WRN helicase inhibitors

Abstract

The present disclosure is directed to compounds of Formula I: wherein A, R1, and R2 are described herein, which are compounds useful for inhibiting Werner Syndrome dsDNA helicase (WRN) and methods of treating disease using these compounds. This disclosure further provides compounds and their use for the treatment of cancers characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR), including colorectal, gastric, endometrial and ovarian cancer.

Description

WRN HELICASE INHIBITORS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application Serial No. 63 / 679,305 filed August 5, 2024, the entire contents of which are incorporated by reference herein. REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The contents of the electronic sequence listing (26014-WO-PCT_SL.xml; Size: 4,684 bytes; and Date of Creation: September 16, 2024) are herein incorporated by reference in their entirety. FIELD

[0003] The present disclosure relates to compounds of Formula I, or pharmaceutically acceptable salts thereof, and their use for inhibiting Werner Syndrome dsDNA helicase (WRN). Another aspect of the disclosure are compounds of Formula I and their use for treating cancer, for example in the treatment of cancer characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR). Still another aspect of the disclosure provides methods of treating cancers, including colorectal, gastric and endometrial cancer using the compounds disclosed herein. BACKGROUND

[0004] Werner syndrome dsDNA helicase (WRN) is one of five RecQ family ATP-dependent motors involved in DNA unwinding for repair. WRN can uniquely process hypermutated DNA structures (e.g., G-quadruplexes, bubble DNA, Holliday junctions, forked DNA). WRN helicase activity is dispensable in WT / MSS cells, but essential when mismatch repair (MMR) genes are mutated (MSI cells).

[0005] A common initial event in cancer development is the loss of DNA mismatch repair (MMR) of colorectal (CRC), endometrial, ovarian and gastric cancers (Aaltonen, L. A. et al., Clues to the pathogenesis of familial colorectal cancer, Science 260, 812-816 (1993), Bonneville R et al., Landscape of Microsatellite Instability Across 39 Cancer Types. JCO Precis Oncol.1: PO.17.00073 (2017)). A high mutational burden and frequent deletion and insertion events in repetitive DNA tracts (known as microsatellite instability (MSI)) occurs in cancers that have lost competence in mismatch repair (MMR). While progress has been made in the treatment ofmicrosatellite instability high (MSI-H) cancers, there is still a significant unmet medical need in CRC and other MSI-H indications (André T., et al. Pembrolizumab in Microsatellite-Instability- High Advanced Colorectal Cancer. N Engl J Med 383(23):2207-2218 (2020)). Through large- scale functional genomics screens across large panels of cell lines, Werner Syndrome RecQ helicase (WRN) has been identified as being selectively required for the survival of cell lines with defective mismatch repair that have become MSI-H (Behan, F. M. et al., Nature 568, 511- 516 (2019); and Chan, E. M. et al., Nature 568, 551-556 (2019). Kategaya, L., Perumal, et al., iScience 13, 488-497 (2019), and Lieb, S. et al., eLife 8, e43333 (2019)). See also US20230046859. Depletion of WRN leads to anti-proliferative effects resulting in activation of multiple DNA damage signaling markers, induction of cell cycle arrest and apoptosis in MMR cancer models but not cancer cells with an intact MMR pathway. This indicates that WRN provides a DNA repair and maintenance function that is essential for cell survival in MSI cancers. Thus, inhibiting the WRN helicase is an attractive strategy for the treatment of mismatch repair defective cancers. There remains a need for new treatments and therapies for the treatment of cancer, particularly in cancers characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR), including colorectal, gastric, ovarian, or endometrial cancer. SUMMARY

[0006] The present disclosure is directed to compounds of Formula I:wherein A, R1, and R2are described below.

[0007] This disclosure provides spirocyclic compounds for inhibiting Werner Syndrome dsDNA helicase (WRN) and methods of treating disease using these compounds. This disclosure further provides compounds and their use for the treatment of cancers characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR), including colorectal, gastric, endometrial and ovarian cancer.

[0008] Another aspect of the disclosure provides substituted spirocyclic compounds, analogues and derivatives thereof and their use for inhibiting Werner Syndrome RecQ DNA helicase(WRN). Another aspect of the disclosure provides methods of treating disease using the compounds described herein, including the use in treating cancer, including treatment of cancers characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR). Still another aspect of the disclosure provides methods of treating cancers, including colorectal, gastric and endometrial cancer using the compounds disclosed herein. Yet another aspect of the disclosure provides the use of the compounds of this disclosure as research chemicals, intermediate compounds, combinations, processes and formulations.

[0009] Another aspect of the disclosure provides orally bioavailable WRN inhibitor compounds with context-selective gH2AX induction activity.

[0010] Another aspect of the disclosure provides compounds of Formula I, pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof and / or combinations thereof, said compounds being inhibitors of Werner Syndrome RecQ DNA Helicase (WRN). Another aspect of the disclosure provides methods of treating, preventing, or ameliorating a disease or condition, comprising administering to a subject in need thereof an effective amount of a WRN inhibitor. Another aspect of the disclosure provides compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof and combinations thereof useful for the treatment of cancer, wherein the cancers are characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR). Also provided in this disclosure are compounds that bind to, and / or inhibit WRN, and are therefore useful as research chemicals, e.g., as a chemical probe, and as tool compounds. Various embodiments of the disclosure are described herein.

[0011] The summary of the technology described above is non-limiting and other features and advantages of the technology will be apparent from the following detailed description, and from the claims. DETAILED DESCRIPTION

[0012] The present disclosure is directed to compounds of Formula I:or a pharmaceutically acceptable salt thereof wherein, A is heteroaryl selected from pyrimidinyl, thiazolyl, and pyrazolyl, optionally substituted with 1 to 3 groups of Ra;R1is selected from C1-3haloalkyl, halogen and C1-6alkyl; R2is selected from heterocycloalkyl, C3-10 cycloalkyl, -(CH2)naryl, -N(R3)2, -NHaryl; said heterocycloalkyl, cycloalkyl, aryl optionally substituted with 1 to 3 groups selected from Ra; R3is selected from heterocycloalkyl and aryl said heterocycloalkyl and aryl optionally substituted with 1 to 3 groups selected from Ra; Rais independently selected from hydroxyl, C1-6alkyl, halogen, C1-3haloalkyl, -(CH2)nCO2H, -C(O)NH- aryl, -C(O)aryl, -C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, aryl, heterocycloalkyl, and heteroaryl optionally substituted with 1 to 3 groups selected from Rb; Rbis independently selected from hydroxyl, C1-6alkyl, -COOH, and halogen and n is 0 to 3.

[0013] An embodiment of structural Formula I is realized when A is unsubstituted or substituted pyrimidinyl. Another embodiment of structural Formula I is realized when A is unsubstituted or substituted thiazolyl. Another embodiment of structural Formula I is realized when A is unsubstituted or substituted pyrazolyl.

[0014] A further embodiment of structural Formula I is realized when A is substituted pyrimidinyl. Another embodiment of structural Formula I is realized when A is substituted thiazolyl. Another embodiment of structural Formula I is realized when A is substituted pyrazolyl.

[0015] A further embodiment of structural Formula I is realized when A is pyrimidinyl substituted with -(CH2)nC(O)OH. Another embodiment of structural Formula I is realized when A is thiazolyl substituted with -(CH2)nC(O)OH. Another embodiment of structural Formula I is realized when A is pyrazolyl substituted with -(CH2)nC(O)OH.

[0016] Another embodiment of structural Formula I is realized when R1is C1-3 haloalkyl. A subembodiment of this aspect is realized when R1is selected from CHF2, CH2F, CF3, CHCl2, CH2Cl, CCl3, CHBr2, CH2Br, and CBr3. Another subembodiment of this aspect is realized when R1is selected from CHF2, CH2F, and CF3. Still another subembodiment of this aspect is realized when R1is CF3.

[0017] Another embodiment of Formula I is realized when R1is halogen selected from chlorine, fluorine, bromine and iodine.

[0018] Another embodiment of Formula I is realized when R1is C1-6alkyl. Still another subembodiment of this aspect is realized when R1is CH3.

[0019] Another embodiment of Formula I is realized when R2is unsubstituted or substituted heterocycloalkyl. An aspect of this embodiment is realized when R2heterocycloalkyl is selectedfrom unsubstituted or substituted piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, and tetrahydrofuranyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted piperidyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted oxetanyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted pyrrolyl. Another subembodiment of this aspect is realized when the heterocycloalkyl is unsubstituted or substituted pyrrolidinyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted piperazinyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted morpholinyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted tetrahydrofuranyl.

[0020] Another embodiment of Formula I is realized when R2is unsubstituted or substituted C3-10 cycloalkyl. An aspect of this embodiment is realized when R2is selected from unsubstituted or substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydroindenyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted cyclopropyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted cyclobutyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted cyclopentyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted cyclohexyl. Another aspect of this embodiment is realized when R2is unsubstituted or substituted dihydro- indenyl.

[0021] Another embodiment of Formula I is realized when R2is -(CH2)nphenyl, said phenyl optionally substituted with 1 to 3 groups selected from Ra.

[0022] Another embodiment of Formula I is realized when R2is -N(R3)2, and R3is heterocycloalkyl, said heterocycloalkyl optionally substituted with 1 to 3 groups selected from Ra. A subembodiment of this aspect is realized when the heterocycloalkyl is selected from unsubstituted or substituted piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, and tetrahydrofuranyl. Another subembodiment of this aspect is realized when the heterocycloalkyl is selected from unsubstituted or substituted piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, and tetrahydrofuranyl. Another subembodiment of this aspect is realized when the heterocycloalkyl is unsubstituted or substituted pyrrolyl. Another subembodiment of this aspect is realized when the heterocycloalkyl is unsubstituted or substituted pyrrolidinyl. Another subembodiment of this aspect is realized when the heterocycloalkyl is unsubstituted or substituted tetrahydrofuranyl.

[0023] Another embodiment of Formula I is realized when R2is -N(R3)2, and R3is aryl, said aryl optionally substituted with 1 to 3 groups selected from Ra. A subembodiment of this aspect is realized when the aryl is unsubstituted or substituted phenyl.

[0024] Another embodiment of Formula I is realized when both R3in -N(R3)2of R2are the same. Another embodiment is realized when the R3s in -N(R3)2 of R2are different. Another embodiment is realized one of R3in -N(R3)2of R2is unsubstituted or substituted phenyl and the other is unsubstituted or substituted pyrrolidinyl.

[0025] Another embodiment of Formula I is realized when Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, CF3, -C(O)NH-phenyl, - C(O)phenyl, -C(O)heterocycloalkyl, and -C(O)heteroaryl, said methyl, ethyl, propyl, butyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb. An aspect of this embodiment is realized when Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, CH2CO2H, and CF3. In another aspect of this embodiment, Rais CH2CO2H. Another aspect of this embodiment is realized when Rais - C(O)NH-phenyl, said phenyl unsubstituted or substituted with 1 to 3 groups of Rb. Another aspect of this embodiment is realized when Rais -C(O)phenyl, said phenyl unsubstituted or substituted with 1 to 3 groups of Rb. Another aspect of this embodiment is realized when Rais - C(O)heterocycloalkyl, said heterocycloalkyl unsubstituted or substituted with 1 to 3 groups of Rb. Another aspect of this embodiment is realized when the heterocycloalkyl in - C(O)heterocycloalkyl of Rais selected from unsubstituted or substituted piperidinyl and pyrrolidinyl. Another aspect of this embodiment is realized when Rais -C(O)heteroaryl, said heteroaryl unsubstituted or substituted with 1 to 3 groups of Rb. Another aspect of this embodiment is realized when the heteroaryl in -C(O)heteroaryl of Rais selected from unsubstituted or substituted pyrimidinyl and pyrrolyl.

[0026] Another embodiment of structural Formula I is realized when A is unsubstituted or substituted pyrimidinyl, R1is C1-3 haloalkyl, halogen, C1-6 alkyl, R2is heterocycloalkyl or C1-10 cycloalkyl selected from unsubstituted or substituted piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydro-indenyl, and Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb.

[0027] Another embodiment of structural Formula I is realized when A is unsubstituted or substituted pyrimidinyl, R1is C1-3haloalkyl, halogen, C1-6alkyl, R2is -(CH2)nphenyl or -N(R3)2, R3is heterocycloalkyl or phenyl said heterocycloalkyl and phenyl optionally substituted with 1 to 3 groups selected from Ra, and Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, -C(O)heterocycloalkyl, -C(O)heteroaryl,said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb.

