancer therapy. However, DNA repair-deficient cancers often become dependent on backup DNA repair pathways, presenting a "weak spot" that can be targeted for cancer cell elimination and the basis for synthetic lethality. Synthetic lethality is exemplified by the success of poly(ADP-ribose) polymerase (PARP) inhibitors in the treatment of BRCA-deficient breast and ovarian cancers (Audeh MW et al., Lancet (2010); 376(9737):245-51).

[0002] DNA damage repair processes are important for genome maintenance and stability, and double-stranded strand breaks (DSBs) are primarily repaired by the non-homologous end joining (NHEJ) pathway during the G1 phase of the cell cycle and homologous recombination (HR) during the S-G2 phase. The lesser-regarded alternative end joining (alt-EJ), also known as microhomology-mediated end joining (MMEJ), is typically considered a "backup" DSB repair pathway when NHEJ or HR are impaired. Studies of multiple genes have highlighted a role for DNA polymerase theta (Polθ, encoded by POLQ) in stimulating MMEJ in higher organisms (Chan SH et al., PLoS Genet. (2010); 6: e1001005; Roerink SF et al., Genome research. (2014); 24: 954-962; Ceccaldi R. et al., Nature (2015); 518: 258-62; and Mateos-Gomez PA et al., Nature (2015); 518: 254-57).

[0003] Polθ is unique among human DNA polymerases in that it exhibits not only a C-terminal DNA polymerase domain but also an N-terminal helicase domain separated by a long, poorly conserved central domain whose function after Rad51 binding is unknown (Seki et al., 2003; Shima et al., 2003; Yousefzadeh and Wood, 2013). The N-terminal ATPase / helicase domain belongs to the HELQ class of the SF2 helicase superfamily. In homologous recombination-deficient (HRD) cells, Polθ can direct error-prone DNA synthesis at DNA damage sites via the alt-EJ pathway. The helicase domain of Polθ has been shown to suppress the HR pathway through disruption of the Rad51 nucleoprotein complex formation, which is involved in the initiation of HR-dependent DNA repair reactions after ionizing radiation. This anti-recombinase activity of Polθ promotes the alt-EJ pathway. Furthermore, the helicase domain of Polθ contributes to microhomology-mediated strand annealing (Chan SH et al., PLoS Genet. (2010); 6: e1001005; and Kawamura K et al., Int. J. Cancer (2004); 109: 9-16). Polθ uses this annealing activity to efficiently promote end-joining in the alt-EJ pathway when ssDNA overhangs contain microhomologies greater than 2 bp (Kent T. et al., Elife (2016); 5: e13740; and Kent T. et al., Nat. Struct. Mol. Biol. (2015); 22: 230-237). This reannealing activity is achieved through the combined action of Rad51 interaction and subsequent ATPase-mediated displacement of Rad51 from DSB damage sites. Once annealed, the primer strand of DNA can be extended by the polymerase domain of Pol θ.

[0004] Pol θ expression is nearly absent in normal cells but is upregulated in breast, lung, and ovarian cancers (Ceccaldi R. et al., Nature (2015); Vol. 518, pp. 258-62). Furthermore, increased Pol θ expression correlates with poor prognosis in breast cancer (Lemee F et al., Proc Natl Acad Sci USA. (2010); Vol. 107: 13390-5). Cancer cells exhibiting deficiencies in HR, NHEJ, or ATM have been shown to be highly dependent on Pol θ expression (Ceccaldi R. et al., Nature (2015); Vol. 518: pp. 258-62; Mateos-Gomez PA et al., Nature (2015); Vol. 518: pp. 254-57; and Wyatt DW et al., Mol. Cell (2016); Vol. 63: pp. 662-73). Pol θ is therefore an attractive target for novel synthetic lethal therapies in cancers that contain DNA repair defects. Summary of the Invention

[0005] Disclosed herein are certain thiadiazolyl derivatives that inhibit Pol θ activity, particularly by inhibiting the ATP-dependent helicase domain activity of Pol θ. Also disclosed are pharmaceutical compositions containing such compounds and methods for treating and / or preventing diseases treatable by inhibiting Pol θ, such as cancers, including homologous recombination (HR)-deficient cancers.

[0006] In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0007] [ka] (In the formula, X, R 1 , R 2 , R 3 , and n has the meaning provided herein below).

[0008] In a related embodiment, a pharmaceutical composition is provided comprising a compound of Formula (I) or Table 1, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0009] In another aspect, provided is a method for treating and / or preventing a disease characterized by overexpression of Pol θ in a patient, comprising administering to the patient a therapeutically effective amount of a compound of Formula (I) or Table 1, or a pharmaceutically acceptable salt thereof (or an embodiment thereof disclosed herein). In one embodiment, the patient is in need of such treatment. In another embodiment, the compound of Formula (I) or Table 1, or a pharmaceutically acceptable salt thereof (or an embodiment thereof disclosed herein), is administered as a pharmaceutical composition. In yet another embodiment, the disease is cancer.

[0010] In yet another aspect, there is provided a method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound of Formula (I) or Table 1, or a pharmaceutically acceptable salt thereof (or an embodiment thereof disclosed herein). In one embodiment, the patient is identified as being in need of such treatment. In another embodiment, the compound of Formula (I) or Table 1, or a pharmaceutically acceptable salt thereof (or an embodiment thereof disclosed herein), is administered as a pharmaceutical composition.

[0011] In another aspect, a method is provided for inhibiting DNA repair by Pol theta in cancer cells, comprising contacting the cells with an effective amount of a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof. In one embodiment, the cancer is an HR-deficient cancer.

[0012] In yet another aspect, there is provided a method for treating and / or preventing cancer in a patient, wherein the cancer is characterized by reduced or absent BRCA1 and / or BRCA2 gene expression, absent or mutated BRCA1 and / or BRCA2 genes, or reduced function of BRCA1 or 2 proteins, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof, optionally as a pharmaceutical composition.

[0013] In yet another aspect, there is provided a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof, for inhibiting DNA repair by Pol θ in a cell. In one embodiment, the cell is an HR-deficient cell.

[0014] In another aspect, there is provided a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof, for use in treating and / or preventing a disease in a patient, wherein the disease is characterized by overexpression of Pol θ.

[0015] In yet another aspect, there is provided a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof, for use in treating and / or preventing cancer in a patient, wherein the cancer is characterized by reduced or absent BRCA1 and / or BRCA2 gene expression, absent or mutated BRCA1 and / or BRCA2 genes, or reduced function of BRCA1 or 2 proteins.

[0016] In yet another aspect, there is provided a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof, for use in treating and / or preventing HR-deficient cancer in a patient.

[0017] In another aspect, provided is a compound of Formula (I) or Table 1 (or an embodiment thereof disclosed herein), or a pharmaceutically acceptable salt thereof, for use in treating and / or preventing cancer that is resistant or has developed resistance to poly(ADP-ribose) polymerase (PARP) inhibitor therapy in a patient. Examples of cancers that are resistant to PARP inhibitors include, but are not limited to, breast cancer, ovarian cancer, lung cancer, bladder cancer, liver cancer, head and neck cancer, pancreatic cancer, gastrointestinal cancer, and colorectal cancer.

[0018] In related embodiments of the above methods, uses and compositions, the cancer depends on polymerase theta for proliferation, examples of which are lymphoma, rhabdoid tumor, multiple myeloma, uterine cancer, gastric cancer, peripheral nervous system cancer, rhabdomyosarcoma, bone cancer, colorectal cancer, mesothelioma, breast cancer, ovarian cancer, lung cancer, fibroblastic cancer, central nervous system cancer, urinary tract cancer, upper aerodigestive tract cancer, white blood cells, kidney cancer, skin cancer, esophageal cancer, and pancreatic cancer (https: / / depmap.org / portal / ).

[0019] In some embodiments, the HR-deficient cancer is breast cancer.Breast cancer includes but is not limited to lobular carcinoma in situ (LCIS), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), inflammatory breast cancer, Paget's disease of the nipple, phyllodes tumor, angiosarcoma, adenoid cystic carcinoma, low-grade adenosquamous carcinoma, medullary carcinoma, mucinous carcinoma, papillary carcinoma, tubular carcinoma, metaplastic carcinoma, micropapillary carcinoma, mixed carcinoma, or other breast cancers, including but not limited to triple-negative, HER-positive, estrogen receptor-positive, progesterone receptor-positive, HER- and estrogen receptor-positive, HER- and progesterone receptor-positive, estrogen and progesterone receptor-positive, and HER- and estrogen and progesterone receptor-positive.In other embodiments, the HR-deficient cancer is ovarian cancer. Ovarian cancers include, but are not limited to, epithelial ovarian carcinoma (EOC), mature teratoma, dysgerminoma, endodermal sinus tumor, granulosa-thecal tumor, Sertoli-Leydig cell tumor, and primary peritoneal carcinoma. In some embodiments, ovarian cancers include ovarian epithelial cancer, fallopian tube cancer, and primary peritoneal cancer.

[0020] Also provided herein is a combination therapy that includes a method of treating cancer in a subject in need thereof, the method comprising administering a therapeutically effective amount of a DNA polymerase theta (Pol θ) inhibitor (e.g., a compound of Formula (I) or Formula (II)) to the subject and administering a therapeutically effective amount of a poly ADP-ribose polymerase (PARP) inhibitor to the subject, thereby treating cancer in the subject.

[0021] In another aspect, a method for treating and / or preventing homologous recombination (HR)-deficient cancer in a patient in need thereof is provided, comprising contacting cancer cells in the patient with an effective amount of a Pol θ inhibitor (e.g., a compound of Formula (I) or Formula (II)) and a poly ADP-ribose polymerase (PARP) inhibitor. ART4215, a Pol θ polymerase domain inhibitor, was developed by Artios Pharma and is currently in a Phase 1 / 2a clinical trial. See "A Study of ART4215 for the Treatment of Advanced or Metastatic Solid Tumors" at clinicaltrials.gov, NCT04991480. Other Pol θ polymerase domain inhibitors, including ART558, have also been reported. See Zatreanu D. et al., "Pol θ inhibitors elicit BRCA-gene synthetic lethality and target PARP inhibitor resistance," NATURE COMMUNICATIONS, 2021, 12(1):3636.

[0022] The compound of formula (II) has the following structure: or a pharmaceutically acceptable salt thereof:

[0023] [ka] (In the formula, R 1 , R 2 , R 3 , and n has the meaning provided herein below).

[0024] In some aspects, provided herein are methods of treating cancer in a subject in need thereof, the method comprising administering to the subject a combination comprising a DNA polymerase theta (Pol θ) inhibitor (e.g., a compound of Formula (I) or Formula (II)) and a poly ADP-ribose polymerase (PARP) inhibitor, together with at least one pharmaceutically acceptable carrier, thereby treating cancer in the subject.

[0025] A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in therapy.

