Tiparp inhibitor compounds

Abstract

The present disclosure provides for compounds of Formula (I), and pharmaceutically acceptable salts thereof, that inhibit the activity of TIPARP, wherein the variables have any of the values defined in the specification.

Description

(19) *EP004644385A1* (11) EP 4 644 385 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: 05.11.2025 Bulletin 2025 / 45 (21) Application number: 25173293.9 (22) Date of filing: 29.04.2025 (51) International Patent Classification (IPC): C07D 401 / 14 (2006.01) A61P 35 / 00 (2006.01) A61K 31 / 506 (2006.01) (52) Cooperative Patent Classification (CPC): C07D 401 / 14; A61P 35 / 00 (84) Designated Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC ME MK MT NL NO PL PT RO RS SE SI SK SM TR Designated Extension States: BA Designated Validation States: GE KH LA MA MD TN (30) Priority: 30.04.2024 US 202463640741 P (71) Applicant: AbbVie Inc. North Chicago, IL 60064 (US) (72) Inventors: • Bai, Wen Ju Vernon Hills, 60061 (US) • Cohen, Daniel Wilmette, 60091 (US) • Dai, Yujia Gurnee, 60031 (US) • De La Rosa, Martha A. Gurnee, 60031 (US) • Dubovyk, Igor San Jose, 95126 (US) • Frey, Robin Libertyville, 60048 (US) • Ji, Zhiqin Libertyville, 60048 (US) • Judd, Andrew Grayslake, 60030 (US) • Liu, Dachun Vernon Hills, 60061 (US) • Mandal, Debashis Hayward, 94544 (US) • Punna, Sreenivas Sunnyvale, 94085 (US) (74) Representative: J A Kemp LLP 80 Turnmill Street London EC1M 5QU (GB) (54) TIPARP INHIBITOR COMPOUNDS (57) The present disclosure provides for compounds of Formula (I), and pharmaceutically acceptable salts thereof, that inhibit the activity of TIPARP, wherein the variables have any of the values defined in the specifica- tion. EP 4 64 4 38 5 A 1 Processed by Luminess, 75001 PARIS (FR) Description FIELD

[0001] The present disclosure pertains to compounds that inhibit the activity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)‑inducible poly[adenosine diphosphate (ADP)‑ribose] polymerase (TIPARP). BACKGROUND

[0002] The poly(ADP-ribose) polymerase (PARP) family of enzymes regulates fundamental cellular processes, including transcription, metabolism, and multiple cellular stress responses, through ADP-ribosylation. TIPARP, also known as PARP7, is a PARP enzyme that negatively regulates type I interferon (IFN) signaling. The gene responsible for encoding TIPARP is located in a region on chromosome 3q that frequently has copy number gains in some tumors, resulting in increased expression and restriction of anti-tumor immune responses. It is hypothesized that the inhibition of TIPARP in such tumors can restore type I IFN signaling and selectively activate anti-tumor immune responses in the tumor microenvironment, avoiding systematic cytokine production. (Joseph M. Gozgit, et al., PARP7 Negatively Regulates the Type I Interferon Response in Cancer Cells and Its Inhibition Triggers Antitumor Immunity, 39 CANCER CELL 1214 (2021)). However, off-target activity may prevent full engagement of the immune-mediatedmechanism of action by some compounds that inhibit TIPARP.Compounds that lack sufficient selectivity for TIPARPmay functionally inhibit other PARP enzymes (e.g., PARP1) at concentrations relevant for the inhibition of TIPARP, resulting in application-limiting side effects.

[0003] There remains a need in the art for improved compounds that selectively inhibit TIPARP. In particular, there remains a need in the art for compounds that exhibit high potency and high selectivity for TIPARP inhibition, while also exhibiting low inhibition of other PARP enzymes. SUMMARY

[0004] In one aspect, the invention provides for a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein: L is selected from the group consisting of a bond and CH2; R1, R2, R3, and R4 are independently selected from the group consisting of H and CH3, wherein at least one and no more than two of R1, R2, R3, and R4 is CH3; and Z is selected from the group consisting of CF3 and C(CH3)2OH.

[0005] Another aspect of the invention provides for the compound of the first aspect, or a pharmaceutically acceptable salt thereof, wherein L is a bond.

[0006] Another aspect of the invention provides for the compound of the second aspect, or a pharmaceutically acceptable salt thereof, wherein one of R1 or R4 is CH3; both R1 and R4 are CH3; one of R2 or R3 is CH3; or both R2 and R3 are CH3.

[0007] Another aspect of the invention provides for the compound of the second or third aspect, or a pharmaceutically acceptable salt thereof, wherein Z is CF3.

[0008] Another aspect of the invention provides for the compound of the first aspect, or a pharmaceutically acceptable salt thereof, wherein L is CH2.

[0009] Another aspect of the invention provides for the compound of the fifth aspect, or a pharmaceutically acceptable 2 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 salt thereof, wherein one of R1 or R4 is CH3; both R1 and R4 are CH3; one of R2 or R3 is CH3; or both R2 and R3 are CH3.

[0010] Another aspect of the invention provides for the compound of the fifth or sixth aspect, or a pharmaceutically acceptable salt thereof, wherein Z is C(CH3)2OH.

[0011] Another aspect of the invention provides for the compound of the first aspect, wherein the compound is selected from the group consisting of: 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one; 5‑[(1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluor- omethyl)pyridin‑2(1H)‑one; 5‑[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one; 5‑[(1s,3s)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-ylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluor- omethyl)pyridin‑2(1H)‑one; 5‑[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyl}cyclobu- tyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one; 5‑[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluor- omethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one; 5‑[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluor- omethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluor- omethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluor- omethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1s,3s)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl- methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; and 5‑{[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one.

[0012] Another aspect of the invention provides for 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl] piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one, or a pharmaceutically acceptable salt thereof.

[0013] Another aspect of the invention provides for 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl] piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one.

[0014] Another aspect of the invention provides for a pharmaceutically acceptable salt of 5‑[(1S,3s)‑3‑{(2R)‑2- methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one.

[0015] Another aspect of the invention provides for a pharmaceutical composition comprising the compound of any of 3 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 the aspects above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0016] Another aspect of the invention provides for a method for treating head and neck squamous cell carcinoma (HNSCC), the method comprising administering the compound of the first aspect, or a pharmaceutically acceptable salt thereof, to a human patient in need thereof. The invention also provides a compound of any of the aspects above, or a pharmaceutically acceptable salt thereof, for use in a method for treating head and neck squamous cell carcinoma (HNSCC). The invention also provides a pharmaceutical composition comprising the compound of any of the aspects above, or a pharmaceutically acceptable salt thereof, for use in a method for treating head and neck squamous cell carcinoma. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] FIG. 1 depicts the in vivo efficacy of Example 1 in reducing tumor volume in the MC‑38 (Ker) syngeneic model. FIG. 2 depicts the induction of IFNβ in plasmaand tumors in theMC‑38 (Ker) syngeneicmodel after the administration of Example 1. FIG. 3 depicts the in vivo efficacy of Example 1 in reducing tumor volume in an NCI-H1373 xenograft model. DETAILED DESCRIPTION

[0018] The present invention provides for compounds that inhibit the activity of TIPARP.

[0019] Compounds disclosed herein, including any intermediates,may contain one ormore variable(s) that occurmore than one time in any substituent or in the Formulae herein. Definition of a variable on each occurrence is independent of its definition at another occurrence.

[0020] Compounds disclosed herein, including any intermediates, were named by using ACD / Name 2023.1.2 (File VersionN25E41, Build 134315, 12 July 2023) software programand / or by usingStruct=Namenaming algorithmas part of CHEMDRAW® Professional v. 20.1.1.125.

[0021] Compounds disclosed herein, including any intermediates, may possessmultiple tautomeric forms and exist as equilibrium mixtures thereof. The formulae and structures found herein represent only one of the possible tautomeric forms but should be understood to encompass both individual tautomeric forms and mixtures thereof. DEFINITIONS

[0022] As used in the specification and the appended claims, unless specified to the contrary, the following terms have the meaning indicated:

[0023] The phrase "pharmaceutical composition" refers to a composition suitable for administration in medical use.

[0024] The phrase "pharmaceutically acceptable salt" refers to those salts which are, within the scope of soundmedical judgement, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit / risk ratio.

[0025] The phrase "therapeutically effective amount" refers to an amount of a compound, or a pharmaceutically acceptable salt thereof, sufficient to prevent thedevelopment of or to alleviate to someextent oneormoreof the symptoms of the condition or disorder being treated when administered for treatment in a particular human patient or human patient population.

[0026] The terms "treat," "treating," and "treatment," as used herein, refer to a method of alleviating or abrogating a disease and / or its attendant symptoms. COMPOUNDS

[0027] The present disclosure provides for compounds of Formula (I), or pharmaceutically acceptable salts thereof, 4 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 wherein: L is selected from the group consisting of a bond and CH2; R1, R2, R3, and R4 are independently selected from the group consisting of H and CH3, wherein at least one and no more than two of R1, R2, R3, and R4 is CH3; and Z is selected from the group consisting of CF3 and C(CH3)2OH.

[0028] Exemplary compounds of Formula (I) are shown in Table 1 below. It is to be understood that when there is a discrepancy between the name of any compounds disclosed herein and the structures found in Table 1, the structures in Table 1 shall prevail. Table 1 Ex. # Structure Ex. # Structure 1 12 2 13 5 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (continued) Ex. # Structure Ex. # Structure 3 14 4 15 5 16 6 17 6 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (continued) Ex. # Structure Ex. # Structure 7 18 8 19 9 20 10 21 7 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (continued) Ex. # Structure Ex. # Structure 11 22

[0029] Compounds of Formula (I) may be used in the form of pharmaceutically acceptable salts. Such compoundsmay contain either a basic or an acidic functionality, or both, andmaybe converted to a pharmaceutically acceptable salt, when desired, by using a suitable acid or base.

[0030] The compounds of Formula (I) exhibit a combination of functional properties, including but not limited to, high potency for inhibiting TIPARP, high selectivity for inhibiting TIPARP compared to other PARP family enzymes, and low inhibition of PARP1 and / or PARP2. The compounds of Formula (I) may exhibit high potency and selectivity for inhibiting TIPARP. Additionally, the compounds of Formula (I) may exhibit low inhibition of PARP enzymes other than TIPARP. As used herein, low inhibition of PARP enzymes other than TIPARP means the compounds of Formula (I) lack sufficient potency for inhibiting PARP enzymes other than TIPARP, For example, the compounds of Formula (I) may exhibit low inhibition of PARP1, PARP2, or combinations thereof. METHODS OF MAKING EXEMPLARY COMPOUNDS

[0031] The compounds of the present disclosure may be better understood in connection with the following synthetic schemes andmethodswhich illustrate ameans bywhich the compounds can be prepared. The compounds of the present disclosure can be prepared by a variety of synthetic procedures. Representative synthetic procedures are shown in Schemes 1‑8. The variables have any of the values defined herein, e.g., in the Summary. SYNTHETIC SCHEMES

[0032] 8 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55

[0033] As shown in Scheme 1, methyl 3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylate can be prepared by reacting 5-bromo‑2-methoxy‑3‑(trifluoromethyl)pyridine with methyl 3-bromocyclobutanecarboxylate. The reactionmay be performed in the presence of a cross coupling catalyst, such as (4,4’-dtbbpy)NiCl2, a photocatalyst, such as (Ir[dF(CF3)ppy]2(dtbpy))PF6, 1,1,1,3,3,3-hexamethyl‑2‑(trimethylsilyl)trisilane, 2,6-lutidine, and LED blue light. The reaction may be performed in an inert atmosphere in a solvent, such as 1,2-dimethoxyethane. The trans-isomer may be separated from the cis-isomer via flash chromatography.

[0034] As shown in Scheme 2, 5-bromo‑2-methoxy‑3‑(trifluoromethyl)pyridine can be reacted under Suzuki coupling conditions withmethyl 3‑((4,4,5,5-tetramethyl‑1,3,2-dioxaborolan‑2-yl)methylene)cyclobutane‑1-carboxylate, in the pre- sence of a catalyst, such as Pd(dppf)Cl2·CH2Cl2, and a base, such as potassium carbonate, to provide methyl 3‑((6- methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)methylene)cyclobutane‑1-carboxylate. The reaction is typically performed in an inert atmosphere at an elevated temperature and in a solvent, such as dioxane, water, or mixtures thereof.

[0035] Methyl 3‑((6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)methylene)cyclobutane‑1-carboxylate can be treated with hydrogen gas in the presence of a catalyst, such as Pd / C, to providemethyl 3‑((6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl) methyl)cyclobutane‑1-carboxylate, which may contain a mixture of separable cis‑ and trans-isomers. The trans-isomer may be separated from the cis-isomer via flash chromatography. The reaction is typically performed at ambient temperature in a solvent, such as tetrahydrofuran.

[0036] Scheme 3 shows the synthesis of compounds of Formula (3), wherein R1, R2, R3, R4 and Z are as described herein. Compounds of Formula (1) can be reacted with compounds of Formula (2) in the presence of a base, such as triethylamine, to provide compounds of Formula (3). The reaction is typically performed in a solvent, such as acetonitrile. 9 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55

[0037] Scheme 4 shows an alternate synthesis of compounds of Formula (3), wherein R1, R2, R3, R4, and Z are as described herein. Compounds of Formula (4), wherein PG is a protecting group such as aBOCgroup, can be reactedwith compounds of Formula (2) in the presence of a base, such as triethylamine or N,N-diisopropylethylamine, to provide compounds of Formula (5). The reaction is typically performed in a solvent such as acetonitrile or dimethylacetamide. WhenPG is a BOCgroup, compounds of Formula (5) can be treatedwith an acid, such as trifluoroacetic acid or hydrogen chloride, in a solvent, such as dichloromethane, acetonitrile, dioxane, or mixtures thereof, to provide compounds of Formula (3).

[0038] Scheme5shows thesynthesisof compoundsofFormula (8),whereinR1,R2,R3, andR4areasdescribedherein. Compounds of Formula (4) can be reacted with methyl 2-chloropyrimidine‑5-carboxylate in the presence of a base, such as potassium carbonate, to provide compounds of Formula (6). The reaction is typically performed at an elevated temperature in a solvent, such as N,N-dimethylformamide.

[0039] Alternatively, compounds of Formula (4) can be reacted with methyl 2-chloropyrimidine‑5-carboxylate in N- methyl‑2-pyrrolidinone to provide compounds of Formula (6). The reaction is typically performed at an elevated temperature.