[0028] Another embodiment of structural Formula I is realized when A is unsubstituted or substituted thiazolyl, R1is C1-3 haloalkyl, halogen, C1-6 alkyl, R2is heterocycloalkyl or C1-10 cycloalkyl selected from unsubstituted or substituted piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydro-indenyl, and Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl, and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb

[0029] Another embodiment of structural Formula I is realized when A is unsubstituted or substituted thiazolyl, R1is C1-3haloalkyl, halogen, C1-6alkyl, R2is -(CH2)nphenyl or -N(R3)2, R3is heterocycloalkyl or phenyl said heterocycloalkyl and phenyl optionally substituted with 1 to 3 groups selected from Ra, and Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb

[0030] Another embodiment of structural Formula I is realized when A is unsubstituted or substituted pyrazolyl, R1is C1-3 haloalkyl, halogen, C1-6 alkyl, R2is heterocycloalkyl or C1-10 cycloalkyl selected from unsubstituted or substituted piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydro-indenyl, and Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb.

[0031] Another embodiment of structural Formula I is realized when A is unsubstituted or substituted pyrazolyl, R1is C1-3haloalkyl, halogen, C1-6alkyl, R2is -(CH2)nphenyl or -N(R3)2, R3is heterocycloalkyl or phenyl said heterocycloalkyl and phenyl optionally substituted with 1 to 3 groups selected from Ra, and Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb.

[0032] Another embodiment is realized when R2is -NHphenyl, said phenyl optionally substituted with 1 to 3 groups selected from Ra.26014

[0033] Non-limiting examples of compounds of the disclosure are listed in Table 1 below. Table 1 Compd Structure Name MS ATPase (EI) EC50(nM)

[0034] The terms used herein have their ordinary meaning and the meaning of such terms is independent at each occurrence thereof. That notwithstanding and except where stated otherwise, the following definitions apply throughout the specification and claims. Chemical names, common names and chemical structures may be used interchangeably to describe that same26014 structure. These definitions apply regardless of whether a term is used by itself or in combination with other terms, unless otherwise indicated. Hence the definition of “alkyl” applies to “alkyl” as well as the “alkyl” portion of “hydroxyalkyl”, “haloalkyl”, arylalkyl, alkylaryl, “alkoxy” etc.

[0035] Listed below are definitions of various terms used herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0036] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well-known and commonly employed in the art.

[0037] As used herein, the articles “a” and “an” refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0038] As used herein, the term “about” in quantitative terms refers to plus or minus 10% of the value it modifies (rounded up to the nearest whole number if the value is not sub-dividable, such as a number of molecules or nucleotides).

[0039] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 50 mg to 500 mg” is inclusive of the endpoints, 50 mg and 500 mg, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and / or values.

[0040] As used herein, the term “comprising” may include the embodiments “consisting of” and “consisting essentially of.” The terms “comprise(s),” “include(s),” “having,” “has,” “may,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named ingredients / steps and permit the presence of other ingredients / steps. However, such description should be construed as also describing compositions or processes as “consisting of” and “consisting essentially of” the enumerated components, which allows the presence of only the named components or compounds, along with any acceptable carriers or fluids, and excludes other components or compounds.

[0041] In each of the various embodiments of the disclosure, in the compounds used in the methods herein, each variable (including those in each of Formula I, and the various26014 embodiments thereof) it shall be understood that each variable is to be selected independently of the others unless otherwise indicated.

[0042] In each of the various embodiments of the disclosure, the compounds described herein, including those in each of Formula I and the various embodiments thereof, may exit in different forms of the compounds such as, for example, any solvates, hydrates, stereoisomers, and tautomers of said compounds and of any pharmaceutically acceptable salts thereof.

[0043] It shall be understood that, in the various embodiments of the disclosure described herein, any variable not explicitly defined in the context of the embodiment is as defined in Formula I.

[0044] In the various embodiments described herein, each variable is selected independently of the others unless otherwise indicated.

[0045] "Patient" includes both human and non-human animals. Non-human animals include those research animals and companion animals such as mice, rats, primates, monkeys, chimpanzees, great apes, dogs, and house cats.

[0046] "Pharmaceutical composition" (or “pharmaceutically acceptable composition”) means a composition suitable for administration to a patient. Such compositions may contain the neat compound (or compounds) of the disclosure or mixtures thereof, or salts, solvates, prodrugs, isomers, or tautomers thereof, and one or more pharmaceutically acceptable carriers or diluents. The term “pharmaceutical composition” is also intended to encompass both the bulk composition and individual dosage units comprised of one or more (e.g., two) pharmaceutically active agents such as, for example, a compound of the present disclosure and an additional agent selected from the lists of the additional agents described herein, along with any pharmaceutically inactive excipients. The bulk composition and each individual dosage unit can contain fixed amounts of the afore-said "more than one pharmaceutically active agents". The bulk composition is material that has not yet been formed into individual dosage units. An illustrative dosage unit is an oral dosage unit such as tablets, pills and the like. Similarly, the herein-described method of treating a patient by administering a pharmaceutical composition of the present disclosure is also intended to encompass the administration of the afore-said bulk composition and individual dosage units.

[0047] “Pharmaceutically acceptable carrier” refers to a substance useful in the preparation or use of a pharmaceutical composition and includes, for example, suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonic agents, buffering agents, emulsifiers, absorption delaying agents, salts, drug stabilizers, binders, excipients, disintegration agents, lubricants, wetting agents, sweetening agents, flavoring agents, dyes, and combinations26014 thereof, as would be known to those skilled in the art (see, for example, Remington The Science and Practice of Pharmacy, 22nd Ed. Pharmaceutical Press, 2013, pp.1049-1070).

[0048] The term “effective amount” or “therapeutically effective amount” of a compound of the present disclosure refers to an amount of the compound of the present disclosure that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc.

[0049] In one embodiment, the term “a therapeutically effective amount” refers to the amount of the compound of the present disclosure that, when administered to a subject, is effective to (1) at least partially alleviate, prevent and / or ameliorate a condition, or a disorder or a disease (i) mediated by WRN, or (ii) associated with WRN activity, or (II) characterized by activity (normal or abnormal) of WRN; or (2) reduce or inhibit the activity of WRN.

[0050] In another embodiment, the term “a therapeutically effective amount” refers to the amount of the compound of the present disclosure that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reducing or inhibiting the activity of WRN, or reducing WRN protein levels.

[0051] The term “cancer” refers to a disease characterized by the rapid and uncontrolled growth of aberrant cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers are described herein and include but are not limited to colorectal, gastric, endometrial, prostate, adrenocortical, uterine, cervical, esophageal, breast, kidney, ovarian cancer and the like.

[0052] The terms “tumor” and “cancer” are used interchangeably herein, e.g., both terms encompass solid and liquid, e.g., diffuse or circulating, tumors. As used herein, the term “cancer” or “tumor” includes premalignant, as well as malignant cancers and tumors.

[0053] ‘WRN inhibitor’ or ‘WRN helicase inhibitor’ as used herein means a compound that inhibits Werner Syndrome RecQ DNA helicase (WRN). The term “WRN” as used herein refers to the protein of Werner Syndrome RecQ DNA helicase. The term “WRN” includes mutants, fragments, variants, isoforms, and homologs of full-length wild-type WRN.

[0054] ‘Disease or condition mediated by WRN’ includes a disease or condition, such as cancer, which is treated by WRN inhibition. In particular this can include cancers characterized as microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR).

[0055] ‘Microsatellite unstable cancer’, microsatellite instability-high cancer’, ‘microsatellite high cancer’ and ‘MSI-high cancer’ ‘MSIhi’ and ‘MSI-H’ when used herein, are used26014 interchangeably, and describe cancers that have a high number of alterations in the length of simple repetitive genomic sequences within microsatellites.

[0056] The determination of MSI-H or dMMR tumor status for patients can be performed using, e.g., polymerase chain reaction (PCR) tests for MSI-H status or immunohistochemistry (IHC) tests for dMMR. Methods for identification of MSI-H or dMMR tumor status are described, e.g., in Ryan et al. Crit Rev Oncol Hematol.2017; 116:38-57; Dietmaier and Hofstadter. Lab Invest 2001, 81:1453-1456; and Kawakami et al. Curr Treat Options Oncol. 2015; 16(7): 30).

[0057] Microsatellite instability can be found in colorectal cancer, gastric cancer and endometrial cancer in particular, but also in adrenocortical, uterine, cervical, esophageal, breast, kidney, prostate and ovarian cancers. Examples of microsatellite high cancers include uterine corpus endometrial carcinoma, colon adenocarcinoma, stomach adenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, endocervical adenocarcinoma, esophageal carcinoma, breast carcinoma, kidney renal clear cell carcinoma and ovarian serous cystadenocarcinoma.

[0058] A cancer that has “defective mismatch repair” (dMMR) or “dMMR character” includes cancer types associated with documented MLH1, PMS2, MSH2, MSH3, MSH6, MLH3, and PMS1 mutations or epigenetic silencing, microsatellite fragile sites, or other gene inactivation mechanisms, including but not limited to cancers of the lung, breast, kidney, large intestine, ovary, prostate, upper aerodigestive tract, stomach, endometrium, liver, pancreas, haematopoietic and lymphoid tissue, skin, thyroid, pleura, autonomic ganglia, central nervous system, soft tissue, pediatric rhabdoid sarcomas, melanomas and other cancers. A cell or cancer with “defective” mismatch repair has a significantly reduced (e.g., at least about 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90% decrease) amount of mismatch repair. In some cases, a cell or cancer which is defective in mismatch repair will perform no mismatch repair.

[0059] The term “subject” refers to primates (e.g., humans, male or female), dogs, rabbits, guinea pigs, pigs, rats and mice. In certain embodiments, the subject is a primate, a rat or a mouse. In yet other embodiments, the subject is a human.

[0060] “Inhibit”, “inhibition” or “inhibiting” refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

[0061] The term “treat”, “treating” or “treatment” of any disease or disorder refers to alleviating or ameliorating the disease or disorder (i.e., slowing or arresting the development of the disease or at least one of the clinical symptoms thereof); or alleviating or ameliorating at least26014 one physical parameter or biomarker associated with the disease or disorder, including those which may not be discernible to the patient.

[0062] “Halogen” and "halo" mean fluorine, chlorine, bromine, or iodine. Preferred are fluorine, chlorine and bromine.

[0063] “Alkylene," by itself or as part of another substituent means a divalent hydrocarbon chain radical having the stated number of carbon atoms. For example, -(C1-C5)alkylene, would include, e.g., -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH(CH3)CH2- or - CH2CH2CH2CH2CH2-. A straight alkylene means a divalent straight hydrocarbon chain radical having the stated number of carbon atoms. A branched alkylene means a divalent branched hydrocarbon chain radical having the stated number of carbon atoms. A saturated alkylene means a divalent saturated hydrocarbon chain radical having the stated number of carbon atoms. An unsaturated alkylene means a divalent hydrocarbon chain radical having the stated number of carbon atoms and one or more double or triple covalent bonds within the chain. A cycloalkylene means a divalent hydrocarbon chain radical having the stated number of carbon atoms and a cycloalkyl moiety within the chain.

[0064] "Alkyl" means an aliphatic hydrocarbon group which may be straight or branched and comprising about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups contain about 1 to about 12 carbon atoms in the chain. More preferred alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. "Lower alkyl" means a group having about 1 to about 6 carbon atoms in the chain which may be straight or branched. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl and t-butyl.

[0065] “Haloalkyl” means an alkyl as defined above wherein one or more hydrogen atoms on the alkyl is replaced by a halo group defined above, for example CHF2, CH2F and CF3.

[0066] "Aryl" means an aromatic monocyclic or multicyclic ring system comprising about 6 to about 14 carbon atoms, preferably about 6 to about 10 carbon atoms. The aryl group can be optionally substituted with one or more "ring system substituents" which may be the same or different and are as defined herein. Non-limiting examples of suitable aryl groups include phenyl and naphthyl. "Monocyclic aryl" means phenyl.

[0067] Cycloalkyl" means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 12 carbon atoms, preferably about 3 to about 10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 10 ring atoms. The cycloalkyl can be optionally substituted with one or more substituents, which may be the same or different, as described herein. Monocyclic cycloalkyl refers to monocyclic versions of the cycloalkyl moieties described herein. Non-limiting examples of suitable monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. The term "C3-C10cycloalkyl" encompasses bridged, saturated or unsaturated cycloalkyl groups having 3 to 10 carbons. "Cycloalkyl" also includes non-aromatic rings as well as monocyclic, non-aromatic rings fused to a saturated cycloalkyl group and aromatic rings fused to a saturated cycloalkyl group. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and the like. Examples described by structure.The term "heteroaryl" refers to an aromatic 5-8 membered monocyclic, 8-bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms for monocyclic, 1-6 heteroatoms for bicyclic, or 1-9 heteroatoms for tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S for monocyclic, bicyclic, or tricyclic, respectively). Non-limiting examples of heteroaryls are pyridyl, pyrazolyl, pyrimidinyl, furanyl, oxazolyl, triazolyl, oxadiazolyl, and thiophenyl. The heteroaryl groups herein described may also contain fused rings that share a common carbon-carbon bond.

