Bicyclic tetrahydrothiazepine derivatives

Abstract

The present invention relates to a compound represented by general formula (I) [Formula 1] TIFF2025526683000149.tif39170 (in the formula, R 1 , R 2 and R 4 is as defined herein), compositions comprising the compounds, processes for making the compounds, and methods for using the compounds.

Description

[Technical Field]

[0001] The present invention relates to bicyclic tetrahydrothiazepine compounds that inhibit diacylglycerol kinases (DGK) α and ζ and are useful as T cell activators, their preparation, and pharmaceutical compositions containing said compounds.

[0002] The compounds can be useful as immunotherapeutic agents for the treatment of human diseases. More specifically, the compounds can be used alone or in combination with other immunotherapeutic agents to enhance anti-cancer immunity. [Background technology]

[0003] Cancer immunity is a multistep process regulated by a series of negative immune checkpoints and positive costimulatory receptors and associated intracellular signaling cascades that, when effectively elicited, can achieve antitumor responses (Mellman, I., et al. (2011) Cancer Immunotherapy Comes of Age, Nature 480(7378),480-489). Indeed, PD1 / PDL1 targeting and other immune checkpoint inhibitors have revolutionized cancer immunotherapy, yet more than 70% of patients still do not benefit from immune checkpoint blockade. Similarly, in the case of T cell bispecific antibodies, even in the most promising indication (non-Hodgkin's lymphoma), these T cell binders (TCBs) achieve complete remission in fewer than 50% of patients. T cell exhaustion appears to play a key role in many of these cases of primary or secondary resistance to cancer immunotherapy. A possible reason for this lack of efficacy is that T cell activation occurs via targeting and crosslinking of CD3 (signal 1) but without costimulation (signal 2), for example, via CD28 or 4-1BB. This hypothesis was clinically validated for CAR T-cell therapy, where it was shown that clinically relevant efficacy was only observed after incorporation of a costimulatory domain.

[0004] Diacylglycerol kinases (DGKs) are lipid kinases that catalyze the conversion of diacylglycerol (DAG) to phosphatidic acid (PA), thus limiting DAG-regulated functions and promoting PA-dependent functions (Merida, I., Avila-Flores, A., and Merino, E. 2008: Diacylglycerol kinases: at the hub of cell signaling. Biochem. J. 409(1), 1-18). The DGK family consists of 10 isoforms that can be classified into five subtypes based on the presence of distinct regulatory domains within their structure. Beyond that, the current lack of structural data still hinders a more complete understanding of the mode of action of DGKs. Furthermore, information on certain prokaryotic DGKs and other lipid kinases, such as sphingosine kinase and phosphatidylinositol-3-kinase (PI3K), has provided limited insight into the DGK catalytic mechanisms, which appear to differ from those of classical kinases (Arranz-Nicolas, J. and Merida, I., 2020. Biological regulation of diacylglycerol kinases in normal and neoplastic tissues: New opportunities for cancer immunotherapy. Advances in Biological Regulation, Volume 75; Ma, Q., Gabelli, S.B., Raben, D.M., 2019: Diacylglycerol kinases: relationship to other lipid kinases. Adv Biol Regul 71, 104-110).

[0005] Although several isoforms within the DGK family have been described to play a role in cancer, the α and ζ isoforms are the most extensively studied in this regard. As PA producers, both enzymes are involved in various processes that promote tumor growth and metastasis. On the other hand, as DAG consumers, DGKα and ζ have been widely characterized as negative regulators of T cell responses (Riese, MJ, Moon, EK, Johnson, BD, Albelda, SM, 2016. Diacylglycerol kinases (DGKs): novel targets for improving T cell activity in cancer. Front Cell Dev Biol 4, 108; Noessner, E., 2017. DGK-alpha: a checkpoint in cancer-mediated immuno-inhibition and target for immunotherapy. Front Cell Dev Biol 5, 16; Sakane, F., Mizuno, S., Komenoi, S., 2016. Diacylglycerol kinases as emerging potential drug targets for a variety of diseases: an update. Front Cell Dev Biol 4, 82; Arranz-Nicolas, J. and Merida, I., 2020. Biological regulation of diacylglycerol kinases in normal and neoplastic tissues: New opportunities for cancer immunotherapy,Advances in Biological Regulation,Volume 75).

[0006] These two isozymes, DGKα and DGKζ, are active downstream of CD28 and other costimulatory receptors as well as the T cell receptor (TCR), and their function is to limit the amount of DAG produced and ultimately T cell activation (Merida, I., Andrada, E., Gharbi, S.I., Avila-Flores, A., 2015. Redundant and specialized roles for diacylglycerol kinases alpha and zeta in the control of T cell functions. Sci. Signal. 8(374); Shulga, Y.V., Topham, M.K., Epand, R.M., 2011. Regulation and functions of diacylglycerol kinases. Chem. Rev. 111(10), 6186-6208.). An overview of representative DGK-regulated signaling pathways is shown in Figure 1 (Sim, J.A.; Kim, J.; Yang, D. Beyond Lipid Signaling: Pleiotropic Effects of Diacylglycerol Kinases in Cellular Signaling. Int. J. Mol. Sci. 2020, 21, 6861): Activated PLC1 cleaves PIP2 in the plasma membrane to generate two secondary messengers, DAG and IP3. DAG activates PKC, Ras / MEK / ERK / AP-1, and NF-kB, while IP3 is involved in activating intracellular Ca2+ flux. Upregulated Ca2+ signaling then activates the transcription factor NFAT. Briefly, the production and levels of DAG determine the duration and strength of Ras / MEK / ERK- and PKC-dependent signaling pathways, which are central to T cell activation. Therefore, DGK acts as an intracellular checkpoint, and inhibition of DGK is expected to enhance T cell signaling pathways and T cell activation.

[0007] Experimental evidence suggests that enhanced DGK function and / or expression in tumor-infiltrating T cells (TILs) limits tumor destruction. Experiments using CAR T cells against human mesothelioma transplanted into nude mice demonstrated that tumor-infiltrating CAR T cells expressed high concentrations of surface inhibitory receptors and the inhibitory enzymes SHIP-1, DGKα, and DGKζ (Moon et al., 2014). Furthermore, high DGKα expression was also observed in TILs isolated from human renal tumors (Prinz et al., 2012). In mouse mesoCAR T cells, double deletion of DGKα and DGKζ resulted in enhanced cytokine expression and cytotoxicity against tumor cells (Riese et al., 2013). Similar results were reported for human CAR T cells in which expression of both DGKα and DGKζ was silenced using CRISPR / Cas9 (Jung et al., 2018). All of these studies support the rationale for targeting DGKα / ζ in the development of anticancer therapies (Arranz-Nicolas, J. and Merida, I., 2020. Biological regulation of diacylglycerol kinases in normal and neoplastic tissues: New opportunities for cancer immunotherapy. Advances in Biological Regulation, Volume 75; Riese, MJ, Moon, EK, Johnson, BD, Albelda, SM, 2016. Diacylglycerol kinases (DGKs): novel targets for improving T cell activity in cancer. Front Cell Dev Biol 4, 108.). Knockout mouse models provide further evidence.Mice lacking either DGKα or DGKζ exhibited a hyperresponsive T cell phenotype and improved antitumor immune activity (Riese, MJ, Grewal, J., Das, J., Zou, T., Patil, V., Chakraborty, AK, Koretzky, GA, 2011. Decreased diacylglycerol metabolism enhances ERK activation and augments CD8+ T cell functional responses. J. Biol. Chem. 286(7), 5254-5265; Zha, Y., Marks, R., Ho, AW, Peterson, AC, Janardhan, S., Brown, I., Praveen, K., Stang, S., Stone, JC, Gajewski, TF, 2006. T cell energy is reversed by active Ras and is regulated by diacylglycerol kinase-alpha.Nat.Immunol.7(11),1166-1173;Olenchock,BA,Guo,R.,Carpenter,JH,Jordan,M.,Topham,MK,Koretzky,GA,Zhong,XP,2006a.Disruption of diacylglycerol metabolism impairs the induction of T cell energy.Nat.Immunol.7(11),1174-1181.).

[0008] Taken together, there is substantial evidence that DGKα and DGKζ are high-value targets for cancer immunotherapy. At the same time, there is a lack of compounds capable of potently inhibiting both DGKα and DGKζ with good selectivity over other diacylglycerol kinases, protein kinases, and / or other lipid kinases.

[0009] The present invention describes such dual DGKα / ζ inhibitors with superior selectivity over other protein kinases across a broad range of safety / off-target profiles and other lipid kinases. These compounds potently activate suboptimally stimulated T cells, thereby acting as intracellular enhancers of the costimulatory signaling cascade. These DGKα / ζ inhibitors have the potential to increase the proliferation, cytotoxicity, and longevity of targeted T cells, which may result in improved anti-cancer activity of CPI, CD3-engaged T cell bispecific, and CAR T cells. Furthermore, by engaging signaling nodes central to both the TCR and costimulatory receptors, it is plausible that these molecules could enhance both signals 1 and 2, thus achieving single-agent activity, for example, in inflammatory tumors.

[0010] There remains a need for novel compounds that can activate and expand T cells, thus enabling the treatment, prevention and / or delay of cancer progression.

[0011] It is therefore an object of the present invention to provide compounds useful as DGK α / ζ inhibitors for the treatment or prevention or amelioration of such diseases, which have improved therapeutic properties, in particular improved pharmacokinetic properties. Summary of the Invention

[0012] A first object of the present invention is to provide a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 is a 6-membered heteroaryl, and R 1 is one or more R, which may be the same or different 10 is a 6-membered heteroaryl optionally substituted with R 2 is hydrogen or halogen, R 4 is one or more R, which may be the same or different11 phenyl or pyridine, optionally substituted with R 10 teeth, i) one or more halogens, hydroxy, -S(O)2(C 1~6 -alkyl), -N(R 10e R 10f ), -S(O)2(C 1~6 -cycloalkyl), optionally substituted with cyano, C 1~10 - alkyl; ii) one or more -S(O)2(C 1~6 -alkyl), C 3~10 -cycloalkyl; iii) one or more halogens, -C(O)O-(R 10q 3-10 membered heterocyclyl optionally substituted with iv) one or more C 1~6 -Alkyl, halogen, or halo-C 1~6 -phenyl, optionally substituted with alkyl; v)-N(R 10e R 10f ); vi)-OR 10g ; vii) halogen; R 10e and R 10f are each independently hydrogen and C 1~6 -alkyl, R 10g is C 1~6 -Alkyl and halo-C 1~6 -alkyl, R 10q is C 1~10 - alkyl or or two R's 10 together with the carbon atoms to which they are attached, form one or more C 1~10 -forms a 3- to 10-membered heterocyclyl optionally substituted by alkyl; R 11 teeth, i) halogens; ii) C, optionally substituted with one or more cyanos1~6 - alkyl; iii) C 1~6 -alkoxy; iv) C 3~7 -cycloalkyl; v) one or more halo-C 1~6 -Alkyl, C 3~10 -5-6 membered heteroaryl optionally substituted with cycloalkyl; vi) one or more halogens, C 1~6 -Alkoxy, Halo-C 1~6 -phenyl, optionally substituted with alkyl; vii)-SO2(R 11d ), selected from R 11d is hydrogen, C 1~6 -Alkyl and halo-C 1~6 -alkyl, or a pharmaceutically acceptable salt thereof.