[0040] Compounds of Formula (6) can be treated with methylmagnesium bromide solution to provide compounds of Formula (7). The reaction is typically performed under nitrogen at low temperature in a solvent, such as tetrahydrofuran. WhenPG is a BOCgroup, compounds of Formula (7) can be treatedwith an acid, such as trifluoroacetic acid or hydrogen chloride, in a solvent, such as dichloromethane, ethyl acetate, acetonitrile, dioxane, or mixtures thereof, to provide compounds of Formula (8). 10 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55

[0041] As shown in Scheme 6, the treatment of compounds of Formula (9), which can be prepared as described in Schemes1‑2,with lithiumhydroxideorsodiumhydroxide inwater canprovidecompoundsofFormula (10).The reaction is typically performed at ambient temperature in a solvent, such as methanol, tetrahydrofuran, or mixtures thereof.

[0042] As shown inScheme 7, compounds of Formula (10) can be treatedwith para-toluenesulfonic acidmonohydrate and lithium chloride to provide compounds of Formula (11). The reaction is typically performed at an elevated temperature in a solvent, such as N,N-dimethylformamide.

[0043] Compounds of Formula (I) can be prepared by reacting compounds of Formula (11), wherein L is a bond or CH2, with compounds of Formula (3) in the presence of propanephosphonic acid anhydride and a base, such as N,N- diisopropylethylamine. The reaction is typically performed at ambient temperature in a solvent such as N,N-dimethyl- formamide.

[0044] Alternatively, when L is a bond or CH2, compounds of Formula (I) can be prepared by reacting compounds of Formula (11)with compounds of Formula (3) in the presence of 1‑[bis(dimethylamino)methylene]‑1H‑1,2,3-triazolo[4,5‑b] pyridinium 3-oxid hexafluorophosphate and a base, such as N,N-diisopropylethylamine. The reaction is typically performed at ambient or elevated temperatures in a solvent such as N,N-dimethylformamide. 11 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55

[0045] As shown in Scheme 8, compounds of Formula (12) can be prepared by reacting compounds of Formula (10), wherein L is a bond or CH2, with compounds of Formula (3) in the presence of propanephosphonic acid anhydride and a base, such asN,N-diisopropylethylamine. The reaction is typically performed at ambient temperature in a solvent such as N,N-dimethylformamide.

[0046] Compounds of Formula (12) can be treated with para-toluenesulfonic acid monohydrate and lithium chloride to providecompoundsofFormula (I). The reaction is typically performed inan inert atmosphereat anelevated temperature in a solvent such as N,N-dimethylformamide.

[0047] Specific procedures are also provided in the Synthetic Examples section. Unless otherwise described, the starting materials and reagents are either commercially available or may be prepared by one skilled in the art from commercially available materials using methods known in the art. PHARMACEUTICAL COMPOSITIONS

[0048] Whenemployedasapharmaceutical, a compoundof thepresent disclosuremaybeadministered in the formof a pharmaceutical composition. Such composition may comprise a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, together with one or more pharmaceutically acceptable excipients. METHODS OF USE

[0049] The compounds of Formula (I), or pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, may be administered to a human patient suffering fromheadandnecksquamouscell carcinoma (HNSCC).The term "administering" refers to themethodof contacting a human patient with a compound.

[0050] Thecompoundsof Formula (I), or pharmaceutically acceptable salts thereof,mayalsobeused in thepreparation of a medicament. Such medicament may be used in the treatment of HNSCC. EXAMPLES SYNTHETIC EXAMPLES Example 1 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one Example 1A 12 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (1s,3s)‑methyl 3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylate

[0051] In Example 1A, 5-Bromo‑2-methoxy‑3‑(trifluoromethyl)pyridine (5.12 g, 20.0 mmol), methyl 3-bromocyclobu- tanecarboxylate (4.63 g, 24.0 mmol), (4,4’-dtbbpy)NiCl2 (398 mg, 1.00 mmol, CAS 1034901‑50‑2), (Ir[dF(CF3)ppy]2 (dtbpy))PF6 (224 mg, 0.200 mmol, CAS 870987‑63‑6), 1,1,1,3,3,3-hexamethyl‑2‑(trimethylsilyl)trisilane (5.97 g, 24.0 mmol) and 2,6-lutidine (5.82mL, 50.0mmol)were combinedwith 1,2-dimethoxyethane (80mL). The reactionmixturewas purged with nitrogen for 5minutes and stirred for 16 hours under irradiation (450 nm LED blue light). The reactionmixture was partitioned between ethyl acetate andwater. The organic layer waswashedwith saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by flash chromatography (silica gel, 0‑30% ethyl acetate in heptanes) to give two fractions. The first eluted fraction was the stereoisomerwith trans-configuration. The second eluted fractionwas the title compoundwith cis-configuration.MS (ESI) m / z 290 (M+H)+. Example 1B (1s,3s)‑3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylic acid

[0052] To a solution of Example 1A (900 mg, 3.11 mmol) in the mixture of tetrahydrofuran (6.0 mL) / methanol (6.0 mL) / water (2.0 mL) was added sodium hydroxide (622 mg, 15.6 mmol). The reaction mixture was stirred at room temperature for 1 hour and dilutedwithwater. The pHwasadjusted to 4with the addition of 1MaqueousHCl and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, and filtered. The filtratewas concentrated under vacuum to provide the title compound.MS (ESI)m / z 276 (M+H)+. Example 1C (1s,3s)‑3‑(6-oxo‑5‑(trifluoromethyl)‑1,6-dihydropyridin‑3-yl)cyclobutanecarboxylic acid

[0053] Example 1B (730 mg, 2.65 mmol), para-toluenesulfonic acid monohydrate (1.01 g, 5.30 mmol), and lithium chloride (562mg, 13.3mmol) were combined inN,N-dimethylformamide (15mL). The reactionmixture was stirred at 100 °C for 10 hours, cooled to room temperature, and partitioned between ethyl acetate and water. The organic layer was washed with saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to provide the title compound. MS (ESI) m / z 262 (M+H)+. Example 1D (R)‑2‑(3-methylpiperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0054] In Example 1D, a solution of (R)‑2-methylpiperazine (2.50 g, 25.0mmol) and triethylamine (6.96mL, 49.9mmol) in acetonitrile (80 mL) was cooled to 0 °C in an ice bath, and treated with 2-chloro‑5‑(trifluoromethyl)pyrimidine (4.56 g, 25.0 mmol). The reaction mixture was stirred at room temperature overnight. The insoluble salt material was filtered and washed with a small amount of acetonitrile. The filtrate was concentrated under vacuum. The corresponding residue was triturated with diethyl ether (25 mL), filtered, and dried under vacuum to provide the title compound. The filtrate was concentrated under vacuum to about 8 mL, placed in a refrigerator overnight, and the mixture was filtered. The material was dried under vacuum to provide additional title compound. MS (APCI) m / z 288 (M+CH3CN+H)+. Example 1E 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one

[0055] To a mixture of Example 1C (20.9 mg, 0.0800 mmol), Example 1D (22.6 mg, 0.0920 mmol) and N,N-diisopro- pylethylamine (0.0840 mL, 0.480 mmol) in N,N-dimethylformamide (1.0 mL) was added 50% propanephosphonic acid anhydride in N,N-dimethylformamide (102 mg, 0.160 mmol). The reaction mixture was stirred at room temperature for 1 hour and partitioned between ethyl acetate and water. The organic layer was washed with saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, filtered, concentrated, and purified by flash chromatography to provide the title compound. 1H NMR (400MHz, DMSO‑d6, 90 °C) δ ppm 11.84 (s, br, 1H), 8.64 (d, J = 0.8 Hz, 2H), 7.77 (d, J = 2.6 Hz, 1H), 7.42 (d, J =2.6Hz, 1H), 4.64 - 4.29 (m,3H), 3.96 (s, br, 1H), 3.38 - 3.24 (m, 3H), 3.23 - 3.04 (m, 2H), 2.54 - 2.45 (m, 2H), 2.25 - 2.09 (m, 2H), 1.08 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 490 (M+H)+. 13 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 2 5‑[(1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0056] Example 2 was prepared according to the procedure used for the preparation of Example 13F, substituting Example 1C for Example 13D, to provide the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δppm11.84 (s, br, 1H), 8.65 (d, J = 0.8Hz, 2H), 7.79 (d, J = 2.6Hz, 1H), 7.43 (d, J = 2.6Hz, 1H), 4.60 - 4.52 (m, 2H), 4.41 (s, br, 2H), 3.37 - 3.20 (m, 4H), 2.54 - 2.45 (m, 2H), 2.25 - 2.16 (m, 2H), 1.17 (d, J = 7.0 Hz, 6H). MS (ESI) m / z 504 (M+H)+. Example 3 5‑[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one Example 3A (R)-tert-butyl 3-methyl‑4‑(5‑(trifluoromethyl)pyrimidin‑2-yl)piperazine‑1-carboxylate

[0057] In Example 3A, a mixture of 2-chloro‑5‑(trifluoromethyl)pyrimidine (5.00 g, 27.4 mmol), (R)‑tert-butyl 3-methyl- piperazine‑1-carboxylate (5.49g,27.4mmol) and triethylamine (7.64mL,54.8mmol) inacetonitrile (100mL)wasstirredat 80 °C for 3hours, andcooled to room temperature. The insoluble saltmaterialwasfilteredandwashedwithasmall amount of acetonitrile. The filtrate was concentrated under vacuum. To the residue was added water, and the mixture was extracted with ethyl acetate twice. The combined organic layers were washed with saturated aqueous sodium chloride, dried with anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash chromatography (silica gel, 0‑25% ethyl acetate in heptanes) to provide the title compound. MS (APCI) m / z 347 (M+H)+. Example 3B (R)‑2‑(2-methylpiperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0058] A solution of Example 3A (8.02 g, 23.2 mmol) in dioxane (50 mL) was treated with 4M hydrogen chloride in dioxane (57.9ml, 232mmol) at 0 °C. The reactionmixturewas stirred at room temperature overnight and filtered. The filter cake waswashed with diethyl ether and dried under vacuum to provide the title compound as a hydrochloric acid salt. MS (ESI) m / z 247 (M+H)+. Example 3C 5‑[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one

[0059] To a mixture of Example 1C (20 mg, 0.077 mmol), Example 3B (22 mg, 0.077 mmol) and N,N-diisopropylethy- lamine (0.080mL, 0.46mmol) inN,N-dimethylformamide (1.0mL) was added 50%propanephosphonic acid anhydride in N,N-dimethylformamide (97mg, 0.15mmol). The reactionmixturewas stirred at room temperature for 1 hour, and purified by reverse phaseHPLC (C18, 20‑100%acetonitrile in water with 0.1% trifluoroacetic acid). The acetonitrile was removed by evaporation, and the mixture was neutralized with saturated aqueous sodium bicarbonate. Themixture was extracted withethyl acetateand theorganic layerwasdriedwithanhydroussodiumsulfate andfiltered.Thefiltratewasconcentrated under vacuum to provide the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δppm11.84 (s, 1H), 8.66 (d, J=0.8Hz, 2H), 7.78 (d, J=2.6Hz, 1H), 7.43 (d, J=2.6Hz, 1H), 4.94 - 4.85 (m, 1H), 4.52 - 4.43 (m, 1H), 4.14 (s, br, 1H), 3.85 (s, br, 1H), 3.39 - 3.23 (m, 3H), 3.07 (s, br, 2H), 2.55 - 2.49 (m, 2H), 2.24 - 2.14 (m, 2H), 1.14 (d, J = 6.7 Hz, 3H). MS (ESI)m / z 490 (M+H)+. Example 4 5‑[(1s,3s)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one 14 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 4A (3R,5S)‑tert-butyl 3,5-dimethyl‑4‑(5‑(trifluoromethyl)pyrimidin‑2-yl)piperazine‑1-carboxylate

[0060] To a solution of tert-butyl (3R,5S)‑3,5-dimethylpiperazine‑1-carboxylate (2.00 g, 9.33 mmol) in dimethylaceta- mide (18.7 mL) was added N,N-diisopropylethylamine (2.45 ml, 14.0 mmol) and 2-chloro‑5‑(trifluoromethyl)pyrimidine (1.70 g, 9.33mmol). The reactionmixturewas stirred at 90 °C for 18 hours, cooled to room temperature, dilutedwithwater, and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium chloride twice, dried with anhydrousmagnesiumsulfate, andfiltered. Thefiltratewasconcentratedunder vacuum.The residuewaspurifiedby flash chromatography (silica gel, 1‑20%ethyl acetate in heptanes) to provide the title compound.MS (ESI)m / z 305 (M‑56+H)+. Example 4B 2‑((2R,6S)‑2,6-dimethylpiperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0061] A solution of Example 4A (2.18 g, 6.05 mmol) in dioxane (20.2 mL) was treated with 4M hydrogen chloride in dioxane (15.1ml, 60.4mmol). The reactionmixture was stirred at room temperature overnight, filtered, and the filter cake was dried under vacuum to provide the title compound as a hydrochloric acid salt. MS (ESI) m / z 261 (M+H)+. Example 4C 5‑[(1s,3s)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0062] Example 4C was prepared according to the procedure used for the preparation of Example 3C, substituting Example 4B forExample 3B, to provide the title compound. 1HNMR(400MHz,DMSO‑d6, 90 °C)δppm11.82 (s, 1H), 8.68 (s, 2H), 7.80 (d, J = 2.6 Hz, 1H), 7.44 (d, J= 2.6Hz, 1H), 4.89 - 4.79 (m, 2H), 4.05 (s, br, 2H), 3.45 - 3.24 (m, 2H), 3.11 (s, br, 2H), 2.57 - 2.50 (m, 2H), 2.29 - 2.16 (m, 2H), 1.19 (d, J = 6.9 Hz, 6H). MS (ESI) m / z 504 (M+H)+. Example 5 5‑[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0063] Example 5 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 22C for Example 1D. Purification by flash chromatography provided the title compound. 1H NMR (400 MHz, DMSO‑d6, 90 °C) δ ppm11.84 (s, br, 1H), 8.44 (s, 2H), 7.79 (d, J= 2.6Hz, 1H), 7.44 (d, J= 2.6Hz, 1H), 4.84 - 4.67 (m, 3H), 3.97 (s, br, 2H), 3.48 - 3.22 (m, 4H), 2.56 - 2.49 (m, 2H), 2.27 - 2.18 (m, 2H), 1.43 (s, 6H), 1.14 (d, J = 6.8 Hz, 6H). MS (ESI) m / z 476 (M-H2O+H)+. Example 6 5‑[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one Example 6A (S)‑tert-butyl 2-methyl‑4‑(5‑(trifluoromethyl)pyrimidin‑2-yl)piperazine‑1-carboxylate

[0064] InExample6A, (S)‑tert‑Butyl 2-methylpiperazine‑1-carboxylate (4.00g, 20.0mmol), 2-chloro‑5‑(trifluoromethyl) pyrimidine (3.65g, 20.0mmol) and triethylamine (5.57mL, 39.9mmol)were combined in acetonitrile (80mL). The reaction mixture was stirred at room temperature overnight. The insoluble salt material was filtered. The filtrate was concentrated under vacuum.The residuewas trituratedwithwater, filtered, rinsedwithwater, anddriedunder vacuum toprovide the title compound. MS (ESI) m / z 291 (M‑56+H)+. Example 6B 15 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (S)‑2‑(3-methylpiperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0065] To a solution of Example 6A (6.20 g, 17.9 mmol) in acetonitrile (50 mL) was added 4M hydrogen chloride in dioxane (22.4 mL, 89.6 mmol). The reaction mixture was stirred at room temperature for 4 hours, diluted with tert-butyl methyl ether, and filtered. Thematerial was dried under vacuum to provide the title compound as a hydrochloric acid salt. MS (ESI) m / z 247 (M+H)+. Example 6C 5‑[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one