[0068] “Heterocycloalkyl” (or "heterocyclyl") means a non-aromatic, saturated or partially saturated monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and / or sulfur atoms present in the ring system. Preferred heterocycloalkyls contain about 5 to about 6 ring atoms. The prefix aza, oxa or thia before the heterocycloalkyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom. Any –NH in a heterocycloalkyl ring may exist protected such as, for example, as an -N(Boc), -N(CBz), -N(Tos) group and the like; such protections are also considered part of this disclosure. The heterocycloalkyl can be optionally substituted by one or more substituents, which may be the same or different, as described herein. The nitrogen or sulfur atom of the heterocycloalkyl can be optionally oxidized to the corresponding N-oxide, S- oxide or S,S-dioxide. Thus, the term “oxide,” when it appears in a definition of a variable in a general structure described herein, refers to the corresponding N-oxide, S-oxide, or S,S-dioxide.“Heterocycloalkyl” also includes rings wherein =O replaces two available hydrogens on the same carbon atom (i.e., heterocycloalkyl includes rings having a carbonyl group in the ring). Such =O groups may be referred to herein as “oxo.” An example of such a moiety is pyrrolidinone (orpyrrolidone): . As used herein, the term “monocyclic heterocycloalkyl” refersmonocyclic versions of the heterocycloalkyl moieties described herein and include a 4- to 7-membered heterocycloalkyl groups comprising from 1 to 4 ring heteroatoms, said ring heteroatoms being independently selected from the group consisting of N, N-oxide, O, S, S- oxide, S(O), and S(O)2. The point of attachment to the parent moiety is to any available ring carbon or ring heteroatom. Non-limiting examples of monocyclic heterocycloalkyl groups include piperidyl, oxetanyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, beta lactam, gamma lactam, delta lactam, beta lactone, gamma lactone, delta lactone, and pyrrolidinone, and oxides thereof. A non-limiting example of a monocyclic heterocycloalkyl group includes the . Non-limiting examples of multicyclic heterocycloalkyl groups include, bicyclicgroups. Specific examples include, but are not limited to, ..means an alkyl-O- group in which the alkyl group is as previously described. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond to the parent moiety is through the ether oxygen.

[0070] The term “substituted” means that one or more hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom’s normal valency under the existing circumstances is not exceeded, and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. By “stable compound’ or “stable structure” is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0071] The term “optionally substituted” means optional substitution with the specified groups, radicals or moieties.26014

[0072] “Combination” refers to either a fixed combination in one dosage unit form, or a combined administration where a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a combination partner (e.g., another drug as explained below, also referred to as “therapeutic agent” or “co-agent”) may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative. The single components may be packaged in a kit or separately. One or both of the components (e.g., powders or liquids) may be reconstituted or diluted to a desired dose prior to administration. The terms “co-administration” or “combined administration” or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g., a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time. The term “pharmaceutical combination” as used herein means a product that results from the mixing or combining of more than one therapeutic agent and includes both fixed and non-fixed combinations of the therapeutic agents. The term “fixed combination” means that the therapeutic agents, e.g., a compound of the present disclosure and a combination partner, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non- fixed combination” means that the therapeutic agents, e.g., a compound of the present disclosure and a combination partner, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g., the administration of three or more therapeutic agents.

[0073] When a variable appears more than once in a group, e.g., R1in -C(R1)2-, or a variable appears more than once in a structure presented herein, the variables can be the same or different at each occurrence.

[0074] A solid line , as a bond generally indicates a mixture of, or either of, the possible isomers, e.g., containing (R)- and (S)-stereochemistry. For example: .

[0075] Systems, such as, for indicates that the indicated line (bond) may be attached to any of the substitutable26014

[0076] As well known in the art, a bond drawn from a particular atom wherein no moiety is depicted at the terminal end of the bond indicates a methyl group bound through that bond to the atom, unless stated otherwise. For example:compounds useful in the methods of the disclosure, and / or compositions comprising them useful in said methods, are present in isolated and / or purified form. The term "purified", "in purified form" or “in isolated and purified form” for a compound refers to the physical state of said compound after being isolated from a synthetic process (e.g., from a reaction mixture), or natural source or combination thereof. Thus, the term "purified", "in purified form" or “in isolated and purified form” for a compound refers to the physical state of said compound (or a tautomer or stereoisomer thereof, or pharmaceutically acceptable salt or solvate of said compound, said stereoisomer, or said tautomer) after being obtained from a purification process or processes described herein or well known to the skilled artisan (e.g., chromatography, recrystallization and the like), in sufficient purity to be suitable for in vivo or medicinal use and / or characterizable by standard analytical techniques described herein or well known to the skilled artisan.

[0078] When a functional group in a compound is termed “protected”, this means that the group is in modified form to preclude undesired side reactions at the protected site when the compound is subjected to a reaction. Suitable protecting groups will be recognized by those with ordinary skill in the art as well as by reference to standard textbooks such as, for example, T. W. Greene et al., Protective Groups in Organic Synthesis (1991), Wiley, New York.

[0079] Another embodiment provides prodrugs and / or solvates of the compounds of the disclosure. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press. The term “prodrug” means a compound (e.g., a drug precursor) that is transformed in vivo to yield a compound of the disclosure or a pharmaceutically acceptable salt, hydrate or solvate of the compound. The transformation may occur by various mechanisms (e.g., by metabolic or chemical processes), such as, for example, through hydrolysis in blood. A discussion of the use of prodrugs is provided by T. Higuchi and W. Stella, “Pro-drugs as Novel26014 Delivery Systems,” Vol.14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

[0080] One or more compounds used in the methods of the disclosure may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the disclosure embrace both solvated and unsolvated forms. "Solvate" means a physical association of a compound of the disclosure with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances, the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate" encompasses both solution-phase and isolatable solvates. Non- limiting examples of suitable solvates include ethanolates, methanolates, and the like. "Hydrate" is a solvate wherein the solvent molecule is H2O.

[0081] One or more compounds used in the methods of the disclosure may optionally be converted to a solvate. Preparation of solvates is generally known. Thus, for example M. Caira et al., J. Pharmaceutical Sci., 1993, 3, 601-611, describe the preparation of the solvates of the antifungal fluconazole in ethyl acetate as well as from water. Similar preparations of solvates, hemisolvate, hydrates and the like are described by E. C. van Tonder et al., AAPS PharmSciTech., 5(1), article 12 (2004); and A. L. Bingham et al., Chem. Commun., 603-604 (2001). A typical, non-limiting, process involves dissolving the inventive compound in desired amounts of the desired solvent (organic or water or mixtures thereof) at a higher than ambient temperature and cooling the solution at a rate sufficient to form crystals which are then isolated by standard methods. Analytical techniques such as, for example I. R. spectroscopy, show the presence of the solvent (or water) in the crystals as a solvate (or hydrate).

[0082] Another embodiment provides pharmaceutically acceptable salts of the compounds to be used in the methods of the disclosure. Thus, reference to a compound used in the methods of the disclosure herein is understood to include reference to salts thereof, unless otherwise indicated. The term "salt(s)", as employed herein, denotes acidic salts formed with inorganic and / or organic acids, as well as basic salts formed with inorganic and / or organic bases. In addition, when a compound of the disclosure contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions ("inner salts") may be formed and are included within the term "salt(s)" as used herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful. Salts of the compounds used in the methods of the disclosure may be formed, for26014 example, by reacting a compound of the disclosure with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.

[0083] Exemplary acid addition salts include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonates (also known as tosylates,) and the like.

[0084] Additionally, acids which are generally considered suitable for the formation of pharmaceutically useful salts from basic pharmaceutical compounds are discussed, for example, by P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food & Drug Administration, Washington, D.C. on their website). These disclosures are incorporated herein by reference thereto.

[0085] Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamines, t-butyl amines, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups may be quarternized with agents such as lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfates), long chain halides (e.g., decyl, lauryl, and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.

[0086] All such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the disclosure and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the disclosure.

[0087] Another embodiment provides pharmaceutically acceptable esters of the compounds used in the methods of the disclosure. Such esters include the following groups: (1) carboxylic acid esters obtained by esterification of the hydroxy groups, in which the non-carbonyl moiety of the carboxylic acid portion of the ester grouping is selected from straight or branched chain alkyl (for example, acetyl, n-propyl, t-butyl, or n-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (for example, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (for example, phenyl optionally substituted with, for example, halogen, C1-4alkyl, or C1-4alkoxy or amino);26014 (2) sulfonate esters, such as alkyl- or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters (for example, L-valyl or L-isoleucyl); (4) phosphonate esters and (5) mono-, di- or triphosphate esters. The phosphate esters may be further esterified by, for example, a C1-20 alcohol or reactive derivative thereof, or by a 2,3-di (C6-24)acyl glycerol.

[0088] As mentioned herein, another embodiment provides tautomers of the compounds of the disclosure to be used in the methods herein, and salts, solvates, esters and prodrugs of said tautomers. It shall be understood that all tautomeric forms of such compounds are within the scope of the compounds used in the methods of the disclosure. For example, all keto-enol and imine-enamine forms of the compounds, when present, are included in the disclosure.

[0089] The compounds used in the methods of the disclosure may contain asymmetric or chiral centers, and, therefore, exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds used in the methods of the disclosure as well as mixtures thereof, including racemic mixtures, form part of the present disclosure. In addition, the present disclosure embraces use of all geometric and positional isomers. For example, if a compound used in the methods of the disclosure incorporates a double bond or a fused ring, both the cis- and trans-forms, (E) and (Z) forms, as well as mixtures, are embraced within the scope of the disclosure.

[0090] Another embodiment provides for diastereomeric mixtures and individual enantiomers of the compounds used in the methods of the disclosure. Diastereomeric mixtures can be separated into their individual diastereomers based on their physical chemical differences by methods well known to those skilled in the art, such as, for example, by chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Also, some of the compounds used in the methods of the disclosure may be atropisomers (e.g., substituted biaryls) and are considered as part of this disclosure. Enantiomers can also be separated by use of chiral HPLC column.

[0091] All stereoisomers (for example, geometric isomers, optical isomers and the like) of the compounds used in the methods of the disclosure (including those of the salts, solvates, esters and prodrugs of the compounds as well as the salts, solvates and esters of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated as embodiments within the scope of26014 this disclosure, as are positional isomers (such as, for example, 4-pyridyl and 3-pyridyl). (For example, if a compound of the disclosure incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the disclosure. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the methods of the disclosure).

[0092] Individual stereoisomers of the compounds of the disclosure may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present disclosure can have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms "salt", "solvate", “ester”, "prodrug" and the like, is intended to equally apply to the salt, solvate, ester and prodrug of enantiomers, stereoisomers, rotamers, tautomers, positional isomers, racemates or prodrugs of the inventive compounds.

[0093] In the compounds used in the methods of the disclosure, the atoms may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present disclosure is meant to include all suitable isotopic variations of the compounds of the disclosure. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium (2H). The presence of deuterium in the compounds of the disclosure is indicated by "D". Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples. Isotopically-enriched compounds of the disclosure can be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the schemes and examples herein using appropriate isotopically-enriched reagents and / or intermediates. Such isotopic compounds are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as2H,3H,13C,14C,15N,18O,17O,31P,32P,35S,18F, and36Cl, respectively.

[0094] In another embodiment, the compounds useful in the methods of the disclosure, and / or compositions comprising them useful in said methods, are present in isolated and / or purified form. The term "purified", "in purified form" or “in isolated and purified form” for a compound26014 refers to the physical state of said compound after being isolated from a synthetic process (e.g., from a reaction mixture), or natural source or combination thereof. Thus, the term "purified", "in purified form" or “in isolated and purified form” for a compound refers to the physical state of said compound (or a tautomer or stereoisomer thereof, or pharmaceutically acceptable salt or solvate of said compound, said stereoisomer, or said tautomer) after being obtained from a purification process or processes described herein or well known to the skilled artisan (e.g., chromatography, recrystallization and the like), in sufficient purity to be suitable for in vivo or medicinal use and / or characterizable by standard analytical techniques described herein or well known to the skilled artisan.

[0095] It shall be understood that any carbon as well as heteroatom with unsatisfied valences in the text, schemes, examples and tables herein is assumed to have the sufficient number of hydrogen atom(s) to satisfy the valences.

[0096] Compounds described herein may be administered to a patient orally or parenterally. As formulated into a dosage form suitable for administration, the compounds described herein can be used as a pharmaceutical composition for the prevention, treatment, or remedy of the above diseases.