[0013] A second object of the present invention is a process for the preparation of a compound of formula (I) above, or a pharmaceutically acceptable salt thereof, of formula (XI) [ka] (In the formula, R 1 , R 2 , and R 4 is as defined herein and PG is an amino protecting group) with a suitable deprotecting agent to form said compound of formula (I).

[0014] A third object of the present invention is a pharmaceutical composition comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0015] A fourth object of the present invention is a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, for use in treating, preventing and / or delaying the progression of cancer.

[0016] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the invention, suitable methods and materials are described below.

[0017] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0018] The nomenclature used in this application is based on IUPAC systematic nomenclature unless otherwise indicated. DETAILED DESCRIPTION OF THE INVENTION

[0019] definition "Alkoxy" refers to an alkyl group, as defined previously, attached to the parent molecular moiety through an oxygen atom. Unless otherwise specified, an alkoxy group contains 1 to 12 carbon atoms ("C 1~12 -alkoxy"), preferably 1 to 10 carbon atoms ("C 1~10 -alkoxy"), more preferably 1 to 6 carbon atoms ("C 1~6 In some preferred embodiments, the alkoxy group contains 1 to 4 carbon atoms. In still other embodiments, the alkoxy group contains 1 to 3 carbon atoms. Some non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and tert-butoxy.

[0020] "Alkoxyalkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group has been replaced with an alkoxy group. Preferably, "alkoxyalkyl" refers to an alkyl group in which one, two, or three hydrogen atoms of the alkyl group, most preferably one hydrogen atom, have been replaced with an alkoxy group. Particularly preferred, but non-limiting, examples of alkoxyalkyl are methoxymethyl and 2-methoxyethyl.

[0021] "Alkyl" refers to an alkyl group having a specified number of carbon atoms (i.e., C1 to 10 refers to a saturated, straight-chain (i.e., unbranched) or branched monovalent hydrocarbon chain, or combinations thereof. Particular alkyl groups are those having 1 to 20 carbon atoms ("C1 to C10"). 20 alkyl"), those having 1 to 12 carbon atoms ("C1 12 alkyl), those having 1 to 10 carbon atoms ("C1 10 alkyl"), those having 1 to 8 carbon atoms ("C1-8 alkyl"), those having 1 to 6 carbon atoms ("C1-6 alkyl"), those having 2 to 6 carbon atoms ("C2-6 alkyl"), or those having 1 to 4 carbon atoms ("C 1~4 Examples of alkyl groups include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl; homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0022] "Alkynyl" refers to an alkyl group having the specified number of carbon atoms (i.e., C 2~10 "C" refers to an unsaturated, linear (i.e., unbranched) or branched monovalent hydrocarbon chain having at least one site of acetylenic unsaturation (i.e., having at least one moiety of the formula C≡C), or combinations thereof. Particular alkynyl groups are those having 2 to 20 carbon atoms ("C 2~20 alkynyl"), those having 2 to 8 carbon atoms ("C 2~8 alkynyl) with 2 to 6 carbon atoms ("C 2~6 alkynyl) with 2 to 4 carbon atoms ("C 2~4 Examples of alkynyl groups include, but are not limited to, groups such as ethynyl (or acetylenyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, but-3-ynyl, homologs and isomers thereof.

[0023] "Amino," alone or in combination with other groups, refers to NH2.

[0024] "Aminoalkyl" refers to an alkyl group in which one or more of the alkyl group's hydrogen atoms has been replaced with an amino moiety.

[0025] "Aromatic" is a well-known term in the literature, especially in the IUPAC Compendium of Chemical Terminology, 2 nd This shows the conventional concept of aromaticity as defined in "Aromaticity in Organic Compounds: A Brief Description of the Invention," Eds. Blackwell Scientific Publications, Oxford (1997).

[0026] "Aryl" means a cyclic aromatic hydrocarbon moiety having a monocyclic, bicyclic, or tricyclic aromatic ring of 5 to 14 carbon ring atoms ("C 5~14 "-aryl"). Bicyclic aryl ring systems include fused bicycles having two fused 5-membered aryl rings (designated 5-5), fused bicycles having a 5-membered aryl ring and a fused 6-membered aryl ring (designated 5-6 and 6-5), and bicycles having two fused 6-membered aryl rings (designated 6-6). Aryl groups may be optionally substituted as defined herein. Examples of aryl substituents include, but are not limited to, phenyl, naphthyl, phenanthryl, fluorenyl, indenyl, pentalenyl, azulenyl, and the like. The term "aryl" also includes partially hydrogenated derivatives of cyclic aromatic hydrocarbon moieties, provided that at least one ring of the cyclic aromatic hydrocarbon moiety is aromatic and each may be optionally substituted.

[0027] "Cancer" refers to a disease characterized by the presence of a neoplasia or tumor, resulting from the abnormal and uncontrolled growth of cells (such cells are "cancer cells"). As used herein, the term cancer expressly includes, but is not limited to, hepatocellular carcinoma, malignant tumors and hyperproliferative disorders of the colon (colon cancer), lung cancer, breast cancer, prostate cancer, melanoma, and ovarian cancer.

[0028] The term "cyano," alone or in combination with other groups, refers to CN (ie, nitrile).

[0029] "Cyanoalkyl" refers to an alkyl group in which one or more of the alkyl group's hydrogen atoms has been replaced with a cyano moiety.

[0030] "Cycloalkyl" refers to a monocyclic, bicyclic (including bridged bicyclic and cycloalkyl spiro substituents), or tricyclic ring and a saturated or partially unsaturated carbocyclic moiety having 3 to 10 carbon atoms, i.e., (C 10 ) cycloalkyl. The cycloalkyl moiety can be optionally substituted with one or more substituents. In certain embodiments, the cycloalkyl contains 3 to 8 carbon atoms (i.e., (C3-C8)cycloalkyl). In other certain embodiments, the cycloalkyl contains 3 to 6 carbon atoms (i.e., (C3-C6)cycloalkyl). Examples of cycloalkyl substituents include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and their partially unsaturated (cycloalkenyl) derivatives (e.g., cyclopentenyl, cyclohexenyl, and cycloheptenyl), bicyclo[3.1.0]hexanyl, bicyclo[3.1.0]hexenyl, bicyclo[3.1.1]heptanyl, bicyclo[3.1.1]heptenyl, and bicyclo[1.1.1]pentane. The cycloalkyl moiety can be attached in a "spirocycloalkyl" or "cycloalkylspiro" fashion, such as "spirocyclopropyl."

[0031] "ECx" refers to the effective concentration of a particular compound, e.g., in culture medium or plasma, required to achieve x% of the maximum specific effect in vitro or in vivo. Examples of "ECx" are EC20, EC50, and EC100, which indicate the concentrations of a particular compound in culture medium or plasma required to achieve 20%, 50%, and 100% of the maximum specific effect in vitro or in vivo, respectively. "Halo" or "halogen" refers to fluoro, chloro, bromo, and / or iodo. If a residue is substituted with more than one halogen, this can be referred to using a prefix corresponding to the number of halogen substituents attached, e.g., dihaloaryl, dihaloalkyl, trihaloaryl, etc., refer to aryl and alkyl substituted with two ("di") or three ("tri") halo groups, which may, but need not, be the same halo. Thus, 4-chloro-3-fluorophenyl is within the scope of dihaloaryl. An alkyl group in which one or more hydrogens have been replaced by a halo group is referred to as "haloalkyl," e.g., "C 1~6 A preferred haloalkyl group is trifluoroalkyl (-CF3).

[0032] "Haloalkoxy" refers to an alkoxy group in which at least one halogen replaces each H in the hydrocarbon that makes up the alkyl portion of the alkoxy group. Examples of haloalkoxy groups are difluoromethoxy (-OCHF2) and trifluoromethoxy (-OCF3).

[0033] "Haloalkyl" refers to an alkyl group as defined above that contains at least one carbon atom substituted with at least one halo group, where halo is as defined herein. Examples of "haloalkyl" groups useful in the present invention include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl, and n-butyl, each independently substituted with one or more halo groups, such as fluoro, chloro, bromo, and iodo.

[0034] "Haloaryl" refers to an aryl in which at least one hydrogen has been replaced with a halogen.

[0035] "Heteroaryl" refers to an aromatic heterocyclic monocyclic, bicyclic, or tricyclic ring system of 5 to 14 ring atoms, preferably 5 to 10 ring atoms, and more preferably 5 to 6 ring atoms, containing 1, 2, 3, or 4 heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. In some embodiments, the monocyclic heteroaryl ring can be 5 to 6-membered. Bicyclic heteroaryl ring systems include fused bicycles having two fused 5-membered heteroaryl rings (designated 5-5), fused bicycles having a 5-membered aryl ring and a fused 6-membered heteroaryl ring (designated 5-6 and 6-5), and bicycles having two fused 6-membered aryl rings (designated 6-6). Heteroaryl groups can be optionally substituted as defined herein. Examples of heteroaryl substituents include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, isoxazolyl, benzofuranyl, isothiazolyl, benzothienyl, benzothiophenyl, indolyl, aza-indolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, Examples include benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, pyrrolopyridinyl, furopyridinyl, thienopyridinyl, pyrrolopyridazinyl, pyrrolopyrimidinyl, pyrrolopyrazinyl, thienopyridazinyl, thienopyrimidinyl, thienopyrazinyl, furopyridazinyl, furopyrimidinyl and furopyrazinyl. Most preferably, "5-membered heteroaryl" refers to the following group: [ka]

[0036] "Heteroatom" refers to an atom other than carbon or hydrogen.

[0037] "Heterocycle" or "heterocyclyl" refers to a 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-membered monocyclic, 7-, 8-, 9-, and 10-membered bicyclic (including bridged bicyclic and cycloalkyl spiro substituents), or 10-, 11-, 12-, 13-, 14-, and 15-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more (e.g., 1, 2, 3, or 4) heteroatoms selected from oxygen, nitrogen, and sulfur within the ring, with the remaining ring atoms being carbon. In some embodiments, a heterocycle is a heterocyclic ring. It is cycloalkyl. In certain embodiments, heterocycle or heterocyclyl refers to a 4-, 5-, 6-, or 7-membered heterocycle. When used in reference to a ring atom of a heterocycle, nitrogen or sulfur may also be in oxidized form, and the nitrogen may be substituted with one or more (C1-C6) alkyl or groups. A heterocycle can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. Any of the heterocycle atoms can be optionally substituted with one or more substituents described herein. Examples of such saturated or partially unsaturated heterocyclyls include tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, pyrrolidine 1-oxide, N-hydroxypiperidine, 1-methylpyrrolidinyl ... Examples of heterocyclic rings include, but are not limited to, ethyl pyrrolidine N-oxide, diazirinyl, and quinuclidinyl. The term heterocycle also includes groups in which a heterocycle is fused to one or more aryl, heteroaryl, or cycloalkyl rings, such as indolinyl, 3H-indolyl, chromanyl, azabicyclo[2.2.1]heptanyl, azabicyclo[3.1.0]hexanyl, azabicyclo[3.1.1]heptanyl, octahydroindolyl, or tetrahydroquinolinyl.