[0026] A combination of a compound of formula (I) or formula (II) with a poly ADP ribose polymerase (PARP) inhibitor for use in therapy. [Brief explanation of the drawings]

[0027] [Figure 1] Figure 1 shows the efficacy study of Compound A in the BRCA1 mutant TNBC MDA-MB-436 model. The dotted line indicates the mean starting tumor volume. Statistics were calculated using a mixed-effects model with Tukey's test, *p<0.05 after multiplicity adjustment. [Figure 2] FIG. 2 shows an efficacy study of Compound A in the BRCA1 mutant MDA-MB-436 model. [Figure 3] Figure 3 shows the efficacy study of Compound A in the 134-T PDX model. Statistics were calculated applying a mixed effects model with Tukey's test, *p<0.05 after multiplicity adjustment. [Figure 4] Figure 4 shows an efficacy study of Compound A and prodrug Example 1 in the 134-T PDX model. Statistics were calculated applying a mixed effects model with Tukey's test, *p<0.05 after multiplicity adjustment. [Figure 5]Figure 5 shows an efficacy study of prodrug example 1 in the BRCA1 mutant TNBC MDA-MB-436CDX model. Statistics were calculated applying a mixed effects model with Tukey's test, *p<0.05 after multiplicity adjustment. [Figure 6] Figure 6 shows an efficacy study demonstrating that prodrug Example 1 can resensitize 134-T PDX tumors progressing on niraparib. Statistics were calculated applying a mixed effects model with Tukey's test, *p<0.05 after multiplicity adjustment. [Figure 7] FIG. 7 shows an efficacy study of Compound A in the DLD1 BRCA2 deletion CDX model. [Figure 8] 8 shows an efficacy study of prodrug Example 1 in the HR-deficient human cell line xenograft model MDA-MB-436. Statistical analysis was performed using a mixed-effects model with Sidak's multiple comparison test. [Figure 9] Figure 9 shows an efficacy study of prodrug example 1 in 031-T ovarian PDX tumors progressing on niraparib. Statistics were calculated applying a mixed effects model with Tukey's test, n=4, multiplicity adjusted, *p<0.05. DETAILED DESCRIPTION OF THE INVENTION

[0028] Before the present invention is further described, it is to be understood that the present invention is not limited to particular embodiments described herein, and the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0029] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. Accordingly, this sentence is intended to serve as a predicate for using exclusive terminology, such as "solely," "only," etc., in connection with the recitation of claim elements or for using a "negative" limitation.

[0030] Where a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of that range, and any other stated or intervening value in that stated range, to the tenth of the unit of the lower limit, is encompassed within the scope of the invention unless the context clearly dictates otherwise. The upper and lower limits of these subranges may independently be included in the subranges, and such subranges are also encompassed within the scope of the invention, subject to any specific excluded limit in the stated range. Where a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0031] Where necessary, any definition herein may be used in combination with other definitions to describe groups of complex structure. By convention, the last element of any such definition is the one attached to the parent moiety. For example, the complex group alkoxyalkyl means that the alkoxy group is attached to the parent molecule via the alkyl group.

[0032] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Further, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0033] Definition: Unless otherwise stated, the following terms used in the specification and claims are defined for purposes of this application and have the following meanings:

[0034] The term "alkyl," by itself or as part of another substituent, means, unless otherwise stated, a saturated, straight or branched chain hydrocarbon radical having the specified number of carbon atoms (i.e., C 1~8 means 1 to 8 carbons). Alkyl can be any number of carbons, e.g., C 1~2 , C 1~3 , C 1~4 , C 1~5 , C 1~6 , C 1~7 , C 1~8 , C 1~9 , C 1~10 , C 2~3 , C 2~4 , C 2~5 , C 2~6 , C 3~4 , C 3~5 , C 3~6 , C 4~5 , C 4~6 and C 5~6 Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0035] The term "alkylene" refers to a straight-chain or branched saturated aliphatic group, i.e., a divalent hydrocarbon group, having the indicated number of carbon atoms and linking at least two other groups. The two moieties attached to the alkylene may be attached to the same atom or different atoms of the alkylene group. For example, a straight-chain alkylene is -(CH2) n -, where n is 1, 2, 3, 4, 5, or 6. Representative alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, sec-butylene, pentylene, hexylene, and the like.

[0036] The term "alkoxy" refers to an alkyl group having an oxygen atom connecting the alkyl group to the point of attachment: alkyl-O-. As with the alkyl group, the alkoxy group may have any suitable number of carbon atoms, e.g., C 1~6 and may be linear or branched. Examples of alkoxy groups include methoxy, ethoxy, propoxy, iso-propoxy, butoxy, 2-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, pentoxy, and hexoxy.

[0037] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic ring having the indicated number of ring vertices (e.g., a 3- to 7-membered ring) and having 1 to 5 heteroatoms as ring vertices selected from N, O, and S. For example, "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic ring having 4 to 6 ring members and 1 to 3 heteroatoms as ring vertices independently selected from N, O, and S. A partially unsaturated heterocycloalkyl group has one or more double or triple bonds in the ring, but the heterocycloalkyl group is not aromatic. A heterocycloalkyl group may contain any number of ring atoms, for example, 3 to 6, 4 to 6, 5 to 6, 3 to 7, 4 to 7, or 5 to 7 ring members. Any suitable number of heteroatoms, for example, 1, 2, 3, or 4, or 1 to 2, 1 to 3, 1 to 4, 2 to 3, 2 to 4, or 3 to 4, may be included in the heterocycloalkyl group. Non-limiting examples of heterocycloalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, and the like. Further non-limiting examples of heterocycloalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, etc. Heterocycloalkyl groups may be attached to the remainder of the molecule through a ring carbon or heteroatom.

[0038] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0039] The term "haloalkyl" refers to an alkyl as defined above in which some or all of the hydrogen atoms have been replaced with halogen atoms. With respect to the alkyl group, the haloalkyl group may be any suitable number of carbon atoms, e.g., C 1~6 For example, the term "C1-4 haloalkyl" is meant to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0040] The term "haloalkoxy" refers to an alkoxy group in which some or all of the hydrogen atoms have been replaced with halogen atoms. As with the alkyl group, the haloalkoxy group may be any suitable number of carbon atoms, e.g., C 1~6 and may be linear or branched and substituted with one, two, three, or more halogens. If all hydrogens are replaced by halogens, such as fluorine, the compound is over-substituted, e.g., perfluorinated. Haloalkoxy includes, but is not limited to, trifluoromethoxy, 2,2,2-trifluoroethoxy, perfluoroethoxy, and the like.

[0041] As used herein, the term "heteroatom" is meant to include oxygen (O), nitrogen (N), and sulfur (S).

[0042] The term "pharmaceutically acceptable salts" is intended to include salts of active compounds prepared with relatively non-toxic acids or bases, depending on the specific substituents found in the compounds described herein. When a compound of the present invention contains a relatively acidic functional group, a base addition salt may be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base, either pure or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganic, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, etc., such as, for example, arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When a compound of the invention contains a relatively basic functional group, an acid addition salt may be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, bicarbonate, phosphoric, monohydrogen phosphate, dihydrogen phosphate, sulfuric, monohydrogen sulfuric, hydroiodic, or phosphorous acids, and the like, as well as salts derived from relatively non-toxic organic acids such as acetic, propionic, isobutyric, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic acid, and the like.Also included are salts of amino acids, such as arginine, and organic acids such as glucuronic acid or galacturonic acid (see, e.g., Berge, SM et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, Vol. 66, pp. 1-19). Certain specific compounds of the present invention contain both basic and acidic functional groups that allow the compounds to be converted into either base or acid addition salts.

[0043] The neutral forms of the compounds may be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but the salts are otherwise equivalent to the parent form of the compound for purposes of the present invention.

[0044] In addition to salt forms, the present invention provides compounds in prodrug form. Prodrugs are compounds that readily undergo chemical changes under physiological conditions to provide the parent compound. Prodrugs can also be converted to the parent compound by chemical or biochemical methods in an ex vivo environment. The term "prodrug moiety" refers to the chemical moiety of a prodrug that is cleaved under physiological conditions to form the active parent compound.

[0045] Certain compounds of the present invention may exist in non-solvated form and solvated form, including hydrated form.Generally, solvated form is equivalent to non-solvated form and is intended to be included in the scope of the present invention.Certain compounds of the present invention may exist in multiple crystalline forms or amorphous forms.Generally, all physical forms are intended to be included in the scope of the present invention.

[0046] Certain compounds of the present invention having asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, and individual isomers (e.g., individual enantiomers) are all intended to be encompassed within the scope of the present invention. When a stereochemical depiction is shown, it is meant to refer to a compound in which one isomer is present and the other isomer is substantially absent. "Substantially free" of another isomer indicates that the two isomers are present in a ratio of at least 80 / 20, more preferably 90 / 10, or 95 / 5 or greater. In some embodiments, one of the isomers will be present in an amount of at least 99%.

[0047] The present invention also includes all suitable isotopic variations of the compounds of formula (I) in Table 1, or a pharmaceutically acceptable salt thereof. An isotopic variation of a compound of formula (I), or a pharmaceutically acceptable salt thereof, is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature. Examples of isotopes that can be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine, e.g., 2 H, 3 H, 13 C. 14 C. 15 N, 17 O. 18 O. 18 F and 36 Certain isotopic variations of the compounds of formula (I) or salts or solvates thereof, such as 3 H or 14 Those incorporating radioactive isotopes such as C are useful in drug and / or substrate tissue distribution studies. Tritiated, i.e. 3 H, and carbon-14, i.e. 14 C isotopes are particularly preferred for their ease of preparation and detectability. Additionally, deuterium, i.e. 2Substitution with isotopes such as H can provide certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances. Accordingly, in one embodiment, the present invention includes compounds of Table 1 (e.g., Example 1) in which one or more hydrogen atoms bonded to a carbon atom are replaced by deuterium. Isotopic variations of compounds of formula (I) or pharmaceutical salts thereof can generally be prepared by conventional procedures, for example, by the exemplary methods or by the preparations described in the Examples below using appropriate isotopic variations of suitable reagents.

[0048] The terms "patient" or "subject" are used interchangeably to refer to a human or a non-human animal (e.g., a mammal). In one embodiment, the patient is a human.

[0049] The terms "administration," "administering," and the like, when applied to, e.g., a subject, cell, tissue, organ, or biological fluid, refer to contact of, e.g., a Pol θ inhibitor, a pharmaceutical composition comprising same, or a diagnostic agent with the subject, cell, tissue, organ, or biological fluid. In the context of a cell, administration includes contact of a reagent with the cell (e.g., in vitro or ex vivo), as well as contact of a reagent with a fluid if the fluid is in contact with the cell.

[0050] The terms "treat," "treating," "treatment," and the like refer to a course of action (e.g., administering a Pol Theta inhibitor or a pharmaceutical composition comprising same) initiated after a disease, disorder, or condition, or a symptom thereof, has been diagnosed, observed, etc., and which results in the temporary or permanent elimination, reduction, suppression, alleviation, or amelioration of at least one underlying cause of the disease, disorder, or condition afflicting the subject, or at least one symptom associated with the disease, disorder, or condition afflicting the subject. Thus, treatment includes inhibiting active disease (e.g., arresting the onset or further development of the disease, disorder, or condition, or clinical symptoms associated therewith).

[0051] The term " in need of treatment " or " in need of it " as used herein refers to the judgment made by a doctor or other caregiver that the subject needs or will benefit from treatment.This judgment is based on various factors within the expertise of the doctor or caregiver.For example, the patient is diagnosed with the disease associated with Polθ overexpression or homologous recombination (HR) defective cancer.

[0052] The phrase "therapeutically effective amount" refers to the administration of an agent to a subject, alone or as part of a pharmaceutical composition, in a single dose or as part of a series of doses, in an amount that, when administered to a subject, can have any detectable positive effect on any symptom, appearance, or characteristic of a disease, disorder, or condition. A therapeutically effective amount may be determined by measuring the appropriate physiological effect, which may be adjusted in conjunction with a dosing regimen, diagnostic analysis of the subject's condition, and the like. For example, measuring serum levels of a Pol θ inhibitor (or, for example, a metabolite thereof) at a specific time after administration may be an indicator of whether a therapeutically effective amount has been used.

[0053] The terms "inhibitor" and "activator" refer to inhibitory or activating molecules, respectively, for example, for activating a ligand, receptor, cofactor, gene, cell, tissue, or organ. An inhibitor is, for example, a molecule that reduces, blocks, prevents, delays, inactivates, desensitizes, or downregulates the activation of a gene, protein, ligand, receptor, or cell. An activator is, for example, a molecule that increases, activates, promotes, enhances, sensitizes, or upregulates the activation of a gene, protein, ligand, receptor, or cell. An inhibitor may also be defined as a molecule that reduces, blocks, or inactivates constitutive activity.