[0097] In clinical use of the compounds described herein, usually, the compound is formulated into various preparations together with pharmaceutically acceptable additives according to the dosage form, and may then be administered. By "pharmaceutically acceptable" it is meant the additive, carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. As such, various additives ordinarily used in the field of pharmaceutical preparations are usable. Specific examples thereof include gelatin, lactose, sucrose, titanium oxide, starch, crystalline cellulose, hydroxypropyl methylcellulose, carboxymethylcellulose, corn starch, microcrystalline wax, white petrolatum, magnesium metasilicate aluminate, anhydrous calcium phosphate, citric acid, trisodium citrate, hydroxypropylcellulose, sorbitol, sorbitan fatty acid ester, polysorbate, sucrose fatty acid ester, polyoxyethylene, hardened castor oil, polyvinylpyrrolidone, magnesium stearate, light silicic acid anhydride, talc, vegetable oil, benzyl alcohol, gum arabic, propylene glycol, polyalkylene glycol, cyclodextrin, hydroxypropyl cyclodextrin, and the like.

[0098] Preparations to be formed with those additives include, for example, solid preparations such as tablets, capsules, granules, powders and suppositories; and liquid preparations such as syrups, elixirs and injections. These may be formulated according to conventional methods known in the field of pharmaceutical preparations. The liquid preparations may also be in such a form that may be dissolved or suspended in water or in any other suitable medium in their use.26014

[0099] Especially for injections, if desired, the preparations may be dissolved or suspended in physiological saline or glucose liquid, and a buffer or a preservative may be optionally added thereto.

[0100] The pharmaceutical compositions may contain the compound of the disclosure in an amount of from 1 to 99.9 % by weight, preferably from 1 to 60 % by weight of the composition. The compositions may further contain any other therapeutically-effective compounds.

[0101] In case where the compounds of the disclosure are used for prevention or treatment for the above-mentioned diseases, the dose and the dosing frequency may be varied, depending on the sex, the age, the body weight and the disease condition of the patient and on the type and the range of the intended remedial effect. In general, when orally administered, the dose may be from 0.001 to 50 mg / kg of body weight / day, and it may be administered at a time or in several times. In specific embodiments, the dose is from about 0.01 to about 25 mg / kg / day, in particular embodiments, from about 0.05 to about 10 mg / kg / day, or from about 0.001 to about 50 mg / kg / day. For oral administration, the compositions are preferably provided in the form of tablets or capsules containing from 0.01 mg to 1,000 mg. In specific embodiments, the dose is 0.01, 0.05, 0.1, 0.2, 0.5, 1.0, 2.5, 5, 10, 15, 20, 25, 30, 40, 50, 75, 100, 125, 150, 175, 200, 225, 250, 500, 750, 850 or 1,000 milligrams of a compound described herein. This dosage regimen may be adjusted to provide the optimal therapeutic response.

[0102] The compounds of the disclosure may be used in combination with one or more other drugs in the treatment, prevention, suppression or amelioration of diseases or conditions for which compounds described herein or the other drugs may have utility, where the combination of the drugs together are safer or more effective than either drug alone. Such other drug(s) may be administered in an amount commonly used therefore, contemporaneously or sequentially with a compound described herein or a pharmaceutically acceptable salt thereof. When a compound described herein is used contemporaneously with one or more other drugs, the pharmaceutical composition may in specific embodiments contain such other drugs and the compound described herein or its pharmaceutically acceptable salt in unit dosage form. However, the combination therapy may also include therapies in which the compound described herein, or its pharmaceutically acceptable salt and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the disclosure and the other active ingredients may be used in lower doses than when each is used singly. Accordingly, the pharmaceutical compositions of the disclosure include those that contain one or more other active ingredients, in addition to a compound described herein or a pharmaceutically acceptable salt thereof.26014

[0103] Examples of other active ingredients that may be administered in combination with a compound of any of the Formulas described herein or a pharmaceutically acceptable salt thereof and either administered separately or in the same pharmaceutical composition, include, but are not limited to pain relieving agents, anti-angiogenic agents, anti-neoplastic agents, anti-diabetic agents, anti-infective agents, or gastrointestinal agents, or combinations thereof.

[0104] Suitable compounds that may be used in combination with a compound according to the disclosure include without limitation sildenafil, vardenafil, tadalafil and alprostadil, epoprostenol, iloprost, bosentan, amlodipine, diltiazem, nifedipine, ambrisentan and warfarin, fluticasone, budesonide, mometasone, flunisolide, beclomethasone, montelukast, zafirlukast, zileuton, salmeterol, formoterol, theophylline, albuterol, levalbuterol, pirbuterol, ipratropium, prednisone, methylprednisolone, omalizumab, corticosteroid and cromolyn, atorvastatin, lovastatin, simvastatin, pravastatin, fluvastatin, rosuvastatin, gemfibrozil, fenofibrate, nicotinic acid, clopidogrel and pharmaceutically acceptable salts thereof.

[0105] Accordingly, the combinations described herein can include compounds of any of the Formulas herein and one or more additional therapeutic agents, e.g., one or more anti-cancer agents, cytotoxic or cytostatic agents, hormone treatment, vaccines, and / or other immunotherapies. In other embodiments, the combination is further administered or used in combination with other therapeutic treatment modalities, including surgery, radiation, cryosurgery, and / or thermotherapy. Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thus avoiding possible toxicities or complications associated with the treatment.

[0106] Additionally, a compound of any of the Formulas disclosed herein may be used in combination with one or more other active agents, including but not limited to, other anti-cancer agents that are used in the prevention, treatment, control, amelioration, or reduction of risk of a particular disease or condition (e.g., cell proliferation disorders). In one embodiment, a compound disclosed herein is combined with one or more other anti-cancer agents for use in the prevention, treatment, control amelioration, or reduction of risk of a particular disease or condition for which the compounds disclosed herein are useful. Such other active agents may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of the disclosure.

[0107] In one embodiment, the other active agent is selected from the group consisting of vascular endothelial growth factor (VEGF) receptor inhibitors, topoisomerase II inhibitors, smoothen inhibitors, alkylating agents, anti-tumor antibiotics, anti-metabolites, retinoids,26014 immunomodulatory agents including but not limited to anti-cancer vaccines, CTLA-4, LAG-3 and PD-1 antagonists.

[0108] The additional therapeutic agent also can be, for example, chemotherapy, a chemical compound such as an anti-cancer agent, peptide, antibody, antibody fragment or nucleic acid, which is therapeutically active or enhances the therapeutic activity when administered to a patient in combination with a compound of the present disclosure.

[0109] Additional therapeutic agents such PD-1 antagonists that are useful in any of the treatment methods, medicaments and uses of the disclosure include a monoclonal antibody (mAb), or antigen binding fragment thereof, which specifically binds to PD-1 or PD-Ll, and preferably specifically binds to human PD-1 or human PD-Ll. The mAb may be a human antibody, a humanized antibody or a chimeric antibody, and may include a human constant region. In some embodiments the human constant region is selected from the group consisting of IgGl, IgG2, IgG3 and IgG4 constant regions, and in some embodiments, the human constant region is an IgGl or IgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab'-SH, F(ab')2, scFv and Fv fragments. Examples of PD-1 antagonists include, but are not limited to, pembrolizumab (KEYTRUDA®, Merck and Co., Inc., Rahway, NJ, USA). “Pembrolizumab” (formerly known as MK-3475, SCH 900475 and lambrolizumab and sometimes referred to as “pembro”) is a humanized IgG4 mAb with the structure described in WHO Drug Information, Vol.27, No.2, pages 161-162 (2013). Additional examples of PD-1 antagonists include nivolumab (OPDIVO®, Bristol-Myers Squibb Company, Princeton, NJ, USA), atezolizumab (MPDL3280A; TECENTRIQ®, Genentech, San Francisco, CA, USA), durvalumab (IMFINZI®, Astra Zeneca Pharmaceuticals, LP, Wilmington, DE), avelumab (BAVENCIO®, Merck KGaA, Darmstadt, Germany and Pfizer, Inc., New York, NY), cemiplimab (LIBTAYO®, Regeneron Pharmaceuticals, Inc., Tarrytown, NY, and Sanofi-Aventis LLC, Bridgewater, NJ, U.S.), and dostarlimab (JEMPERLI®, GlaxoSmithKline LLC, Philadelphia, PA).

[0110] Examples of monoclonal antibodies (mAbs) that bind to human PD-1, and are useful in the treatment methods, medicaments and uses of the disclosure, are described in US7488802, US7521051, US8008449, US8354509, US8168757, WO2004 / 004771, WO2004 / 072286, WO2004 / 056875, and US2011 / 0271358.

[0111] Examples of mAbs that bind to human PD-Ll, and are useful in the treatment methods, medicaments and uses of the disclosure, are described in WO2013 / 019906, W02010 / 077634 Al and US8383796. Specific anti-human PD-Ll mAbs useful as the PD-1 antagonist in the treatment methods, medicaments and uses of the disclosure include MPDL3280A, BMS-936559,MEDI4736, MSB0010718C and an antibody which comprises the heavy chain and light chain variable regions of SEQ ID NO:24 and SEQ ID NO:21, respectively, of WO2013 / 019906.

[0112] Other PD-1 antagonists useful in any of the treatment methods, medicaments and uses of the disclosure include an immunoadhesin that specifically binds to PD-1 or PD- L1, and preferably specifically binds to human PD-1 or human PD-Ll, e.g., a fusion protein containing the extracellular or PD-1 binding portion of PD-Ll or PD-L2 fused to a constant region such as an Fc region of an immunoglobulin molecule. Examples of immunoadhesin molecules that specifically bind to PD-1 are described in WO2010 / 027827 and WO2011 / 066342. Specific fusion proteins useful as the PD-1 antagonist in the treatment methods, medicaments and uses of the disclosure include AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein that binds to human PD-1.

[0113] Thus, one embodiment provides a method of treating cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, in combination with a PD-1 antagonist to a subject in need thereof. In such embodiments, the compounds of the disclosure, or a pharmaceutically acceptable salt thereof, and the PD-1 antagonist are administered concurrently or sequentially.

[0114] Specific non-limiting examples of such cancers in accordance with this embodiment include melanoma (including unresectable or metastatic melanoma), head & neck cancer (including recurrent or metastatic head and neck squamous cell cancer (HNSCC)), classical Hodgkin lymphoma (cHL), urothelial carcinoma, gastric cancer, cervical cancer, primary mediastinal large-B-cell lymphoma, microsatellite instability-high (MSI-H) cancer, non-small cell lung cancer, hepatocellular carcinoma, clear cell kidney cancer, colorectal cancer, breast cancer, squamous cell lung cancer, basal carcinoma, sarcoma, bladder cancer, endometrial cancer, pancreatic cancer, liver cancer, gastrointestinal cancer, multiple myeloma, renal cancer, mesothelioma, ovarian cancer, anal cancer, biliary tract cancer, esophageal cancer, and salivary cancer.

[0115] In one embodiment, there is provided a method of treating cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist, wherein said cancer is selected from unresectable or metastatic melanoma, melanoma following complete resection, recurrent, metastatic, or unresectable head and neck squamous cell cancer (HNSCC), classical Hodgkin lymphoma (cHL), urothelial carcinoma, gastric cancer, Merkel cell carcinoma, renal cell carcinoma, endometrial carcinoma, tumor mutational burden-high (TMB- H) cancer, cervical cancer, primary mediastinal large-B-cell lymphoma, microsatellite instability-high (MSI-H) or mismatch repair deficient cancer, non-small cell lung cancer, esophageal cancer, cutaneous squamous cell carcinoma, triple negative breast cancer, and hepatocellular carcinoma. In one such embodiment, the agent is a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiments, the agent is durvalumab or avelumab. In one embodiment, the agent is cemiplimab. In one embodiment, the agent is dostarlimab.

[0116] Pembrolizumab is approved by the U.S. FDA for the treatment of patients with unresectable or metastatic melanoma and for the adjuvant treatment of melanoma following complete resection, and for the treatment of certain patients with recurrent, metastatic, or unresectable head and neck squamous cell cancer (HNSCC), classical Hodgkin lymphoma (cHL), urothelial carcinoma, gastric cancer, Merkel cell carcinoma, renal cell cancer, endometrial cancer, tumor mutational burden-high (TMB-H) cancer, cervical cancer, primary mediastinal large-B-cell lymphoma, microsatellite instability-high (MSI-H) or mismatch repair deficient cancer, non-small cell lung cancer, esophageal cancer, cutaneous squamous cell carcinoma, triple negative breast cancer, and hepatocellular carcinoma, as described in the Prescribing Information for KEYTRUDA™ (Merck & Co., Inc., Rahway, NJ USA; initial U.S. approval 2014, updated January 2023). In another embodiment, there is provided a method of treating cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, in combination with pembrolizumab to a person in need thereof, wherein said cancer is selected from unresectable or metastatic melanoma, adjuvant melanoma, recurrent, unresectable or metastatic head and neck squamous cell cancer (HNSCC), classical Hodgkin lymphoma (cHL), urothelial carcinoma, gastric cancer, Merkel cell carcinoma, renal cell cancer, endometrial cancer, tumor mutational burden-high (TMB-H) cancer, cervical cancer, primary mediastinal large-B-cell lymphoma, microsatellite instability-high (MSI-H) or mismatch repair deficient cancer, non-small cell lung cancer, esophageal cancer, cutaneous squamous cell carcinoma, triple negative breast cancer, and hepatocellular carcinoma.