[0038] "Hydroxy", alone or in combination with other groups, refers to OH.

[0039] "Hydroxyalkyl" refers to an alkyl group in which one or more of the alkyl group's hydrogen atoms has been replaced by a hydroxy moiety. Examples include alcohols and diols.

[0040] "Moiety" and "substituent" refer to an atom or set of chemically bonded atoms that is attached to another atom or molecule by one or more chemical bonds, thereby forming a part of a molecule.

[0041] When indicating the number of substituents, the term "one or more" refers to a range from one substituent to the highest possible number of substitutions, i.e., from the replacement of one hydrogen to the replacement of all hydrogens by substituents; particularly "one or more" refers to one, two, or three; most particularly "one or more" refers to one or two.

[0042] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes cases where the event or circumstance occurs and cases where it does not occur. For example, "an aryl group optionally substituted with an alkyl group" means that the alkyl may be present but does not have to be present, and that the description includes situations where the aryl group is substituted with an alkyl group and situations where the aryl group is not substituted with an alkyl group.

[0043] "Optionally substituted" is intended to mean unsubstituted or substituted. Generally, the substituents can be the same or different.

[0044] "Oxo", alone or in combination with other groups, refers to =O.

[0045] "Pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the free base or free acid, which are not biologically or otherwise undesirable. Salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, especially hydrochloric acid, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, and the like.

[0046] Particularly preferred pharmaceutically acceptable salts of compounds of formula (I) are the salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and methanesulfonic acid.

[0047] The term "protecting group" (PG) refers to a group that selectively blocks a reactive site in a polyfunctional compound so that a chemical reaction, in the sense conventionally associated with synthetic chemistry, can be carried out selectively at an otherwise unprotected reactive site. The protecting group can be removed when appropriate. Exemplary protecting groups are amino-protecting, carboxy-protecting, or hydroxy-protecting groups. Particular protecting groups are tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), fluorenylmethoxycarbonyl (Fmoc), and benzyl (Bn). Further particular protecting groups are tert-butoxycarbonyl (Boc) and fluorenylmethoxycarbonyl (Fmoc). An even more particular protecting group is tert-butoxycarbonyl (Boc). Exemplary protecting groups and their use in organic synthesis are described, for example, in "Protective Groups in Organic Chemistry" by T.W. Greene and P.G.M. Hutts, 5th Ed., 2014, John Wiley & Sons, NY.

[0048] "Prophylaxis," as used herein, includes preventing or delaying the appearance of clinical symptoms of a condition, disorder or condition that develops in a mammal, particularly in a human suffering from or susceptible to the condition, disorder or condition, but who has not yet experienced or exhibited clinical symptoms or asymptomatic symptoms of the condition, disorder or condition.

[0049] "Substituted" refers to the replacement of at least one of the hydrogen atoms of a compound or moiety with another substituent or moiety. Examples of such substituents include, but are not limited to, halogen, -OH, -CN, oxo, alkoxy, alkyl, alkylene, aryl, heteroaryl, haloalkyl, haloalkoxy, cycloalkyl, and heterocycle. For example, the term "haloalkyl" refers to the fact that one or more hydrogen atoms of an alkyl (defined below) have been replaced with one or more halogen atoms (e.g., trifluoromethyl, difluoromethyl, fluoromethyl, chloromethyl, etc.). In one aspect, as used herein, substitution can refer to at least one hydrogen atom of a compound or moiety described herein being replaced with a halogen or alkyl.

[0050] A "therapeutically effective amount" refers to the amount of a compound or molecule of the invention that, when administered to a subject, (i) treats or prevents a particular disease, condition, or disorder, (ii) reduces, ameliorate, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. A therapeutically effective amount may vary depending on the compound, the disease state being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending physician or veterinarian, and other factors.

[0051] A "therapeutically inert carrier" refers to any non-toxic ingredient, such as a disintegrant, binder, filler, solvent, buffer, isotonicity agent, stabilizer, antioxidant, surfactant, or lubricant, that has no therapeutic activity and is used in formulating a pharmaceutical product.

[0052] In particular, the chemical groups defined above are those specifically exemplified in the examples.

[0053] The following abbreviations are used herein:

[0054] BOP = benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate, Brine = saturated aqueous NaCl solution, CAS = Chemical Abstracts Registry Number, CDI = 1,1'-carbonyldiimidazole, DBU = 1,8-diazabicyclo[5,4,0]undec-7-ene, DCM = dichloromethane, DDQ = 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, DMF = N,N-dimethylformamide, DIPEA = N,N-diisopropyl Ethylamine, EDC = 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, ESI = electrospray ionization, EtOAc = ethyl acetate, EtOH = ethanol, h = hour(s), HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate, HBTU = O-benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluorophosphate phosphate, HFIP = hexafluoroisopropanol, HOBt = hydroxybenzotriazole, HPLC = high performance liquid chromatography, m-CPBA = meta-chloroperoxybenzoic acid, MeCN = acetonitrile, MeI = methyloid, MeOH = methanol, min = min(s), MS = mass spectrum, NBS = N-bromosuccinimide, PE = petroleum ether, PyBroP = bromo-tris-pyrrolidino-phosphonium hexafluorophosphate, RT = room temperature , TBAF = tetrabutylammonium fluoride, TBAOH = tetrabutylammonium hydroxide, TBDMS = tert-butyldimethylsilyl, TEA = triethylamine, TFA = trifluoroacetic acid, THF = tetrahydrofuran, TMSOTF = trifluoromethanesulfonic acid trimethylsilyl ester, TLC = thin layer chromatography, T3P = 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide.

[0055] In the description herein, if there is a discrepancy between a depicted structure and the name given to that structure, the depicted structure shall prevail. Furthermore, if the stereochemistry of a structure or portion of a structure is not indicated, for example, with a bold wedge or a dotted line, the structure or portion of the structure is intended to encompass all of its stereoisomers. However, in some cases where multiple chiral centers are present, the structure and name may be represented as a single enantiomer to aid in describing the relative stereochemistry.

[0056] Unless otherwise indicated, "a compound of the formula" or "a compound of formula" or "compounds of the formula" or "compounds of formula" refers to any compound selected from the genus of compounds defined by that formula (including any pharmaceutically acceptable salt of any such compound, unless otherwise specified).

[0057] Certain compounds may exhibit tautomerism. Tautomeric compounds can exist as two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate individual tautomers usually result in mixtures whose chemical and physical properties match those of the compound. The position of the equilibrium depends on the chemical features within the molecule. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates, whereas in phenols, the enol form predominates. Common protic tautomers include keto / enol (-C(=O)-CH-⇔-C(-OH)=CH-), amide / imidic acid (-C(=O)-NH-⇔-C(-OH)=N-), and amidine (-C(=NR)-NH-⇔-C(-NHR)=N-) tautomers. The latter two are particularly common in heteroaryl and heterocyclic rings, and the present invention encompasses all tautomeric forms of the compounds.

[0058] Furthermore, the present invention includes all optical isomers of the compounds of formula (I), i.e., diastereoisomers, diastereomeric mixtures, racemic mixtures, all corresponding enantiomers and / or tautomers thereof, and solvates thereof.

[0059] Compounds of Formula (I) may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Additional asymmetric centers may be present depending on the nature of the various substituents on the molecule. Each such asymmetric center independently produces two optical isomers, and all possible optical isomers and diastereomers, both as mixtures and as pure or partially purified compounds, are intended to be encompassed by the present invention. The present invention is meant to encompass all such isomeric forms of these compounds. The independent syntheses of these diastereomers or their chromatographic separations may be achieved as known in the art by appropriate modification of the methods disclosed herein. Their absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates, which are derivatized, if necessary, with a reagent containing an asymmetric center of known absolute configuration. If desired, racemic mixtures of the compounds can be separated so that the individual enantiomers are isolated. Resolution can be carried out by methods known in the art, such as coupling a racemic mixture of a compound to an enantiomerically pure compound to form a diastereomeric mixture, followed by separation of the individual diastereomers by standard methods such as fractional recrystallization or chromatography.

[0060] In embodiments in which optically pure enantiomers are provided, optically pure enantiomer means that the compound contains greater than 90% by weight of the desired isomer, specifically greater than 95% by weight of the desired isomer, or more specifically greater than 99% by weight of the desired isomer, the weight percentages being based on the total weight of the isomer(s) of the compound. Chirally pure or chirally enriched compounds can be prepared by chirally selective synthesis or by separation of enantiomers. Separation of enantiomers can be carried out on the final product or on a suitable intermediate.

[0061] In some embodiments, compounds of formula (I) are isotopically labeled by having one or more atoms therein replaced by atoms having a different atomic mass or mass number. Such isotopically labeled (i.e., radiolabeled) compounds of formula (I) are considered within the scope of the present disclosure. Examples of isotopes that can be incorporated into compounds of formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, respectively, e.g., 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 Certain isotopically labeled compounds of formula (I), for example those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The radioactive isotope tritium, i.e., 3 H and carbon-14, i.e., 14C are particularly useful for this purpose given their ease of incorporation and ready means of detection. For example, compounds of formula (I) can be enriched with 1, 2, 5, 10, 25, 50, 75, 90, 95, or 99 percent of a given isotope.

[0062] Heavier isotopes, such as deuterium, i.e. 2 Substitutions such as H may result in greater metabolic stability and may confer certain therapeutic advantages, for example, by increasing in vivo half-life or requiring lower dosages.

[0063] 11 C. 18 F, 15 O and 13 Substitution with positron emitting isotopes, such as N, can be useful in positron emission tomography (PET) studies for examining substrate receptor occupancy. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the Examples set forth below, substituting appropriate isotopically labeled reagents for previously employed non-isotopically labeled reagents.