[0066] Example 6C was prepared according to the procedure used for the preparation of Example 1E, substituting Example 6B for Example 1D, to provide the title compound. 1HNMR (400MHz, DMSO‑d6, 90 °C) δ ppm 11.84 (s, br, 1H), 8.64 (d, J= 0.9Hz, 2H), 7.77 (d, J= 2.6Hz, 1H), 7.42 (d, J = 2.6Hz, 1H), 4.62 - 4.32 (m, 3H), 3.97 (s, br, 1H), 3.37 - 3.24 (m, 3H), 3.21 - 3.06 (m, 2H), 2.54 - 2.44 (m, 2H), 2.26 - 2.11 (m, 2H), 1.08 (d, J= 6.7 Hz, 3H). MS (ESI) m / z 490 (M+H)+. Example 7 5‑[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one Example 7A methyl (R)‑2‑(4‑(tert-butoxycarbonyl)‑2-methylpiperazin‑1-yl)pyrimidine‑5-carboxylate

[0067] To a solution ofmethyl 2-chloropyrimidine‑5-carboxylate (3.00 g, 17.4mmol) inN,N-dimethylformamide (30mL) was added potassium carbonate (7.21 g, 52.2 mmol) and (R)-tert-butyl 3-methylpiperazine‑1-carboxylate (3.48 g, 17.4 mmol). The reactionmixturewas stirred at 80 °C for 12 hours and filtered. The filtratewas slowly poured into icewater (500 mL). The material formed was collected via filtration. The filter cake was washed with water and dried under vacuum to provide the title compound. Example 7B tert-butyl (R)-4‑(5-(2-hydroxypropan‑2-yl)pyrimidin‑2-yl)‑3-methylpiperazine‑1-carboxylate

[0068] To a solution of Example 7A (3.50 g, 10.4 mmol) in tetrahydrofuran (35 mL) was added methylmagnesium bromide solution (31.2 mL, 31.2 mmol) dropwise over 30 minutes at ‑78 °C under nitrogen. The reaction mixture was allowed to slowly warm up to 25 °C overnight. The reaction mixture was quenched with saturated aqueous ammonium chloride and was extracted with ethyl acetate three times. The combined organic layers were washed with saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum. The residuewaspurifiedbyflashchromatography (silica gel, petroleumether / ethyl acetate=100:0 to35:65) toprovide the title compound. MS (ESI) m / z 337 (M+H)+. Example 7C (R)‑2‑(2‑(2-methylpiperazin‑1-yl)pyrimidin‑5-yl)propan‑2-ol

[0069] Asolutionof Example7B (2.60g, 7.73mmol) in dichloromethane (30mL)was treatedwith trifluoroacetic acid (10 mL). The reaction mixture was stirred at room temperature for 30 minutes, quenched by addition of saturated aqueous sodium bicarbonate, and concentrated to remove the organic solvent. The aqueous residue was lyophilized to provide a crude product, which was purified by reversed phase HPLC (C18, 5 - 45% acetonitrile in water containing 10 mM ammonium bicarbonate) to provide the title compound. MS (ESI) m / z 237 (M+H)+. Example 7D 16 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 5‑[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0070] Example 7D was prepared according to the procedure used for the preparation of Example 1E, substituting Example 7C for Example 1D. Purification by flash chromatography provided the title compound. 1H NMR (400 MHz, DMSO‑d6, 90 °C) δ ppm11.83 (s, br, 1H), 8.43 (s, 2H), 7.78 (d, J= 2.6Hz, 1H), 7.42 (d, J= 2.6Hz, 1H), 4.87 - 4.77 (m, 1H), 4.73 (s, 1H), 4.42 - 4.35 (m, 1H), 3.97 (s, br, 3H), 3.40 - 3.06 (m, 4H), 2.55 - 2.47 (m, 2H), 2.24 - 2.13 (m, 2H), 1.43 (s, 6H), 1.07 (d, J = 6.6 Hz, 3H). MS (ESI) m / z 462 (M-H2O+H)+. Example 8 5‑[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one Example 8A (S)‑tert-butyl 3-methyl‑4‑(5‑(trifluoromethyl)pyrimidin‑2-yl)piperazine‑1-carboxylate

[0071] InExample8A, (S)‑tert‑Butyl 3-methylpiperazine‑1-carboxylate (4.00g, 20.0mmol), 2-chloro‑5‑(trifluoromethyl) pyrimidine (3.65 g, 20.0 mmol), and triethylamine (5.57 mL, 39.9 mmol) were combined in acetonitrile (80 mL). The reaction mixture was stirred at 80 °C for 4 hours and cooled to room temperature. The insoluble salt material was filtered. The filtrate was concentrated under vacuum. The residue was purified by flash chromatography (silica gel, 0‑50% ethyl acetate in heptanes) to provide the title compound. MS (ESI) m / z 347 (M+H)+. Example 8B (S)‑2‑(2-methylpiperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0072] To a solution of Example 8A (5.80 g, 16.8 mmol) in acetonitrile (50 mL) was added 4M hydrogen chloride in dioxane (21 mL, 84 mmol). The reaction mixture was stirred at room temperature for 4 hours and filtered. The filter cake was rinsedwith a small amount of acetonitrile and dried under vacuum toprovide the title compoundas ahydrochloric acid salt. The filtratewas concentratedanddriedunder vacuum toprovide additional title compoundasahydrochloric acid salt. MS (ESI) m / z 247 (M+H)+. Example 8C 5‑[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one

[0073] Example 8C was prepared according to the procedure used for the preparation of Example 3C, substituting Example 8B forExample 3B, to provide the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δppm11.83 (s, 1H), 8.66 (d, J = 0.8Hz, 2H), 7.78 (d, J= 2.6Hz, 1H), 7.43 (d, J = 2.6Hz, 1H), 4.94 - 4.84 (m, 1H), 4.53 - 4.43 (m, 1H), 4.14 (s, br, 1H), 3.82 (s, br, 1H), 3.39 -3.23 (m,3H), 3.08 (s, br, 2H), 2.56 -2.49 (m,2H),2.24 -2.14 (m,2H),1.14 (d,J=6.7Hz,3H).MS(ESI) m / z 490 (M+H)+. Example 9 5‑[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one Example 9A methyl (R)‑2‑(4‑(tert-butoxycarbonyl)‑3-methylpiperazin‑1-yl)pyrimidine‑5-carboxylate

[0074] Example 9Awas prepared according to the procedure used for the preparation of Example 7A, substituting tert- butyl (R)‑2-methylpiperazine‑1-carboxylate for (R)‑tert-butyl 3-methylpiperazine‑1-carboxylate, to provide the title com- pound. 1HNMR(400MHz,DMSO-d6)δppm8.79 (s, 2H), 4.58 -4.45 (m,2H),4.28 -4.19 (m,1H), 3.80 (s, 3H), 3.29 (brdd,J = 4.0, 13.4 Hz, 2H), 3.13 - 3.07 (m, 2H), 1.41 (s, 9H), 1.02 (d, J = 6.7 Hz, 3H). 17 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 9B tert-butyl (R)‑4‑(5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl)‑2-methylpiperazine‑1-carboxylate

[0075] Example 9B was prepared according to the procedure used for the preparation of Example 7B, substituting Example 9A for Example 7A, to provide the title compound. MS (ESI) m / z 337 (M+H)+. Example 9C (R)‑2‑(2‑(3-methylpiperazin‑1-yl)pyrimidin‑5-yl)propan‑2-ol

[0076] Example 9C was prepared according to the procedure used for the preparation of Example 7C, substituting Example 9B for Example 7B, to provide the title compound. MS (ESI) m / z 237 (M+H)+. Example 9D 5‑[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0077] Example 9D was prepared according to the procedure used for the preparation of Example 1E, substituting Example 9C for Example 1D. Purification by flash chromatography provided the title compound. 1H NMR (400 MHz, DMSO‑d6, 90 °C) δ ppm 11.83 (s, br, 1H), 8.42 (s, 2H), 7.77 (d, J = 2.6 Hz, 1H), 7.42 (d, J = 2.6 Hz, 1H), 4.73 (s, 1H), 4.57 - 4.29 (m, 3H), 3.96 (s, br, 2H), 3.38 - 3.06 (m, 4H), 2.53 - 2.44 (m, 2H), 2.25 - 2.10 (m, 2H), 1.43 (s, 6H), 1.09 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 462 (M-H2O+H)+. Example 10 5‑[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one Example 10A methyl (S)‑2‑(4‑(tert-butoxycarbonyl)‑2-methylpiperazin‑1-yl)pyrimidine‑5-carboxylate

[0078] Example 10Awas preparedaccording to the procedure used for the preparation of Example 7A, substituting tert- butyl (S)‑3-methylpiperazine‑1-carboxylate for (R)‑tert-butyl 3-methylpiperazine‑1-carboxylate, to provide the title com- pound. MS (ESI) m / z 281 (M‑56+H)+. Example 10B tert-butyl (S)‑4‑(5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl)‑3-methylpiperazine‑1-carboxylate

[0079] Example 10B was prepared according to the procedure used for the preparation of Example 7B, substituting Example 10A for Example 7A, to provide the title compound. MS (ESI) m / z 337 (M+H)+. Example 10C (,S)‑2‑(2‑(2-methylpiperazin‑1-yl)pyrimidin‑5-yl)propan‑2-ol

[0080] Example 10C was prepared according to the procedure used for the preparation of Example 7C, substituting Example 10B for Example 7B, to provide the title compound. MS (ESI) m / z 237 (M+H)+. Example 10D 5‑[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0081] Example 10D was prepared according to the procedure used for the preparation of Example 1E, substituting 18 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 10C for Example 1D. Purification by flash chromatography provided the title compound. 1H NMR (400 MHz, DMSO‑d6, 90 °C) δ ppm11.84 (s, br, 1H), 8.43 (s, 2H), 7.78 (d, J= 2.6Hz, 1H), 7.42 (d, J= 2.6Hz, 1H), 4.88 - 4.78 (m, 1H), 4.73 (s, 1H), 4.43 - 4.35 (m, 1H), 3.97 (s, br, 3H), 3.40 - 3.04 (m, 4H), 2.55 - 2.45 (m, 2H), 2.25 - 2.12 (m, 2H), 1.43 (s, 6H), 1.07 (d, J = 6.6 Hz, 3H). MS (ESI) m / z 462 (M-H2O+H)+. Example 11 5‑[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one Example 11A methyl (S)‑2‑(4‑(tert-butoxycarbonyl)‑3-methylpiperazin‑1-yl)pyrimidine‑5-carboxylate

[0082] Example 11Awas prepared according to the procedure used for the preparation of Example 7A, substituting tert- butyl (S)‑2-methylpiperazine‑1-carboxylate for (R)-tert-butyl 3-methylpiperazine‑1-carboxylate, to provide the title com- pound. 1HNMR(400MHz,DMSO-d6)δppm8.79 (s, 2H), 4.59 -4.45 (m,2H), 4.24 (brd,J=3.0Hz,1H), 3.80 (m,4H), 3.31 - 3.26 (m, 1H), 3.09 (d, J = 9.5 Hz, 2H), 1.41 (s, 9H), 1.02 (d, J = 6.7 Hz, 3H). Example 11B tert-butyl (S)‑4‑(5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl)‑2-methylpiperazine‑1-carboxylate

[0083] Example 11B was prepared according to the procedure used for the preparation of Example 7B, substituting Example 11A for Example 7A, to provide the title compound. MS (ESI) m / z 337 (M+H)+. Example 11C (S)‑2‑(2‑(3-methylpiperazin‑1-yl)pyrimidin‑5-yl)propan‑2-ol

[0084] Example 11C was prepared according to the procedure used for the preparation of Example 7C, substituting Example 11B for Example 7B, to provide the title compound. MS (ESI) m / z 237 (M+H)+. Example 11D 5‑[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0085] Example 11D was prepared according to the procedure used for the preparation of Example 1E, substituting Example 11C for Example 1D. Purification by flash chromatography provided the title compound. 1H NMR (400 MHz, DMSO‑d6, 90 °C) δ ppm 11.85 (s, br, 1H), 8.42 (s, 2H), 7.77 (d, J = 2.6 Hz, 1H), 7.42 (d, J = 2.6 Hz, 1H), 4.73 (s, 1H), 4.57 - 4.27 (m, 3H), 3.97 (s, br, 2H), 3.51 - 3.41 (m, 1H), 3.35 - 3.08 (m, 3H), 2.53 - 2.44 (m, 2H), 2.25 - 2.10 (m, 2H), 1.43 (s, 6H), 1.09 (d, J= 6.6 Hz, 3H). MS (ESI) m / z 462 (M-H2O+H)+. Example 12 5‑{[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0086] Example 12 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D for Example 1C, to provide the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δppm11.79 (s, br, 1H), 8.64 (d, J = 0.9Hz, 2H), 7.71 (d, J = 2.6Hz, 1H), 7.39 (d, J = 2.6Hz, 1H), 4.59 - 4.34 (m, 3H), 3.95 (s, br, 1H), 3.32 - 3.03 (m, 4H), 2.45 (d, J = 7.0 Hz, 2H), 2.42 - 2.34 (m, 1H), 2.25 - 2.15 (m, 2H), 1.93 - 1.79 (m, 2H), 1.06 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 504 (M+H)+. Example 13 19 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 5‑{[(1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one Example 13A methyl 3‑((6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)methylene)cyclobutanecarboxylate

[0087] In Example 13A, 5-Bromo‑2-methoxy‑3‑(trifluoromethyl)pyridine (1.02 g, 4.00 mmol), methyl 3‑((4,4,5,5-tetra- methyl‑1,3,2-dioxaborolan‑2-yl)methylene)cyclobutanecarboxylate (1.01 g, 4.00 mmol), Pd(dppf)Cl2 CH2Cl2 ([1,1’- bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex with dichloromethane, 163 mg, 0.200 mmol) and po- tassium carbonate (1.11 g, 8.00 mmol) were combined in a mixture of dioxane (9.0 mL) and water (3.0 mL). The reaction mixture was purged with nitrogen for 5minutes, stirred at 90 °C for 16 hours, cooled to room temperature, and partitioned between ethyl acetate and water. The organic layer was dried with anhydrous sodium sulfate, treated with 3-mercapto- propyl functionalized silica gel, filtered, and the filtrate was concentrated under vacuum. The residuewas purified by flash chromatography (silica gel, 0‑20% ethyl acetate in heptanes) to provide the title compound. MS (ESI) m / z 302 (M+H)+. Example 13B (1r,3s)‑methyl 3‑((6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)methyl)cyclobutanecarboxylate