[0117] In another embodiment, there is provided a method of treating cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, in combination with a PD-1 antagonist to a person in need thereof, wherein said cancer is selected from melanoma, non-small cell lung cancer, head and neck squamous cell cancer (HNSCC), Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, urothelial carcinoma, microsatellite instability-high cancer, gastric cancer, Merkel cell carcinoma, hepatocellular carcinoma, esophageal cancer and cervical cancer. In one such26014 embodiment, the agent is a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In another such embodiment, the agent is durvalumab. In another such embodiment, the agent is avelumab. In other such embodiment, the agent is durvalumab or avelumab.

[0118] In another embodiment, there is provided a method of treating cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist, wherein said cancer is selected from melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, bladder cancer, breast cancer, gastrointestinal cancer, multiple myeloma, hepatocellular cancer, lymphoma, renal cancer, mesothelioma, ovarian cancer, esophageal cancer, anal cancer, biliary tract cancer, colorectal cancer, cervical cancer, thyroid cancer, and salivary cancer. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In another such embodiment, the agent is durvalumab. In another such embodiment, the agent is avelumab. In other such embodiment, the agent is durvalumab or avelumab.

[0119] In one embodiment, there is provided a method of treating unresectable or metastatic melanoma comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0120] In one embodiment, there is provided a method of treating recurrent or metastatic head and neck squamous cell cancer (HNSCC) comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0121] In one embodiment, there is provided a method of treating classical Hodgkin lymphoma (cHL) comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment,26014 the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0122] In one embodiment, there is provided a method of treating urothelial carcinoma comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0123] In one embodiment, there is provided a method of treating gastric cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0124] In one embodiment, there is provided a method of treating cervical cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0125] In one embodiment, there is provided a method of treating primary mediastinal large-B- cell lymphoma comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD- 1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0126] In one embodiment, there is provided a method of treating microsatellite instability-high (MSI-H) cancer comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0127] In one embodiment, there is provided a method of treating non-small cell lung cancer comprising administering an effective amount of a compound of the disclosure, or a26014 pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0128] In one embodiment, there is provided a method of treating hepatocellular carcinoma comprising administering an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, to a person in need thereof, in combination with a PD-1 antagonist. In one such embodiment, the agent is pembrolizumab. In another such embodiment, the agent is nivolumab. In another such embodiment, the agent is atezolizumab. In other such embodiment, the agent is durvalumab or avelumab.

[0129] In another embodiment, there is provided a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a chemotherapy, and a PD-1 inhibitor. In particular, the chemotherapy and PD-1 inhibitor are selected from those described above. More particularly, the chemotherapy is irinotecan (Camptosar®) and the PD-1 inhibitor is pembrolizumab.

[0130] Examples of vascular endothelial growth factor (VEGF) receptor inhibitors include, but are not limited to, bevacizumab (sold under the trademark AVASTIN by Genentech / Roche), axitinib, (N-methyl-2-[[3-[([pound])-2-pyridin-2-ylethenyl]-l H-indazol-6-yl]sulfanyl]benzamide, also known as AG013736, and described in PCT Publication No. WO01 / 002369), Brivanib Alaninate ((S)-((R)-l-(4-(4-Fluoro-2-methyl-lH-indol-5-yloxy)-5-methylpyrrolo[2,l- f][l,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate, also known as BMS-582664), motesanib (N-(2,3-dihydro-3,3-dimethyl-l H-indoi-6-yl)-2-[(4-pyridinyimethy)amino]-3- pyridinecarboxamide. and described in PCT Publication No. WO 02 / 068470), pasireotide (also known as SO 230, and described in PCT Publication No. WO02 / 010192), and sorafenib (sold under the tradename NEXAVAR).

[0131] Examples of topoisomerase II inhibitors include but are not limited to, etoposide (also known as VP-16 and Etoposide phosphate, sold under the tradenames TOPOSAR, VEPESID and ETOPOPHOS), and teniposide (also known as VM-26, sold under the tradename VUMON).

[0132] Examples of alkylating agents include but are not limited to, 5-azacytidine (sold under the trade name VIDAZA), decitabine (sold under the trade name of DECOGEN), temozolomide (sold under the trade names TEMODAR and TEMODAL by Schering-Plough / Merck), dactinomycin (also known as actinomycin-D and sold under the tradename COSMEGEN), melphalan (also known as L-PAM, L-sarcolysin, and phenylalanine mustard, sold under the tradename ALKERAN), altretamine (also known as hexamethylmelamine (HMM), sold under26014 the tradename HEXALEN), carmustine (sold under the tradename BCNU), bendamustine (sold under the tradename TREANDA), busulfan (sold under the tradenames BUSULFEX and MYLERAN), carboplatin (sold under the tradename PARAPLATIN), lomustine (also known as CCNU, sold under the tradename CeeNU), cisplatin (also known as CDDP, sold under the tradenames PLATINOL and PLATINOL-AQ), chlorambucil (sold under the tradename LEUKERAN), cyclophosphamide (sold under the tradenames CYTOXAN and NEOSAR), dacarbazine (also known as DTIC, DIC and imidazole carboxamide, sold under the tradename DTIC-DOME), altretamine (also known as hexamethylmelamine (HMM) sold under the tradename HEXALEN), ifosfamide (sold under the tradename IFEX), procarbazine (sold under the tradename MATULANE), mechlorethamine (also known as nitrogen mustard, mustine and mechloroethamine hydrochloride, sold under the tradename MUSTARGEN), streptozocin (sold under the tradename ZANOSAR), thiotepa (also known as thiophosphoamide, TESPA and TSPA, and sold under the tradename THIOPLEX).

[0133] Examples of anti-tumor antibiotics include, but are not limited to, doxorubicin (sold under the tradenames ADRIAMYCIN and RUB EX), bleomycin (sold under the tradename LENOXANE), daunorubicin (also known as dauorubicin hydrochloride, daunomycin, and rubidomycin hydrochloride, sold under the tradename CERUBIDINE), daunorubicin liposomal (daunorubicin citrate liposome, sold under the tradename DAUNOXOME), mitoxantrone (also known as DHAD, sold under the tradename NOVANTRONE), epirubicin (sold under the tradename ELLENCE), idarubicin (sold under the tradenames IDAMYCIN, IDAMYCIN PFS), and mitomycin C (sold under the tradename MUTAMYCIN).

[0134] Examples of anti-metabolites include, but are not limited to, claribine (2- chlorodeoxyadenosine, sold under the tradename LEUSTATIN), 5-fluorouracil (sold under the tradename ADRUCIL), 6-thioguanine (sold under the tradename PURINETHOL), pemetrexed (sold under the tradename ALIMTA), cytarabine (also known as arabinosylcytosine (Ara-C), sold under the tradename CYTOSAR-U), cytarabine liposomal (also known as Liposomal Ara-C, sold under the tradename DEPOCYT), decitabine (sold under the tradename DACOGEN), hydroxyurea (sold under the tradenames HYDREA, DROXIA and MYLOCEL), fludarabine (sold under the tradename FLUDARA), floxuridine (sold under the tradename FUDR), cladribine (also known as 2-chlorodeoxyadenosine (2-CdA) sold under the tradename LEUSTATIN), methotrexate (also known as amethopterin, methotrexate sodium (MTX), sold under the tradenames RHEUMATREX and TREXALL), and pentostatin (sold under the tradename NIPENT).26014

[0135] Examples of retinoids include, but are not limited to, alitretinoin (sold under the tradename PANRETIN), tretinoin (all-trans retinoic acid, also known as ATRA, sold under the tradename VESANOID), Isotretinoin (13-c / s-retinoic acid, sold under the tradenames ACCUTANE, AMNESTEEM, CLARAVIS, CLARUS, DECUTAN, ISOTANE, IZOTECH, ORATANE, ISOTRET, and SOTRET), and bexarotene (sold under the tradename TARGRETIN).

[0136] In such combinations a compound of the invention and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).

[0137] In certain embodiments of the methods, kits and uses provided herein, the anti-human compounds of Formula I and the anti-human PD-1 monoclonal antibody are administered on the same day. In some embodiments, the compounds of Formula I and the anti-human PD-1 monoclonal antibody are administered sequentially. In some embodiments, the compounds of Formula I and the anti-human PD-1 monoclonal antibody are administered concurrently. In some embodiments, the compounds of Formula I and the anti-human PD-1 monoclonal antibody are co-formulated.

[0138] In one embodiment, the disclosure provides a product comprising a compound of the present disclosure and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is the treatment of a disease or condition mediated by WRN. Products provided as a combined preparation include a composition comprising the compound of formula (I) and the other therapeutic agent(s) together in the same pharmaceutical composition, or the compound of the present disclosure and the other therapeutic agent(s) in separate form, e.g., in the form of a kit.

[0139] In one embodiment, the disclosure provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of the present disclosure. In one embodiment, the kit comprises means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet. An example of such a kit is a blister pack, as typically used for the packaging of tablets, capsules and the like.

[0140] The kit of the disclosure may be used for administering different dosage forms, for example, oral and parenteral, for administering the separate compositions at different dosage intervals, or for titrating the separate compositions against one another. To assist compliance, the kit of the disclosure typically comprises directions for administration.

[0141] In the combination therapies of the disclosure, the compound of the present disclosure and the other therapeutic agent may be manufactured and / or formulated by the same or different26014 manufacturers. Moreover, the compound of the present disclosure and the other therapeutic may be brought together into a combination therapy: (i) prior to release of the combination product to physicians (e.g., in the case of a kit comprising the compound of the present disclosure and the other therapeutic agent); (ii) by the physician themselves (or under the guidance of the physician) shortly before administration; (II) in the patient themselves, e.g., during sequential administration of the compound of the present disclosure and the other therapeutic agent.

[0142] Accordingly, the disclosure provides the use of a compound of the present disclosure for treating a disease or condition mediated by WRN, wherein the medicament is prepared for administration with another therapeutic agent. The disclosure also provides the use of another therapeutic agent for treating a disease or condition mediated by WRN, wherein the medicament is administered with a compound of the present disclosure.

[0143] The disclosure also provides a compound of the present disclosure for use in treating a disease or condition mediated by WRN, wherein the compound of the present disclosure is prepared for administration with another therapeutic agent. The disclosure also provides another therapeutic agent for use in treating a disease or condition mediated by WRN, wherein the other therapeutic agent is prepared for administration with a compound of the present disclosure. The disclosure also provides a compound of the present disclosure for use in treating a disease or condition mediated by WRN, wherein the compound of the present disclosure is administered with another therapeutic agent. The disclosure also provides another therapeutic agent for use in a method of treating a disease or condition mediated by WRN, wherein the other therapeutic agent is administered with a compound of the present disclosure.

[0144] The disclosure also provides the use of a compound of the present disclosure for treating a disease or condition mediated by WRN, wherein the patient has previously (e.g., within 24 hours) been treated with another therapeutic agent. The disclosure also provides the use of another therapeutic agent for treating a disease or condition mediated by WRN, wherein the patient has previously (e.g., within 24 hours) been treated with compound of the present disclosure.

[0145] The following examples are intended to illustrate the disclosure and are not to be construed as being limitations thereon. Temperatures are given in degrees Celsius. If not mentioned otherwise, all evaporations are performed under reduced pressure, typically between about 15 mm Hg and 100 mm Hg (=20-133 mbar). Abbreviations used are those conventional in the art.

[0146] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts utilized to synthesize the compounds of the present disclosure are either26014 commercially available or can be produced by organic synthesis methods known to one of ordinary skill in the art. Further, the compounds of the present disclosure can be produced by organic synthesis methods known to one of ordinary skill in the art as shown in the following examples.

[0147] The structures of all final products, intermediates and starting materials are confirmed by standard analytical spectroscopic characteristics, e.g., MS, IR or NMR. The absolute stereochemistry of certain isomers has been determined by analyses of X-ray crystal structures of complexes in which the respective compounds are bound to WRN or by small molecule X-ray crystal structures of a precursor of the final compound. The meanings of the abbreviations in Examples are shown below. ADP Adenosine diphosphate ATP Adenosine triphosphate 3-26014 MgSO4 Magnesium sulfate MS (EI) Mass spectroscopy electron ionization data MSSe mean ngs o e abbreva ons n e nuc ear magne c resonance spec ra are s own be ow: s = singlet, d = doublet, dd = double doublet, dt = double triplet, ddd = double double doublet, Sept = septet, t = triplet, m = multiplet, br = broad, brs = broad singlet, q = quartet J = coupling constant and Hz = hertz.