[0064] Compounds of the Invention In one embodiment, a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 is a 6-membered heteroaryl, and R 1 is one or more R, which may be the same or different 10 is a 6-membered heteroaryl optionally substituted with R 2 is hydrogen or halogen, R 4 is one or more R, which may be the same or different 11 and selected from phenyl and pyridine, optionally substituted with R 10 teeth, i) one or more halogens, hydroxy, -S(O)2(C 1~6 -alkyl), -N(R 10e R 10f ), -S(O)2(C 1~6 -cycloalkyl), optionally substituted with cyano, C 1~10 - alkyl; ii) one or more -S(O)2(C 1~6 -alkyl), C 3~10 -cycloalkyl; iii) one or more halogens, -C(O)O-(R 10q 3-10 membered heterocyclyl optionally substituted with iv) phenyl; v)-N(R 10e R 10f ); vi)-OR 10g ; vii) halogen; R 10e and R 10f are each independently hydrogen and C 1~6 -alkyl, R 10g is C 1~6 -Alkyl and halo-C 1~6 -alkyl, R 10q is C 1~10 - alkyl or or two R's 10 together with the carbon atoms to which they are attached, form one or more C 1~10 -forms a 3- to 10-membered heterocyclyl optionally substituted by alkyl; R 11 teeth, i) halogens; ii) C, optionally substituted with one or more cyanos 1~6 - alkyl; iii) C 1~6 -alkoxy; iv) C 3~7 -cycloalkyl; v) one or more halo-C 1~6 -Alkyl, C3~10 -5-6 membered heteroaryl optionally substituted with cycloalkyl; vi) one or more halogens, C 1~6 -Alkoxy, Halo-C 1~6 -phenyl, optionally substituted with alkyl; vii)-SO2(R 11d ), selected from R 11d is hydrogen, C 1~6 -Alkyl and halo-C 1~6 -alkyl, or a pharmaceutically acceptable salt thereof.

[0065] In another embodiment, R 1 is one or more R 10 Provided are compounds of formula (I) as described herein that are pyridyl, pyrimidinyl, pyridazinyl, triazinyl, optionally substituted with:

[0066] In another embodiment, R 1 is one or more R 10 Provided are compounds of formula (I) as described herein that are pyridyl, pyrimidinyl, triazinyl, optionally substituted with:

[0067] In another embodiment, R 1 is one or more R 10 Provided are compounds of formula (I) as described herein which are pyridyl, triazinyl, optionally substituted with

[0068] In another embodiment, R 2 is hydrogen or fluorine.

[0069] In another embodiment, R 2 is fluorine.

[0070] In another embodiment, R 4 But one R11 Provided are compounds of formula (I) as described herein, wherein the compound is phenyl substituted with

[0071] In another embodiment, R 10 is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy, or 10 and R 1 taken together to form trimethyl-6,8-dihydro-1,7-naphthyridinyl, dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl.

[0072] In another embodiment, R 10 is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile.

[0073] In another embodiment, R 11 is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propanenitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridonyl.

[0074] In another embodiment, R 11is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy.

[0075] In another embodiment, there is provided a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein: R 1 is one or more R, which may be the same or different 10 pyridyl, pyrimidinyl, pyridazinyl, triazinyl, optionally substituted by R 2 is hydrogen or halogen, R 4 is one R 11 is phenyl substituted with R 10 teeth, i) one or more halogens, hydroxy, -S(O)2(C 1~6 -alkyl), -N(R 10e R 10f ), -S(O)2(C 1~6 -cycloalkyl), optionally substituted with cyano, C 1~10 - alkyl; ii) one or more -S(O)2(C 1~6 -alkyl), C 3~10 -cycloalkyl; iii) one or more halogens, -C(O)O-(R 10q 3-10 membered heterocyclyl optionally substituted with iv) phenyl; v)-N(R 10e R 10f ); vi)-OR 10g ; vii) halogen; R 10e and R 10f are each independently hydrogen and C 1~6 -alkyl, R 10g is C 1~6 -Alkyl and halo-C 1~6-alkyl, R 10q is C 1~10 - alkyl or R 11 teeth, i) halogens; ii) C, optionally substituted with one or more cyanos 1~6 - alkyl; iii) C 1~6 -alkoxy; iv) C 3~7 -cycloalkyl; v) one or more halo-C 1~6 -Alkyl, C 3~10 -5-6 membered heteroaryl optionally substituted with cycloalkyl; vi) one or more halogens, C 1~6 -Alkoxy, Halo-C 1~6 -phenyl, optionally substituted with alkyl; vii)-SO2(R 11d ), selected from R 11d is hydrogen, C 1~6 -Alkyl and halo-C 1~6 -alkyl, or a pharmaceutically acceptable salt thereof.

[0076] In another embodiment, there is provided a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein: R 1 is one or more R, which may be the same or different 10 pyridyl, pyrimidinyl, triazinyl, optionally substituted by R 2 is hydrogen or fluorine, R 4 is one R 11 is phenyl substituted with R 10is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy or Or, R 10 and R 1 together to form trimethyl-6,8-dihydro-1,7-naphthyridinyl, dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl, R 11 is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propanenitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridine, or a pharmaceutically acceptable salt thereof.

[0077] In another embodiment, there is provided a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein: R 1 is one or more R, which may be the same or different 10 pyridyl, triazinyl, optionally substituted by R 2 is fluorine, R 4 is one R 11 is phenyl substituted with R 10 is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile, R 11is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, or a pharmaceutically acceptable salt thereof.

[0078] In another preferred embodiment, (3R)-3-amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(2-tert-butyl-4-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butylpyrimidin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-(4-chlorobenzyl)-8-fluoro-1,1-diketo-7-[5-(trifluoromethoxy)-3-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-isopropoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[6-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)pyrimidin-2-yl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butyl-1,2,4-triazin-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-6-methyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; Methyl 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylate; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-[5-(3-chloro-1-methylsulfonyl-propyl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-diketo-7-(5-morpholino-3-pyridyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-triketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-3-pyridyl]-2-methyl-propionitrile; (3R)-3-amino-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-7-[5-(trifluoromethyl)-3-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-diketo-7-[5-(1-mesyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5,6-dimethyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-dioxo-7(3R)-3-amino-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-dioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; and (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one;

[0013] Provided is a compound of formula (I) as described herein selected from:

[0079] In another preferred embodiment, (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-6-methyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-triketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-3-pyridyl]-2-methyl-propionitrile; (3R)-3-amino-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-7-[5-(trifluoromethyl)-3-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-diketo-7-[5-(1-mesyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5,6-dimethyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; and (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one;

[0013] Provided is a compound of formula (I) as described herein selected from:

[0080] Manufacturing process Processes for preparing the compounds of formula (I) described herein, or pharmaceutically acceptable salts thereof, are also an object of the present invention.

[0081] The present invention provides a process for the preparation of the compounds described herein, or a pharmaceutically acceptable salt thereof, of formula (XI) [ka] (In the formula, R 1 , R 2 , and R 4is as defined herein and PG is an amino protecting group) with a suitable deprotecting agent to form said compound of formula (I).

[0082] The preparation of the compounds of formula (I) of the present invention can be carried out by sequential or convergent synthetic routes. The synthesis of the present invention is shown in the following general scheme. The skills required to carry out the reactions and purification of the resulting products are known to those skilled in the art. The substituents and indices used in the following process descriptions have the meanings given herein unless indicated to the contrary.

[0083] If one of the starting materials, intermediates, or compounds of formula (I) contains one or more functional groups that are not stable or reactive under the reaction conditions of one or more reaction steps, suitable protecting groups (such as those described in "Protective Groups in Organic Chemistry," by T.W. Greene and P.G.M. Hutts, 5th Edition, 2014, John Wiley & Sons, NY) can be introduced prior to a critical step by applying methods well known in the art. Such protecting groups can be removed at a later stage of the synthesis using standard methods described in the literature.

[0084] When the starting material or intermediate contains a stereocenter, the compound of formula (I) can be obtained as a mixture of diastereomers or enantiomers, which can be separated by methods well known in the art, such as chiral HPLC, chiral SFC, or chiral crystallization. Racemates can be separated into their antipodes via diastereomeric salts, for example, by crystallization with an optically pure acid, or by separating the antipodes by specific chromatographic methods using either a chiral adsorbent or a chiral eluent. Starting materials and intermediates containing a stereocenter can also be separated to obtain diastereomerically / enantiomerically enriched starting materials and intermediates. The use of such diastereomerically / enantiomerically enriched starting materials and intermediates in the synthesis of the compound of formula (I) generally results in the respective diastereomerically / enantiomerically enriched compounds of formula (I).

[0085] Those skilled in the art will recognize that in the synthesis of compounds of formula (I), unless otherwise desired, an "orthogonal protecting group strategy" can be applied to cleave some protecting groups one at a time without affecting other protecting groups in the molecule. The principle of orthogonal protection is well known in the art and has been described in the literature (e.g., Barany and R.B. Merrifield, J. Am. Chem. Soc. 1977, 99, 7363; H. Waldmann et al., Angew. Chem. Int. Ed. Engl. 1996, 35, 2056).

[0086] Those skilled in the art will recognize that the reaction sequence may vary depending on the reactivity and nature of the intermediates.

[0087] More specifically, compounds of formula (I) can be prepared by the methods described below, the methods described in the Examples, or similar methods. Suitable reaction conditions for the individual reaction steps are known to those skilled in the art. For literature-described reaction conditions affecting the described reactions, see, for example, Comprehensive Organic Transformations: A Guide to Functional Group Preparations, 2nd Edition, Richard C. Larock, John Wiley & Sons, New York, NY, 1999. The reactions could be easily carried out with or without a solvent. There are no particular restrictions on the nature of the solvent used, so long as it does not adversely affect the reaction or the reagents involved and is capable of dissolving the reagents to at least some extent. The described reactions can occur over a wide range of temperatures, and the exact reaction temperature is not critical to the present invention. The described reactions are conveniently carried out at a temperature range from -78°C to reflux. The reaction time required for the reaction can also vary widely, depending on many factors, particularly the reaction temperature and the nature of the reagents. However, a period of 0.5 hours to several days is usually sufficient to obtain the intermediates and compounds described. The reaction sequence is not limited to the sequence shown in the scheme, but the order of the reaction steps can be freely changed depending on the starting materials and their respective reactivities.

[0088] If the starting materials or intermediates are not commercially available or their synthesis is not described in the literature, they can be prepared analogously to existing procedures for similar analogs or as outlined in the experimental section.