[0088] Example13A (740mg, 2.46mmol) and tetrahydrofuran (10mL)wereadded to210mgwet 5%Pd / C inapressure reactor. The reaction mixture was stirred at 25 °C for 8 hours under hydrogen (50 psi), filtered, washed with tetrahy- drofuran, and concentrated under vacuum. The residue was purified by flash chromatography (silica gel, 0‑20% ethyl acetate in heptanes) to provide the title compound,which containedabout 20%corresponding trans-isomer.MS (ESI)m / z 304 (M+H)+. Example 13C (1r,3s)‑3‑((6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)methyl)cyclobutanecarboxylic acid

[0089] To a solution of Example 13B (710 mg, 2.34 mmol) in a mixture of tetrahydrofuran (6.0 mL) / methanol (6.0 mL) / water (2.0 mL) was added sodium hydroxide (468 mg, 11.7 mmol). The reaction mixture was stirred at room temperature for 1 hour and dilutedwithwater. The pHwasadjusted to 4with the addition of 1MaqueousHCl and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, andfiltered. Thefiltratewasconcentratedunder vacuum toprovide the title compoundwhich containedabout 20% corresponding trans-isomer. MS (ESI) m / z 290 (M+H)+. Example 13D (1r,3s)‑3‑((6-oxo‑5‑(trifluoromethyl)‑1,6-dihydropyridin‑3-yl)methyl)cyclobutanecarboxylic acid

[0090] Example 13C (676 mg, 2.34 mmol), para-toluenesulfonic acid monohydrate (889 mg, 4.67 mmol), and lithium chloride (495mg, 11.7 mmol) were combined inN,N-dimethylformamide (10mL). The reactionmixture was stirred at 100 °C for 10 hours, cooled to room temperature, and partitioned between ethyl acetate and water. The organic layer was washed with saturated aqueous sodium chloride, dried with anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to provide the tittle compound which contained about 20% corresponding trans-isomer. The material was further purifiedby chiral SFC (Chiralpak®AD-H, 5µm,30×250mm;mobile phaseA: carbondioxide;mobile phaseB: isopropanol; flow rate: 80g / minute).Thefirst eluted fractionwascollected toprovide the title compound.MS(ESI) m / z 276 (M+H)+. Example 13E 2‑((3R,5S)‑3,5-dimethylpiperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0091] In Example 13E, a mixture of (2R,6S)‑2,6-dimethylpiperazine (1.00 g, 8.76 mmol), 2-chloro‑5‑(trifluoromethyl) pyrimidine (1.60g, 8.76mmol) and triethylamine (4.88mL, 35.0mmol)were combined in acetonitrile (35mL). The reaction mixturewasstirredat room temperatureovernight. The insolublesaltmaterialwasfilteredandwashedwithasmall amount of acetonitrile. The combined filtrate was concentrated under vacuum to provide the title compound. MS (ESI) m / z 261 20 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (M+H)+. Example 13F 5‑{[(1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0092] Example 13D (27.5 mg, 0.100 mmol), 1‑[bis(dimethylamino)methylene]‑1H‑1,2,3-triazolo[4,5-b]pyridinium 3- oxidhexafluorophosphate (76.0mg,0.200mmol) andN,N-diisopropylethylamine (0.070mL,0.400mmol)were combined inN,N-dimethylformamide (1.0mL) and the reactionmixturewas stirredat room temperature for 5minutes. To the reaction mixturewasaddedExample13E (26.0mg, 0.100mmol), and the resultingmixturewasstirredat 50 °C for 24hours, cooled to room temperature, and purified by reverse phase HPLC (C18, 20‑100% acetonitrile / in water with 0.1% trifluoroacetic acid). The acetonitrile was removed by evaporation, and the mixture was neutralized with saturated aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate, dried with anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to provide the title compound. 1H NMR (400 MHz, DMSO‑d6, 90 °C) δ ppm 11.79 (s, br, 1H), 8.64 (d, J = 0.8 Hz, 2H), 7.71 (d, J = 2.6, 1H), 7.39 (d, J = 2.6 Hz, 1H), 4.54 (d, J = 13.7 Hz, 2H), 4.36 (s, br, 2H), 3.26 - 3.12 (m,3H), 2.46 (d, J=7.3Hz,2H),2.43 -2.33 (m,1H),2.24 -2.15 (m,2H),1.95 -1.84 (m,2H), 1.14 (d, J=6.9Hz,6H).MS (ESI) m / z 518 (M+H)+. Example 14 5‑{[(1s,3s)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0093] Example 14 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13DandExample 4B forExample 1CandExample 1D, respectively, to provide the title compound. 1HNMR (400 MHz,DMSO‑d6, 90 °C)δppm11.65 (s, br, 1H), 8.68 (d,J=0.9Hz,2H), 7.71 (d,J=2.6Hz, 1H), 7.40 (d,J=2.6Hz, 1H), 4.87 - 4.78 (m, 2H), 3.98 (s, br, 2H), 3.42 - 2.90 (m, 3H), 2.47 (d, J = 7.5 Hz, 2H), 2.45 - 2.35 (m, 1H), 2.27 - 2.18 (m, 2H), 1.96 - 1.85 (m, 2H), 1.17 (d, J = 6.8 Hz, 6H). MS (ESI) m / z 518 (M+H)+. Example 15 5‑{[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutylmethyl}‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0094] Example 15 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13DandExample 3B forExample 1CandExample 1D, respectively, to provide the title compound. 1HNMR (400 MHz,DMSO‑d6, 90 °C)δppm11.76 (s, br, 1H), 8.65 (d,J=0.9Hz,2H), 7.71 (d,J=2.6Hz, 1H), 7.39 (d,J=2.6Hz, 1H), 4.93 - 4.81 (m, 1H), 4.51 - 4.41 (m, 1H), 3.97 (s, br, 2H), 3.38 - 3.00 (m, 4H), 2.45 (d, J = 7.7 Hz, 2H), 2.43 - 2.34 (m, 1H), 2.28 - 2.15 (m, 2H), 1.93 - 1.81 (m, 2H), 1.12 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 504 (M+H)+. Example 16 5‑{[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0095] Example 16 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13DandExample 8B forExample 1CandExample 1D, respectively, to provide the title compound. 1HNMR (400 MHz,DMSO‑d6, 90 °C)δppm11.78 (s, br, 1H), 8.65 (d,J=0.9Hz,2H), 7.71 (d,J=2.6Hz, 1H), 7.39 (d,J=2.6Hz, 1H), 4.92 -4.82 (m,1H),4.49 -4.42 (m,1H),3.98 (s, br, 2H), 3.35 -3.01 (m,4H),2.45 (d,J=7.5Hz,2H), 2.43 -2.34 (m,1H), 2.28 -2.15 (m, 2H), 1.93 - 1.82 (m, 2H), 1.12 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 504 (M+H)+. Example 17 5‑{[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0096] Example 17 was prepared according to the procedure used for the preparation of Example 1E, substituting 21 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 13DandExample 6B forExample 1CandExample 1D, respectively, to provide the title compound. 1HNMR (400 MHz,DMSO‑d6, 90 °C)δppm11.78 (s, br, 1H), 8.64 (d,J=0.9Hz,2H), 7.70 (d,J=2.6Hz, 1H), 7.39 (d,J=2.6Hz, 1H), 4.60 - 4.30 (m, 3H), 3.95 (s, br, 1H), 3.32 - 3.04 (m, 4H), 2.45 (d, J = 7.6 Hz, 2H), 2.42 - 2.33 (m, 1H), 2.24 - 2.16 (m, 2H), 1.93 - 1.79 (m, 2H), 1.05 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 504 (M+H)+. Example 18 5‑{[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0097] Example 18 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 7C for Example 1C and Example 1D, respectively. Purification by flash chromatography provided the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δ ppm11.79 (s, br, 1H), 8.42 (s, 2H), 7.71 (d, J=2.6Hz, 1H), 7.39 (d, J=2.6Hz, 1H), 4.86 - 4.66 (m, 2H), 4.41 - 4.33 (m, 1H), 3.94 (s, br, 2H), 3.29 - 3.02 (m, 4H), 2.45 (d, J=7.2Hz, 2H), 2.43 - 2.34 (m, 1H), 2.28 - 2.14 (m, 2H), 1.92 - 1.82 (m, 2H), 1.42 (s, 6H), 1.05 (d, J=6.7Hz, 3H).MS (ESI)m / z476 (M- H2O+H)+. Example 19 5‑{[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0098] Example 19 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 9C for Example 1C and Example 1D, respectively. Purification by flash chromatography provided the title compound. 1HNMR (400MHz, DMSO‑d6, 90 °C) δ ppm11.78 (s, br, 1H), 8.41 (s, 2H), 7.70 (d, J= 2.6Hz, 1H), 7.39 (d, J= 2.6 Hz, 1H), 4.73 (s, 1H), 4.55 - 4.21 (m, 3H), 3.95 (s, br, 1H), 3.22 - 2.85 (m, 4H), 2.45 (d, J= 7.4 Hz, 2H), 2.42 - 2.33 (m, 1H), 2.24 - 2.15 (m, 2H), 1.93 - 1.79 (m, 2H), 1.42 (s, 6H), 1.06 (d, J= 6.7 Hz, 3H). MS (ESI)m / z 476 (M- H2O+H)+. Example 20 5‑{[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0099] Example 20 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 10C for Example 1C and Example 1D, respectively. Purification by flash chromatography provided the title compound. 1HNMR(400MHz,DMSO‑d6, 90 °C)δppm11.79 (s, br, 1H), 8.42 (s, 2H), 7.71 (d,J=2.6,1H), 7.39 (d, J=2.6Hz, 1H), 4.86 - 4.66 (m, 2H), 4.41 - 4.33 (m, 1H), 3.95 (s, br, 2H), 3.29 - 3.00 (m, 4H), 2.45 (d, J=7.2Hz, 2H), 2.42 - 2.34 (m, 1H), 2.28 - 2.14 (m, 2H), 1.93 - 1.82 (m, 2H), 1.42 (s, 6H), 1.05 (d, J= 6.6 Hz, 3H). MS (ESI)m / z 476 (M- H2O+H)+. Example 21 5‑{[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one

[0100] Example 21 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 11C for Example 1C and Example 1D, respectively. Purification by flash chromatography provided the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δ ppm11.78 (s, br, 1H), 8.41 (s, 2H), 7.71 (d, J=2.6Hz, 1H), 7.39 (d, J = 2.6 Hz, 1H), 4.73 (s, 1H), 4.54 - 4.23 (m, 3H), 3.96 (s, br, 1H), 3.22 - 2.85 (m, 4H), 2.45 (d, J = 7.3 Hz, 2H), 2.42 - 2.33 (m, 1H), 2.24 - 2.15 (m, 2H), 1.94 - 1.79 (m, 2H), 1.42 (s, 6H), 1.06 (d, J = 6.7 Hz, 3H). MS (ESI)m / z 476 (M- H2O+H)+. Example 22 5‑{[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one 22 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 22A methyl 2‑((2S,6R)‑4‑(tert-butoxycarbonyl)‑2,6-dimethylpiperazin‑1-yl)pyrimidine‑5-carboxylate

[0101] In Example 22A, methyl 2-chloropyrimidine‑5-carboxylate (1.50 g, 8.69 mmol) and tert-butyl (3S,5R)‑3,5- dimethylpiperazine‑1-carboxylate (3.73 g, 17.4 mmol) were combined inN-methyl‑2-pyrrolidinone (20 mL). The reaction mixturewas stirred at 140 °C for 12 hours, cooled to room temperature, dilutedwithwater, and extractedwith ethyl acetate three times. The combined organic layers were concentrated under vacuum. The residue was purified by flash chromatography (silica gel, 0‑100% ethyl acetate in petroleum ether) to provide the title compound. MS (ESI) m / z 351 (M+H)+. Example 22B tert-butyl (3S,5R)‑4‑(5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl)‑3,5-dimethylpiperazine‑1-carboxylate

[0102] To a solution of Example 22A (1.60 g, 4.57mmol) in tetrahydrofuran (15mL) at ‑78 °C under nitrogenwas added 3.0Mmethylmagnesiumbromide (7.61mL, 22.8mmol). The reactionmixturewas stirred at room temperature for 2 hours, quenched with aqueous ammonium chloride, diluted with water, and extracted with ethyl acetate three times. The combined organic layerswerewashedwith saturated aqueous sodiumchloride, driedwith anhydrous sodiumsulfate, and filtered. The filtrate was concentrated under vacuum. The residuewas purified by flash chromatography (silica gel, 0‑50% ethyl acetate in petroleum ether) to provide the title compound. MS (ESI) m / z 351 (M+H)+. Example 22C 2‑(2‑((2S,6R)‑2,6-dimethylpiperazin‑1-yl)pyrimidin‑5-yl)propan‑2-ol

[0103] AmixtureofExample 22B (800mg, 2.28mmol) in 4MHCl in ethyl acetate (2mL)was stirredat room temperature for 2 hours and concentrated under vacuum. The pHwas adjusted to about 8 by progressively adding saturated aqueous sodium bicarbonate. Themixture was purified by preparative reversed phaseHPLC (C18, 5 - 95%acetonitrile in water) to provide the title compound. MS (ESI) m / z 251 (M+H)+. Example 22D 5‑{[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one

[0104] Example 22D was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 22C for Example 1C and Example 1D, respectively. Purification by flash chromatography provided the title compound. 1HNMR (400MHz,DMSO‑d6, 90 °C) δ ppm11.78 (s, br, 1H), 8.44 (s, 2H), 7.71 (d, J=2.6Hz, 1H), 7.40 (d, J=2.6Hz, 1H), 4.80 - 4.67 (m, 3H), 3.95 (s, br, 2H), 3.34 - 3.02 (m, 3H), 2.46 (d, J=7.5Hz, 2H), 2.44 - 2.35 (m, 1H), 2.27 - 2.18 (m, 2H), 1.96 - 1.86 (m, 2H), 1.43 (s, 6H), 1.12 (d, J= 6.8 Hz, 6H). MS (ESI) m / z 490 (M-H2O+H)+. Example 23 3‑(trifluoromethyl)‑5‑[(1s,3s)‑3‑{4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]pyridin‑2(1H)‑one Example 23A methyl 3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylate

[0105] Example 23A was prepared according to the procedure used for the preparation of Example 1A. Purification provided the title compound as themixture of stereoisomers with both trans configuration and cis configuration. MS (ESI) m / z 290 (M+H)+. Example 23B 23 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylic acid

[0106] Example 23B was prepared according to the procedure used for the preparation of Example 1B, substituting Example 23A for Example 1A, to provide the title compound. MS (ESI) m / z 276 (M+H)+. Example 23C tert-butyl 4‑(5‑(trifluoromethyl)pyrimidin‑2-yl)piperazine‑1-carboxylate