[0054] The "activity" of a molecule may describe or refer to the binding of the molecule to a ligand or receptor; catalytic activity; the ability to stimulate gene expression or cell signaling, differentiation, or maturation; antigenic activity; modulation of the activity of other molecules; and the like.

[0055] "Pharmaceutically acceptable carrier or excipient" means a carrier or excipient that is generally safe, non-toxic, and not biologically or otherwise undesirable and is useful in preparing pharmaceutical compositions, and includes carriers or excipients that are acceptable for veterinary use as well as for human pharmaceutical use. "Pharmaceutically acceptable excipient," as used in the specification and claims, includes both one such excipient and more than one such excipient.

[0056] As used herein, a wavy line "" that crosses a single bond, double bond, or triple bond in any chemical structure depicted herein.

[0057] [ka] " represents the point of attachment of a single, double, or triple bond to the rest of the molecule. Additionally, a bond extending to the center of a ring (e.g., a phenyl ring) is meant to indicate a bond at any of the available ring vertices. One of skill in the art will understand that multiple substituents shown attached to a ring will provide a stable compound and will occupy any otherwise sterically compatible ring vertices.

[0058] As used herein, "about" is intended to modify the numerical value it modifies, and indicates such a value as a variable within a margin of error. When a specific margin of error, such as a standard deviation, for the average value given in a chart or table of data is not stated, the term "about" refers to a range that will encompass ±10%, preferably ±5%, and should be understood to include the recited value and range.

[0059] "Disease," as used herein, is intended to be used interchangeably with the terms "disorder," "syndrome," and "condition" (similar to medical condition), which are generally synonymous in that all reflect an abnormal condition of the human or animal body or one of its parts that affects normal function, is typically manifested by distinguishing signs and symptoms, and results in a reduction in the lifespan or quality of life of the human or animal.

[0060] "Patient" is generally synonymous with the term "subject" and, as used herein, includes all mammals, including humans. Preferably, the patient is human.

[0061] "Inhibiting," "reducing," or any variation of these terms in reference to Pol Theta includes any measurable decrease or complete inhibition to achieve a desired result. For example, there may be about, or at least about, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or more of a decrease in Pol Theta activity compared to its normal activity, or any range derivable therein.

[0062] The term "homologous recombination" refers to the cellular process of genetic recombination in which nucleotide sequences are exchanged between two similar or identical DNAs.

[0063] The term "homologous recombination (HR)-deficient cancer" refers to cancers characterized by a reduction or absence of a functional HR repair pathway. HR deficiency can result from the absence of one or more HR-associated genes or the presence of one or more mutations in one or more HR-associated genes. Examples of HR-associated genes include BRCA1, BRCA2, RAD54, RAD51B, Ct1P (choline transporter-like protein), PALB2 (partner and localizer of BRCA2), XRCC2 (X-ray complementation defective repair in Chinese hamster cells 2), RECQL4 (RecQ protein-like 4), BLM (Bloom syndrome, RecQ helicase-like), WRN (Werner syndrome, one or more HR-associated genes), Nbs1 (nibrin), and genes encoding Fanconi anemia (FA) proteins or FA-like genes, such as FANCA, FANCB, FANCC, FANCD1 (BRCA2), FANCD2, FANCE, FANCF, FANCG, FANCI, FANJ (BRIP1), FANCL, FANCM, FANCN (RALB2), FANCP (SLX4), FANCS (BRCA1), RAD51C, and XPF.

[0064] The term "Pol θ overexpression" refers to increased expression or activity of Pol θ in diseased cells, e.g., cancerous cells, relative to expression or activity of Pol θ in normal cells (e.g., non-diseased cells of the same type). The amount of Pol θ may be at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, or more relative to Pol θ expression in normal cells. Examples of Pol θ cancers include, but are not limited to, breast cancer, ovarian cancer, cervical cancer, lung cancer, colorectal cancer, gastric cancer, bladder cancer, and prostate cancer.

[0065] As used herein, "poly ADP-ribose polymerase (PARP) inhibitor" refers to an agent that inhibits PARP activity, including PARP 1 and PARP 2. Examples of PARP inhibitors include, but are not limited to, niraparib, rucaparib, olaparib, talazoparib, and veliparib.

[0066] Compound: In some embodiments, the compound of formula (I)

[0067] [ka] (In the formula, R 1 H, halo, C 1~4 Alkyl, C 1~4 Alkoxy, C 1~4 Haloalkyl, or C 1~4 haloalkoxy; Each R 2 independently, halo, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Alkoxy, or C 1~4 haloalkoxy; X is a prodrug moiety; R 3 is C 1~4 Alkyl, C 1~4 Haloalkyl, C 3~6 cycloalkyl, or heterocycloalkyl, 3~6 The cycloalkyl and the heterocycloalkyl may be selected from 1 to 4 R 3a Optionally substituted with substituents, each of which is halo, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Haloalkoxy, -L 3 -OC 1~4 Alkyl, -L 3 independently selected from —OH, and oxo; Each L 3 is a bond and C 1~4 independently selected from alkylene; Each heterocycloalkyl has 4 to 6 ring members and 1 to 3 heteroatoms as ring vertices independently selected from N, O, and S; n is 0, 1, 2, or 3. or a pharmaceutically acceptable salt thereof is provided herein.

[0068] In some embodiments, X in formula (I) or subembodiments thereof is —CH(R c )-OP(O)(OR a )(OR b ), -CH(R c )-OC(O)-C 1-6 Alkylene -COH, -CH(R c )-OC(O)-C 1~6 Alkylene-OP(O)(OR a )(OR b ), -CH(R c )-OC(O)-C 1~6 Alkylene-P(O)(OR a )(OR b ), -CH(R c )-OC(O)-C 1~6 Alkylene-NR a R b , or -CH(R c )-OC(O)-C 1~6 alkylene-heterocycloalkyl, wherein R a and R b are each independently H or C 1~4 alkyl, and R c is independently selected from hydrogen and methyl.

[0069] In some embodiments, X in formula (I) or subembodiments thereof is —CH2O—P(O)(OR a )(OR b ), -CH2-OC(O)-C 1~6 Alkylene -CO2H, -CH2-OC(O)-C 1~6 Alkylene-OP(O)(OR a )(OR b ), -CH2-OC(O)-C1~6 Alkylene-P(O)(OR a )(OR b ), -CH2-OC(O)-C 1~6 Alkylene-NR a R b , or -CH2-OC(O)-C 1~6 alkylene-heterocycloalkyl, wherein R a and R b are each independently H or C 1~4 It is alkyl.

[0070] In some embodiments, X in formula (I) or subembodiments thereof is —CH2O—P(O)(OR a )(OR b ), -CH2-OC(O)-C 1~6 Alkylene -CO2H, or -CH2-OC(O)-C 1~6 Alkylene-P(O)(OR a )(OR b )

[0071] In some embodiments, X in formula (I) or subembodiments thereof is —CH2O—P(O)(OR a )(OR b ) or -CH2-OC(O)-C 1~6 It is alkylene-CO2H.

[0072] In some embodiments, X in formula (I) or subembodiments thereof is —CH—OC(O)—C 1~6 It is alkylene-piperidinyl.

[0073] In some embodiments, X in formula (I) or subembodiments thereof is —CH2O—P(O)(OR a )(OR b )

[0074] In some embodiments, X in formula (I) or subembodiments thereof is —CH—OC(O)—C 1~6 It is alkylene-CO2H.

[0075] In some embodiments, X in formula (I) or subembodiments thereof is:

[0076] [ka]

[0077] In some embodiments, X in formula (I) or subembodiments thereof is:

[0078] [ka]

[0079] In some embodiments, X in formula (I) or subembodiments thereof is:

[0080] [ka]

[0081] In some embodiments, X in formula (I) or subembodiments thereof is:

[0082] [ka]

[0083] In some embodiments, X in formula (I) or subembodiments thereof is:

[0084] [ka]

[0085] In some embodiments, X in formula (I) or subembodiments thereof is:

[0086] [ka]

[0087] In some embodiments, R in formula (I) or subembodiments thereof 1 is C 1~4 In some embodiments, R in formula (I) or subembodiments thereof is alkyl. 1 is methyl.

[0088] In some embodiments, each R in Formula (I) or subembodiments thereof 2 independently, halo, C 1~4 Alkoxy, or C 1~4 In some embodiments, each R in formula (I) or subembodiments thereof is haloalkoxy. 2 are independently halo or C 1~4 In some embodiments, each R in formula (I) or subembodiments thereof is alkoxy. 2 is independently F, Cl, or methoxy. In some embodiments, n in Formula (I) or subembodiments thereof is 2, and two R 2 is Cl and methoxy.

[0089] In some embodiments, n in Formula (I) or subembodiments thereof is 1, 2, or 3. In some embodiments, n in Formula (I) or subembodiments thereof is 1. In some embodiments, n in Formula (I) or subembodiments thereof is 2. In some embodiments, n in Formula (I) or subembodiments thereof is 3.

[0090] In some embodiments, n in Formula (I) or subembodiments thereof is 2, and two R 2 is halo and methoxy.

[0091] In some embodiments, R in formula (I) or subembodiments thereof 3 is C 1~4 Alkyl or C 1~4In some embodiments, R in formula (I) or subembodiments thereof is haloalkyl. 3 is C 1~4 In some embodiments, R in formula (I) or subembodiments thereof is alkyl. 3 is methyl.

[0092] In some embodiments, R in formula (I) or subembodiments thereof 3 is C 3~6 cycloalkyl, or heterocycloalkyl, 3~6 The cycloalkyl and the heterocycloalkyl may be selected from 1 to 4 R 3a Optionally substituted with substituents, each of which is halo, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Haloalkoxy, -L 3 -OC 1~4 Alkyl, -L 3 -OH, and oxo.

[0093] In some embodiments, the compound of formula (I) has the following structure:

[0094] [ka] (In the formula, X, R 1 , R 2 , and R 3 has the meaning provided herein). In some embodiments, R 1 is C 1~4 alkyl, and each R 2 is independently halo, C 1~4 Alkoxy, or C 1~4 haloalkoxy, R 3 is C 1~4 It is alkyl.

[0095] In some embodiments, the compound of formula (I) has the following structure:

[0096] [ka] wherein X has the meaning provided herein.

[0097] Compounds of the present disclosure can exist in tautomeric forms, and it will be understood that any reference to a named or structurally depicted compound is intended to encompass all tautomeric forms of such compound.

[0098] Representative compounds of formula (I) are listed in Table 1 below.

[0099] [Table 1]

[0100] In some embodiments, the compound of the invention is a compound from Table 1.

[0101] The compounds of formula (I) are depicted as (Z) isomers relative to the double bond between the thiadiazole moiety and the nitrogen in the amide group.

[0102] [ka]

[0103] Compounds of formula (I) are also intended to include compounds of formula (Ic), the (E) isomer.

[0104] [ka]

[0105] Pharmaceutical Composition The compounds of Formula (I) or Table 1 provided herein or pharmaceutically acceptable salts thereof may be in the form of a composition suitable for administration to a subject. Generally, such compositions are pharmaceutical compositions comprising a compound of Formula (I) or Table 1 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. The pharmaceutical compositions may be used in any of the methods disclosed herein; thus, for example, the pharmaceutical compositions may be administered to a subject ex vivo or in vivo to perform the therapeutic methods and uses described herein.

[0106] Pharmaceutical compositions may be formulated to be compatible with the intended method or route of administration; exemplary routes of administration are provided herein. Additionally, pharmaceutical compositions may be used in combination with other therapeutically active agents or compounds described herein to treat the diseases, disorders, and conditions contemplated by this disclosure.