[0089] The present compounds of formula (I), or pharmaceutically acceptable salts thereof, can be prepared by the process described below (Scheme 1), together with synthetic methods known in the art of organic chemistry, or modifications and derivatizations familiar to those skilled in the art. [ka]

[0090] Suitable starting materials for the preparation of compounds of formula (I) are compounds of formula (II) 2 is F or Cl, and X 1 is already R 1 or Br) or a nitro compound followed by R 1 The -CO2 alkyl can be synthesized to the following: Compounds of formula (II) can be reacted with a suitably protected cysteine derivative (III) in a solvent such as 1,2-dichloroethane or DMF in the presence of a base such as DIPEA or potassium carbonate at elevated temperature to give compounds of formula (IV). A preferred protecting group (PG) for cysteine derivatives (III) is Boc. The nitro group in compounds of formula (IV) can be reduced using iron in the presence of either hydrogen chloride or ammonium chloride in a solvent mixture of water and ethanol at elevated temperature to give compounds of formula (V). Alternatively, this conversion can be achieved with other reducing agents, such as tin(II) chloride, or by catalytic hydrogenation. Compounds of formula (V) can be cyclized to compounds of formula (VI) using standard amide coupling conditions. Preferably, this cyclization is carried out using 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (50% solution in EtOAc) with a base such as DIPEA in a solvent such as DMF at room temperature. 1 is Cl, Br, I, or a sulfonate group in the presence of a base such as potassium carbonate, optionally with an additive such as potassium iodide, in a solvent such as DMSO or DMF at room temperature to obtain a compound of formula (VIII). Oxidation of the sulfur atom in the compound of formula (VIII) with an appropriate amount of an oxidizing agent such as m-CPBA in a solvent such as DCM at room temperature gives a compound of formula (IX). 1In the case of compounds of formula (VIII) where is Br, this group can be converted to the boronic acid derivative (X) at this stage as described in the scheme below by reaction with bis(pinacolato)diboron and a base such as potassium acetate in the presence of a palladium(0) catalyst, for example 1,1'-bis(diphenylphosphino)ferrocenedichloropalladium(II), in an aprotic solvent such as 1,4-dioxane at elevated temperature to give the boronic acid derivative (X). 1 Compounds of formula (X1) can be obtained by cross-coupling a boronic acid derivative (X) with a heteroaryl halide in an aprotic solvent, such as 1,4-dioxane or THF, at elevated temperatures in the presence of a palladium(0) catalyst, such as 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium(II) or tetrakis(triphenylphosphine)palladium(0) or Xphos Pd G4, and a base, such as potassium carbonate, sodium carbonate, potassium phosphate, or potassium acetate. Alternatively, compounds of formula (XI) can be prepared from the corresponding boronic acid derivative (X) by reacting with a heteroaryl methyl sulfide to produce compounds of formula (X1) in the presence of a palladium(0) precursor catalyst, such as [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium(II), a copper salt, such as copper(I) thiophene-2-carboxylate hydrate, and an additive, such as zinc acetate, in an aprotic solvent, such as THF, at elevated temperatures. Final deprotection provides compounds of formula (I). When the N-protecting group (PG) is Boc, typical conditions for this deprotection step include TFA in a solvent such as DCM at room temperature, hydrogen chloride in a solvent such as dioxane or ethyl acetate at room temperature, or hexafluoroisopropanol at reflux temperature. When the N-protecting group (PG) is Alloc, typical conditions are palladium(0)-catalyzed saponification in an aprotic solvent in the presence of either a nucleophile such as morpholine or a reducing agent such as phenylsilane.

[0091] Furthermore, the substituent R 1 and R 4may contain functional groups that may be modified before removal of the N-protecting group (PG) or that require the use of appropriate protecting groups during synthesis. These protecting groups may be removed before removal of the N-protecting group (PG) or may be removed simultaneously using appropriate methods [Peter G.M.Wuts, Greene's Protective Groups in Organic Synthesis, 5th Edition, Hoboken, NJ: Wiley-Interscience].

[0092] Alternatively, compounds of formula (I) where R1 is triazole can be prepared as shown in Scheme 2. [ka]

[0093] Esters of formula (XI) can be converted to acids of formula (XII) by treatment with MeOH, such as LiOH, in a protic solvent such as MeOH, water, and THF. Coupling of acid (XII) with hydrazine in the presence of a coupling agent such as CDI provides hydrazide (XIII), which is coupled with a 1,2-diketo compound in the presence of ammonium acetate at elevated temperature to provide arylheterosyls of formula (XIV). Oxidation as described in Scheme 1 provides sulfones of formula (XV), and final deprotection as described in Scheme 1 yields the title compounds of formula (I).

[0094] Diketones / ketoaldehydes can be prepared by oxidation of the corresponding methyl ketones with, for example, selenodioxide at elevated temperatures.

[0095] R may contain a boc protecting group, which can be deprotected and acylated with, for example, chloroformic acid as methyl chloroformate chemoselectively in the presence of the unprotected ring amine in formula (I).

[0096] Pharmaceutical Compositions and Administration Another object of the present invention is a pharmaceutical composition comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0097] The compounds of formula (I) and their pharmaceutically acceptable salts can be used as medicines in the form of pharmaceutical preparations.The pharmaceutical preparations can be administered to the body orally (for example, in the form of tablets, coated tablets, sugar-coated tablets, hard and soft gelatin capsules, solutions, emulsions, or suspensions), nasally (for example, in the form of nasal drops), or rectally (for example, in the form of suppositories).However, administration can also be carried out parenterally, such as intramuscularly or intravenously (for example, in the form of injections).Administration can also be carried out topically, for example, transdermally, or in the form of eye drops or ear drops.

[0098] The compounds of formula (I) and their pharmaceutically acceptable salts can be processed with pharmaceutically inert inorganic or organic carriers for the manufacture of pharmaceutical preparations such as tablets, coated tablets, sugar-coated tablets, hard gelatin capsules, injectable solutions or topical preparations, etc. Lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc. can be used as such carriers for tablets, coated tablets, sugar-coated tablets and hard gelatin capsules, for example.

[0099] Suitable carriers for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols, etc. However, depending on the nature of the active substance, no carrier is usually required for soft gelatin capsules.

[0100] Suitable carriers for the production of solutions and syrups are, for example, water, alcohols, polyols, saccharose, glucose, invert sugar, vegetable oil and the like.

[0101] Suitable carriers for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils and the like.

[0102] Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

[0103] Suitable carriers for topical ophthalmic formulations are, for example, cyclodextrin, mannitol, or many other carriers and excipients known in the art.

[0104] Furthermore, the pharmaceutical preparations may contain preservatives, solubilizers, viscosity-increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorings, salts for varying osmotic pressure, buffers, masking agents or antioxidants. The pharmaceutical formulations of the present invention may further contain other therapeutically valuable substances.

[0105] Pharmaceutical products comprising compounds of formula (I), or pharmaceutically acceptable salts thereof, and therapeutically inactive excipients are also an object of the present invention, as are processes for their preparation, which involve bringing one or more compounds of formula (I) and / or their pharmaceutically acceptable salts, and, if desired, one or more other therapeutically valuable substances, together with one or more pharmaceutically acceptable salt excipients, into a galenical dosage form.

[0106] The dosage can vary within a wide range and, of course, must be adjusted to the individual requirements in each specific case. Generally, for oral administration, a daily dosage of about 0.1 mg to 20 mg / kg body weight, preferably 0.5 mg to 4 mg / kg body weight (e.g., about 300 mg / person), preferably divided into 1 to 3 individual doses, each of which may consist of the same amount, would be appropriate. For topical administration, the formulation may contain 0.001% to 15% by weight of the drug, and the required dose, which may be between 0.1 and 25 mg, may be administered as a single dose per day, a single dose per week, multiple doses (2 to 4 times per day), or multiple doses per week. However, it is clear that, where indicated, the upper or lower limits set forth herein may be exceeded.

[0107] The pharmaceutical composition of the present invention can be prepared as follows.

[0108] Preparation of Pharmaceutical Compositions Containing Compounds of the Invention TIFF2025526683000009.tif80170

[0109] Manufacturing Procedure: 1. Mix ingredients 1, 2, 3 and 4 and granulate with purified water. 2. Dry the granules at 50°C. 3. Pass the granules through suitable grinding equipment. 4. Add ingredient 5, mix for 3 minutes and compress in a suitable press. TIFF2025526683000010.tif80170

[0110] Manufacturing Procedure: 1. Mix ingredients 1, 2 and 3 in a suitable mixer for 30 minutes. 2. Add ingredients 4 and 5 and mix for 3 minutes. 3. Fill into suitable capsules. TIFF2025526683000011.tif53170

[0111] Manufacturing Procedure: A portion of a compound of formula (I) is dissolved in a mixture of polyethylene glycol 400 and water for injection. The pH is adjusted to 5.0 with acetic acid. The remaining amount of water is added to adjust the volume to 1.0 ml. The solution is filtered, filled into vials using an appropriate overage, and sterilized.

[0112] Indications The compounds of formula (I) can be used in an effective amount to treat subjects, particularly humans, suffering from cancer.

[0113] In one aspect, the present invention provides a compound of formula (I), as described herein, or a pharmaceutically acceptable salt thereof, for use as a therapeutically active substance.

[0114] In a further aspect, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, for use in treating, preventing and / or delaying the progression of cancer.

[0115] In a further aspect, the present invention provides the use of a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, for the treatment, prevention and / or delay of progression of cancer.

[0116] In a further aspect, the present invention provides the use of a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment, prevention and / or delay of progression of cancer.

[0117] In a further aspect, the present invention provides a method for the treatment, prevention and / or delay of progression of cancer, comprising administering a therapeutically effective amount of a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof.

[0118] As used herein, the terms "treatment" or "treating" and grammatical variations thereof refer to therapeutic therapy. With respect to a particular condition, treating means: (1) ameliorating the condition or one or more biological manifestations of the condition; (2) (a) interfering with one or more points in the biological cascade that leads to or causes the condition, or (b) one or more biological manifestations of the condition; (3) alleviating one or more symptoms, effects, or side effects associated with the condition or its treatment; or (4) slowing the progression of the condition or one or more biological manifestations of the condition. Prophylactic therapy using the methods and / or compositions of the present invention is also contemplated. Those skilled in the art will understand that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially reduce the likelihood or severity of a condition or its biological manifestations, or to delay the onset of such a condition or its biological manifestations. Prophylactic therapy is appropriate when a subject is considered to be at high risk for developing cancer, for example, when the subject has a strong family history of cancer or when the subject has been exposed to a carcinogen.

[0119] As immunotherapeutics that act on immune cells rather than directly on cancer cells, the present disclosure may also be envisioned for use as an anti-cancer vaccine, including approaches in which immune cells are cultured and engineered ex vivo and the molecules disclosed herein are used as a means of costimulating the ex vivo engineered cells.

[0120] In one embodiment, the cancer is a blood cancer, such as lymphoma, leukemia, or myeloma. Blood cancers contemplated herein include one or more leukemias, such as B-cell acute lymphoblastic leukemia ("BALL"), T-cell acute lymphoblastic leukemia ("TALL", acute lymphoblastic leukemia (ALL); one or more chronic leukemias, including but not limited to, chronic myeloid leukemia (CML) and chronic lymphocytic leukemia (CLL); B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, These include, but are not limited to, small cell or large cell follicular lymphoma, malignant lymphoproliferative conditions, mucosa-associated lymphoid tissue (MALT) lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, and "preleukemia," which are a diverse collection of hematological conditions united by the ineffective production (or dysplasia) of myeloid blood cells.

[0121] In further embodiments, the cancer is a non-hematological cancer, such as a sarcoma, carcinoma, or melanoma. Non-hematological cancers contemplated herein include, but are not limited to, neuroblastoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, gastric cancer, brain cancer, lung cancer (e.g., non-small cell lung cancer - NSCLC), pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, uterine cancer, adrenal cancer, and head and neck cancer.