[0107] In Example 23C, a 4-neck 1-L jacketed reactor was equippedwith overhead stirring, aHuber temperature probe, and a nitrogen inlet. Under a nitrogen atmosphere, the reactor was charged with 2-chloro‑5‑(trifluoromethyl)pyrimidine (59.6 g, 326 mmol), which was dissolved in acetonitrile (600 mL). The solution was cooled to 5 °C internal and then tert- butyl piperazine‑1-carboxylate (60.8g,326mmol)wasadded followedbyN,N-diisopropylethylamine (142mL,816mmol). The reaction mixture was stirred at room temperature for 1 hour, transferred to a round bottom flask, and concentrated under vacuum. The residue was dissolved in dichloromethane, washed with saturated aqueous ammonium chloride, washed with saturated aqueous sodium chloride, dried with anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under vacuum to provide the title compound. MS (APCI) m / z 277 (M‑56+H)+. Example 23D 2‑(piperazin‑1-yl)‑5‑(trifluoromethyl)pyrimidine

[0108] In Example 23D, a 4-neck 2-L jacketed reactor was equippedwith overhead stirring, a Huber temperature probe andanitrogen inlet.Under anitrogenatmosphere, the reactorwas chargedwithExample 23C (108g, 325mmol), followed by addition of dioxane (1000mL) and 4MHCl (300mL, 1200mmol). The reactionmixturewas stirred at room temperature for 18 hours, filtered, rinsed with tert-butyl methyl ether, and dried under vacuum. The residue was suspended in dichloromethane and cooled in an ice bath. Saturated aqueous sodium bicarbonate was added slowly until a neutral pH was reached, and the mixture was separated. The aqueous layer was extracted with dichloromethane twice. The combined organic layers were washed with saturated aqueous sodium chloride, dried with anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under vacuum to provide the title compound. To the aqueous layer was addedNaOHpellets to adjust the pH>10, and themixturewasextractedwith dichloromethane three times. The combined organic layers were washed with saturated aqueous sodium chloride, dried with anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under vacuum to provide additional title compound. MS (APCI) m / z 233 (M+H)+. Example 23E (3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutyl)(4‑(5‑(trifluoromethyl)pyrimidin‑2-yl)piperazin‑1-yl)methanone

[0109] To a mixture of Example 23B (84.0 mg, 0.305 mmol), Example 23D (70.9 mg, 0.305 mmol), and N,N- diisopropylethylamine (0.320 mL, 1.83 mmol) in N,N-dimethylformamide (2.0 mL) was added 50% propanephosphonic acid anhydride inN,N-dimethylformamide (388mg,0.610mmol). The reactionmixturewasstirredat room temperature for 1 hour and partitioned with ethyl acetate and water. The organic layer was washed with saturated aqueous sodium chloride, driedwith anhydrous sodiumsulfate, and filtered. The filtratewas concentrated under vacuum to provide the title compound. MS (ESI) m / z 490 (M+H)+. Example 23F 3‑(trifluoromethyl)‑5‑[(1s,3s)‑3‑{4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]pyridin‑2(1H)‑one

[0110] Example 23E (134mg, 0.274mmol), para-toluenesulfonic acidmonohydrate (104mg, 0.548mmol), and lithium chloride (58.0mg, 1.37mmol)were combined inN,N-dimethylformamide (2.0mL). The reactionmixturewasstirredat 100 °C for 10hours, cooled to room temperature, andpurifiedby reversephaseHPLC(C18,10‑100%acetonitrile / inwaterwith 0.1% trifluoroacetic acid). The acetonitrile was removed by evaporation under vacuum, and the resulting mixture was neutralizedwith saturatedaqueous sodiumbicarbonate. The resultingmixturewasextractedwith ethyl acetate, driedwith anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum. The residue was further purified by flash chromatography to provide the two fractions. The first eluted fraction was the stereoisomer with trans-configuration. The second eluted fraction was the title compound with cis-configuration. 1H NMR (500 MHz, DMSO‑d6) δ ppm 12.21 (s, 1H), 8.73 (d, J = 0.8Hz, 2H), 7.83 (d, J = 2.6Hz, 1H), 7.50 (d, J = 2.6Hz, 1H), 3.86 - 3.79 (m, 4H), 3.58 - 3.54 (m, 2H), 3.52 - 24 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 3.48 (m, 2H), 3.37 - 3.24 (m, 2H), 2.49 - 2.41 (m, 2H), 2.23 - 2.13 (m, 2H). MS (ESI) m / z 476 (M+H)+. Example 24 5‑[(1r,3r)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one

[0111] Example 24 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 25CandExample 4B forExample 1CandExample 1D, respectively, to provide the title compound. 1HNMR (400 MHz,DMSO‑d6, 90 °C) δ ppm11.61 (s, 1H), 8.68 (d, J= 0.9Hz, 2H), 7.85 (d, J = 2.6Hz, 1H), 7.51 (d, J = 2.6Hz, 1H), 4.89 - 4.79 (m,2H), 3.86 (s, br, 2H), 3.52 - 3.36 (m, 2H), 3.30 - 3.02 (m, 2H), 2.68 - 2.51 (m, 2H), 2.34 - 2.24 (m, 2H), 1.19 (d, J=6.9 Hz, 6H). MS (ESI) m / z 504 (M+H)+. Example 25 5‑[(1R,3r)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one Example 25A (1r,3r)‑methyl 3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylate

[0112] Example 25A was prepared as described in Example 1A. The first eluted fraction was the title compound with trans configuration. MS (ESI) m / z 290 (M+H)+. Example 25B (1r,3r)‑3‑(6-methoxy‑5‑(trifluoromethyl)pyridin‑3-yl)cyclobutanecarboxylic acid

[0113] Example 25B was prepared according to the procedure used for the preparation of Example 1B, substituting Example 25A for Example 1A, to provide the title compound. MS (ESI) m / z 276 (M+H)+. Example 25C (1r,3r)‑3‑(6-oxo‑5‑(trifluoromethyl)‑1,6-dihydropyridin‑3-yl)cyclobutanecarboxylic acid

[0114] Example 25C was prepared according to the procedure used for the preparation of Example 1C, substituting Example 25B for Example 1B, to provide the title compound. MS (ESI) m / z 262 (M+H)+. Example 25D 5‑[(1R,3r)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl) pyridin‑2(1H)‑one

[0115] Example 25D was prepared according to the procedure used for the preparation of Example 1E, substituting Example 25C for Example 1C, to provide the title compound. 1H NMR (400 MHz, DMSO-d6, 90 °C) δ ppm 11.87 (s, 1H), 8.65 (d, J= 0.8Hz, 2H), 7.84 (d, J= 2.6Hz, 1H), 7.49 (d, J= 2.7Hz, 1H), 4.69 - 4.27 (m, 3H), 3.82 (s, br, 1H), 3.42 - 3.28 (m, 3H), 3.26 - 3.03 (m, 2H), 2.64 - 2.50 (m, 2H), 2.31 - 2.20 (m, 2H), 1.08 (d, J = 6.7 Hz, 3H). MS (ESI) m / z 490 (M+H)+. Example 26 3‑(trifluoromethyl)‑5‑[(1r,3r)‑3‑{4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]pyridin‑2(1H)‑one

[0116] Example26waspreparedasdescribed inExample23F.Thefirst eluted fractionwas the title compoundwith trans configuration. 1HNMR(600MHz,DMSO‑d6)δppm12.22 (s, 1H), 8.73 (d,J=0.9Hz, 2H), 7.93 (d,J=2.6Hz,1H), 7.58 (d,J =2.6Hz, 1H), 3.86 - 3.81 (m, 4H), 3.62 - 3.58 (m, 2H), 3.44 - 3.34 (m, 4H), 2.55 - 2.51 (m, 2H), 2.31 - 2.23 (m, 2H).MS (ESI) m / z 476 (M+H)+. 25 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Example 27 5‑{[(1s,3s)‑3‑{4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(trifluoromethyl) pyridin‑2(1H)‑one Example 27A 2‑(2‑(piperazin‑1-yl)pyrimidin‑5-yl)propan‑2-ol

[0117] In Example 27A, 2‑(2-Chloropyrimidin‑5-yl)propan‑2-ol (1.00 g, 5.79mmol), piperazine (1.50 g, 17.4mmol), and N,N-diisopropylethylamine (5.06mL, 29.0mmol) were combined in acetonitrile (10mL). The reactionmixture was stirred at 80 °C for 2 hours, cooled to room temperature, and concentrated under vacuum. The residue was purified by flash chromatography to provide the title compound. MS (ESI) m / z 223 (M+H)+. Example 27B 5‑{[(1s,3s)‑3‑{4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}‑3‑(trifluoromethyl) pyridin‑2(1H)‑one

[0118] Example 27B was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 27A for Example 1C and Example 1D, respectively. Purification by flash chromatography provided the title compound. 1HNMRppm (400MHz,DMSO‑d6)δ12.15 (s, 1H), 8.44 (s, 2H), 7.80 (d, J=2.5Hz, 1H), 7.48 (d, J = 2.5Hz, 1H), 5.08 (s, 1H), 3.71 - 3.63 (m, 4H), 3.53 - 3.46 (m, 2H), 3.43 - 3.36 (m, 2H), 3.24 - 3.11 (m, 1H), 2.43 (d, J= 7.3 Hz, 2H), 2.40 - 2.30 (m, 1H), 2.20 - 2.10 (m, 2H), 1.89 - 1.79 (m, 2H), 1.41 (s, 6H). MS (ESI) m / z 462 (M-H2O+H)+. Example 28 5‑[(1s,3s)‑3‑{4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyri- din‑2(1H)‑one

[0119] Example 28 was prepared according to the procedure used for the preparation of Example 1E, substituting Example27A forExample1D.The reactionmixturewaspurifiedby reversephaseHPLC(C18,20‑100%acetonitrile / water with 0.1% trifluoroacetic acid). The acetonitrile was removed by evaporation under vacuum, and the resultingmixturewas neutralized with saturated aqueous sodium bicarbonate, extracted with ethyl acetate, dried with anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum. The residue was further purified by flash chromato- graphy to provide the title compound. 1HNMR (400MHz, DMSO‑d6) δ ppm12.20 (s, 1H), 8.45 (s, 2H), 7.83 (d, J = 2.6 Hz, 1H), 7.50 (d, J = 2.6 Hz, 1H), 5.08 (s, 1H), 3.75 - 3.65 (m, 4H), 3.55 - 3.49 (m, 2H), 3.47 - 3.42 (m, 2H), 3.38 - 3.23 (m, 2H), 2.48 - 2.40 (m, 2H), 2.24 - 2.11 (m, 2H), 1.41 (s, 6H). MS (ESI) m / z 448 (M-H2O+H)+. Example 29 3‑(trifluoromethyl)‑5‑{[(1s,3s)‑3‑{4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]methyl}pyri- din‑2(1H)‑one

[0120] Example 29 was prepared according to the procedure used for the preparation of Example 1E, substituting Example 13D and Example 23D for Example 1C and Example 1D, respectively, to provide the title compound. 1H NMR (600MHz,DMSO‑d6)δppm12.15 (s, br, 1H), 8.72 (d,J=0.8Hz, 2H), 7.80 (d,J=2.5Hz, 1H), 7.49 (d,J=2.5Hz, 1H), 3.83 - 3.77 (m, 4H), 3.56 - 3.51 (m, 2H), 3.47 - 3.41 (m, 2H), 3.22 - 3.15 (m, 1H), 2.43 (d, J=7.5Hz, 2H), 2.40 - 2.30 (m, 1H), 2.20 - 2.12 (m, 2H), 1.89 - 1.80 (m, 2H). MS (ESI) m / z 490 (M+H)+. TIPARP GST‑(TEV) FUSION PROTEIN EXPRESSION AND PURIFICATION Expression

[0121] The full-length cDNAencoding the humanTIPARP (NP_001171647.1,UniProtKB -Q7Z3E1) of 657 aminoacids was the template to generate the expression construct. The nucleotide sequences were synthesized with codon optimization for expression in Sf9 insect cells. The TIPARP fusion protein includes GST, glutathione S-transferase, a 26 kDa protein whose DNA sequence is frequently integrated into expression vectors for production of recombinant 26 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 proteins.

[0122] A truncatedTIPARPprotein sequence, consisting of aminoacids 441 to657,was cloned into vector pFastBac™1 (Thermo Fisher Scientific) with an N-terminal GST tag followed by a TEV protease site to produce the TIPARP fusion protein, TIPARP GST‑(TEV).

[0123] The TIPARP fusion protein was heterologously expressed in Sf9 insect cell cultures seeded with baculovirus- infected insect cells (BIICs) infectedwith the pFastBac™1recombinant baculovirus. The baculovirus-infected cell cultures were grown in aWave Bioreactor (Cytiva, Marlborough, MA, USA) on a 20-liter scale. The Sf9 insect cells were grown in Sf‑900™ IISFMmedia (ThermoFisherScientific,Waltham,MA,USA) toadensityof 2.4x106cells / mLbefore infectionwith baculovirus. For the infection, BIICswere added, and cellswere harvested after 72 hours of incubation by centrifugation at 4,000 x g for 10 minutes at 5 °C and transferred to ‑80 °C until further processing.