[0148] Compounds of this invention can be prepared using the intermediates and processes outlined below. The following exemplified compounds of the invention may contain one or more asymmetric centers and can thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. The invention is meant to comprehend all such isomeric forms of these compounds. In the examples, where a compound has one or more stereocenters, the stereocenters are indicated with an asterisk, as shown below:

[0149] Typically, the compounds of Formula (I) can be prepared according to the Schemes provided infra. The examples which outline specific synthetic routes, and the generic schemes below provide guidance to the synthetic chemist of ordinary skill in the art, who will readily appreciate that the solvent, concentration, reagent, protecting group, order of synthetic steps, time, temperature, and the like can be modified as necessary.26014

[0150] The examples are meant to be illustrative and should not be construed as further limiting. The contents of the figures and all references, patents, and published patent applications cited throughout this application are expressly incorporated herein by reference. EXAMPLES

[0151] The WRN inhibitors described in this invention were prepared in accordance with the non-limiting examples below. Example 1 2-(2-(phenylamino)-5-((4-(trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetic acid (1) Step a– synthesis of ethyl 4-(2-ethoxy-2-oxoethyl)-2-(phenylamino)thiazole-5-carboxylate (8) To ag, 9.31 mmol), Cs2CO3(6.07 g, 18.62 mmol), Xantphos (0.718 g, 1.24 mmol) and Pd2(dba)3(0.568 g, 0.621 mmol) under N2, and the mixture was stirred at 100 °C for 16 h. The reaction mixture was treated with water (50 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by reverse phase HPLC to afford ethyl 4-(2-ethoxy-2-oxoethyl)-2- (phenylamino)thiazole-5-carboxylate (8). MS (ESI): m / z (M+H)+335.0. Step b – synthesis of 4-(carboxymethyl)-2-(phenylamino)thiazole-5-carboxylic acid (9) To a(8) (0.40 g, 1.12 mmol) in a mixture of MeOH (5 mL) and water (0.25 mL) was added LiOH (0.086 g, 3.59 mmol). The reaction mixture was stirred at 25 °C for 3 h, and then was neutralized and purified by reverse phase HPLC to give 4-(carboxymethyl)-2-(phenylamino)thiazole-5-carboxylic acid (9). MS (ESI): m / z (M+H)+278.9.26014 Step c – Synthesis of 4-(2-methoxy-2-oxoethyl)-2-(phenylamino)thiazole-5-carboxylic acid (10) A mixture (0.20 g, 0.719mmol) and conc. was °C for 8 h. The mixture was treated with 20 mL of water and extracted with EtOAc (15 mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give crude 4-(2-methoxy-2-oxoethyl)-2- (phenylamino)thiazole-5-carboxylic acid (10), which was used in the next step directly without further purification. MS (ESI): m / z (M+H)+293.0. Step d- Synthesis of methyl 2-(2-(phenylamino)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (12) F3C F3C10 12 To a solution of 4-(2-methoxy-2-oxoethyl)-2-(phenylamino)thiazole-5-carboxylic acid (10) (0.095 g, 0.33 mmol) in DCM (1.5 mL) was added 4-(trifluoromethyl)aniline 11 (0.052 g, 0.33 mmol), Pyridine (0.04 mL, 0.49 mmol) and POCl3(0.036 mL, 0.390 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. Then the solvent was removed in vacuum. The residue was treated with water (10 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by reversed phase HPLC to give methyl 2-(2-(phenylamino)-5- ((4-(trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (12). MS (ESI): m / z (M+H)+436.0.1H NMR (CDCl3, 400 MHz) δ 10.02 (s, 1H), 7.77 (d, J = 8.5 Hz, 2H), 7.61 (d, J = 8.6 Hz, 2H), 7.40-7.47 (m, 2H), 7.34 (d, J = 7.6 Hz, 2H), 7.18-7.25 (m, 1H), 3.99 (s, 2H), 3.90 (s, 3H).26014 Step e – Synthesis of 2-(2-(phenylamino)-5-((4-(trifluoromethyl)phenyl)carbamoyl)thiazol- 4-yl)acetic acid (3) To a thiazol-4-yl) g, was mmol). The reaction mixture wasstirred at 25 °C for 16 h, and then was purified by reverse phase HPLC to give 2-(2-(phenylamino)-5-((4-(trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetic acid. MS (ESI): m / z (M+H)+422.0.1H (DMSO-d6, 400 MHz) δ 10.70 (s, 1H), 10.15 (s, 1H), 7.87 (d, J = 8.6 Hz, 2H), 7.69 (d, J = 8.6 Hz, 2H), 7.60 (d, J = 7.9 Hz, 2H), 7.37 (t, J = 8.0 Hz, 2H), 7.05 (t, J = 7.4 Hz, 1H), 3.98 (s, 2H). Example 2 2-(2-(2-(phenylcarbamoyl)-2,3-dihydro-1H-inden-4-yl)-5-((4-(trifluoromethyl)phenyl)- carbamoyl)thiazol-4-yl)acetic acid (2) Scheme 2Step a. To a solution methyl 2-(2-bromo-5-((4-(trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (13) (0.155 g, 0.37 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-26014 dihydro-1H-indene-2-carboxylate (14) (0.151 g, 0.44 mmol), Pd(dppf)Cl2^CH2Cl2(0.03 g, 0.037 mmol) in 1,4-Dioxane (3.0 mL) was added Cs2CO3 (0.298 g, 0.46 mL, 0.92 mmol). The resulting mixture was degassed, refilled with nitrogen and heated at 60oC for 2 h. Then the reaction mixture was diluted with ethyl acetate, washed with water, dried with MgSO4, filtered, concentrated and purified by silica gel column chromatography using 0–20% EtOAc in hexanes to afford tert-butyl 4-(4-(2-methoxy-2-oxoethyl)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-2-yl)-2,3-dihydro-1H-indene-2-carboxylate (15). Step b. A mixture of hydrogen chloride (1.00 mL, 4.00 mmol, 4M in dioxane)) and tert-butyl 4-(4-(2- methoxy-2-oxoethyl)-5-((4-(trifluoromethyl)phenyl)carbamoyl)thiazol-2-yl)-2,3-dihydro-1H- indene-2-carboxylate (15) (0.115 g, 0.21 mmol) was stirred for 2 h at rt. The reaction mixture was evaporated to dryness and taken to the next step without purification. Step c. To a solution of 4-(4-(2-methoxy-2-oxoethyl)-5-((4-(trifluoromethyl)phenyl)carbamoyl)thiazol- 2-yl)-2,3-dihydro-1H-indene-2-carboxylic acid (16) (0.030 g, 0.059 mmol) in DCM (1.0 mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (0.009 g, 0.064 mmol). The mixture was stirred at rt for 15 min. To the resulting brown solution was added aniline 11 (0.006 g, 0.064.4 mmol) followed by DIEA (0.023 mL, 0.129 mmol). The reaction mixture was stirred at rt for 1 h, then was quenched with a solution of aqueous 1 N HCl and diluted with DCM. The organic layer was separated, and the precipitate from the aqueous layer was filtered and washed with water to afford methyl 2-(2-(2-(phenylcarbamoyl)-2,3-dihydro-1H-inden-4-yl)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (17), which was taken to the next step without purification. Step d. To a solution of methyl 2-(2-(2-(phenylcarbamoyl)-2,3-dihydro-1H-inden-4-yl)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (17) (0.014.g, 0.024 mmol) in 1,2- Dichloroethane (1 mL) was added trimethyltin(IV) hydroxide (0.009 g, 0.048 mmol) and heated at 50 °C for 6 h. Then the reaction mixture was concentrated to dryness and purified by reverse phase HPLC to obtained 2-(2-(2-(phenylcarbamoyl)-2,3-dihydro-1H-inden-4-yl)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetic acid (2). Example 3 2-(2-(4-(pyrimidine-4-carbonyl)piperazin-1-yl)-4-((4-(trifluoromethyl)phenyl)- carbamoyl)pyrimidin-5-yl)acetic acid26014 Scheme 3.To a solution of 5-bromo-2-(methylthio)pyrimidine-4-carboxylic acid (18) (2.5 g, 10 mmol) in DCM (25 ml) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (1.99 mL, 15 mmol) at rt. The mixture was stirred for 15 min. The resulting clear light brown solution was cooled to 0 °C, followed by the addition of a solution of 4-(trifluoromethyl)aniline (1.94 g, 12 mmol) in DCM (2 mL) followed N-ethyl-N-isopropylpropan-2-amine (5.24 mL, 30.1 mmol). The ice bath was removed, and the resulting mixture was stirred at rt for 10 min, diluted with DCM and washedwith an aqueous solution of 1 N HCl (50 mL). The organic layer was dried with Na2SO4,filtered, and concentrated. The crude was purified by silica gel column chromatography using26014 10—20% EtOAc in hexanes to afford 5-bromo-2-(methylthio)-N-(4- (trifluoromethyl)phenyl)pyrimidine-4-carboxamide (19). Step b. A mixture of 5-bromo-2-(methylthio)-N-(4-(trifluoromethyl)phenyl)pyrimidine-4-carboxamide (19) (0.75 g, 1.91 mmol), 2nd Generation P(t-Bu)3Pd pre-catalyst [Chloro[(tri-tert- butylphosphine)-2-(2-aminobiphenyl)] palladium(II)) (0.196 g, 0.38 mmol)], and potassium acetate (0.563 g, 5.74 mmol) was degassed and then to this mixture was added a solution of ((2- (tert-butoxy)-2-oxoethyl)zinc(II) bromide 20 (13.3 mL, 6.69 mmol) in THF. The resulting mixture was mixture was degassed and heated at 100 °C in a microwave reactor for 1 h. The reaction mixture was cooled to rt, carefully quenched with an aqueous solution of HCl (0.5 M), and concentrated to remove THF. Then the resulting mixture was transferred to a separatory funnel and extracted with EtOAc. The organic layer was washed successively with water andbrine, dried with Na2SO4, filtered, and concentrated. The crude product was purified by silicagel column chromatography using 20—50% EtOAc in hexanes to afford tert-butyl 2-(2- (methylthio)-4-((4-(trifluoromethyl)phenyl)carbamoyl)pyrimidin-5-yl)acetate (21). Step c. To a solution of tert-butyl 2-(2-(methylthio)-4-((4-(trifluoromethyl)phenyl)carbamoyl)pyrimidin- 5-yl)acetate (21) (0.136 g, 0.32 mmol) in DCM (1 mL) was added 3-chloroperoxybenzoic acid (0.165 g, 0.96 mmol) at rt. The reaction mixture was stirred at rt for 1 h, then was quenched with a saturated aqueous solution of NaHCO3 (1 mL) and extracted with DCM (2 x 20 mL). Theorganic layer was dried with Na2SO4, filtered and concentrated. The crude tert-butyl 2-(2-(methylsulfonyl)-4-((4-(trifluoromethyl)phenyl)carbamoyl)pyrimidin-5-yl)acetate (22) was taken to the next step without purification. Step d. A mixture of tert-butyl 2-(2-(methylsulfonyl)-4-((4- (trifluoromethyl)phenyl)carbamoyl)pyrimidin-5-yl)acetate (22) (0.050 g, 0.11 mmol), piperazin- 1-yl(pyrimidin-4-yl)methanone hydrochloride (0.027 g, 0.12 mmol) and N-ethyl-N- isopropylpropan-2-amine (0.076 mL, 0.435 mmol) in NMP (1 mL) was heated under microwave irradiation at 150 °C for 1 h. The reaction mixture was diluted with water and extracted withextracted with EtOAc (2 x 15 mL). The organic layer was separated, dried with Na2SO4, filtered,26014 and concentrated. The crude product was purified by silica gel column chromatography using 30—60% EtOAc in hexanes to afford methyl 2-(2-(4-(pyrimidine-4-carbonyl)piperazin-1-yl)-4- ((4-(trifluoromethyl)phenyl)carbamoyl)pyrimidin-5-yl)acetate (24). Step e. To a solution of methyl 2-(2-(4-(pyrimidine-4-carbonyl)piperazin-1-yl)-4-((4- (trifluoromethyl)phenyl)carbamoyl)pyrimidin-5-yl)acetate (24) (0.025 g, 0.04 mmol) in DCM (1 mL) was added TFA (0.33 mL). The mixture wasstirred for 30 min. The reaction was concentrated and the crude product was purified by reverse phase HPLC to afford 2-(2-(4- (pyrimidine-4-carbonyl)piperazin-1-yl)-4-((4-(trifluoromethyl)phenyl)carbamoyl)pyrimidin-5- yl)acetic acid (3). Example 4 2-(2-((1-benzoylpyrrolidin-3-yl)(phenyl)amino)-5-((4-(trifluoromethyl)phenyl)- carbamoyl)thiazol-4-yl)acetic