[0107] Pharmaceutical compositions containing the active ingredient (e.g., a compound of Formula (I) or Table 1, or a pharmaceutically acceptable salt thereof) may be in a form suitable for oral use, such as a tablet, capsule, troche, lozenge, aqueous or oily suspension, dispersible powder or granule, emulsion, hard or soft capsule, or syrup, solution, microbeads, or elixir. Pharmaceutical compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents, such as sweeteners, flavoring agents, coloring agents, and preservatives, to provide pharmaceutically elegant and palatable preparations. Tablets, capsules, etc. contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets, capsules, etc. These excipients may be, for example, diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid; binders, such as starch, gelatin or acacia, and lubricants, such as magnesium stearate, stearic acid or talc.

[0108] Pharmaceutical compositions typically contain a therapeutically effective amount of a compound of Formula (I) or Table 1 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. Suitable pharmaceutically acceptable excipients include, but are not limited to, antioxidants (e.g., ascorbic acid and sodium bisulfate), preservatives (e.g., benzyl alcohol, methylparaben, ethyl or n-propyl, p-hydroxybenzoate), emulsifiers, suspending agents, dispersing agents, solvents, fillers, bulking agents, surfactants, buffers, vehicles, diluents, and / or adjuvants. For example, a suitable vehicle may be physiological saline solution or citrate-buffered saline, optionally supplemented with other materials commonly used in pharmaceutical compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. Those skilled in the art will readily recognize various buffers that can be used in the pharmaceutical compositions and dosage forms discussed herein. Typical buffer solutions include, but are not limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. For example, the buffer solution component may be a water-soluble material, such as phosphoric acid, tartaric acid, lactic acid, succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, and salts thereof. Acceptable buffering agents include, for example, Tris buffer, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), 2-(N-morpholino)ethanesulfonic acid sodium salt (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), and N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS).

[0109] All compounds and pharmaceutical compositions provided herein can be used in all methods provided herein.For example, compounds and pharmaceutical compositions provided herein can be used in all methods for treating and / or preventing all diseases or disorders provided herein.Therefore, compounds and pharmaceutical compositions provided herein are for use as medicines.

[0110] Administration route The compounds of Formula (I) or Table 1 or pharmaceutically acceptable salts thereof and compositions comprising them may be administered in any suitable manner. Suitable routes of administration include oral, parenteral (e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implant), intraperitoneal, intracisternal, intraarticular, intracerebral (intraparenchymal) and intracerebroventricular), nasal, intravaginal, sublingual, intraocular, intrarectal, topical (e.g., transdermal), buccal, and inhalation. Depot injections, typically administered subcutaneously or intramuscularly, may also be utilized to administer the compounds of Formula (I) or Table 1 or pharmaceutically acceptable salts thereof over a defined period of time. Certain embodiments of the present invention contemplate oral administration.

[0111] dosage The compounds of Formula (I) or Table 1 provided herein or their pharmaceutically acceptable salts may be administered to a subject in an amount that depends, for example, on the goal of administration (e.g., the desired degree of recovery); the age, weight, sex, and health and physical condition of the subject to whom the formulation is administered; the route of administration; and the nature of the disease, disorder, condition, or symptoms thereof. The dosing regimen may also take into account the presence, nature, and extent of any adverse effects associated with the administered drug.

[0112] Generally, dosage parameters determine the dosage amount that is less than the amount that would cause irreversible toxicity to the subject (maximum tolerated dose (MTD)) and greater than or equal to the amount required to produce a measurable effect on the subject. Such amount is determined, for example, by pharmacokinetic and pharmacodynamic parameters related to ADME, taking into account the route of administration and other factors.

[0113] The effective dose (ED) is the dose or amount of a drug that produces a therapeutic response or desired effect in a proportion of subjects who take it. 50 ED is the dose or amount of a drug that produces a therapeutic response or desired effect in 50% of the population to which it is administered. 50 The effective dose is usually used as a measure of a reasonable expectation of a drug's effectiveness, but it is not necessarily the dose that a clinician would consider appropriate, taking into account all relevant factors. Therefore, in some situations, the effective dose may be calculated as the ED 50 In other situations, the effective dose is calculated as ED 50 and in still other situations, the effective dose is less than the calculated ED 50 is the same as

[0114] Combination of Polθ inhibitors with poly (ADP-ribose) polymerase (PARP) inhibitors The combination of drugs described in this section may exhibit synergistic effects. The term "synergistic effect" or "synergistic action" as used herein refers to the action of two drugs, such as a DNA polymerase theta (Polθ) inhibitor (e.g., a compound of Formula (I) or Formula (II)) and a poly ADP-ribose polymerase (PARP) inhibitor, to produce an effect, for example, to delay the symptomatic progression of cancer or its symptoms, that exceeds the simple addition of the effects of each drug administered by itself. Synergistic effects may be calculated using suitable methods, such as the sigmoid-Emax formula (Holford, NHG, and Scheiner, LB, Clin. Pharmacokinet. 6:429-453 (1981)), the Loewe summation formula (Loewe, S., and Muischnek, H., Arch. Exp. Pathol Pharmacol. 114:313-326 (1926)), and the median effect formula (Chou, TC, and Talalay, P., Adv. Enzyme Regul. 22:27-55 (1984)). Each of the above formulas may be applied to experimental data to generate corresponding graphs that aid in assessing the effect of drug combinations. The corresponding graphs associated with the above formulas are concentration-effect curves, isobologram curves, and combination index curves, respectively.

[0115] In some embodiments, the present invention provides a combination therapy comprising a therapeutically effective amount of a Pol Theta inhibitor (e.g., a compound of Formula (I) or Formula (II)) and a PARP inhibitor.The "therapeutically effective amount" of the combination of a drug (i.e., a Pol Theta inhibitor of Formula (I) or Formula (II)) and a PARP inhibitor is an amount sufficient to provide an observable improvement over the baseline of the clinically observable signs and symptoms of the disorder treated with the combination.The observable improvement can be visually confirmed by a clinician, or can be visually confirmed by biological tests, biopsies, and assays.

[0116] In some embodiments, provided herein are methods of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a Pol Theta inhibitor of Formula (I) or Formula (II), and administering to the subject a therapeutically effective amount of a PARP inhibitor, thereby treating cancer in the subject.

[0117] In another aspect, there is provided a method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient, comprising administering to the patient an effective amount of a compound of Formula (I) and administering to the patient an effective amount of a poly ADP-ribose polymerase (PARP) inhibitor. In one embodiment, the method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient in need thereof comprises administering to the patient an effective amount of the compound of Example 1 and administering to the patient an effective amount of niraparib.

[0118] In one embodiment, a method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient in need thereof comprises administering to the patient an effective amount of the compound of Example 1 and administering to the patient an effective amount of olaparib.

[0119] In another aspect, there is provided a method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient, comprising contacting cancer cells in the patient with an effective amount of a compound of formula (II) and an effective amount of a poly ADP-ribose polymerase (PARP) inhibitor. In one embodiment, the method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient in need thereof comprises contacting cancer cells in the patient with an effective amount of Compound A and an effective amount of niraparib.

[0120] In one embodiment, a method for treating and / or preventing homologous recombination (HR) deficient cancer in a patient in need thereof comprises contacting cancer cells in the patient with an effective amount of Compound A and an effective amount of olaparib.

[0121] In some embodiments, provided herein are methods of treating cancer in a subject in need thereof, the method comprising administering to the subject a combination comprising a Pol Theta inhibitor of Formula (I) or Formula (II) and a PARP inhibitor, together with at least a pharmaceutically acceptable carrier, thereby treating cancer in the subject.

[0122] In some embodiments, there is provided the use of a combination of a Pol Theta inhibitor of Formula (I) or Formula (II) and a PARP inhibitor for the manufacture of a medicament.

[0123] In another embodiment, there is provided the use of a combination of a Pol Theta inhibitor of Formula (I) or (II) and a PARP inhibitor to treat cancer.

[0124] In some embodiments, the cancer is characterized as a homologous recombination (HR) deficient cancer.

[0125] In some embodiments, the Pol θ inhibitor is an inhibitor of the ATPase domain of Pol θ.

[0126] In some embodiments, the cancer is characterized by reduced or absent BRCA1 and / or BRCA2 gene expression, the absence of BRCA1 and / or BRCA2 genes, the absence or mutation of BRCA1 and / or BRCA2 proteins, reduced function of BRCA1 and / or BRCA2 proteins, or a combination thereof.

[0127] Pol θ inhibitors for combination therapy with PARP inhibitors In some embodiments, a suitable Pol θ inhibitor for combination therapy treatment with a PARP inhibitor described in this section is a compound of formula (II):

[0128] [ka] (In the formula, R1 H, halo, C 1~4 Alkyl, C 1~4 Alkoxy, C 1~4 Haloalkyl, or C 1~4 haloalkoxy; Each R 2 independently, halo, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Alkoxy, or C 1~4 haloalkoxy; R 3 is C 1~4 Alkyl, C 1~4 Haloalkyl, C 3~6 cycloalkyl, or heterocycloalkyl, 3~6 The cycloalkyl and the heterocycloalkyl may be selected from 1 to 4 R 3a Optionally substituted with substituents, each of which is halo, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Haloalkoxy, -L 3 -OC 1~4 Alkyl, -L 3 independently selected from —OH, and oxo; Each L 3 is a bond and C 1~4 independently selected from alkylene; Each heterocycloalkyl has 4 to 6 ring members and 1 to 3 heteroatoms as ring vertices independently selected from N, O, and S; n is 0, 1, 2, or 3. or a pharmaceutically acceptable salt thereof.

[0129] In some embodiments, R in formula (II) or subembodiments thereof 1 is C 1~4 In some embodiments, R in formula (II) or subembodiments thereof is alkyl. 1 is methyl.

[0130] In some embodiments, each R in Formula (II) or subembodiments thereof 2 is halo, C 1~4 Alkoxy, or C 1~4 In some embodiments, each R in formula (II) or subembodiments thereof is haloalkoxy. 2 are independently halo or C 1~4 In some embodiments, each R in formula (I) or subembodiments thereof is alkoxy. 2 is independently F, Cl, or methoxy. In some embodiments, n in Formula (I) or subembodiments thereof is 2, and two R 2 is Cl and methoxy.

[0131] In some embodiments, n in Formula (II) or subembodiments thereof is 1, 2, or 3. In some embodiments, n in Formula (II) or subembodiments thereof is 1. In some embodiments, n in Formula (II) or subembodiments thereof is 2. In some embodiments, n in Formula (II) or subembodiments thereof is 3.

[0132] In some embodiments, n in Formula (II) or subembodiments thereof is 2, and two R 2 is halo and methoxy.

[0133] In some embodiments, R in formula (II) or subembodiments thereof 3 is C 1~4 Alkyl or C 1~4 In some embodiments, R in formula (II) or subembodiments thereof is haloalkyl. 3 is C 1~4 In some embodiments, R in formula (II) or subembodiments thereof is alkyl. 3 is methyl.

[0134] In some embodiments, R in formula (II) or subembodiments thereof 3 is C 3~6cycloalkyl, or heterocycloalkyl, 3~6 The cycloalkyl and the heterocycloalkyl may be selected from 1 to 4 R 3a Optionally substituted with substituents, each of which is halo, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 Haloalkoxy, -L 3 -OC 1~4 Alkyl, -L 3 -OH, and oxo.

[0135] In some embodiments, the Pol Theta inhibitor of formula (II) is:

[0136] [ka]

[0137] In some embodiments, the Pol θ inhibitor of formula (II) is compound A

[0138] [ka] or a pharmaceutically acceptable salt thereof.

[0139] In some embodiments, a Pol θ inhibitor suitable for combination therapy treatment with a PARP inhibitor described in this section is a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the Pol θ inhibitor of Formula (I) is the compound of Example 1 ("Example 1").

[0140] [ka]

[0141] In some embodiments, the Pol Theta inhibitor for the combination therapy is ART558, which has the following structure:

[0142] [ka]

[0143] In some embodiments, the Pol Theta inhibitor for the combination therapy is ART4215.