[0124] TheTIPARP fusion protein expressed in insect cells was purified using the following protocol: cells were thawed, homogenized and lysed in 20 mM HEPES (4‑(2-hydroxyethyl)‑1-piperazineethanesulfonic acid), pH 7.5, 250 mM NaCl, 5% glycerol (w / v), 0.1 % Triton X‑100, 0.5 mM MgCl2, 0.5 mM Tris (2-carboxyethyl) phosphine hydrochloride (TCEP), using a Dounce homogenizer. Benzonase® Nuclease (MilliporeSigma, Burlington, MA, USA) and SIGMAFAST™ Protease Inhibitor Cocktail Tablets (MilliporeSigma) were added to the lysate according to the manufacturer’s recom- mendations. The cell lysate was clarified by centrifugation at 35,000 x g for 60 minutes at 5 °C. Supernatant was sterile filtered through a 0.45 µm filter. Purification

[0125] TheTIPARP fusion proteinwas purified from the supernatant in batchmode usingGlutathioneSepharose 4Fast FlowGST-taggedproteinpurification resin (Cytiva)equilibrated incolumnbuffer (20mMHEPESpH7.5,150mMNaCl, 5% glycerol (v / v), 0.5mMTCEP). Supernatant and resinwere gentlymixed and allowed to settle into a columnby gravity flow. Resin was washed with 20 column volumes of column buffer (20mMHEPES pH 7.5, 150mMNaCl, 5% glycerol, 0.5 mM TCEP).Proteinwaseluted from thecolumnusing columnbufferwith theaddition of 10mMglutathione.Proteinwas further diluted 3-fold with 20mMMES (2‑(N-morpholino)ethanesulfonic acid) pH 6.1, 5% glycerol (v / v), and 0.5 mMTCEP buffer and loaded onto HiTrap® SP Sepharose™ Fast Flow resin (MilliporeSigma) equilibrated in 20 mM MES pH 6.1, 50 mM NaCl, 5%glycerol, and0.5mMTCEP.TheTIPARP fusionproteinwaselutedwithasodiumchloridegradient from0.05Mto 1 M in 10 column volumes, with the fusion protein eluting at 400 mM NaCl. The molecular weight of the purified fusion protein was confirmed by mass spectroscopy. SYNTHESIS OF PROBE 1 Probe 1 N‑{2‑[2‑(2‑{[1‑(5-cyanopyridin‑2-yl)‑4‑{3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]benzoyl}piperazin‑2-yl]methoxy} ethoxy)ethoxy]ethyl}‑2’,7’-difluoro‑3’,6’-dihydroxy‑3-oxo‑3H-spiro[[2]benzofuran‑1,9’-xanthene]‑5-carboxamide Probe 1A 3‑[(E)‑(3-oxo‑2-benzofuran‑1(3H)‑ylidene)methyl[benzonitrile

[0126] To amixture of isobenzofuran‑1(3H)‑one (5 g, 37.3mmol) and 3-formylbenzoic acid (4.89 g, 37.3 mmol) in ethyl acetate (20mL)was added sodiummethoxide (30.2 g, 168mmol) dropwise at 0 °C.Methanol (10mL)was added, and the mixture was stirred at 80 °C for 1 hour. LCMS showed that the starting material was consumed. The solvent was evaporated, and the residuewas dilutedwithwater (1.5 L). Themixturewas acidified using 2Naqueous hydrochloric acid at 20 °C to give pH = 1~2. The mixture was filtered, and the filter cake was washed with water (500 mL), and dried under high vacuum to give the title compound. 1HNMR (400MHz, DMSO‑d6) δ ppm7.45 - 7.61 (m, 7H) 8.24 - 8.36 (m, 2H). ESI- MS m / z 246.0 (M-H). Probe 1B 3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]benzoic acid

[0127] Toamixture of 3‑[(E)‑(3-oxo‑2-benzofuran‑1(3H)‑ylidene)methyl]benzonitrile (6 g, 24.27mmol) inwater (36mL) wasaddeda solution of sodiumhydroxide (5.82g, 146mmol) inwater (7.2mL) at 20 °C. The reactionmixturewaswarmed to 90 °C and stirred for 1 hour. The reactionmixture was cooled to 70 °C and hydrazine (10.88mL, 340mmol) was added. The resulting mixture was stirred at 70 °C for another 18 hours. Thin layer chromatography (tetrahydrofuran / petroleum 27 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 ether = 1 / 1) showed the startingmaterial was consumed and a new spotwas formed. Themixturewas extractedwith ethyl acetate (3 × 60 mL). The organic phase was discarded, and the aqueous phase was acidified with 1 N aqueous hydrochloric acid at 20 °C to pH = 4. The precipitate was collected by filtration. The filter cake was washed with water (60 mL)andpetroleumether (60mL), anddriedunderhighvacuumtogive the title compound. 1HNMR(400MHz,DMSO‑d6)δ ppm 4.38 (s, 2H) 7.42 (t, J=7.70 Hz, 1H) 7.58 (br d, J=7.58 Hz, 1H) 7.80 (br dd, J=16.44, 7.89 Hz, 2H) 7.85 - 7.93 (m, 2H) 7.93 - 7.99 (m, 1H) 8.26 (d, J=7.58 Hz, 1H) 12.60 (s, 1H) 12.67 - 13.07 (m, 1H). ESI-MS m / z 281.1 (M+H)+. Probe 1C tert-butyl 4‑(5-cyanopyridin‑2-yl)‑3‑(hydroxymethyl)piperazine‑1-carboxylate

[0128] In Probe 1C, tert-Butyl 3‑(hydroxymethyl)piperazine‑1-carboxylate (300mg, 1.387mmol) was combinedwith 6- chloronicotinonitrile (192 mg, 1.387 mmol) in dimethylacetamide (4 mL). N,N‑Diisopropylethylamine (0.727 mL, 4.16 mmol)wasaddedand the reactionwasheatedat 90 °C for 2.25hours. The reactionmixturewascooled, dilutedwithwater, and extracted with ethyl acetate. The organic layer was dried over sodium sulfate and filtered. The filtrate was concentrated, and the residue was purified using a silica gel cartridge and was eluted with 0‑40% ethyl acetate / heptanes to provide the title compound. 1HNMR(600MHz,CDCl3)δppm8.40 (dd, J=2.4, 0.8Hz, 1H), 7.65 (dd, J=9.0, 2.3Hz, 1H), 6.65 (d, J = 9.0 Hz, 1H), 4.60 (s, 2H), 4.27 (s, 1H), 4.06 (d, J = 12.6 Hz, 1H), 3.98 (s, 1H), 3.63 (s, 1H), 3.49 (s, 2H), 3.33 (s, 1H), 3.27 (td, J = 12.2, 3.7 Hz, 1H), 3.16 (s, 1H), 1.50 (s, 8H). ESI-MS m / z 219.67 (M+H-Boc)+. Probe 1D tert-butyl 3‑({2‑[2‑(2-azidoethoxy)ethoxy]ethoxy}methyl)‑4‑(5-cyanopyridin‑2-yl)piperazine‑1-carboxylate

[0129] In Probe 1D, tert‑Butyl 4‑(5-cyanopyridin‑2-yl)‑3‑(hydroxymethyl)piperazine‑1-carboxylate (58.3 mg, 0.183 mmol) and 2‑(2‑(2-azidoethoxy)ethoxy)ethyl methanesulfonate (55.7 mg, 0.220 mmol) were combined in anhydrous N,N-dimethylformamide (1 mL). Sodium hydride (60% inmineral oil, 11 mg, 0.275mmol) was added and themixture was stirred at ambient temperature for 1 hour, and then at 50 °C for 2 hours. Additional 2‑(2‑(2-azidoethoxy)ethoxy)ethyl methanesulfonate (55.7mg,0.220mmol)wasaddedand the reactionmixturewasstirredat 50 °C for2hours.The reaction mixture was diluted to 3 mL using 90% DMSO / water and was purified in one injection on a Gilson® RP-HPLC running uniPoint softwarewithaWaters™DeltapakC18200x25mmcolumn (15µmparticle size, 100Angstromporosity) andwas eluted with a gradient of 5% A (0.1% TFA-water):B (acetonitrile) to 100% A:B [0‑5 minutes: 5% A; 5‑45 minute linear gradient to 100% B 2.375% / minute gradient] with a 20 mL / minute flowrate. Fractions containing the title compound were combined and lyophilized to give the title compound as the trifluoroacetate salt. ESI-MS m / z 476.1 (M+H)+. Probe 1E 6‑[2‑({2‑[2‑(2-azidoethoxy)ethoxy]ethoxy}methyl)piperazin‑1-yl]pyridine‑3-carbonitrile

[0130] In Probe 1E, tert‑Butyl 3‑({2‑[2‑(2-azidoethoxy)ethoxy]ethoxy}methyl)‑4‑(5-cyanopyridin‑2-yl)piperazine‑1-car- boxylate trifluoroacetate salt (47.2mg,0.080mmol)wasdissolved in 1mL trifluoroacetic acidand immediately evaporated to dryness under a stream of dry nitrogen gas to give the title compound as the bis trifluroacetate salt. ESI-MSm / z 376.16 (M+H)+. Probe 1F 6‑[2‑({2‑[2‑(2-azidoethoxy)ethoxy]ethoxy}methyl)‑4‑({3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]phenyl}methyl)piper- azin‑1-yl]pyridine‑3-carbonitrile

[0131] In Probe 1F, 6‑[2‑({2‑[2‑(2-Azidoethoxy)ethoxy]ethoxy}methyl)piperazin‑1-yl]pyridine‑3-carbonitrile bis trifluor- oacetate salt (48.2 mg, 0.080 mmol) and 3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]benzoic acid (24.63 mg, 0.088 mmol) were combined in anhydrous N,N-dimethylformamide (1 mL). PyAOP ((7-azabenzotriazol‑1-yloxy)tripyrrolidino- phosphonium hexafluorophosphate, 45.8 mg, 0.088 mmol) and N,N-diisopropylethylamine (69.7 µl, 0.399 mmol) were added and the reactionwas shaken at ambient temperature for one hour. An additional 24mgPyAOPwas added, and the reactionmixture was shaken for an additional hour at ambient temperature. The reactionmixture was diluted to 3mLwith 90% DMSO / water and was purified in one injection on a Gilson® RP-HPLC running uniPoint software with a Waters™ Deltapak C18 200 x 25 mm column (15 um particle size, 100 Angstrom porosity) and was eluted with a gradient of 5% A (0.1% TFA-water):B (acetonitrile) to 100% A:B [0‑5 minute: 5% A; 5‑45 minute linear gradient to 100% B 2.375% / minute 28 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 gradient] with a 20mL / minute flowrate. Fractions containing the title compoundwere combined and lyophilized to give the title compound as a trifluoroacetate salt. 1HNMR (600MHz,DMSO‑d6) δ ppm12.61 (s, 1H), 8.51 (dd, J = 2.4, 0.7 Hz, 1H), 8.27 (dd, J = 7.8, 1.4 Hz, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.92 - 7.80 (m, 4H), 7.44 (d, J = 7.8 Hz, 1H), 7.39 (t, J = 7.6 Hz, 3H), 7.28 (s, 2H), 6.90 (d, J = 9.2 Hz, 1H), 4.80 (s, 1H), 4.50 (s, 1H), 4.36 (s, 3H), 4.31 (s, 1H), 4.19 (s, 2H), 3.74 (s, 1H), 3.56 - 3.51 (m, 3H), 3.37 - 3.32 (m, 2H), 3.26 (s, 1H), 3.16 (s, 4H). ESI-MS m / z 637.81 (M+H)+. Probe 1G 6‑[2‑({2‑[2‑(2-aminoethoxy)ethoxy]ethoxy}methyl)‑4‑{3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]benzoyl}piperazin‑1- yl]pyridine‑3-carbonitrile

[0132] To 6‑[2‑({2‑[2‑(2-azidoethoxy)ethoxy]ethoxy}methyl)‑4‑({3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]phenyl} methyl)piperazin‑1-yl]pyridine‑3-carbonitrile trifluoroacetate salt (31 mg, 0.049 mmol) in tetrahydrofuran was added palladium on carbon (25.9 mg, 0.024 mmol) and the reaction mixture was stirred under H2 at 1 atm for 16 hours. The reaction mixture was filtered and the filtrate was purified on a silica gel cartridge, eluting with NH4OH / CH3OH / CH2Cl2 to provide the title compoundAnalytical LCMSTFAmethod [gradient of 5‑100%acetonitrile (A) and 0.1% trifluoroacetic acid inwater (B)wasused,at aflowrateof1.5mL / minute (0‑0.05minute5%A,0.05‑1.2minute5‑100%A,1.2‑1.4minute100% A, 1.4‑1.5 min 100‑5% A. 0.25 min post-run delay]: Rt = 0.73 min, ESI-MS m / z 612.5 (M+H)+. Probe 1H N‑{2‑[2‑(2‑{[1‑(5-cyanopyridin‑2-yl)‑4‑{3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl)methyl]benzoyl}piperazin‑2-yl]methoxy} ethoxy)ethoxy]ethyl }‑2’, 7’-difluoro‑3’, 6’-dihydroxy‑3-oxo‑3H-spiro[[2]benzofuran‑1,9’-xanthene]‑5-carboxamide

[0133] In Probe 1H, 6‑[2‑({2‑[2‑(2-Aminoethoxy)ethoxy]ethoxy}methyl)‑4‑{3‑[(4-oxo‑3,4-dihydrophthalazin‑1-yl) methyl]benzoyl}piperazin‑1-yl]pyridine‑3-carbonitrile and 2,5-dioxopyrrolidin‑1-yl 2’,7’-difluoro‑3’,6’-dihydroxy‑3 -oxo‑3H-spiro[isobenzofuran‑1,9’-xanthene]‑5-carboxylate (8.79 mg, 0.017 mmol) were combined in anhydrous N,N- dimethylformamide containing 2% N,N-diisopropylethylamine (v / v) and the mixture was shaken at ambient temperature overnight. The reactionmixturewasdiluted to3mLwith90%dimethyl sulfoxide / waterandwaspurified inone injectionona Gilson®RP-HPLC running uniPoint software with aWaters™Deltapak C18 200 x 25mmcolumn (15 umparticle size, 100 Angstromporosity) andwaselutedwithagradient of 5%A (0.1%TFA-water):B (acetonitrile) to100%A:B [0‑5minutes: 5% A;5‑45minutes lineargradient to100%B2.375% / minutesgradient]witha20mL / minuteflowrate.Fractionscontaining the title compound were combined and lyophilized to give the title compound as a trifluoroacetate salt. Analytical LCMS FA method [A gradient of 5‑100%acetonitrile (A) and 0.1% formic acidwater: acetonitrile (98:2) (B) was used, at a flow rate of 1.5 mL / minute (0‑0.05 minute 0% A, 0.05‑2.8 minute 0‑100% A, 2.8‑3.0 minute 100‑0% A, 160‑1500 amu positive / ne- gative ESI-MS ionization]: Rt = 1.493 minute, ESI-MS m / z 503.8 (M+2H)2+, 1006.4 (M+H)+, 1005.0 (M-H)-; analytical LCMSAAmethod [A gradient of 5‑100% acetonitrile (A) and 10mM ammonium acetate water: acetonitrile (98:2) (B) was used, at a flow rate of 1.5mL / minute (0‑0.05minute 0%A, 0.05‑2.8minute 0‑100%A, 2.8‑3.0minute 100‑0%A, 160‑1500 amu positive / negative ESI-MS ionization]: Rt = 1.016 minute, ESI-MSm / z 503.4 (M+2H)2+, 1028.0 (M+Na)+, 1003.8 (M- H)-. SYNTHESIS OF PROBE 2 Probe 2 2’,7’-difluoro-N‑[2‑(2‑{2‑[{[4‑(8-fluoro‑6-oxo‑3,4,5,6-tetrahydro‑1H-azepino[5,4,3‑cd]indol‑2-yl)phenyl]methyl}(methyl) amino]ethoxy}ethoxy)ethyl]‑3’,6’-dihydroxy‑3-oxo‑3H-spiro[[2]benzofuran‑1,9’-xanthene]‑5-carboxamide Probe 2A tert-butyl [2‑(2‑{2‑[{[4‑(8-fluoro‑6-oxo‑3,4,5,6-tetrahydro‑1H-azepino[5,4,3-cd]indol‑2-yl)phenyl]methyl}(methyl)amino] ethoxy}ethoxy)ethyl]carbamate