acidStep a. A mixture of ethyl 2-bromo-4-(2-ethoxy-2-oxoethyl)thiazole-5-carboxylate (25) (4 g, 12.4 mmol), tert-butyl 3-(phenylamino)pyrrolidine-1-carboxylate (26) (3.91 g, 14.90 mmol), Pd- PEPPSI-IpentCl (Pd-Pyridine Enhanced Precatalyst Preparation Stabilization and Initiation- pentCl) (2.14 g, 2.5 mmol) and Cs2CO3 (12.14 g, 37.2 mmol) in dioxane (80 mL) was stirred at 110 °C for 8 h under N2atmosphere. The crude product was concentrated and directly loaded into column and purified by silica gel chromatography using 0–50% EtOAc in hexanes to give ethyl 2-((1-(tert-butoxycarbonyl)pyrrolidin-3-yl)(phenyl)amino)-4-(2-ethoxy-2- oxoethyl)thiazole-5-carboxylate (27). MS (ESI): m / z (M+H)+504.3.26014To a stirred solution of ethyl 2-((1-(tert-butoxycarbonyl)pyrrolidin-3-yl)(phenyl)amino)-4-(2- ethoxy-2-oxoethyl)thiazole-5-carboxylate (27) (2.5 g, 4.96 mmol) in DCM (25 mL) was added TFA (5 mL). The mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated in vacuo to give ethyl 4-(2-ethoxy-2-oxoethyl)-2-(phenyl(pyrrolidin-3-yl)amino)thiazole-5- carboxylate (28). The crude product was used in the next step directly. MS (ESI): m / z (M+H)+404.3Step c. To a stirred solution of ethyl 4-(2-ethoxy-2-oxoethyl)-2-(phenyl(pyrrolidin-3-yl)amino)thiazole- 5-carboxylate (28) (1 g, 2.48 mmol) in DMF (15 mL) was added benzoic acid (0.908 g, 7.43 mmol), TCFH (1.043 g, 3.72 mmol) and NMI (0.407 g, 4.96 mmol) at 25 °C. The resulting reaction mixture was stirred at 25 °C for 16 h, diluted with water (50 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layer was washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography using 75% EtOAc in hexanes to give 2-((1-benzoylpyrrolidin-3- yl)(phenyl)amino)-4-(2-methoxy-2-oxoethyl)thiazole-5-carboxylic acid (29). MS (ESI): m / z (M+H)+508.3.26014 Step d. To a mixture of ethyl 2-((1-benzoylpyrrolidin-3-yl)(phenyl)amino)-4-(2-ethoxy-2- oxoethyl)thiazole-5-carboxylate (29) (0.90 g, 1.78 mmol) in a mixture of MeOH (10 mL) and water (5 mL) was added LiOH (0.425 g, 17.73 mmol). The resulting mixture wasstirred at 25 °C for 16 h and then diluted with water (20 mL) and EtOAc (30 mL). The organic layer was separated, and the aqueous layer was acidified with 1 N HCl to pH < 7. The aqueous layer was re-extracted with EtOAc (30 mL x 3), the combined organic layers were washed with brine (30 mL x 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the corresponding bis acid (0.50 g, 0.89 mmol, 50.0 % yield) as white oil. The crude product was used in the next step directly. MS (ESI): m / z (M+H)+452.2. A mixture of this bis acid (0.50 g, 1.11 mmol) and HCl (2 mL, 24.36 mmol) in MeOH (10 mL) was stirred at 20 °C for 8 h. The mixture was purified by reverse phase HPLC to give 2-((1-benzoylpyrrolidin-3- yl)(phenyl)amino)-4-(2-methoxy-2-oxoethyl)thiazole-5-carboxylic acid (30). MS (ESI): m / z (M+H)+466.2Step e. To a stirred solution of 2-((1-benzoylpyrrolidin-3-yl)(phenyl)amino)-4-(2-methoxy-2- oxoethyl)thiazole-5-carboxylic acid (30) (0.33 g, 0.71 mmol) in DMF (5 mL) was added 4- (trifluoromethyl)aniline 11 (0.343 g, 2.18 mmol), TCFH (0.298 g, 1.06 mmol) and NMI (0.116 g, 1.42 mmol) at 25 °C. The mixture wasstirred at 25 °C for 16 h, and was purified by reverse HPLC to give methyl 2-(2-((1-benzoylpyrrolidin-3-yl)(phenyl)amino)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (31). MS (ESI): m / z (M+H)+609.2.1H NMR (Methanol-d4, 400 MHz): δ 7.66-7.75 (m, 2H), 7.53-7.64 (m, 5H), 7.33-7.51 (m, 7H), 5.12 (br t, J = 5.8 Hz, 1H), 3.94-4.13 (m, 3H), 3.38-3.88 (m, 6H), 2.02-2.45 (m, 2H).26014 F3C F F O F O Step f.To a - (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetate (31) in DCE (2 mL) was added trimethylstannanol (0.107 g, 0.59 mmol). The resulting mixture wasstirred at rt for 3 h, and then purified by reverse phase HPLC to give 2-(2-((1-benzoylpyrrolidin-3-yl)(phenyl)amino)-5-((4- (trifluoromethyl)phenyl)carbamoyl)thiazol-4-yl)acetic acid (4)as a solid. MS (ESI): m / z (M+H)+595.2.1H NMR (DMSO-d6, 400 MHz): δ 9.82-9.93 (m, 1H), 7.73-7.84 (m, 2H), 7.49-7.66 (m, 6H), 7.32-7.47 (m, 6H), 5.04-5.28 (m, 1H), 3.89-4.01 (m, 4H), 3.41 (d, J = 8.0 Hz, 2H), 2.19- 2.36 (m, 1H), 1.89 (dd, J = 12.9, 7.6 Hz, 1H). Example 5 2-(1-(4-fluoro-3-hydroxybenzyl)-3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4- yl)acetic acid Step 1 -To a solution of dimethyl 2-oxopentanedioate (32) (10 g, 57.4 mmol) in toluene (100 mL) was added N,N-Dimethylformamide dimethyl acetal (10.26 g, 86 mmol) at 20 °C. The mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated to give crude dimethyl (Z)-3- ((dimethylamino) methylene)-2-oxopentanedioate (13.16 g) as an oil, which was used in the next step without further purification. To a solution of dimethyl (Z)-3-((dimethylamino)methylene)-2- oxopentanedioate (13.16 g, 57.4 mmol) in HOAc (100 mL) was added hydrazine hydrate (11.50 g, 230 mmol) at 20 °C. The mixture was stirred at 20 °C for 16 h. The reaction mixture was26014 concentrated to remove most of HOAc. The residue was treated with EtOAc (300 mL), washed successively with saturated aqueous solution of NaHCO3 (100 mL) and brine (50 mL), dried over Na2SO4, filtered and concentrated to give crude methyl 4-(2-methoxy-2-oxoethyl)-1H-pyrazole- 3-carboxylate (33) as a solid, which was used in the next step without further purification.1H NMR (CDCl3, 400 MHz) δ 7.76 (s, 1H), 3.95 (s, 3H), 3.86 (s, 2H), 3.73 (s, 3H). Step 2 To a solution of (33) (3 g, 15.14mmol) in DMF (30 mL) was added PMB-Cl (2.61 g, 16.65 mmol) and K2CO3 (6.28 g, 45.4 mmol) at 25 °C. The mixture was stirred at 25 °C for 16 h. The reaction mixture was diluted with water (250 mL) and extracted with EtOAc (150 mL x 4). The combined organic layers were washed successively with water (150 mL) and brine (150 mL), dried over Na2SO4, filtered and concentrated to give crude product, which was purified by silica gel chromatography using 0—10% EtOAc in hexanes to give methyl 4-(2-methoxy-2-oxoethyl)-1-(4-methoxybenzyl)-1H-pyrazole-3- carboxylate (34) as the major product.: m / z (M+H)+318.9. Step 3 To a solution ofpyrazole-3- carboxylate (34) (2.34 g, 7.34 mmol) in THF (69 mL) and water (23 mL) was added lithium hydroxide hydrate (1.54 g, 36.7 mmol) at 25 °C. Then the reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated to removed most of THF. The residue was diluted with water (200 mL), and pH of the resulting solution was adjusted to ~ 4 with 1 M HCl. The aqueous layer was extracted with EtOAc (40 mL x 4). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered and concentrated to give 4- (carboxymethyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylic acid (35).26014 Step 4.To a solution of di- - - 3-carboxylic acid 35 (2.13 g, 7.33 mmol) in MeOH (50 mL) was added conc. HCl (0.60 mL, 7.33 mmol) at 25 °C. Then the reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated to remove MeOH. The residue was treated with EtOAc (100 mL), washed with water (50 mL) and then with brine (20 mL), dried over Na2SO4, filtered and concentrated to give 4-(2-methoxy-2- oxoethyl)-1-(4-methoxybenzyl)-1H-pyrazole-3-carboxylic acid (36), which was used in the next step without further purification. MS (ESI): m / z (M+H)+304.8. Step 53-carboxylic acid (36) (2.23 g, 7.33 mmol) in DMF (50 mL) was added 4-(trifluoromethyl)aniline 11 (1.536 g, 9.54 mmol), TCFH (3.09 g, 11.00 mmol) and NMI (1.807 g, 22.00 mmol) at 25 °C. Then the reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was purified by reverse phase HPLC to give methyl 2-(1-(4-methoxybenzyl)-3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4- yl)acetate (37). MS (ESI): m / z (M+H)+448.026014 Step 6. To a solution of methyl 2-(1-(4-methoxybenzyl)-3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H- pyrazol-4-yl)acetate (37) (1.58 g, 3.54 mmol) in MeCN (19.5 mL) and water (19.5 mL) was added CAN (5.82 g, 10.62 mmol) at 25 °C. Then the reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (30 mL x 4). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated to give crude product, which was purified by silica gel chromatography using 0– 22% EtOAc in hexanes to give methyl 2-(3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol- 4-yl)acetate (38). MS (ESI): m / z (M+H)+327.9.To a solution of methyl 2-(3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4-yl)acetate (38) (0.15 g, 0.49 mmol) in DMF (2.5 mL) was added 4-(bromomethyl)-1-fluoro-2- methoxybenzene (39) (0.11 g, 0.50 mmol) and K2CO3(0.19 g, 1.38 mmol) at 25 °C. The mixture was stirred at 25 °C for 36 h. The reaction mixture was diluted with water (25 mL) and extracted with EtOAc (15 mL x 4). The combined organic layers were washed with water (20 mL) and then with brine (20 mL), dried over Na2SO4, filtered and concentrated to give crude product, which was purified by silica gel chromatography using 0–15% EtOAc in hexanes to give methyl 2-(1-(4-fluoro-3-methoxybenzyl)-3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4- yl)acetate (40).1H NMR of 40 (CDCl3, 400 MHz) δ 8.87 (s, 1H), 7.79 (d, J = 8.5 Hz, 2H), 7.60 (d, J = 8.6 Hz, 2H), 7.51 (s, 1H), 7.13 - 7.04 (m, 1H), 6.86 - 6.75 (m, 2H), 5.28 (s, 2H), 3.96 (s, 2H), 3.88 (s, 3H), 3.73 (s, 3H). MS (ESI): m / z (M+H)+466.0 Step 826014(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4-yl)acetate (40) (0.040 g, 0.09 mmol) in DCE (1.2 mL) was added Me3SnOH (0.047g, 0.258 mmol) at 25 °C. Then the reaction mixture was stirred at 25 °C for 16 h, and at 45 °C for another 8 h. The reaction mixture was concentrated to give crude product, which was purified by reverse to give 2-(1-(4-fluoro-3-methoxybenzyl)-3- ((4-(trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4-yl)acetic acid (41). MS (ESI): m / z (M+H)+452.11H NMR (CDCl3, 400 MHz) δ 8.95 (s, 1H), 7.82 (d, J = 8.6 Hz, 2H), 7.65 (d, J = 8.6 Hz, 2H), 7.47 (s, 1H), 7.10 (dd, J = 10.9, 8.3 Hz, 1H), 6.76-6.89 (m, 2H), 5.28 (s, 2H), 3.89 (s, 3H), 3.82 (s, 2H).To a solution of 2-(1-(4-fluoro-3-methoxybenzyl)-3-((4-(trifluoromethyl)phenyl)carbamoyl)-1H- pyrazol-4-yl)acetic acid (41) (0.100 g, 0.22 mmol) in DCM (2 mL) was added BBr3(0.67 mL, 0.67 mmol) dropwise at 0 °C under N2. Then the reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was diluted with DCM (20 mL), washed with ice water (15 mL) and then with brine (15 mL), dried over Na2SO4, filtered and concentrated to give crude product, which was purified by reverse phase HPLC to give 2-(1-(4-fluoro-3-hydroxybenzyl)-3-((4- (trifluoromethyl)phenyl)carbamoyl)-1H-pyrazol-4-yl)acetic acid (5). Observed m / z 438.1 (M+H)+.1H NMR (DMSO-d6, 400 MHz) δ 12.21 (s, 1H), 10.41 (s, 1H), 9.97 (s, 1H), 8.03 (d, J =26014 8.5 Hz, 2H), 7.85 (s, 1H), 7.67 (d, J = 8.7 Hz, 2H), 7.13 (dd, J = 11.2, 8.3 Hz, 1H), 6.85 (dd, J = 8.5, 2.0 Hz, 1H), 6.74 (ddd, J = 8.3, 4.2, 2.1 Hz, 1H), 5.34 (s, 2H), 3.74 (s, 2H).