[0122] Co-administration of Compounds of Formula (I) and Other Agents The compound of formula (I) or its salt, or the compound disclosed herein or its pharmaceutically acceptable salt, may be used alone or in combination with other drugs for treatment. For example, the second drug in a combined pharmaceutical formulation or administration regimen may have complementary activity to the compound of formula (I), so that they do not adversely affect each other. The compounds may be administered together in a single pharmaceutical composition or separately. In one embodiment, the compound, or a pharmaceutically acceptable salt, may be co-administered with a cytotoxic agent to treat proliferative diseases and cancer.

[0123] The term "co-administering" refers to simultaneous administration or separate, sequential administration in any manner of a compound of formula (I) or a salt thereof, or a compound disclosed herein or a pharmaceutically acceptable salt thereof, and additional active pharmaceutical ingredient(s), including cytotoxic agents and radiation therapy. If not administered simultaneously, the compounds are administered in close temporal proximity to each other. Furthermore, it does not matter whether the compounds are administered in the same dosage form; for example, one compound may be administered topically and the other compound may be administered orally.

[0124] Typically, any drug with anti-cancer activity can be co-administered.Examples of such drugs can be found in Cancer Principles and Practice of Oncology by V.T.Devita and S.Heilman (editors), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers.Those skilled in the art will be able to identify which combination of drugs is useful based on the specific characteristics of the drug and the disease involved.

[0125] In one aspect, the present invention provides a pharmaceutical composition as described herein, further comprising an additional therapeutic agent.

[0126] In one embodiment, the additional therapeutic agent is a chemotherapeutic agent.

[0127] In one embodiment, the additional therapeutic agent is a cytotoxic agent.

[0128] In one embodiment, the additional therapeutic agent is a cancer immunotherapeutic agent.

[0129] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents the function of cells and / or causes cell death or destruction. Cytotoxic agents include radioisotopes (At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 , and radioactive isotopes of Lu); chemotherapeutic agents; growth inhibitory agents; enzymes and fragments thereof, such as nucleases; and toxins (including fragments and / or variants thereof), such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin.

[0130] Exemplary cytotoxic agents may be selected from anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, pro-apoptotic agents, LDH-A inhibitors, fatty acid biosynthesis inhibitors, cell cycle signaling inhibitors, HDAC inhibitors, proteasome inhibitors, and inhibitors of cancer metabolism.

[0131] "Chemotherapeutic agents" include chemical compounds useful in the treatment of cancer. Examples of chemotherapeutic agents include erlotinib (TARCEVA®, Genentech / OSI Pharm.), bortezomib (VELCADE®, Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX®, AstraZeneca), sunitib (SUTENT®, Pfizer / Sugen), letrozole (FEMARA®, Novartis), and rivaroxaban. is), imatinib mesylate (GLEEVEC®, Novartis), finasunate (VATALANIB®, Novartis), oxaliplatin (ELOXATIN®, Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib (TYKERB®, GSK572016, GlaxoSmithKline), lonafamib (SCH 66336), sorafenib (NEXAVAR®, Bayer Labs), gefitinib (IRESSA®, AstraZeneca), AG1478, alkylating agents such as thiotepa and CYTOXAN® cyclophosphamide, alkylsulfonates such as busulfan, improsulfan, and piposulfan, aziridines such as benzodopa, carboquone, mesuredopa, and uredopa, altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramine ethylenimines and methylameramines, including methylameramine, acetogenins (especially bullatacin and bullatacinone), camptothecins (including topotecan and irinotecan), bryostatin, kallistatin, CC-1065 (including its azozelesin, carzelesin, and bizelesin synthetic analogs), cryptophycins (especially cryptophycin 1 and cryptophycin 8), corticosteroids (including prednisone and prednisolone),cyproterone acetate, finasteride, and dutasteride (5a-reductase inhibitors), vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dolastatins, aldesleukin, talc, duocarmycin (synthetic analogs KW-2189 and CBI-TM I), eleutherobin, pancratistatin, sarcodictyin, spongistatin, nitrogen mustards such as chlorambucil, chromafazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembicine, fenesterine, prednimustine, trofosfamide, uracil mustard, nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine, antibiotics such as enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γII and calicheamicin chol (Angew Chem. Inti. Ed. Engl. 1994) 33:183-186); dynemicins, e.g., dynemicin A, bisphosphonates, e.g., clodronate, esperamicin, and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores, aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), morpholino-doxo rubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, chelamycin, rhodolubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, antimetabolites such as methotrexate and 5-fluorouracil (5-FU), folic acid analogues such as denopterin, methotrexate,Pteropterin, trimetrexate, purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, antiadrenal agents such as aminoglutethimide, mitotane, trilostane, folic acid replacement solutions such as furoic acid, aceglatone, aldophosphamide glycosides , aminolevulinic acid, eniluracil, amsacrine, bestravcil, bisantrene, edatrexate, defofamine, demecolcine, diaziquone, elfomitin, elliptinium acetate, epothilone, etoglucide, gallium nitrate, hydroxyurea, lentinan, lonidynin, maytansinoids such as maytansine and ansamitocins, mitoguazone, mitoxantrone, mopidamol, nitraelin, pentostatin, phenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.), razoxane, rhizoxin, schizofuran, spirogermanium, tenuazonic acid, triaziquone, 2,2',2"-trichlorotriethylamine, trichothecenes (especially T-2 toxin, veracrine A, roridin A, and anguidine), urethane, vindesine, dacarbazine, mannomustine, mitobronitol, mitolactol, pipobroman, gacytosine, arabinoside ("Ara-C"), cyclophosphamide, thiotepa, taxoids such as TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE® (cremophor-free), albumin-engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, 111.), and TAXOTERE® (docetaxel; Sanofi-Aventis), chlorambucil, GEMZAR® (gemcitabine),6-thioguanine, mercaptopurine, methotrexate, platinum analogs such as cisplatin and carboplatin, vinblastine, etoposide (VP-16), ifosfamide, mitoxantrone, vincristine, NAVELBINE® (vinorelbine), novantrone, teniposide, edatrexate, daunomycin, aminopterin, capecitabine (XELODA®), ibandronate, CPT-II, the topoisomerase inhibitor RFS2000, difluoromethylomithine (DMFO), retinoids such as retinoic acid, and pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0132] Chemotherapeutic agents also include: (i) antihormonal agents that act to regulate or inhibit hormone action on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), for example, tamoxifen (including NOLVADEX (registered trademark) and tamoxifen citrate), raloxifene, droxifene, iodoxifene, 4-hydroxytamoxifen, trihydroxyphene, ketoxifene, LY1; 17018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as 4(5)-imidazole, aminoglutethimide, MEGASE® (megstrol acetate), AROMASIN® (exemestane; Pfizer), formestany, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole; Novartis), and ARIMIDEX® (anastrozole; AstraZeneca); (iii) antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, tripterelin, medroxyprogesterone acetate, diethylstilbestrol, Premarin, fluoxymesterone, all-trans-lethiol, (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly agents that inhibit the expression of genes in signal transduction pathways involved in abnormal cell growth, such as PKC-alpha, Ralf, and H-Ras; (vii) ribozymes such as VEGF expression inhibitors (e.g., ANGIOZYME®), HER2 expression inhibitors; (viii) vaccines such as gene therapy vaccines, for example, ALLOVECTIN®, LEUVECTIN®, VAXID®; PROLEUKIN®, rIL-2; topoisomerase I inhibitors such as LURTOTECAN®; ABARELIX® rmRH; and (ix) pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0133] Chemotherapeutic agents also include antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech), cetuximab (ERBITUX®, Imclone), panitumumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech / Biogen Idee), pertuzumab (OMNITARG®, 2C4, Genentech), trastuzumab (HERCEPTIN®, Genentech), tositumomab (Bexxar, Corixia), and the antibody-drug conjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Additional humanized monoclonal antibodies with therapeutic potential as agents in combination with the compounds of the invention include apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, celizumab pegol, cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, nat ... tuzumab, nimotuzumab, norobizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab, pexelizumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, reslizumab, reslivizumab, rovelizumab, lupizumab, sibrotuzumab, siplizumab, sontuzumab, tacatatuzumab tetraxetan, tadoxizumab, talizumab, tefibazumab, tocilizumab, toralizumab, tucotuzumab celmoreukin, tuxituuzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab, and interleukin-12 and anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), an all-human sequence, full-length IgGi lambda antibody engineered to recognize the p40 protein.

[0134] Chemotherapeutic agents also include "EGFR inhibitors," which refer to compounds that bind to or otherwise directly interact with EGFR and inhibit or reduce the signaling activity of EGFR, alternatively referred to as "EGFR antagonists." Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb579 (ATCC CRL HB8506), MAb455 (ATCC CRL HB8507), MAb225 (ATCC CRL8508), MAb528 (ATCC CRL8509) (see U.S. Pat. No. 4,943,533, Mendelsohn et al.), and variants thereof, such as chimerized 225 (C225 or cetuximab, ERBUTIX®) and reconstituted human 225 (H225) (see WO 96 / 40210, Imclone Systems, Inc.). Inc.), IMC-11F8, a fully human EGFR-targeting antibody (Imclone), antibodies that bind to type II mutant EGFR (U.S. Pat. No. 5,212,290), humanized and chimeric antibodies that bind to EGFR as described in U.S. Pat. No. 5,891,996, and human antibodies that bind to EGFR, such as ABX-EGF or panitumumab (WO 98 / 50433, Abgenix / Amgen), EMD55900 (Stragliotto et al. Eur. J. Cancer 32A:636-640 (1996)), EMD7200 (matuzumab) (a humanized EGFR antibody against EGFR that competes for EGFR binding of both EGF and TGF-alpha (EMD / Merck)), the human EGFR antibody, HuMax-EGFR (GenMab), the fully human antibodies known as E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3, and described in U.S. Pat. No. 6,235,883, MDX-447 (Medarex Inc), and mAb806 or humanized mAb806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)).Anti-EGFR antibodies can be conjugated to cytotoxic agents to produce immunoconjugates (see, for example, European Patent Application Publication No. 659,439 A2, Merck Patent GmbH). EGFR antagonists include those described in U.S. Patent Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, 5,866,572, and small molecules such as compounds described in PCT Publications WO 98 / 14451, WO 98 / 50038, WO 99 / 09016, and WO 99 / 24037.Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA®, Genentech / OSI Pharmaceuticals), PD183805 (CI 1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.), ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca), ZM105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca), BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim), Ingelheim), PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide), EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7 -ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butynamide) (Wyeth), AG1478 (Pfizer); AG1571 (SU5271, Pfizer), and dual EGFR / HER2 tyrosine kinase inhibitors, such as lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine).