[0134] In Probe 2A, 8-Fluoro‑2‑{4‑[(methylamino)methyl]phenyl}‑1,3,4,5-tetrahydro‑6H-azepino[5,4,3‑cd]indol‑6-one hydrochloride (100.45mg, 0.253mmol) was combinedwith 2,2-dimethyl‑4-oxo‑3,8,11-trioxa‑5-azatridecan‑13-yl metha- nesulfonate (Julie Moreau and Jacqueline Marchand-Brynaert, Eur. J. Org. Chem.,2011, 1641‑1644; 124 mg, 0.380 mmol) in 1 mL anhydrous N,N-dimethylformamide. Potassium carbonate (105 mg, 0.760 mmol) was added, and the reaction mixture was stirred at 70 °C for 16 hours. The reaction mixture was diluted with 25 mL saturated aqueous NH4Cl 29 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 and was extracted with 3 x 15 mL dichloromethane. The combined organic layers were dried over MgSO4, filtered, and concentrated. The residue was diluted to 4 mL using 90% DMSO / water and was purified in 2 injections on a Gilson® RP- HPLC running uniPoint software with a Waters™ Deltap C18 200 x 25 mm column (15 um particle size, 100 Angstrom porosity) and was eluted with a gradient of 5% A (0.1% TFA-water):B (acetonitrile) to 100% A:B [0‑5 minutes: 5% A; 5‑45 minute linear gradient to 100% B 2.375% / minute gradient] with a 20 mL / minute flowrate. Fractions containing the title compound were combined and lyophilized to give the title compound as the trifluoroacetate salt. 1H NMR (600 MHz, DMSO‑d6) δ ppm 11.77 (s, 1H), 9.71 (s, 1H), 8.28 (t, J = 5.8 Hz, 1H), 7.76 - 7.72 (m, 2H), 7.69 - 7.64 (m, 2H), 7.45 (dd, J = 10.9, 2.4 Hz, 1H), 7.36 (dd, J = 9.1, 2.5 Hz, 1H), 6.76 (t, J = 5.7 Hz, 1H), 4.52 - 4.45 (m, 1H), 4.34 (dd, J = 12.8, 5.3 Hz, 1H), 3.80 (t, J=5.3Hz, 2H), 3.63 - 3.53 (m, 8H), 3.40 (q, J=5.0Hz, 4H), 3.34 (s, 1H), 3.28 - 3.22 (m, 1H), 3.07 (q, J=6.0Hz, 4H), 2.78 (d, J = 4.4 Hz, 3H), 1.37 (d, J = 1.9 Hz, 1H), 1.35 (s, 8H). ESI-MS m / z 555.51 (M+H)+. Probe 2B 2‑(4‑{[{2‑[2‑(2-aminoethoxy)ethoxy]ethyl}(methyl)amino]methyl}phenyl)‑8-fluoro‑1,3,4,5-tetrahydro‑6H-azepino [5,4,3‑cd]indol‑6-one

[0135] In Probe 2B, tert‑Butyl [2‑(2‑{2‑[{[4‑(8-fluoro‑6-oxo‑3,4,5,6-tetrahydro‑1H-azepino[5,4,3‑cd]indol‑2-yl)phenyl] methyl}(methyl)amino]ethoxy}ethoxy)ethyl]carbamate trifluoroacetate salt (21.63 mg, 0.032 mmol) was dissolved in trifluoroacetic acid (1 mL) and immediately evaporated to dryness to give the title compound as the trifluoroacetate salt. ESI-MS m / z 455.04 (M+H)+. Probe 2C 2’,7’-difluoro-N‑[2‑(2‑{2‑[{[4‑(8-fluoro‑6-oxo‑3,4,5,6-tetrahydro‑1H-azepino[5,4,3‑cd]indol‑2-yl)phenyl]methyl}(methyl) amino]ethoxy}ethoxy)ethyl]‑3’,6’-dihydroxy‑3-oxo‑3H-spiro[[2]benzofuran‑1,9’-xanthene]‑5-carboxamide

[0136] In Probe 2C, 2‑(4‑ {[{2‑[2‑(2-aminoethoxy)ethoxy]ethyl}(methyl)amino]methyl}phenyl)‑8-fluoro‑1,3,4,5-tetrahy- dro‑6H-azepino[5,4,3-cd]indol‑6-one bis trifluoroacetate salt (22 mg, 0.032 mmol) and 2,5-dioxopyrrolidin‑1-yl 2’,7’- difluoro‑3’,6’-dihydroxy‑3-oxo‑3H-spiro[[2]benzofuran‑1,9’-xanthene]‑5-carboxylate (ThermoFisher Scientific, 18.06 mg, 0.035 mmol) were combined in 1 mL anhydrous dimethylformamide containing 2% diisopropylethylamine (v / v) and shaken at ambient temperature for 24 hours. The reaction mixture was diluted to 3 mL with 90% dimethyl sulfoxide / - water and was purified in 1 injection with time collection on a Phenomenex® Gemini® 5 µMNX-C18 110 Angstrom 250 x 21.2 mm column and was eluted with a gradient A (0.1% TFA-water):B (CH3CN) [0‑5 minutes: 5% A; 5‑35 minute linear gradient to 100% B 3.16% / minutes gradient] with a 20 mL / minute flowrate. Fractions containing the title compound were combined and lyophilized to give the title compound as a trifluoroacetate salt. Analytical LCMS TFA method [gradient of 5‑100% acetonitrile (A) and 0.1% trifluoroacetic acid in water (B) was used, at a flow rate of 1.5 mL / minute (0‑0.05minute 5%A, 0.05‑1.2 minutes 5‑100%A, 1.2‑1.4minutes 100%A, 1.4‑1.5 minutes 100‑5%A. 0.25minute post-run delay]: Rt = 0.68 minute, ESI-MS m / z 425.4 (M+2H)2+, 849.6 (M+H)+. TIME RESOLVED-FLUORESCENCE RESONANCE ENERGY TRANSFER (TR-FRET) ASSAY

[0137] TR-FRETassays were used to measure the binding of compounds to TIPARP and PARP1, and to characterize biochemical selectivity. Compound stock solutions (10mM)were serially diluted in 3-fold increments in dimethyl sulfoxide (DMSO). Diluted compounds were dispensed (30 nL) into white ProxiPlate‑384-well plates (PerkinElmer, Waltham, MA, USA) using an ECHO® 550 acoustic Labcyte dispenser (Beckman-Coulter Life Sciences, Indianapolis, IN, USA) to achieve a final starting concentration of 30 µM to 0.5 nM in the assay. This was followed by addition of 5 µL of 2X protein / probe mix (Table 2) made in assay buffer (Tris-HCl 50 mM pH 8.0; 5 mM MgCl2; 5 µM ZnCl2; 1 mM DTT (dithiothreitol) ; 0.15% BSA (bovine serum albumin) (w / v); and 0.01% Triton X‑100 (w / v)) at final concentrations listed in Table 2. Subsequently, 5µL of 2X antibody (Tb-labeled anti-GSTantibody, Thermo Fisher Scientific,Waltham,MA, USA) in HEPES buffered saline was added to each well at a final concentration of 1 nM. The samples were then incubated for 2 hours under ambient conditions. Fluorescence was measured and TR-FRET ratios were determined on an EnVision multimode plate reader (PerkinElmer) using a 520 nanometer (nm) excitation wavelength and collecting fluorescence emission at 495 nm. Results are shown in Table 3. As shown in Table 3, compounds in keeping with Formula (I) as described herein exhibited high potency for inhibiting TIPARP and comparatively lower inhibition of PARP1 (i.e., high selectivity for TIPARP inhibition). 30 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Table 2 Protein Probe Protein (nM) Probe (nM) TIPARP GST‑(TEV) Probe 1 1 100 PARP1 GST* Probe 2 4 10 *BPS BioScience Inc., San Diego, CA, USA Table 3 Ex. # TIPARP GST‑(TEV) IC50 (µM) PARP1 GST IC50 (µM) PARP1 / TIPARP IC50 Ratio Ex. # TIPARP GST‑(TEV) IC50 (µM) PARP1 GST IC50 (µM) PARP1 / TIPARP IC50 Ratio 1 0.00240 2.74 1140 16 0.00334 1.16 347 2 0.00375 5.02 1340 17 0.00500 0.206 41.2 3 0.00474 0.504 106 18 0.00586 0.871 149 4 0.00570 3.12 547 19 0.00579 6.75 1170 5 0.00230 2.24 974 20 0.00695 2.13 306 6 0.00343 0.283 82.5 21 0.0129 0.331 25.7 7 0.00379 1.58 417 22 0.00280 2.73 975 8 0.00879 1.20 137 23 0.00274 0.136 49.6 9 0.00432 4.36 1010 24 1.87 3.08 1.65 10 0.0101 3.08 305 25 2.00 2.03 1.02 11 0.0109 0.616 56.5 26 1.62 1.28 0.790 12 0.00316 3.65 1160 27 0.00234 0.231 98.7 13 0.00379 7.38 1950 28 0.00569 0.507 89.1 14 0.00217 4.11 1890 29 0.00175 0.263 150 15 0.00215 0.417 194 CELLTITER-GLO® CELL ASSAY FOR TIPARP INHIBITION

[0138] NCI-H1373 cells have been shown to undergo cell cycle arrest following inhibition of TIPARP. The NCI-H1373 CellTiter-Glo® assay was used to determine cell numbers after TIPARP inhibition-induced cell cycle arrest, and enabled screening of compounds for cellular TIPARP inhibition potency.

[0139] Compound stock solutions (10 mM) were serially diluted in 3-fold increments in dimethyl sulfoxide. Sample aliquots (120 nL) were dispensed into Corning® 384-well Flat Clear Bottom White Polystyrene TC-treated Microplates (Corning Life Sciences, Corning, NY, USA) using an ECHO® 550 acoustic Labcyte dispenser (Beckman-Coulter Life Sciences, Indianapolis, IN, USA) to achieve final concentrations between 30 µM and 0.5 nM in the assay. This was followed by the addition of 40 µL of 1000 NCI-H1373 cells (ATCC®, Manassas, VA, USA) in complete growth medium RPMI 1640 growth medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 20 mg / L L-glutamate, 25 mM HEPES (4‑(2-hydroxyethyl)‑1-piperazineethanesulfonic acid), 10% (v / v) heat inactivated FBS (fetal bovine serum), 1% (w / v) penicillin-streptomycin, and 100 nM 10-Cl-BBQ (10-chloro‑7H-benzimidazo[2,1-a]benz[de]isoquinolin‑7-one, (Tocris, Minneapolis, MN, USA). Duplicate cell plates with 40 µL cells / well along with various concentrations of compounds were kept in Nunc™ Square BioAssay Dishes (Thermo Fisher Scientific) at 98% relative humidity in a 5% CO2 incubator at 37 °C for 6 days. On day 7, plates were moved to ambient lab conditions for 10 minutes until the temperature equilibrated. Following equilibration, CellTiter-Glo® Luminescent Cell Viability Assay reagent (Promega™, Madison, WI, USA) was added (15 µL) to each well of the plates. After a 20 minutes incubation, luminescence was detected using a ViewLux™ microplate imager (PerkinElmer, Waltham, MA, USA). Data analysis was performed using GraphPad Prism (Dotmatics, Boston, MA, USA). Results are shown in Table 4. 31 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 Table 4 Ex. # NCI-H1373 CTG EC50 (µM) Ex. # NCI-H1373 CTG EC50 (µM) Ex. # NCI-H1373 CTG EC50 (µM) 1 0.0172 11 0.337 21 0.160 2 0.0322 12 0.0292 22 0.0270 3 0.00965 13 0.0333 23 0.0442 4 0.0111 14 0.0109 24 6.49 5 0.0344 15 0.0148 25 10.2 6 0.0454 16 0.0191 26 9.10 7 0.0717 17 0.0341 27 0.0387 8 0.0559 18 0.0667 28 0.174 9 0.0677 19 0.0712 29 0.0182 10 0.284 20 0.0817 CELLTITER-GLO® DLD‑1 BRCA2 (‑ / -) AND DLD‑1 PARENT CELL ASSAY FOR PARP 1 / 2 INHIBITION

[0140] The DLD‑1 BRCA2 (‑ / ‑) CellTiter-Glo® assay measured the synthetic lethality of PARP1 / 2 inhibitors and was usedasasurrogate toallowmeasurement of activity of bothPARP1andPARP2 incells. If PARP1andPARP2are inhibited and are trapped onto chromatin duringDNA replication, there is replication fork stalling and accumulation of double strand breaks. In BRCAwild-type cells, the resulting double strand DNA breaks are repaired via homologous recombination. In cells withmutation of BRCA1 or BRCA2, homologous recombination repair of DNA double strand breaks is defective, and cell death occurs after PARP1 / 2 inhibitor treatment (Lord CJ, Ashworth A. PARP inhibitors: synthetic lethality in the clinic. Science. 2017 March 17; 355(6330):1152‑1158). Testing compounds in viability assays with the DLD‑1-BRCA2 (‑ / ‑) and DLD‑1 parent cell enabled assessment on PARP1 / 2-dependent (cytotoxicity in the BRCA2 (‑ / ‑) cells only) vs. non- selective cytotoxicity in both cell lines. Accordingly, viability assays with DLD‑1 parent and DLD‑1 BRCA2 cells demon- strated low PARP 1 / 2 inhibitory activity for compounds of Formula (I).

[0141] Compound stock solutions (10 mM) were serially diluted in 3-fold increments in dimethyl sulfoxide (DMSO). Sample aliquots (120 nL) were dispensed into Corning® 384-well Flat Clear Bottom White Polystyrene TC-treated Microplates (Corning Life Sciences, Corning, NY, USA) using an ECHO® 550 acoustic Labcyte dispenser (Beckman- Coulter Life Sciences, Indianapolis, IN, USA) to achieve concentrations between 30 µM and 0.5 nM in the assay. In the assay, 40µLof500DLD‑1parent cells (HORIZON®Therapeutics,Deerfield, IL,USA)or40µLof 1200DLD‑1-BRCA2 (‑ / ‑) cells (HORIZON® Therapeutics) in complete growth medium containing RPMI 1640 growth medium (ThermoFisher Scientific, Waltham, MA, USA) supplemented with 20 mg / L L-glutamate, 25 mM HEPES (4‑(2-hydroxyethyl)‑1-piper- azineethanesulfonic acid), 10% (v / v) heat inactivated FBS (fetal bovine serum), and 1% (w / v) Penicillin-Streptomycin were added.