[0144] PARP inhibitors for combination therapy with Polθ inhibitors The combination therapy described herein provides a PARP inhibitor for use with a Pol θ inhibitor (e.g., a compound of formula (I) or formula (II)). Numerous agents with PARP inhibitory activity and methods for making them are known in the art. Each of these is encompassed by the present disclosure. In some embodiments, the PARP inhibitor is

[0145] [ka] or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

[0146] In some embodiments, the PARP inhibitor is

[0147] [ka] or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

[0148] Selective Combination Therapy Embodiments Embodiment 1. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a Pol Theta inhibitor, or a pharmaceutically acceptable salt thereof, and administering to the subject a therapeutically effective amount of a PARP inhibitor, or a pharmaceutically acceptable salt thereof.

[0149] Embodiment 1a. The method of embodiment 1, wherein the Pol θ inhibitor is an inhibitor of the ATPase domain of Pol θ.

[0150] Embodiment 2. The method of embodiment 1, wherein the Pol Theta inhibitor is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof.

[0151] Embodiment 3. The Pol θ inhibitor has the structure

[0152] [ka] or a pharmaceutically acceptable salt thereof.

[0153] Embodiment 4. The method of embodiment 1, wherein the Pol Theta inhibitor is a compound of formula (II), as defined herein, or a pharmaceutically acceptable salt thereof.

[0154] Embodiment 5. The Pol θ inhibitor is Compound A

[0155] [ka] or a pharmaceutically acceptable salt thereof.

[0156] Embodiment 6. The method of embodiment 1, wherein the Pol inhibitor is ART558 or ART4215, or a pharmaceutically acceptable salt thereof.

[0157] Embodiment 7. The method of any one of embodiments 1-6, wherein the PARP inhibitor is selected from the group consisting of niraparib, rucaparib, olaparib, talazoparib, and veliparib, or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

[0158] Embodiment 8. The method of any one of embodiments 1-6, wherein the PARP inhibitor is niraparib, preferably niraparib tosylate monohydrate.

[0159] Embodiment 9. A PARP inhibitor

[0160] [ka] or a pharmaceutically acceptable salt thereof.

[0161] Embodiment 10. The method of any one of embodiments 1 to 9, wherein the cancer is a homologous recombination (HR) deficient cancer.

[0162] Embodiment 11. The method of any one of embodiments 1 to 9, wherein the cancer is characterized by reduced or absent BRCA1 and / or BRCA2 gene expression, absent or mutated BRCA1 or BRCA2 genes, or reduced function of BRCA1 or 2 proteins.

[0163] Embodiment 12. The method of any one of embodiments 1 to 9, wherein the cancer is a solid tumor.

[0164] Embodiment 13. The method of any one of embodiments 1-9, wherein the cancer is lymphoma, rhabdoid tumor, multiple myeloma, uterine cancer, gastric cancer, peripheral nervous system cancer, rhabdomyosarcoma, bone cancer, colorectal cancer, mesothelioma, breast cancer, ovarian cancer, lung cancer, fibroblastic carcinoma, central nervous system cancer, urinary tract cancer, upper aerodigestive tract cancer, leukemia, kidney cancer, skin cancer, esophageal cancer, and pancreatic cancer.

[0165] Embodiment 14. The method of any one of embodiments 1 to 13, wherein the cancer is PARP inhibitor resistant or has developed resistance to PARP inhibitor therapy.

[0166] Embodiment 15. The method of any one of embodiments 1-14, wherein the Pol Theta inhibitor and the PARP inhibitor are in separate dosage forms.

[0167] Embodiment 16. The method of any one of embodiments 1 to 14, wherein the Pol Theta inhibitor and the PARP inhibitor are in the same dosage form.

[0168] Embodiment 17. A combination comprising a compound of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof and a PARP inhibitor or a pharmaceutically acceptable salt thereof.

[0169] Embodiment 18. The combination of embodiment 17, wherein the PARP inhibitor is niraparib, rucaparib, olaparib, talazoparib, and veliparib, AZD5305, or AZD9574, or a pharmaceutically acceptable salt thereof.

[0170] Embodiment 19. A compound of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, wherein the compound of Formula (I) or Formula (II) is to be administered simultaneously or sequentially with a PARP inhibitor.

[0171] Embodiment 20. The use of embodiment 19, wherein the PARP inhibitor is niraparib, rucaparib, olaparib, talazoparib, and veliparib, AZD5305, or AZD9574, or a pharmaceutically acceptable salt thereof.

[0172] Embodiment 21. Use of a compound of Formula (I) or Formula (II) in the manufacture of a medicament for treating cancer, wherein the compound of Formula (I) or Formula (II) is to be administered simultaneously or sequentially with a PARP inhibitor.

[0173] Embodiment 22. The use of embodiment 21, wherein the PARP inhibitor is niraparib, rucaparib, olaparib, talazoparib, and veliparib, AZD5303, or AZD9574, or a pharmaceutically acceptable salt thereof. [Example]

[0174] The following examples and references (intermediates) are set forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below have been performed or that they are all that may be performed. It should be understood that the illustrative descriptions set forth in the present tense have not necessarily been performed, but rather the descriptions may have been performed to generate data of the nature described therein, etc. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for.

[0175] Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius (°C), and pressure is at or near atmospheric. Standard abbreviations are used, including: THF = tetrahydrofuran; EtOAc = ethyl acetate; TFA = trifluoroacetic acid; DCM = dichloromethane; DMSO = dimethyl sulfoxide; DMF = dimethylformamide; BOC = tert-butoxycarbonyl; TCFH = N,N,N,N'-tetramethylchloroformamidinium hexafluorophosphate; NMI = N-methylimidazole; TBAF = tetrabutylammonium fluoride; DIBAL-H = diisobutylaluminum hydride; LDA = lithium diisopropylamide.

[0176] Automated preparative HPLC (MDAP) directly connected to a mass spectrometer Mass-coupled automated preparative HPLC was used to prepare the compounds of the present invention under the following conditions: UV detection was the average signal from wavelengths of 210 nm to 350 nm, and mass spectra were recorded on a mass spectrometer using alternating scan positive and negative mode electrospray ionization.

[0177] MDAP method B Method B is Xselect CSH C 18The run was carried out on a column (typically 150 mm x 30 mm internal diameter, 5 μm packing diameter) at ambient temperature. The solvents used were as follows: A = 0.1% v / v formic acid solution in water B = 0.1% v / v formic acid solution in acetonitrile. The gradient used was as follows:

[0178] [Table A]

[0179] MDAP method C Method C is Xselect CSH C 18 The run was carried out on a column (typically 150 mm x 30 mm internal diameter, 5 μm packing diameter) at ambient temperature. The solvents used were as follows: A = 0.1% v / v formic acid solution in water B = 0.1% v / v formic acid solution in acetonitrile. The gradient used was as follows:

[0180] [Table B]

[0181] Synthesis Example: Compound of formula (I)

[0182] [Example 1] (Z)-(2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl dihydrogen phosphate

[0183] [ka]

[0184] Step 1: 2-Chloro-5-methoxypyridin-4-ylboronic acid

[0185] [ka]

[0186] To a stirred solution of 2-chloro-5-methoxypyridine (10.0 g, 69.65 mmol) in THF (500 mL) was added dropwise LDA (14.9 g, 139.30 mmol) under a N2 atmosphere at -78 °C. The resulting mixture was stirred at -78 °C for 2 hours. Triisopropyl borate (26.2 g, 139.30 mmol) was then added to the above mixture at -78 °C. The resulting mixture was stirred at -78 °C for 2 hours. The resulting mixture was then stirred at room temperature for 16 hours. The resulting mixture was quenched with HCl (2N) and stirred at room temperature for 30 minutes. The resulting mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. 2-Chloro-5-methoxypyridin-4-ylboronic acid (9 g, 68.9%) was obtained as a brown solid. MS (ESI) (C6H7BClNO3) (M+1) + Calculated value: 188.0; Measured value: 188.0.

[0187] Step 2: Methyl 2-chloro-5-methoxy-6-methyl-(4,4-bipyridine)-3-carboxylate

[0188] [ka]

[0189] To a degassed solution of methyl 4-chloro-6-methylpyridine-3-carboxylate (700 mg, 3.77 mmol) and 2-chloro-5-methoxypyridin-4-ylboronic acid (918 mg, 4.90 mmol) in dioxane (6 mL) and HO (2 mL), Pd(dppf)Cl (275 mg, 0.37 mmol) and KCO (1563 mg, 11.31 mmol) were added portionwise under a nitrogen atmosphere at 80 °C. The resulting mixture was stirred at 80 °C for 16 h under a nitrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (3 × 20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography using 0-60% ethyl acetate in petroleum ether to give methyl 2-chloro-5-methoxy-6-methyl-(4,4-bipyridine)-3-carboxylate (220 mg, 19.9%) as a white solid. MS (ESI) (C 14 H 13 ClN2O3) (M+1) + Calculated value, 293.1; measured value, 293.1.

[0190] Step 3: 2-chloro-5-methoxy-6-methyl-(4,4-bipyridine)-3- Carboxylic Acid

[0191] [ka]

[0192] To a stirred solution of methyl 2-chloro-5-methoxy-6-methyl-(4,4-bipyridine)-3-carboxylate (220 mg, 0.75 mmol) in THF (3 mL) and water (1 mL) was added LiOH.HO (126 mg, 3.01 mmol). The resulting mixture was stirred at room temperature for 2 hours. The mixture was acidified to pH 3 using citric acid. The resulting mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give 2-chloro-5-methoxy-6-methyl-(4,4-bipyridine)-3-carboxylic acid (160 mg, 76.3%) as a white solid. MS (ESI) (C13 H 11 ClN2O3) (M+1) + Calculated value: 279.0; Measured value: 279.0.

[0193] Step 4: 2'-chloro-5'-methoxy-N-(5-methoxy-1,3,4-thiadiazol-2-yl)-6-methyl-(4,4'-bipyridine)-3-carboxamide

[0194] [ka]

[0195] To a solution of 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (50 g, 169 mmol) in MeCN (500 mL) was added 1-methylimidazole (69.2 g, 843 mmol) and 5-methoxy-1,3,4-thiadiazol-2-amine (24.31 g, 185 mmol) in one portion at 25° C. A solution of TCFH (52 g, 185 mmol) in MeCN (500 mL) was added dropwise at 25° C. under nitrogen. After stirring at 25° C. for 4 hours, the mixture was filtered and the filter cake was washed with a 1:1 MeCN:water solution (200 mL). The filter cake was dried under vacuum at 45°C to give 2'-chloro-5'-methoxy-N-(5-methoxy-1,3,4-thiadiazol-2-yl)-6-methyl-(4,4'-bipyridine)-3-carboxamide (57 g, 79% yield, adjusted to 96% purity) as an off-white solid. MS (ESI) (C 16 H 14 ClN5O3S) (M+1) + Calculated value: 392.0, measured value: 392.1. 1 H NMR (400 MHz, DMSO-d6) δ 12.93 (s, 1H), 8.81 (s, 1H), 8.17 (s, 1H), 7.55 (s, 1H), 7.44 (s, 1H), 4.08 (s, 3H), 3.63 (s, 3H), 2.59 (s, 3H).