[0135] Chemotherapeutic agents include "tyrosine kinase inhibitors," such as the EGFR-targeted drugs described in the previous paragraph; small molecule HER2 tyrosine kinase inhibitors, such as TAK165 available from Takeda; CP-724,714, an oral selective inhibitor of ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual HER inhibitors, such as EKB-569 (available from Wyeth), which preferentially binds to EGFR but inhibits both HER2 and EGFR-overexpressing cells; lapatinib (GSK572016, available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033, Pharmacia); Raf-1 inhibitors, such as ISIS, which inhibits Raf-1 signaling. antisense drug ISIS-5132 available from GlaxoSmithKline Pharmaceuticals; non-HER-targeted TK inhibitors such as imatinib mesylate (GLEEVEC®, available from GlaxoSmithKline); multi-targeted tyrosine kinase inhibitors such as sunitinib (SUTENT®, available from Pfizer); VEGF receptor tyrosine kinase inhibitors such as vatalanib (PTK787 / ZK222584, available from Novartis / Schering AG); the MAPK extracellular regulated kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines such as CGP 59326, CGP 60261, and CGP 62706; pyrazolopyrimidine, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrphostins containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those that bind to HER-encoding nucleic acids); quinoxalines (U.S. Patent No. 5,804,396); tryphostins (U.S. Patent No. 5,804,396); ZD6474 (Astra Zeneca);PTK-787 (Novartis / Schering AG); pan-HER inhibitors, such as CI-1033 (Pfizer); Affinitac (ISIS 3521, Isis / Lilly); imatinib mesylate (GLEEVEC®); PKI 166 (Novartis); GW2016 (GlaxoSmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); INC-ICl I (Imclone), rapamycin (sirolimus, RAPAMUNE®); or the following patent publications: U.S. Pat. No. 5,804,396, WO 1999 / 09016 (American Cyanamid), WO 1998 / 43960 (American Cyanamid), 1997 / 38983 (Warner Lambert), 1999 / 06378 (Warner Lambert), 1999 / 06396 (Warner Lambert), 1996 / 30347 (Pfizer, Inc.), 1996 / 33978 (Zeneca), 1996 / 3397 (Zeneca), and 1996 / 33980 (Zeneca).

[0136] Chemotherapeutic agents include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, BCG (raw), bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, and interferon alfa- 2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium, quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, valrubicin, zoledronate, and zoledronic acid, and pharmaceutically acceptable salts thereof.

[0137] Chemotherapeutic agents include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, and hydrocortisone-17- butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasone-17-propionate, fluocortolone caproate, fluocortolone pivalate, and fluprednidene acetate; phenylalanine-glutamine-glycine (PEG) and its D-form (feG) (IMULAN) Immunoselective anti-inflammatory peptides (ImSAIDs) such as BioTherapeutics, LLC; antirheumatic drugs such as azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomide, minocycline, and sulfasalazine; etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), cetolithumab pegol (Cimzia), and golimumab (Symptoms); tumor necrosis factor alpha (TNFα) blockers such as lebrikizumab (Kineret); interleukin 1 (IL-1) blockers such as anakinra (Kineret); T-cell costimulation blockers such as abatacept (Orencia); interleukin 6 (IL-6) blockers such as tocilizumab (ACTEMERA®); interleukin 13 (IL-13) blockers such as lebrikizumab; interferon alpha (IFN) blockers such as rontalizumab; beta 7 integrin blockers such as rhuMAb Beta7; IgE pathway blockers such as anti-M1 prime; secreted homotrimeric LTa3 and membrane-bound heterotrimeric LTa1 / β2 blockers such as anti-lymphotoxin alpha (LTa); radioisotopes (e.g., At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212、P 32 、Pb 212, and radioactive isotopes of Lu); various investigational drugs such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3, and farnesyltransferase inhibitors (L-739749, L-744832); polyphenols such as quercetin, resveratrol, piceatannol, epigallocatechin gallate, theaflavins, flavanols, procyanidins, betulinic acid and its derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin; podophyllotoxin; tegafur (UFTORAL®); bexarotene (TARGRETIN®); clodronate (e.g., BONEFOS®) ) or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), pamidronate (AREDIA®), tiludronate (SKELID®), or risedronate (ACTONEL®); as well as epidermal growth factor receptor (EGF-R); vaccines such as the THERATOPE® vaccine; perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); CCI-779; tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors such as oblimersen sodium (GENASENSE®); pixantrone; lonafarnib (SCH 6636, SARASAR™); and pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of CHOP, which is an abbreviation for the combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, and FOLFOX, which is an abbreviation for the treatment regimen with oxaliplatin in combination with 5-FU and leucovorin (ELOXATIN™).

[0138] In another embodiment, the compound of formula (I) can be co-formulated with a cancer immunotherapeutic agent. Cancer immunotherapeutic agents include, for example, small molecule drugs, antibodies, or other biological or small molecules. Examples of biological cancer immunotherapeutic agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In one aspect, the antibody is a monoclonal antibody. In another aspect, the monoclonal antibody is humanized or human. In another aspect, the antibody is a bispecific antibody.

[0139] In one aspect, the cancer immunotherapeutic agent is either (i) an agonist of a stimulatory (including costimulatory) receptor or (ii) an antagonist of an inhibitory (including co-inhibitory) signal on T cells, both of which result in amplification of antigen-specific T cell responses (often referred to as immune checkpoint modulators).

[0140] Certain stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or coinhibitory receptors are CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fnl4, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTfiR, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin α / TNPβ, TNFR2, TNFα, LT R, lymphotoxin α It is a TNF family molecule that binds to cognate TNF receptor family members, including 1β2, FAS, FASL, RELT, DR6, TROY, and NGFR.

[0141] In one aspect, a T cell response may be stimulated by a combination of a compound of Formula (I) and (i) an antagonist of a protein that inhibits T cell activation (e.g., an immune checkpoint inhibitor), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, Galectin-9, CEACAM-1, BTLA, CD69, Galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and (ii) one or more of B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0142] Other drugs that can be combined with the compound of formula (I) for the treatment of cancer include the antagonist of inhibitory receptors on NK cells or the agonist of activating receptors on NK cells.For example, the compound of formula (I) can be combined with the antagonist of KIR, such as lirilumab.

[0143] Still other agents for combination therapy include agents that inhibit or deplete macrophages or monocytes, including CSF-1R antagonists such as CSF-1R antagonist antibodies, including, but not limited to, RG7155 or FPA-008.

[0144] In another embodiment, the compounds of formula (I) can be used in conjunction with one or more agents that include agonistic agents that ligate positive costimulatory receptors, blocking agents that attenuate signaling through inhibitory receptors, antagonists, and one or more agents that systemically increase the frequency of anti-tumor T cells, agents that overcome different immunosuppressive pathways within the tumor microenvironment (e.g., blocking inhibitory receptor binding (e.g., PD-L1 / PD-1 interactions), depleting or inhibiting Tregs (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), inhibiting metabolic enzymes such as IDO, or reversing / preventing T cell anergy or exhaustion), and innate immune activation and / or inflammation at the tumor site.

[0145] In some embodiments, the cancer immunotherapeutic agent is a CTLA-4 antagonist, such as an antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, YERVOY (ipilimumab) or tremelimumab. In another aspect, the cancer immunotherapeutic agent is a PD-1 antagonist, such as an antagonist PD-1 antibody. Suitable PD-1 antibodies include, for example, OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO 2012 / 145493). Cancer immunotherapeutic agents may also include pidilizumab (CT-011), although its specificity for PD-1 binding has been questioned. Another approach to targeting the PD-1 receptor is a recombinant protein called AMP-224, which consists of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1.

[0146] In another embodiment, the cancer immunotherapeutic agent is a PD-L1 antagonist, such as an antagonist PD-L1 antibody. Suitable PD-L1 antibodies include, for example, TECENTRIQ (atezolizumab) (RG7446; WO 2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO 2007 / 005874), and MSB0010718C (WO 2013 / 79174).

[0147] In another embodiment, the cancer immunotherapeutic agent is a LAG-3 antagonist, for example, an antagonist LAG-3 antibody.Suitable LAG-3 antibodies include, for example, BMS-986016 (WO 2010 / 19570, WO 2014 / 08218), or IMP-731 or IMP-321 (WO 2008 / 132601, WO 2009 / 44273).

[0148] In another embodiment, the cancer immunotherapeutic agent is a CD137 (4-1BB) agonist, such as an agonist CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566 (WO 2012 / 32433).

[0149] In another embodiment, the cancer immunotherapeutic agent is a GITR agonist, for example, an agonistic GITR antibody. Suitable GITR antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (WO2006 / 105021, WO2009 / 009116) and MK-4166 (WO2011 / 028683).

[0150] In another embodiment, the cancer immunotherapeutic agent is IDO antagonist.Suitable IDO antagonist includes, for example, INCB-024360 (WO2006 / 122150, WO2007 / 75598, WO2008 / 36653, WO2008 / 36642), indoximod or NLG-919 (WO2009 / 73620, WO2009 / 1156652, WO2011 / 56652, WO2012 / 142237).

[0151] In another embodiment, the cancer immunotherapeutic agent is an OX40 agonist, for example, an agonistic OX40 antibody.Suitable OX40 antibody includes, for example, MEDI-6383 or MEDI-6469.In another embodiment, the cancer immunotherapeutic agent is an OX40L antagonist, for example, an antagonistic OX40 antibody.Suitable OX40L antagonist includes, for example, RG-7888 (WO06 / 029879).

[0152] In another aspect, the cancer immunotherapeutic agent is a CD40 agonist, e.g., an agonist CD40 antibody. In yet another embodiment, the cancer immunotherapeutic agent is a CD40 antagonist, such as an antagonist CD40 antibody. Suitable CD40 antibodies include, for example, lucatumumab or dacetuzumab.

[0153] In another embodiment, the cancer immunotherapeutic agent is a CD27 agonist, such as an agonist CD27 antibody. Suitable CD27 antibodies include, for example, valilumab.

[0154] In another embodiment, the cancer immunotherapeutic agent is MGA271 (directed against B7H3) (WO 2011 / 109400). [Example]

[0155] The present invention will be more fully understood by reference to the following examples, which, however, should not be construed as limiting the scope of the claims to the examples.

[0156] 1) Preparation Examples Unless otherwise stated, all reactions and intermediates were prepared under an argon atmosphere.

[0157] 1.1) General Procedure Alkylation: General Procedure 1 To a solution of the intermediate of formula (VI) (2.74 mmol) in DMSO (10 mL) at room temperature was added potassium carbonate (1.14 g, 8.23 mmol), potassium iodide (228 mg, 1.37 mmol), and the reagent of formula (VII) (3.29 mmol). The reaction was stirred at room temperature for 2 hours, quenched with water, and extracted twice with EtOAc. The combined organic layers were washed with water, saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The desired product (VIII) was used crude in the next step or purified by flash column chromatography on silica gel or reverse-phase preparative HPLC.