[0152] The disclosed subject matter is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0153] All references (e.g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the following claims. Biological Assays and Data WRN Helicase Assay_High ATP

[0154] To measure WRN helicase unwinding activity, a 384 well fluorescent plate-based assay was used following the separation of a fluor / quencher labeled forked double stranded DNA substrate. For construction of assay ready plates, 200 nL compounds were dispensed in DMSO via 3-fold serial dilution, into NBS DEFINE low volume all black assay plates (Corning 3820) using an Echo acoustic dispenser. All assay reaction solutions were prepared in 1X Assay Buffer (25 mM HEPES pH 7.3, 50 mM NaCl, 2 mM MgCl2, 0.01% Tween-20, 0.5 mM TCEP, 0.5% DMSO). Solution preparation: a 2X enzyme mix was generated containing 500 pM of recombinant WRN helicase core (500-942) and 60 nM annealed fluor / quencher labeled double stranded DNA oligo (IDT Custom synthesis (sequence 1) TAMRA- 5’GAACGAACACATCGGGTACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT-‘3 (SEQ ID NO: 1) (sequence 2): 5’- TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGTACCCGATGTGTTCGTTC-BHQ2-‘3 (SEQ ID NO: 2). and a 2X substrate mix was made consisting of 4 mM ATP (Fisher #R1441). Reactions were assembled first by dispensing 10 µL of 60 nM annealed fluorescent quenched labeled double stranded DNA oligo in 1X assay buffer to a single column to serve as the min control. Following this, 10 µL of 2X enzyme mix was added to all remaining wells. The reaction plate was covered and incubated at room temperature for 1 hour. After preincubation, the unwinding reaction was initiated by the addition of 10 μL of 2X substrate mix across all wells of26014 the assay plate. The plate was gently mixed and incubated at room temperature for 1 hour, prior to quenching by the addition of 2 µL of 10% SDS. Fluorescence intensity was measured using excitation and emission wavelengths of 544 nm and 590 nm, respectively. High fluorescence intensity (enzyme + substrates + DMSO) represents the signal from max unwinding (no helicase inhibition), while low florescence intensity (substrates + DMSO) represents no helicase unwinding activity / complete inhibition. Assay data was normalized to max and min signals and reported as % inhibition. Compound concentration response curves were fit by a 4P inhibition model to extract IC50, hill slope, and % maximum inhibition. WRN ATPase Assay

[0155] To measure WRN ATPase activity, the accumulation of ADP was quantitated via ADP- Glo Assay kit (Promega) in all black Non-binding Surface(NBS ) low volume 384 well microplates (Corning, 3820). For construction of assay ready plates, 200 nL compounds were dispensed in DMSO via 3-fold serial dilution (100 μM top concentration), into assay plates using an Echo acoustic dispenser. All assay reaction solutions were prepared in 1X assay buffer (25 mM HEPES pH 7.3, 50 mM NaCl, 2 mM MgCl2, 0.01% Tween-20, 0.5 mM TCEP, 1% DMSO) and a final assay volume of 20 μL was used. For determination of the ATP Km value for WRN helicase using ADP-Glo, 250 pM WRN helicase core (500-942), 100 nM ssDNA (IDT; 5’- TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGTACCCGATGTGTTCGTTC-‘3 (SEQ ID NO: 3)) allowed to react for 1, 2.5, 5, 10, 15, 20, 40 and 60 minutes in the presence of varying ATP concentrations (Fisher #R1441; 0-1000 μM). At each ATP concentration a control containing no WRN was included to subtract the assay background. The initial rate of ATP hydrolysis was determined for each ATP concentration and plotted with respect to ATP, fit by non-linear least squares regression (GraphPad Prism V9.4.1); and the Km extracted. Compound IC50s were determined through reacting 250 pM WRN and 1X Km ATP and respective compound at indicated concentrations for 1 hour before quenching by the addition of the ADP-Glo kit reagents. High luminescence (enzyme + substrates + DMSO) represents the signal from max ATP hydrolysis (no ATPase inhibition), while low luminescence (substrates + DMSO or WRN + Substrates + 20 μM positive inhibition control) represents no ATPase activity / complete inhibition. Compound concentration response curves were fit by 4P (Spotfire) inhibition model to extract IC50, hill slope, and % maximum inhibition.

[0156] The compounds of this disclosure have an IC50(nM) range between 458 – 12034 nM. IC50 values for compounds 1 through 5 are listed in Table 2 below.26014 Table 2 Compound ATPase EC50(nM) 1 5514

Claims

1. 26014 WHAT IS CLAIMED IS:

1. A compound of Formula I:salt thereof wherein, A is heteroaryl selected from pyrimidinyl, thiazolyl, and pyrazolyl, unsubstituted or substituted with 1 to 3 substituents selected from Ra; R1is selected from C1-3haloalkyl, halogen and C1-6alkyl; R2is selected from heterocycloalkyl, C3-10 cycloalkyl, -(CH2)naryl, -N(R3)2, -NHaryl; said heterocycloalkyl, cycloalkyl, aryl unsubstituted or substituted with 1 to 3 substituents selected from Ra; R3is selected from heterocycloalkyl and aryl said heterocycloalkyl and aryl unsubstituted or substituted with 1 to 3 substituents selected from Ra; Rais independently selected from hydroxyl, C1-6alkyl, halogen, C1-3haloalkyl, - (CH2)nC(O)OH, -C(O)NH- aryl, -C(O)aryl, -C(O)heterocycloalkyl, and - C(O)heteroaryl, said alkyl, aryl, heterocycloalkyl and heteroaryl, unsubstituted or substituted with 1 to 3 substituents selected from Rb; Rbis independently selected from hydroxyl, C1-6alkyl, -COOH, and halogen; and n is 0, 1, 2 or 3.

2. The compound according to claim 1 wherein A is pyrimidinyl, unsubstituted or substituted with 1 to 3 substituents selected from Ra, or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1 wherein A is thiazolyl, unsubstituted or substituted with 1 to 3 substituents selected from Ra, or a pharmaceutically acceptable salt thereof.

4. The compound according to claim 1 wherein A is pyrazolyl, unsubstituted or substituted with 1 to 3 substituents selected from Ra.26014 5. The compound according to claim 1 wherein R1is selected from CHF2, CH2F, CF3, CHCl2, CH2Cl, CCl3, CHBr2, CH2Br, and CBr3, halogen, methyl, ethyl, propyl, butyl, pentyl and hexyl, or a pharmaceutically acceptable salt thereof.

6. The compound according to any one of claims 1 to 5 wherein R1is selected from CHF2, CH2F, and CF3, or a pharmaceutically acceptable salt thereof.

7. The compound according to any one of claims 1 to 6 wherein R2is selected from piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydroindenyl, said piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or dihydroindenyl unsubstituted or substituted with 1 to 3 substituents selected from Ra, or a pharmaceutically acceptable salt thereof.

8. The compound according to any one of claims 1 to 7 wherein R2is selected from piperidyl, pyrrolyl, pyrrolidinyl, and dihydroindenyl, said piperidyl, pyrrolyl, pyrrolidinyl, or dihydroindenyl unsubstituted or substituted with 1 to 3 substituents selected from Ra, or a pharmaceutically acceptable salt thereof.

9. The compound according to any one of claims 1 to 6 wherein R2is -(CH2)nphenyl, wherein said phenyl is unsubstituted or substituted with 1 to 3 substituents selected from Ra, or a pharmaceutically acceptable salt thereof.

10. The compound according to any one of claims 1 to 6 wherein R2is -N(R3)2, or a pharmaceutically acceptable salt thereof.

11. The compound according to claim 10 wherein R3is selected from piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, and phenyl, said piperidyl, oxetanyl, pyrrolyl, pyrrolidinyl, piperazinyl, morpholinyl,26014 tetrahydrofuranyl, and phenyl unsubstituted or substituted with 1 to 3 substituents selected from Ra, or a pharmaceutically acceptable salt thereof.

12. The compound according to any one of claims 1, 10 and 11, wherein each occurrence of R3in -N(R3)2of R2is different, or a pharmaceutically acceptable salt thereof.

13. The compound according to any one of claims 1 to 12 wherein Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl, and heteroaryl, optionally substituted with 1 to 3 groups selected from Rb, or a pharmaceutically acceptable salt thereof.

14. The compound according to any one of claims 1, 2, and 5-13 wherein A is pyrimidinyl, unsubstituted or substituted with 1 to 3 substitutents selected from Ra; R1is C1-3haloalkyl, halogen, or C1-6alkyl; R2is -(CH2)nphenyl, -N(R3)2, heterocycloalkyl selected from piperidyl, oxetanyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, or C1-10cycloalkyl selected from unsubstituted or substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydro-indenyl, said phenyl, heterocycloalkyl, or cycloalkyl unsubstituted or substituted with 1 to 3 Rasubstituents; R3is heterocycloalkyl or phenyl, said heterocycloalkyl or phenyl unsubstituted or substituted with 1 to 3 substituents selected from Ra; each Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 substituents selected from Rb; and n is 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.

15. The compound according to any one of claims 1, 3, and 5-13 wherein A is thiazolyl, unsubstituted or substituted with 1 to 3 substitutents selected from Ra;26014 R1is C1-3haloalkyl, halogen, or C1-6alkyl; R2is -(CH2)nphenyl, -N(R3)2, heterocycloalkyl selected from piperidyl, oxetanyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, or C1-10cycloalkyl selected from unsubstituted or substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydro-indenyl, said phenyl, heterocycloalkyl, or cycloalkyl unsubstituted or substituted with 1 to 3 Rasubstituents; R3is heterocycloalkyl or phenyl, said heterocycloalkyl or phenyl unsubstituted or substituted with 1 to 3 substituents selected from Ra; each Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 substituents selected from Rb; and n is 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.

16. The compound according to any one of claims 1, 3, and 5-13 wherein A is pyrazolyl, unsubstituted or substituted with 1 to 3 substitutents selected from Ra; R1is C1-3haloalkyl, halogen, or C1-6alkyl; R2is -(CH2)nphenyl, -N(R3)2, heterocycloalkyl selected from piperidyl, oxetanyl, pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, or C1-10cycloalkyl selected from unsubstituted or substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and dihydro-indenyl, said phenyl, heterocycloalkyl, or cycloalkyl unsubstituted or substituted with 1 to 3 Rasubstituents; R3is heterocycloalkyl or phenyl, said heterocycloalkyl or phenyl unsubstituted or substituted with 1 to 3 substituents selected from Ra; each Rais independently selected from hydroxyl, methyl, ethyl, propyl, butyl, chlorine, fluorine, bromine, -CH2CO2H, CF3, -C(O)NH-phenyl, -C(O)phenyl, - C(O)heterocycloalkyl, and -C(O)heteroaryl, said alkyl, phenyl, heterocycloalkyl and heteroaryl, optionally substituted with 1 to 3 substituents selected from Rb; and n is 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof.

17. The compound according to claim 1 selected from:26014or a pharmaceutically acceptable salt thereof.

18. A pharmaceutical composition comprising a compound of any one of claims 1 through 17 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

19. A method of modulating or inhibiting WRN activity in a subject, comprising administering to the subject in need of such modulation or inhibition a therapeutically effective amount of the compound according to any one of claims 1 through 17, or a pharmaceutically acceptable salt thereof.26014 20. A method of treating a disorder or disease which can be treated by WRN inhibition in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 through 17, or a pharmaceutically acceptable salt thereof.

21. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound according to any one of claims 1 through 17, or a pharmaceutically acceptable salt thereof.

22. The method according to claim 21 wherein the cancer is selected from cervical, adrenocortical, kidney, breast, esophageal, colorectal, gastric, prostate, ovarian, endometrial and uterine. 23 The compound of any one of claims 1 through 17, or a pharmaceutically acceptable salt thereof, for use as a medicament.

24. The compound of any one of claims 1 through 17, or a pharmaceutically acceptable salt thereof, for use as a medicament for the treatment of a disease that is treated by WRN inhibition.

25. The compound of any one of claims 1 through 17, or a pharmaceutically acceptable salt thereof, for use as a medicament for the treatment of cancer.

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