[0196] Step 5: (Z)-Di-tert-butyl((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl)phosphate

[0197] [ka]

[0198] A mixture of 30 g (77.0 mmol) of 2'-chloro-5'-methoxy-N-(5-methoxy-1,3,4-thiadiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide and 300 mL of DMF was cooled to 20 °C. To this mixture, 31.7 g (230 mmol) of K2CO3, 12.7 g (77.0 mmol) of KI, and then 39.6 g (153 mmol) of di-tert-butyl(chloromethyl)phosphate were added over approximately 10 minutes. The mixture was stirred at 20 °C for 24 hours. The reaction was warmed to 25 °C, diluted with 300 mL of EtOAc, and quenched with 375 mL of water. The layers were separated, and the organic layer was washed successively with 300 mL of water and 300 mL of 10 wt / w% aqueous NaCl solution. After cooling to 10°C, the solution was treated with 300 mL of heptane over approximately 3 hours. The slurry was stirred overnight, filtered, and washed successively with 60 mL of 60% heptane in EtOAc and 2 x 90 mL of heptane. The wet cake was dried overnight under vacuum at 50°C to give 27.0 g (57% yield) of (Z)-di-tert-butyl((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl)phosphate as an off-white solid. MS (ESI) (C 25 H 33 ClN5O7PS) (M+1) + Calculated value, 614.2; measured value, 614.2. 1H NMR (DMSO-d6, 400 MHz) δ 9.29 (s, 1H), 8.14 (s, 1H), 7.42 (s, 1H), 7.29 (s, 1H), 5.85 (br d, 2H, J=10.3 Hz), 4.07 (s, 3H), 3.62 (s, 3H), 2.57 (s, 3H), 1.39 (s, 18H).

[0199] Step 6: (Z)-(2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl dihydrogen phosphate

[0200] [ka]

[0201] A mixture of 5 g (8.14 mmol) of (Z)-di-tert-butyl ((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl)phosphate and 20 mL of water was heated to 35°C. To this slurry was added 15 mL (398 mmol) of formic acid over a period of about 10 minutes. The mixture was stirred for about 4 hours, followed by the addition of 70 mL of water over a period of about 2 hours. After cooling to 10°C, the slurry was stirred overnight, filtered, and washed successively with 2 x 25 mL of water and 2 x 25 mL of THF. The wet mass was dried overnight under vacuum at 50° C. to give 3.23 g (79% yield before drying, 72% yield after drying) of (Z)-(2-((2′-chloro-5′-methoxy-6-methyl-[4,4′-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl dihydrogen phosphate as an off-white solid. MS (ESI) (C 17 H 17 ClN5O7PS) (M+1) + Calculated value: 502.0; measured value: 502.1. 1H NMR (DMSO-d6, 400 MHz) δ 9.30 (s, 1H), 8.15 (s, 1H), 7.42 (s, 1H), 7.30 (s, 1H), 5.81 (d, 2H, J=9.3 Hz), 4.07 (s, 3H), 3.64 (s, 3H), 2.58 (s, 3H).

[0202] [Example 2] (Z)-4-((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methoxy)-4-oxobutanoic acid

[0203] [ka]

[0204] Step 1: tert-Butyl (chloromethyl) succinate

[0205] [ka]

[0206] To a solution of 4-(tert-butoxy)-4-oxobutanoic acid (1.507 g, 8.65 mmol) in a solvent mixture of ethanol (37.5 mL) and water (5.5 mL) was added cesium carbonate (1.409 g, 4.33 mmol). The mixture was sonicated briefly and then stirred for 15 min. The solvent was evaporated and the residue was dried under high vacuum overnight. Dry 4-(tert-butoxy)-4-oxobutanoic acid Cs-salt was dissolved in N,N-dimethylformamide (DMF) (27.5 mL), bromochloromethane (36.6 mL, 562 mmol) was added, and the solution was stirred at room temperature overnight. The precipitate was filtered off, and excess bromochloromethane was evaporated. To the remaining DMF solution, 50 mL of brine and 50 mL of water were added. The mixture was then extracted with EtOAc (40 + 20 + 20 mL), and the combined organic layers were washed with brine, dried over magnesium sulfate, and evaporated. The crude product was used in the next step without further purification. 1 H NMR (400 MHz, DMSO-d6) δ 5.86 (s, 2H), 2.58-2.66 (m, 2H), 2.48-2.53 (m, J=1.70, 1.70, 3.50 Hz, 2H), 1.39 (s, 9H).

[0207] Step 2: (Z)-tert-butyl ((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl)succinate

[0208] [ka]

[0209] A mixture of 2'-chloro-5'-methoxy-N-(5-methoxy-1,3,4-thiadiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide (250 mg, 0.638 mmol), tert-butyl(chloromethyl)succinate (320 mg, 1.436 mmol), KI (106 mg, 0.638 mmol), and KCO (265 mg, 1.914 mmol) in N,N-dimethylformamide (DMF) (5 mL) was stirred for 3 h at 50° C. The mixture was cooled, then 10 mL of brine and 10 mL of water were added, extracted with EtOAc, and the organic layer was washed with brine, dried over sodium sulfate, and evaporated. The residue was purified by silica gel chromatography (12 g Isco RediSep Rf gold column, eluting with 0-50% hexanes (3 / 1 EtOAc / EtOH)) to give (Z)-tert-butyl ((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl)succinate (216 mg, 0.374 mmol, 58.6% yield) as a solid foam. MS (ESI) (C 25 H 28 ClN5O7S) (M+1) + Calculated value, 578.1; measured value, 578.1. 1 H NMR (DMSO-d6, 400 MHz) δ 9.16 (s, 1H), 8.14 (s, 1H), 7.42 (s, 1H), 7.28 (s, 1H), 5.94 (s, 2H), 4.05 (s, 3H), 3.63 (s, 3H), 2.6-2.6 (m, 6H), 2.49 (br s, 1H), 1.34 (s, 9H).

[0210] Step 3: (Z)-4-((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methoxy)-4-oxobutanoic acid

[0211] [ka]

[0212] (Z)-tert-Butyl ((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl)succinate (216 mg, 0.374 mmol) was dissolved in dichloromethane (DCM) (10 mL). TFA (3 mL, 38.9 mmol) was added and the mixture was allowed to stand at room temperature for 50 minutes, at which point it was evaporated to dryness. The product was isolated by preparative HPLC (MDAP method B, TFA) to give (Z)-4-((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methoxy)-4-oxobutanoic acid (136 mg, 0.261 mmol, 69.7% yield) as a white lyophilizate. The lyophilizate was crystallized by suspending in 5 mL of ether, sonicating for a few minutes, and then slurrying overnight at room temperature. The solid was filtered off and dried to give 106 mg of white crystalline material. MS (ESI) (C 21 H 20 ClN5O7S) (M+1) + Calculated value: 522.1, measured value: 522.1. 1 H NMR (DMSO-d6, 400 MHz) δ 9.16 (s, 1H), 8.15 (s, 1H), 7.43 (s, 1H), 7.29 (s, 1H), 5.94 (s, 2H), 4.05 (s, 3H), 3.64 (s, 3H), 3.4-3.5 (m, 1H), 2.6-2.7 (m, 6H), 2.53 (d, 1H, J=4.4 Hz)

[0213] [Example 3] (Z)-(2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl 2-(piperidin-4-yl)acetate, formate

[0214] [ka]

[0215] Step 1: tert-butyl 4-(2-(chloromethoxy)-2-oxoethyl)piperidine-1-carboxylate

[0216] [ka]

[0217] To a solution of 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)acetic acid (1.5 g, 6.04 mmol) in a solvent mixture of ethanol (37.5 mL) and water (5.5 mL) was added cesium carbonate (0.994 g, 3.02 mmol). The mixture was sonicated briefly and then stirred for 1 h. The solvent was evaporated and the residue was dried under high vacuum overnight. Dry 2-(1-(tert-butoxycarbonyl)piperidin-4-yl)acetic acid Cs-salt was dissolved in N,N-dimethylformamide (DMF) (27.5 mL) and bromochloromethane (25.5 mL, 393 mmol) was added. The solution was stirred at room temperature overnight. Excess bromochloromethane was evaporated. To the remaining slurry was added 100 mL of ice water. The mixture was extracted with EtOAc (30 + 25 mL), and the combined organic layers were washed with brine (1×), dried over Mg sulfate, and evaporated to give tert-butyl 4-(2-(chloromethoxy)-2-oxoethyl)piperidine-1-carboxylate (1.735 g, 5.95 mmol, 98% yield) as a pale yellow oil. 1 H NMR (DMSO-d6, 400 MHz) δ 5.85 (s, 2H), 3.90 (br d, 2H, J=12.2 Hz), 2.37 (d, 2H, J=6.8 Hz), 1.86 (dtd, 1H, J=3.7, 7.6, 15.0 Hz), 1.6-1.7 (m, 2H), 1.3-1.5 (m, 11H), 1.06 (dq, 2H, J=4.4, 12.2 Hz).

[0218] Step 2: tert-Butyl (Z)-4-(2-((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methoxy)-2-oxoethyl)piperidine-1-carboxylate

[0219] [ka]

[0220] A mixture of 2'-chloro-5'-methoxy-N-(5-methoxy-1,3,4-thiadiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide (100 mg, 0.255 mmol), tert-butyl 4-(2-(chloromethoxy)-2-oxoethyl)piperidine-1-carboxylate (0.124 mL, 0.510 mmol), KI (42.4 mg, 0.255 mmol), and KCO (106 mg, 0.766 mmol) in N,N-dimethylformamide (DMF) (2 mL) was heated at 50 °C for 40 min. The mixture was diluted with 2 mL of DMF, filtered, and acidified with 50 μL of formic acid (FA). The product was isolated by preparative HPLC (MDAP Method C, FA) to give tert-butyl (Z)-4-(2-((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methoxy)-2-oxoethyl)piperidine-1-carboxylate (153 mg, 0.236 mmol, 93% yield). This material was used in the next step without further purification. MS (ESI) (C 29 H 35 ClN6O7S) (M+1) + Calculated value: 647.2, measured value: 647.3.

[0221] Step 3: (Z)-(2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl 2-(piperidin-4-yl)acetate, formate

[0222] [ka]

[0223] To a solution of tert-butyl (Z)-4-(2-((2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methoxy)-2-oxoethyl)piperidine-1-carboxylate (150 mg, 0.232 mmol) in dichloromethane (DCM) (4 mL) was added trifluoroacetic acid (TFA) (4.00 mL) and the mixture was stirred for 15 minutes, at which point it was evaporated. The product was isolated by preparative HPLC (MDAP method B, FA) to give (Z)-(2-((2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carbonyl)imino)-5-methoxy-1,3,4-thiadiazol-3(2H)-yl)methyl 2-(piperidin-4-yl)acetate, formate (74 mg, 0.125 mmol, 53.8% yield) as a white lyophilizate. MS (ESI) (C 24 H 27 ClN6O5S) (M+1) + Calculated value: 547.1, measured value: 547.2. 1H NMR (DMSO-d6, 400 MHz) δ 9.15 (s, 1H), 8.36 (s, 1H), 8.15 (s, 1H), 7.42 (s, 1H), 7.29 (s, 1H), 5.94 (s, 2H), 4.04 (s, 3H), 3.63 (s, 3H), 3.03 (br d, 2H, J=12.2 Hz), 2.6-2.7 (m, 2H), 2.57 (s, 3H), 2.35 (d, 2H, J=6.8 Hz), 1.8-2.0 (m, 1H), 1.70 (br d, 2H, J=11.7 Hz), 1.1-1.3 (m, 3H).

[0224] Biological Examples 1. Hydrolysis Rate of Prodrugs (Example 1) The compound of Example 1 ("Example 1") is a highly soluble phosphate prodrug of the active parent molecule, Compound A. Example 1 is cleaved by alkaline phosphatase in the brush border of the intestinal microvilli to produce Compound A.

[0225] The hydrolysis kinetics, kcat, Km and kcat / Km of Example 1 were determined with recombinant human, mouse and rat alkaline phosphatase from commercial sources using the phosphate sensor protein from Invitrogen.

[0226] Example 1 was hydrolyzed with similar efficiency by recombinant human, mouse, and rat alkaline phosphatases (Table 2). Three commercially available alkaline phosphatases (APases) were tested for the rate of hydrolysis of Example 1. The human and rat enzymes are intestinal alkaline phosphatases, while the mouse APase is a tissue-nonspecific isozyme, which may explain some of the differences in kinetic parameters.