[0142] Duplicate cell plates with 40 µL cells / well along with various concentrations of compounds were kept in Nunc™ SquareBioAssayDishes (ThermoFisherScientific) at 98% relative humidity in a 5%CO2 incubator at 37 °C for 6 days.On day 7, plates were moved to ambient lab conditions for 10 minutes until the temperature equilibrated. Following equilibration, CellTiter-Glo® Luminescent Cell Viability Assay reagent (Promega™, Madison, WI, USA) was added (15 µL) to each well of the plates. After a 20 minutes incubation, luminescence was detected using a ViewLux™ microplate imager (PerkinElmer,Waltham,MA, USA). Data analysis was performed usingGraphPadPrism (Dotmatics, Boston,MA, USA).Results are shown inTable 5. As shown in Table 5,DLD‑1parent andDLD‑1BRCA2 (‑ / ‑) cells showedgood viability after incubation with compounds in keeping with Formula (I) as described herein (i.e., low inhibition of PARP1 / 2). Table 5 Ex. # DLD‑1 CTG EC50 (µM) DLD‑1 BRCA2 (‑ / -) CTG EC50 (µM) Ex. # DLD‑1 CTG EC50 (µM) DLD‑1 BRCA2 (‑ / -) CTG EC50 (µM) Ex. # DLD‑1 CTG EC50 (µM) DLD‑1 BRCA2 (‑ / ‑) CTG EC50 (µM) 1 >30 18.5 11 16.0 1.18 21 >30 1.06 2 22.1 16.5 12 11.1 5.48 22 >30 9.19 32 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 (continued) Ex. # DLD‑1 CTG EC50 (µM) DLD‑1 BRCA2 (‑ / -) CTG EC50 (µM) Ex. # DLD‑1 CTG EC50 (µM) DLD‑1 BRCA2 (‑ / -) CTG EC50 (µM) Ex. # DLD‑1 CTG EC50 (µM) DLD‑1 BRCA2 (‑ / ‑) CTG EC50 (µM) 3 >30 5.03 13 11.1 6.96 23 13.3 0.785 4 23.9 7.17 14 14.3 4.49 24 >30 23.4 5 >30 20.5 15 10.3 1.38 25 >30 >30 6 14.4 1.13 16 15.2 0.997 26 >30 >30 7 >30 17.2 17 9.75 0.217 27 >30 0.894 8 >30 5.73 18 12.0 1.05 28 >30 5.45 9 >30 >30 19 >30 18.3 29 5.96 0.23 10 >30 >30 20 >30 3.00 MC‑38 (KER) SYNGENEIC MODEL EFFICACY

[0143] The murine cell line MC‑38 derived from C57BL6 murine colon adenocarcinoma cells was obtained from Kerafast (Boston, MA, USA). A suspension of 2.5 x 105 cells in culture medium mixed with Matrigel® (Corning Life Sciences, Lowell, MA, USA; 1: 1, volume:volume) was injected subcutaneously in the right hind flank of female C57BL / 6 mice to generate tumor formation. Treatment started when the size of the flank tumors was approximately 120 mm3. Vehicleand fourdifferentdosesofExample1wereadministeredoncedaily toeachofeightmiceateachdose level for up to 21 days. Each administration consisted of a single 0.2 mL oral (PO) dose in a vehicle formulation (10% ethanol, 30% PEG300, and 60% PHOSAL® 50 PG).

[0144] Results are depicted in FIG. 1. As shown, the administration of Example 1 inhibited the growth of MC‑38 (Ker) tumors compared to vehicle. The maximum tumor growth inhibition (TGIMax) achieved by Doses A-D were 46%, 77%, 98%, and 99%, respectively. TUMOR SELECTIVE INDUCTION OF IFNβ IN MC‑38 (KER) SYNGENEIC MODEL

[0145] The murine cell line MC‑38 derived from C57BL / 6 murine colon adenocarcinoma cells was obtained from Kerafast (Boston, MA, USA). A suspension of 2.5 x 105 cells in culture medium mixed with Matrigel® (Corning Life Sciences, Lowell, MA, USA; 1: 1, volume:volume) was injected subcutaneously in the right hind flank of female C57BL / 6 mice to generate tumor formation. Treatment started when the size of the flank tumors was approximately 500 mm3. Vehicle and three different doses of Example 1were administered once daily to each of four to fivemice at each dose level for 2 days. Each administration consisted of a single 0.2mLPOdose in a vehicle formulation (10%ethanol, 30%PEG300, and 60% PHOSAL® 50 PG).

[0146] Bloodand tumorswere collected 3hours after the last dose. Bloodwas transferred tomicrofuge tubes containing EDTA, centrifuged at 2000 x g for 8minutes at 4 °C, and the upper plasma layerwas transferred to new tubes and frozen at ‑80 °C. Tumors were weighed and 200 to 500 mg of tumor tissue was transferred to microfuge tubes, which were then centrifuged at 15000 x g for 10 minutes at 4 °C. The supernatant was transferred to a fresh 96 well plate. 20 µL of cold Dulbecco’s Phosphate Buffered Saline (DPBS) was added to the tubes containing the tumors and the tumors were dislodged with pipette tips. The tubes were then centrifuged at 500 x g for 10 minutes at 4 °C, and the supernatant was combined with the corresponding supernatant from the previous step in the 96 well plate. The plate containing the combined supernatants (tumor fluid) was centrifuged at 500 x g for 3 minutes at 4 °C, and clean supernatant was transferred to a new 96 well plate which was then frozen at ‑80 °C. The concentration of IFNβ in the thawed plasma and tumor fluid was determined using a Milliplex MAP mouse IFNβ assay (EMD Millipore, Burlington, MA, USA).

[0147] Results are depicted in FIG. 2. As shown, the administration of Example 1 induced IFNβ in MC‑38 (Ker) tumors compared to vehicle but not in plasma, indicating a selective induction of cytokines in the tumor microenvironment. NCI-H1373 XENOGRAFT TUMOR MODEL EFFICACY

[0148] The human xenograft line NCI-H1373 was obtained from ATCC (American Type Culture Collection, Manassas, Virginia, USA). A suspension of 5 x 106 cells in culture mediummixed with Matrigel® (Corning Life Sciences, Lowell, MA, USA; 1: 1, volume:volume) was injected subcutaneously in the right hind flank of female SCID-beige mice to generate 33 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 tumor formation. Treatment started when the size of the flank tumors was approximately 120mm3. The TIPARP inhibition activity of Example 1 was evaluated by dosing five separate treatment groups with vehicle and four different doses of Example 1, administered twice a day (BID) for 14 days. Each administration consisted of a 0.2 mL PO dose in a vehicle formulation (10% ethanol, 30% PEG300, and 60% PHOSAL® 50).

[0149] Results are depicted in FIG. 3. As shown, the administration of Example 1 inhibited the growth of NCI-H1373 tumors compared to vehicle. The TGIMax achieved by Doses A-D were 26%, 46%, 73%, and 92%, respectively. Claims 1. A compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein: L is selected from the group consisting of a bond and CH2; R1,R2,R3,andR4are independently selected from thegroupconsistingofHandCH3,whereinat least oneandno more than two of R1, R2, R3, and R4 are CH3; and Z is selected from the group consisting of CF3 and C(CH3)2OH. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L is a bond. 3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein one of R1 or R4 is CH3; both R1 and R4 are CH3; one of R2 or R3 is CH3; or both R2 and R3 are CH3. 4. The compound of claim 2 or 3, or a pharmaceutically acceptable salt thereof, wherein Z is CF3. 5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L is CH2. 6. The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein one of R1 or R4 is CH3; both R1 and R4 are CH3; one of R2 or R3 is CH3; or both R2 and R3 are CH3. 7. The compound of claim 5 or 6, or a pharmaceutically acceptable salt thereof, wherein Z is C(CH3)2OH. 8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl }cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(14)‑one; 5‑[{1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(tiifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl }cyclobutyl]‑3‑(tri- fluoromethyl)pyridin‑2(14)‑one; 5‑[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl }cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(14)‑one; 5‑[(1s,3s)‑3-f (3R,5S)‑3,5-dimethyl‑4‑[5‑(tiifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(tri- fluoromethyl)pyridin‑2(14)‑one; 5‑[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyl}cyclobu- tyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 34 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 5‑[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl }cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(14)‑one; 5‑[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoro- methyl)pyridin‑2(1H)‑one; 5‑[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl]‑3‑(tri- fluoromethyl)pyridin‑2(1H)‑one; 5‑[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl } cyclobu- tyl]‑3‑(trifluoromethyl)pyridin‑2(14)‑one; 5‑{[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1s,3s)‑3‑{(2R,6S)‑2,6-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1s,3s)‑3‑{(3R,5S)‑3,5-dimethyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(3R)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(3S)‑3-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(2S)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(3R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1S,3s)‑3‑{(2R)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(3S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3-methylpiperazine‑1-carbonyl}cyclobutyl] methyl}‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; 5‑{[(1R,3s)‑3‑{(2S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑2-methylpiperazine‑1-carbonyl }cyclobutyl] methyl }‑3‑(trifluoromethyl)pyridin‑2(1H)‑one; and 5‑{[(1s,3s)‑3‑{(3R,5S)‑4‑[5‑(2-hydroxypropan‑2-yl)pyrimidin‑2-yl]‑3,5-dimethylpiperazine‑1-carbonyllcyclobu- tyl]methyl}‑3‑(tiifluoromethyl)pyiidin‑2(1H)‑one. 9. The compound of claim 8, wherein the compound is 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl] piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one, or a pharmaceutically acceptable salt there- of. 10. The compound of claim 9, wherein the compound is 5‑[(1S,3s)‑3‑{(2R)‑2-methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl] piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one. 11. The compound of claim 9, wherein the compound is a pharmaceutically acceptable salt of 5‑[(1S,3s)‑3‑{(2R)‑2- methyl‑4‑[5‑(trifluoromethyl)pyrimidin‑2-yl]piperazine‑1-carbonyl}cyclobutyl]‑3‑(trifluoromethyl)pyridin‑2(1H)‑one. 12. Apharmaceutical composition comprising the compoundof anyprevious claim, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 13. Acompound according to any oneof claims 1 to 11or a pharmaceutical composition according to claim12, for use in a method for treating head and neck squamous cell carcinoma (HNSCC), the method comprising administering the compound, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, to a human patient in need thereof. 35 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 36 EP 4 644 385 A1 37 EP 4 644 385 A1 38 EP 4 644 385 A1 39 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 40 EP 4 644 385 A1 5 10 15 20 25 30 35 40 45 50 55 41 EP 4 644 385 A1 REFERENCES CITED IN THE DESCRIPTION This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard. Non-patent literature cited in the description • JOSEPH M. GOZGIT et al. PARP7 Negatively Regulates the Type I Interferon Response in Cancer Cells and Its Inhibition Triggers Antitumor Immunity. CANCER CELL, 2021, vol. 39, 1214

[0002] • CHEMICAL ABSTRACTS, 1034901‑50‑2

[0051] • CHEMICAL ABSTRACTS, 870987‑63‑6

[0051] • JULIE MOREAU ; JACQUELINE MARCHAND- BRYNAERT. Eur. J. Org. Chem., 2011, 1641-1644

[0134] • LORD CJ ; ASHWORTH A. PARP inhibitors: synthetic lethality in the clinic. Science, 17 March 2017, vol. 355 (6330), 1152-1158

[0140] 摘要 本公開提供式(I)化合物及其藥學上可接受的鹽，這些化合物能夠抑制 TIPARP 的活性，其中式(I)中的變數具有說明書中定義的任意值。

Claims

1. A compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein: L is selected from the group consisting of a bond and CH2; R1, R2, R3, and R4 are independently selected from the group consisting of H and CH3, wherein at least one and no more than two of R1, R2, R3, and R4 are CH3; and Z is selected from the group consisting of CF3 and C(CH3)2OH.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L is a bond.

3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein one of R1 or R4 is CH3; both R1 and R4 are CH3; one of R2 or R3 is CH3; or both R2 and R3 are CH3.

4. The compound of claim 2 or 3, or a pharmaceutically acceptable salt thereof, wherein Z is CF3.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L is CH2.

6. The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein one of R1 or R4 is CH3; both R1 and R4 are CH3; one of R2 or R3 is CH3; or both R2 and R3 are CH3.

7. The compound of claim 5 or 6, or a pharmaceutically acceptable salt thereof, wherein Z is C(CH3)2OH.

8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: 5-[(1S,3s)-3-{(2R)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl }cyclobutyl]-3-(trifluoromethyl)pyridin-2(14)-one; 5-[{1s,3s)-3-{(2R,6S)-2,6-dimethyl-4-[5-(tiifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl }cyclobutyl]-3-(trifluoromethyl)pyridin-2(14)-one; 5-[(1S,3s)-3-{(3R)-3-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl }cyclobutyl]-3-(trifluoromethyl)pyridin-2(14)-one; 5-[(1s,3s)-3-f (3R,5S)-3,5-dimethyl-4-[5-(tiifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(14)-one; 5-[(1s,3s)-3-{(3R,5S)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-3,5-dimethylpiperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one; 5-[(1R,3s)-3-{(2S)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl }cyclobutyl]-3-(trifluoromethyl)pyridin-2(14)-one; 5-[(1S,3s)-3-{(3R)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-3-methylpiperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one; 5-[(1R,3s)-3-{(3S)-3-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one; 5-[(1S,3s)-3-{(2R)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-2-methylpiperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one; 5-[(1R,3s)-3-{(3S)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-3-methylpiperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one; 5-[(1R,3s)-3-{(2S)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-2-methylpiperazine-1-carbonyl } cyclobutyl]-3-(trifluoromethyl)pyridin-2(14)-one; 5-{[(1S,3s)-3-{(2R)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1s,3s)-3-{(2R,6S)-2,6-dimethyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1s,3s)-3-{(3R,5S)-3,5-dimethyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1S,3s)-3-{(3R)-3-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1R,3s)-3-{(3S)-3-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1R,3s)-3-{(2S)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1S,3s)-3-{(3R)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-3-methylpiperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1S,3s)-3-{(2R)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-2-methylpiperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1R,3s)-3-{(3S)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-3-methylpiperazine-1-carbonyl}cyclobutyl]methyl}-3-(trifluoromethyl)pyridin-2(1H)-one; 5-{[(1R,3s)-3-{(2S)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-2-methylpiperazine-1-carbonyl }cyclobutyl]methyl }-3-(trifluoromethyl)pyridin-2(1H)-one; and 5-{[(1s,3s)-3-{(3R,5S)-4-[5-(2-hydroxypropan-2-yl)pyrimidin-2-yl]-3,5-dimethylpiperazine-1-carbonyllcyclobutyl]methyl}-3-(tiifluoromethyl)pyiidin-2(1H)-one.

9. The compound of claim 8, wherein the compound is 5-[(1S,3s)-3-{(2R)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one, or a pharmaceutically acceptable salt thereof.

10. The compound of claim 9, wherein the compound is 5-[(1S,3s)-3-{(2R)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one.

11. The compound of claim 9, wherein the compound is a pharmaceutically acceptable salt of 5-[(1S,3s)-3-{(2R)-2-methyl-4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazine-1-carbonyl}cyclobutyl]-3-(trifluoromethyl)pyridin-2(1H)-one.

12. A pharmaceutical composition comprising the compound of any previous claim, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

13. A compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12, for use in a method for treating head and neck squamous cell carcinoma (HNSCC), the method comprising administering the compound, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, to a human patient in need thereof.

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