[0227] [Table 2]

[0228] 2. Biochemical activity of compound A Compound A is a potent inhibitor (pIC50 = 7.8) of the ATPase activity of the helicase domain of Pol θ, as demonstrated using a Pol θ ATPase biochemical assay. Consistent with the high sequence homology of Pol θ across species, similar inhibition of Pol θ was observed with Compound A when tested across species (rat, pIC50 = 8.0; dog, pIC50 = 7.8; mouse, pIC50 = 7.8; monkey, pIC50 = 7.9). The ATPase activity of the Pol θ recombinant helicase domain was measured in an enzyme activity assay. The production of ADP from the ATP substrate was determined in a coupled enzyme assay related to NAD+ production. NAD+ production was measured by monitoring the absorbance change at 340 nm in a kinetic manner. The slope of the rate plot was determined for each concentration of Compound A and expressed as % inhibition of enzyme activity. Data represent the mean ± SD.

[0229] Furthermore, compound A was demonstrated to have over 1000-fold selectivity for Polθ versus Hel308, which share 26% sequence identity and 40% sequence similarity.

[0230] 3. Cellular activity of compound A The steady-state cellular concentration of Compound A was measured in HeLa cells after a 4-hour exposure to 20 mM Compound A (Table 3). Based on a calculated release rate of 5.12%, the free cellular concentration of Compound A is calculated to be 3.087 μM (Table 3).

[0231] [Table 3]

[0232] 4. Solubility and PK Assays The solubility of Compound A in FASSIF (fasted state simulated small intestinal fluid) is 13 μg / mL. The FASSIF solubilities for the prodrugs (Examples 1, 2, and 3) are greater than 1000 μg / mL, 72 μg / mL, and 839 μg / mL, respectively.

[0233] Pharmacokinetic data were measured for the prodrug compounds of Examples 1 and 2 and the parent compound (Compound A). Wistar-Han rats were orally dosed with 1% methylcellulose formulations containing each compound at doses as listed in Table 4 below. Blood samples were collected up to 24 hours and analyzed by LC-MS / MS for Compound A concentration in all three groups.

[0234] [Table 4A]

[0235] 5. Evaluation of in vitro combination synergy index between Compound A and PARP inhibitors The cellular efficacy of Compound A, Niraparib, and the combination of Compound A and Niraparib was assayed as % cell viability in a 7-day CellTiter-Glo® (CTG) assay using the BRCA1 mutant breast cancer cell line MDA-MB-436 and the BRCA2 mutant ovarian cancer cell line PEO1. The single agent EC50 value of Compound A was greater than 30 μM in both cell lines.

[0236] Synergy between Compound A and Niraparib was evaluated in the BRACA1 mutant breast cancer cell line MDA-MB-436. Cells were treated with an 8x5 drug matrix containing Compound A in 8-point, 3-fold dilutions ranging from 30 μM to 0.014 μM and Niraparib in 5-point, 3-fold dilutions ranging from 100 nM to 1.2 nM. After 7 days, cell viability was assessed using a CTG assay. Dose-response curves were interpolated using GraphPad Prism 9ART, and the synergy of the drug combination using data from the cell viability assay was analyzed with ComBenefit 2.02 (Table 4). Synergy scores for the cotreatment of Compound A and Niraparib were calculated using Combenefit. Synergy scores for the combination matrix calculated using the Bliss, HAS (lowest criteria), and Loewe (highest criteria) models are shown. The results showed that niraparib synergized with Compound A and reduced the EC50 value of Compound A in MDA-MB-436 cells.

[0237] Using the same experimental setup, the synergy between Compound A and Niraparib was also evaluated in PEO1 cells, a BRCA2 mutant ovarian cancer cell line. Dose-response curves were interpolated using GraphPad Prism 9 (Figure 4A), and the synergy of the drug combination was analyzed with ComBenefit 2.02 (Table 4). The results showed that Niraparib synergized with Compound A, increasing the EC 50 It was shown to decrease the value.

[0238] [Table 4B]

[0239] Further cell line screening confirmed the synergistic effect of the combination of Compound A and niraparib in several homologous recombination deficient (HR-D) in vitro models, as summarized in Table 5.

[0240] [Table 5]

[0241] Synergy between ART558 and niraparib was evaluated in MDA-MB-436 cells. See Table 6.

[0242] [Table 6]

[0243] The synergistic effect of Compound A and ART558 was confirmed with talazoparib. See Table 7.

[0244] [Table 7]

[0245] 6. Efficacy study of Compound A in HR-deficient human cell line xenograft model MDA-MB-436 The effect of Compound A on tumor growth in vivo was evaluated in mice bearing BRCA1 mutant MDA-MB-436 cell line xenografts. 7 ) live MDA-MB-436 cells were implanted into the flanks of 5-7 week-old female NSG mice (Jax) with 50% Matrigel. Tumor volumes were approximately 200 mm 3 Upon reaching a tumor size of 100 mg / kg, animals were randomized into an efficacy study. MDA-MB-436 tumor-bearing animals were dosed with vehicle (0.5% methylcellulose) BID PO, or with Compound A at 10 mg / kg BID or 30 mg / kg BID PO alone, or in combination with niraparib at 35 mg / kg QD PO.

[0246] The effect of Compound A on tumor growth in vivo was evaluated in 5-7 week-old female NSG mice bearing the BRCA1 mutant MDA-MB-436 CDX model. A dose of 30 mg / kg Compound A, bid, PO, induced a significant but modest 24% tumor growth inhibition (p<0.05). A dose of 35 mg / kg niraparib, qd, also induced a significant tumor volume change compared to vehicle (72% TGI) on day 43 (p<0.05). Complete tumor regression was observed when niraparib was administered at doses of 10 mg / kg and 30 mg / kg in combination with Compound A, bid (Figure 1). These in vivo studies suggest that Compound A is synergistic in combination with niraparib in the HR-deficient TNBC CDX model.

[0247] To explore the lowest effective dose of Compound A with a clinically relevant 25 mg / kg niraparib (corresponding to the 200 mg clinical dose), the efficacy of BID and QD regimens of Compound A was evaluated over a study range of 0.2 mg / kg to 6 mg / kg. While there was limited monotherapy response to a 1 mg / kg BID dose of Compound A, a 25 mg / kg QD dose of niraparib delayed tumor growth by approximately 40 days (1200 mm ) compared to vehicle. 3 Tumor volumes reached 21 days with vehicle vs. 61 days with niraparib (Figure 2). Combination of 25 mg / kg niraparib with Compound A administered at 0.1, 0.3, 1, or 3 mg / kg BID resulted in tumor shrinkage in 10, 30, 50, and 90% of mice, respectively. Combination of 25 mg / kg niraparib with Compound A administered at 0.2, 0.6, 2, or 6 mg / kg QD resulted in tumor shrinkage in 20, 20, 80, and 100% of mice, respectively. Compound A administered at 1 mg / kg BID with 25 mg / kg niraparib resulted in tumor shrinkage in 50% of mice. The frequency of tumor shrinkage was enhanced by increasing the dose of Compound A (Figure 2).

[0248] 7. Efficacy study of Compound A in BRCA mutant ovarian PDX models In vivo efficacy studies were conducted using an HR-deficient (BRCA1 frameshift (FS) mutation) PARP inhibitor advanced ovarian carcinoma patient-derived xenograft (PDX) model, 134-T.

[0249] 4mm x 4mm fragments of the HR-deficient (BRCA1 mutant) 134-T ovarian cancer PDX model were implanted into female NOD SCID gamma (NSG) mice, resulting in a mean tumor volume of approximately 150mm. 3 Upon reaching 1 mg / kg BID, animals were randomized and treated with either niraparib (25 mg / kg QD) or 1 mg / kg BID dose of Compound A, or a combination of 25 mg / kg niraparib and doses of Compound A ranging from 0.1 mg / kg BID to 6 mg / kg. While there was no significant monotherapy response to the 1 mg / kg BID dose of Compound A (p=0.77), 25 mg / kg QD niraparib alone resulted in 58% tumor growth inhibition (p=0.005) (Figure 3). Both 1 mg / kg and 3 mg / kg BID doses of Compound A with niraparib resulted in statistically significant increases in tumor volume at day 28 compared to both vehicle and niraparib (TGI% 97 and 105, respectively) (p=0.03 for both) (Figure 3).

[0250] 8. Efficacy Study of Prodrug Example 1 and Compound A in a BRCA Mutant Ovarian PDX Model The effects of prodrug Example 1 and Compound A on tumor growth were evaluated in mice bearing an ovarian PDX model, the 134-T model. A dose of 25 mg / kg niraparib QD resulted in a significant difference in tumor volume compared to vehicle (60% TGI, p<0.0001), and a dose of 1 mg / kg Compound A BID combined with niraparib resulted in a significant difference in tumor volume of 92% TGI (p<0.001) compared to vehicle and niraparib (Fig. 4). Additionally, a dose of 1 mg / kg Example 1 BID combined with niraparib showed a statistically significant difference in tumor volume compared to vehicle and niraparib, with a TGI of 103% (p<0.001) (Fig. 4). These results suggest that prodrug Example 1 demonstrated potent combination efficacy with niraparib in the 134-T HR-deficient PDX model.

[0251] 9. Efficacy Study of Prodrug Example 1 in the MDA-MB-436 TNBC CDX Model Efficacy of Prodrug Example 1 in Combination with Niraparib and Olaparib in the TNBC MDA-MB 436 CDX Model. 7 ) live MDA-MB-436 cells were implanted with 50% Matrigel into the dorsal flank of 5-7 week-old NSG mice (Jax). Tumor volumes were approximately 200 mm 3 Upon reaching a tumor size of 100 mg / kg, animals were randomized into an efficacy study. MDA-MB-436 tumor-bearing animals were dosed for 42 days with either the vehicle 0.5% methylcellulose BID PO, or with 1 mg / kg of Example BID or 2 mg / kg QD PO alone or in combination with 25 mg / kg niraparib QD PO or 100 mg / kg olaparib QD PO.

[0252] 1 mg / kg Example 1 BID induced a significant but modest monotherapy 31% tumor growth inhibition compared to vehicle at day 42 (p=0.0371), 25 mg / kg niraparib alone QD induced 76% tumor growth inhibition (p<0.0001), and 100 mg / kg olaparib alone QD dose induced 84% tumor growth inhibition compared to vehicle at day 42 (p<0.0001) (FIG. 5). Both the 1 mg / kg BID and 2 mg / kg QD doses of Example 1 induced statistically significant differences in tumor volume compared to vehicle at day 42 (101% and 94% TGI, respectively, p<0.0001), and also compared to niraparib at day 42 (p=0.00233). Dosing of 1 mg / kg Example 1 BID in combination with olaparib induced a significant tumor volume difference at 96% TGI on day 42 compared to vehicle (p<0.0001) and olaparib (p=0.002) (Figure 5).

[0253] The efficacy of the combination of prodrug Example 1 with niraparib was also studied in the homologous recombination (HR)-deficient (BRCA2 mutant) ovarian cancer cell line xenograft CDX PEO1 model and the HR-deficient (BRCA1 mutant) 134-T ovarian cancer PDX. In both studies, dosing of 1 mg / kg Example 1 BID in combination with niraparib induced statistically significant differences in tumor volume compared to vehicle and compared to niraparib.

[0254] 10. Resensitization of 134-T PDX tumors progressing on niraparib Animals bearing 134-T PDX tumors were treated with either vehicle or 25 mg / kg niraparib QD. Vehicle-treated animals reached the maximum tolerated tumor volume on day 42, while niraparib treatment progressed slowly (Figure 6). On day 21, the mean tumor volume reached 500 mm 3When tumor size reached 1400 mm, the animals treated with niraparib were randomized into three groups: the first group continued to receive 25 mg / kg niraparib alone; the second group switched to a dose of 1 mg / kg POLQ inhibitor, Example 1, BID; and the third group received a combination of 25 mg / kg niraparib and 1 mg / kg Example 1, BID. Tumors treated with niraparib alone progressed further, reaching approximately 1400 mm on day 45. 3 The group of Example 1 single agent grew slowly and reached a size of 945 mm 3 In addition, the combination group of 1 mg / kg niraparib BID and Example 1 significantly controlled disease progression, with a mean tumor volume of 525 mm 3 This data suggests that inhibiting POLQ may resensitize tumors progressing on PARP inhibitors.

[0255] The efficacy of the combination of Example 1 and niraparib is studied in a PARP inhibitor-resistant ovarian PDX model. See Parmar et al., "The CHK1 Inhibitor Prexasertib Exhibits Monotherapy Activity in High-Grade Serous Ovarian Cancer Models and Sensitizes to PARP Inhibition," Clin Cancer Res; 25(20), 6127-6140 (2019).

[0256] 11. Efficacy study in the DLD1 BRCA2 deletion CDX model The efficacy of Compound A or Niraparib as monotherapy, as well as the efficacy of the combination of Compound A and Niraparib, was evaluated in the BRCA2 deletion model DLD1 (DLD1 BRCA2 del ) was studied. Furthermore, this study investigated the durability and response rate of the combination of Compound A and niraparib. The efficacy of each treatment group was compared with a vehicle control group or niraparib administered alone at 35 mg / kg QD (once daily). Furthermore, the efficacy of the treatment response for the combination group was compared with niraparib alone.

[0257] DLD1 BRCA2 del Cells were grown in RPMI containing 10% fetal bovine serum. The mean tumor volume at the start of dosing was approximately 148 mm 3 Seven mice were randomized to each treatment group. The study consisted of eight treatment groups. TGI was calculated by comparing the treatment groups on day 24 with the control group on day 24. Compound A administered at 3 mg / kg, 10 mg / kg, or 30 mg / kg BID resulted in TGIs of -50%, -64%, and -12%, respectively. Niraparib resulted in a TGI of -12%. The combination of 35 mg / kg niraparib QD with Compound A administered at 3 mg / kg, 10 mg / kg, or 30 mg / kg resulted in mean TGIs of 112%, 111%, and 117%, respectively. The vehicle, Compound A, and Niraparib monotherapy groups were terminated on day 24, and the Compound A and Niraparib combination group continued the study. All treatments were discontinued on day 54, and tumor regrowth was evaluated until day 122. The combination group did not progress after treatment ended (see Figure 7). The combination of Compound A and niraparib enhanced complete responses and tumor shrinkage, and the responses were maintained throughout the observation period.

[0258] 12. Efficacy study in HR-deficient human cell line xenograft model MDA-MB-436 The effect of Example 1 on tumor growth in vivo was evaluated in 6-8 week old female NOD SCID mice bearing the BRCA1 mutant MDA-MB-436 CDX model.

[0259] Ten mice were assigned to each treatment group in the efficacy study. The study consisted of four treatment groups. Mice were orally administered Vehicle A, BID (Vehicle of Example 1; 1% 400 cps methylcellulose in sterile water) and Vehicle B, QD (Vehicle of Niraparib, 0.5% 400 cps methylcellulose in sterile water) (Group 1), 1 mg / kg of Example 1, BID (Group 2), 25 mg / kg of Niraparib, QD (Group 3), and 25 mg / kg of Niraparib combined with 1 mg / kg of Example 1, BID, QD (Group 4).

[0260] The duration for the vehicle and Example 1 treatment groups was 29 days or earlier if the tumor volume endpoint criteria were met. The tumor volume endpoint for each treatment group was 50% of the mice within the treatment group currently or previously exceeding 2000 mm 3 If an individual mouse reached the endpoint before the group, it was euthanized and the remaining mice continued until they reached the group endpoint criterion.

[0261] The vehicle control group reached the endpoint tumor volume on study day 29, and both groups 1 and 2 were euthanized on study day 30. The study continued for groups 3 and 4 until study day 78. Figure 8 shows the mean tumor volumes including only mice that remained in the study on the day of measurement. Mice were euthanized when they reached the tumor volume endpoint or on day 78, whichever came first. No significant weight loss was observed at any time during the study.

[0262] Compared to the vehicle group, each treatment group resulted in a statistically significant TGI. Example 1, niraparib, and the combination resulted in a TGI of 35%, 79%, and 88%, respectively. At 78 days, the efficacy of the combination of Example 1 and niraparib was compared to the efficacy of niraparib alone. The combination of niraparib and Example 1 resulted in a TGI of 23.7 mm 3 All tumors responded with 100% tumor shrinkage. In addition to tumor shrinkage, complete responses were also observed in 70% of mice.

[0263] 13. Efficacy study in 031-T ovarian PDX tumors progressing on niraparib In vivo efficacy studies were conducted using an HR-deficient (BRCA1 nonsense mutation) carboplatin-treated advanced ovarian carcinoma patient-derived xenograft (PDX) model, 031-T.

[0264] 4mm x 4mm fragments of the HR-deficient (BRCA1 mutant) 031-T ovarian cancer PDX model were implanted into female NOD SCID gamma (NSG) mice, resulting in a mean tumor volume of approximately 100mm. 3 When tumor volume reached 500 mm, animals were randomized and treated with either vehicle or niraparib (35 mg / kg, QD). Tumors did not respond to 35 mg / kg niraparib QD and progressed within 27 days. On day 27, the mean tumor volume was 500 mm. 3 When tumor size reached 59 days, animals receiving niraparib were randomized into two groups: the first group continued to receive 35 mg / kg niraparib QD alone; the second group received a dose combination of 35 mg / kg niraparib QD and 1 mg / kg Example 1 BID (n=4). Both vehicle and niraparib-only tumors progressed further, reaching approximately 2000 mm on day 59. 3 However, the combination group of niraparib and Example 1 administered at 1 mg / kg BID significantly delayed disease progression, with a mean tumor volume of 230 mm 3 (27% tumor shrinkage at day 27 vs. 123% TGI) (Figure 9). This data suggests that the addition of a POLQ inhibitor delays tumor progression to a PARP inhibitor, while niraparib monotherapy results in continued tumor growth.

[0265] Certain embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. It is anticipated that variations on the disclosed embodiments may become apparent to those skilled in the art upon reading the foregoing description, and that such variations may be employed by those skilled in the art as desired. Accordingly, the present invention may be carried out otherwise than as specifically described herein, and the present invention is intended to include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all such possible variations is encompassed by the present invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0266] All publications, patent applications, accession numbers, and other references mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

Claims

1. Compounds of formula (I) 【Chemistry 1】 (In the formula, R １ is H, halo, C １～４ Alkyl, C １～４ Alkoxy, C １～４ haloalkyl, or C １～４ haloalkoxy; Each R ２ are independently halo, C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Alkoxy, or C １～４ haloalkoxy; X is a prodrug moiety; R ３ is C １～４ Alkyl, C １～４ Haloalkyl, C ３～６ cycloalkyl, or heterocycloalkyl, ３～６ The cycloalkyl and the heterocycloalkyl may each be selected from 1 to 4 R ３ａ Optionally substituted with substituents, each of which is halo, C １～４ Alkyl, C １～４ Haloalkyl, C １～４ Haloalkoxy, -L ３ -O-C １～４ Alkyl, -L ３ independently selected from —OH, and oxo; Each L ３ is a bond and C １～４ independently selected from alkylene; each heterocycloalkyl has 4 to 6 ring members and 1 to 3 heteroatoms as ring vertices independently selected from N, O, and S; n is 0, 1, 2, or 3. or a pharmaceutically acceptable salt thereof.

2. X is —CH(R ｃ )-OP(O)(OR ａ ) (OR ｂ ), —CH(R ｃ )-O-C(O)-C １～６ Alkylene CO ２ H, —CH(R ｃ )-O-C(O)-C １～６ Alkylene O-P(O)(OR ａ ) (OR ｂ ), —CH(R ｃ )-O-C(O)-C １～６ Alkylene-P(O)(OR ａ ) (OR ｂ ), —CH(R ｃ )-O-C(O)-C １～６ Alkylene NR ａ R ｂ , or —CH(R ｃ )-O-C(O)-C １～６ alkyleneheterocycloalkyl, and R ａ and R ｂ are each independently H or C １～４ alkyl, and R ｃ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is independently selected from hydrogen and methyl.

3. X is -CH ２ O-P(O)(OR ａ ) (OR ｂ ) or -CH ２ —O—C(O)—C １～６ Alkylene-CO ２ 2. The compound of claim 1, wherein R is H, or a pharmaceutically acceptable salt thereof.

4. X is -CH ２ O-P(O)(OR ａ ) (OR ｂ 2. The compound of claim 1, wherein:

5. X is -CH ２ —O—C(O)—C １～６ Alkylene-CO ２ 2. The compound of claim 1, wherein R is H, or a pharmaceutically acceptable salt thereof.

6. X is, 【Chemistry 2】 2. The compound of claim 1, wherein:

7. X is, 【Transformation 3】 2. The compound of claim 1, wherein:

8. X is, 【Chemistry 4】 2. The compound of claim 1, wherein:

9. X is, 【Transformation 5】 2. The compound of claim 1, wherein:

10. R １ is C １～４ 10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein R is alkyl.

11. R １ The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein is methyl.

12. Each R ２ But independently, Halo, C １～４ Alkoxy, or C １～４ 12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, which is haloalkoxy.

13. Each R ２ are independently halo or C １～４ 12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, which is alkoxy.

14. Each R ２ The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein is independently F, Cl, or methoxy.

15. The compound according to any one of claims 1 to 14, wherein n is 1, 2, or 3, or a pharmaceutically acceptable salt thereof.

16. R ３ But C １～４ Alkyl or C １～４ 16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, which is haloalkyl.

17. R ３ But C １～４ 16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein R is alkyl.

18. R ３ The compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein is methyl.

19. R ３ But C ３～６ 16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, which is cycloalkyl or heterocycloalkyl.

20. The compound of claim 1 selected from the group consisting of: 【Transformation 6】 or a pharmaceutically acceptable salt thereof.

21. A pharmaceutical composition comprising a compound according to any one of claims 1 to 20 and at least one pharmaceutically acceptable excipient.

22. A method for treating a disease characterized by overexpression of Pol θ in a patient, comprising administering to the patient a therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 21.

23. 23. The method of claim 22, wherein the patient is in recognized need of such treatment and the disease is cancer.

24. 22. A method for treating homologous recombination (HR) deficient cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 21.

25. 25. The method of claim 24, wherein the patient is in recognized need of such treatment.

26. A method for treating cancer in a patient, wherein the cancer is characterized by reduced or absent expression of BRCA1 and / or BRCA2 genes, the absence or mutation of BRCA1 and / or BRCA2 genes, or reduced function of BRCA1 and / or BRCA2 proteins, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 21.

27. 27. The method of any one of claims 22 to 26, wherein the cancer is lymphoma, rhabdoid tumor, multiple myeloma, uterine cancer, gastric cancer, peripheral nervous system cancer, rhabdomyosarcoma, bone cancer, colorectal cancer, mesothelioma, breast cancer, ovarian cancer, lung cancer, fibroblastic cancer, central nervous system cancer, urinary tract cancer, upper aerodigestive tract cancer, leukemia, kidney cancer, skin cancer, esophageal cancer, and pancreatic cancer.

28. 28. The method of any one of claims 22 to 27, further comprising administering to the patient a therapeutically effective amount of a PARP inhibitor or a pharmaceutically acceptable salt thereof.

29. 29. The method of claim 28, wherein the PARP inhibitor is niraparib.

30. 30. The method of claim 29, wherein the PARP inhibitor is niraparibut tosylate monohydrate.

31. 29. The method of claim 28, wherein the PARP inhibitor is olaparib.

32. A Pol θ inhibitor for use in the treatment of cancer, wherein the Pol θ inhibitor is a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 21.

33. Use of a Pol θ inhibitor in the manufacture of a medicament for treating cancer, wherein the Pol θ inhibitor is a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 21.

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