[0158] Oxidation: General Procedure 2 A solution of intermediate of formula (VIII) (2.74 mmol) and m-CPBA (1.18 g, 6.85 mmol) in DCM (10 mL) was stirred at room temperature for 1 day. The reaction was diluted with ethyl acetate and THF, washed with 2N aqueous sodium hydroxide, 1N aqueous HCl, and saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and the solvent removed under reduced pressure. The desired product (IX) was used crude in the next step or purified by flash column chromatography on silica gel or reverse-phase preparative HPLC.

[0159] Formation of boron pinacol esters: General procedure 3a Argon was bubbled through a mixture of the compound of formula (IX) (1 equivalent), bis(pinacolato)diboron (CAS: 73183-34-3) (2 equivalents), and potassium acetate (3 equivalents) in 1,4-dioxane (0.2 M) for 5 minutes. 1,1'-Bis(diphenylphosphino)ferrocenedichloropalladium(II) (0.1 equivalents) was added at room temperature, and the reaction mixture was heated to 80°C for 6 hours. The reaction mixture was cooled to room temperature, diluted with EtOAc and THF, washed with saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and the solvent was removed under reduced pressure. The desired product (X) was used crude in the next step or purified by flash column chromatography on silica gel.

[0160] Boronic Acid Formation: General Procedure 3b Argon was bubbled through a mixture of the compound of formula (IX) (1 equivalent), bis(pinacolato)diboron (CAS: 73183-34-3) (2.5 equivalents), and potassium acetate (3.1 equivalents) in 1,4-dioxane (0.05 M) for 5 minutes. 1,1'-Bis(diphenylphosphino)ferrocenedichloropalladium(II) (0.12 equivalents) was added at room temperature, and the reaction mixture was heated to reflux for 3 hours. The reaction mixture was cooled to room temperature, filtered through celite, and the solvent was removed under reduced pressure. The desired product (X) was purified by preparative HPLC.

[0161] Cross-coupling reactions: General procedure 4a Argon was bubbled through a mixture of the compound of formula (X) (1 equivalent), a heteroaryl halide (1.3 equivalents), and KCO or KOAc (3 equivalents) in 1,4-dioxane and water (0.1 M) for 5 minutes. 1,1'-Bis(diphenylphosphino)ferrocenedichloropalladium(II) (0.2 equivalents) was added, and the reaction mixture was heated to 80 °C for 6 hours and stirred for 5 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc. The catalyst and solids were removed by filtration through decalite. The filtrate was washed with water, saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The desired product (XI) was used crude in the next step and purified by flash column chromatography on silica gel or reverse-phase preparative HPLC.

[0162] Cross-coupling reactions: General procedure 4b Argon was bubbled through a mixture of the compound of formula (X) (1 equivalent), a heteroaryl halide (1.3 equivalents), and Na2CO3 (2.5 equivalents) in 1,4-dioxane and water (0.1 M) for 5 minutes. Tetrakis(triphenylphosphine)palladium(0) (CAS: 14221-01-3) (0.1 equivalents) was added, and the reaction mixture was heated to 90 °C and stirred for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc. The catalyst and solids were removed by filtration through decalite. The filtrate was washed with water, saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The desired product (XI) was used crude in the next step and purified by flash column chromatography on silica gel, preparative TLC, or reverse-phase preparative HPLC.

[0163] Cross-coupling reactions: General procedure 4c Argon was bubbled through a mixture of the compound of formula (X) (1.2 equiv.), heteroaryl(methyl sulfide) (1 equiv.), copper(I) thiophene-2-carboxylate hydrate (CAS: 1292766-17-6) (2.25 equiv.), and zinc acetate (1.2 equiv.) in THF (0.5 M) for 5 minutes. [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.1 equiv.) was added, and the reaction mixture was heated to 70 °C for 6 hours. After cooling, the mixture was filtered through celite, and the filtrate was diluted with EtOAc, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The desired product (XI) was used crude in the next step or purified by flash column chromatography on silica gel, preparative TLC, or reverse-phase preparative HPLC.

[0164] ·Boc deprotection: General procedure 5a To a solution of the intermediate of formula (XI) (38.9 μmol) in DCM (1 mL) was added 4M HCl in dioxane (200 μl, 800 μmol, 20.6 equivalents), and the reaction was stirred at room temperature overnight. The solvent was evaporated, and the residue was suspended in DCM and diethyl ether. The solid was filtered off, washed with diethyl ether, and dried in vacuo to give the desired product (I).

[0165] Boc deprotection: General procedure 5b To a solution of the intermediate of formula (XI) (0.25 mmol) in EtOAc (4 mL) was added HCl / EtOAc (4.0 mL, 16 mmol, 63 equiv) at 0° C. The reaction mixture was stirred at 20° C. for 3 hours and then concentrated in vacuo. The remaining residue was purified by preparative HPLC and dried by lyophilization to give the desired product (I).

[0166] Boc deprotection: General procedure 5c A solution of intermediate (XI) (22.7 μmol) in 1,1,1,3,3,3-hexafluoropropan-2-ol (1.5 mL) was stirred at reflux for 5 days, the solvent was evaporated, and the remaining residue was dried under high vacuum to give the desired product (I).

[0167] Boc deprotection: General procedure 5d To a solution of the intermediate of formula (XI) (0.25 mmol) in 1,1,1,3,3,3-hexafluoropropan-2-ol (4 mL) was added HCl / dioxane or HCl / EtO (0.5 mmol, 2 equivalents) at 0 °C. The reaction mixture was stirred at 20 °C for 2 h. The solvent was evaporated, and the resulting solid was dissolved in DCM and concentrated again to remove traces of 1,1,1,3,3,3-hexafluoropropan-2-ol. This process was repeated twice, followed by drying under high vacuum to give the desired product (I).

[0168] Boc deprotection: General procedure 5e A solution of the intermediate of formula (XI) (0.2 mmol) in DCM (1.5 mL) and TFA (0.5 mL, 0.02 mmol, 1.0 equiv.) was stirred at 20 °C for 0.3 h. The reaction was carefully basified with a saturated solution of NaHCO3 to pH = 8 at 0-5 °C. The mixture was extracted with DCM (10 mL × 3). The organic layer was washed with brine (10 mL), dried over Na2SO4, and concentrated under vacuum below 35 °C. The residue was dissolved in EtOAc (30 mL), and 4 M HCl (0.1 mL) was added dropwise to the mixture with stirring at 0-5 °C. The mixture was concentrated under vacuum below 35 °C, dissolved in deionized water (20 mL), and lyophilized to give the desired product (I).

[0169] 1.2) Synthesis of Examples Example 1 (3R)-3-Amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) (2R)-3-(4-bromo-5-fluoro-2-nitro-phenyl)sulfanyl-2-(tert-butoxycarbonylamino)propanoic acid [ka] To a solution of N-Boc-L-cysteine (CAS: 20887-95-0) (23.3 g, 105 mmol, 1.0 equiv.) and 1-bromo-2,4-difluoro-5-nitrobenzene (CAS: 345-24-4) (25.0 g, 105 mmol, 1.0 equiv.) in THF (150 mL) was added Na2CO3 (22.3 g, 210.3 mmol, 2.0 equiv.). The mixture was stirred at 50 °C for 12 h and then filtered. The filtrate was diluted with HO (200 mL) and extracted with EtOAc (3 × 300 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude title compound (32.6 g, 74.2 mmol, 71% yield) was obtained as a yellow oil, which was used in the next step without further purification. MS(ESI):340.8[M-isobutene-CO2+H]+.

[0170] Step b) (2R)-3-(2-amino-4-bromo-5-fluoro-phenyl)sulfanyl-2-(tert-butoxycarbonylamino)propanoic acid [ka] To a solution of (2R)-3-(4-bromo-5-fluoro-2-nitro-phenyl)sulfanyl-2-(tert-butoxycarbonylamino)propanoic acid (32.6 g, 74.2 mmol, 1.0 equiv) in THF (150 mL) and HO (100 mL) was added Fe (12.43 g, 222.6 mmol, 3.0 equiv) and NHCl (11.91 g, 222.6 mmol, 3.0 equiv), and the mixture was stirred at 50 °C for 16 h. The reaction mixture was filtered, and the filtrate was extracted with EtOAc (3 × 300 mL). The combined organic layers were washed with brine (500 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give the remaining crude title compound (28.1 g, 68.66 mmol, 93% yield) as a brown oil, which was used in the next step without further purification. MS(ESI):353.0[M-isobutene+H] +

[0171] Step c) tert-butyl N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepin-3-yl]carbamate [ka] To a solution of (2R)-3-(2-amino-4-bromo-5-fluoro-phenyl)sulfanyl-2-(tertbutoxycarbonylamino)propanoic acid (28.1 g, 68.6 mmol, 1.0 equiv) in THF (150 mL) was added T3P (52.4 g, 82.3 mmol, 49.00 mL, 50% in EtOAc, 1.2 equiv) and DPIEA (17.7 g, 137.3 mmol, 23.92 mL, 2.0 equiv), and the mixture was stirred at 65° C. for 2 h. The reaction mixture was quenched by the addition of HO (100 mL) and then extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure, and the remaining residue was triturated with MTBE at 25 °C for 30 min to give the title compound (5.4 g, 13.8 mmol, 20% yield) as a white solid. MS (ESI): 334.9 [M-isobutene + H] + .

[0172] Step d) tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepin-3-yl]carbamate (2.7 g, 6.9 mmol) and 1-(bromomethyl)-4-chloro-benzene (CAS: 622-95-7) in analogy to general procedure 1 and obtained as a white solid (5.4 g, 76% yield). MS (ESI): 517.1 [M+H] + .

[0173] Step e) tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate (1.2 g, 2.33 mmol, 1 equiv.) in analogy to general procedure 2 and obtained as a white solid (1.26 g, 97.8% yield). MS (ESI): 491.1 / 493.1 [M-isobutene + H] + .

[0174] Step f) tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (1.2 g, 2.33 mmol, 1 equiv.) in analogy to general procedure 3a and obtained as a light brown solid (699 mg, 43% yield). MS (ESI): Boronic ester cleavage observed in MS: 457.2 ([M-(2,3-dimethyl-butadiene)-isobutene + H] + .

[0175] Step g) tert-butyl N-[(3R)-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (60 mg, 0.1 mmol, equivalents: 1) with 2-bromo-4-tert-butyl-pyridine (CAS: 50488-34-1) in analogy to general procedure 4a and obtained as a white solid (21.8 mg, 35% yield). MS (ESI): 602.4 [M+H] + .

[0176] Step h) (3R)-3-amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (21.8 mg, 0.036 mmol, equivalents: 1) in analogy to general procedure 5a and obtained as a light brown powder (16.8 mg, 85% yield). MS (ESI): 502.3 [M + H] + .

[0177] Examples 2-7 and 31 in the table below were prepared similarly to Example 1 using the appropriate bromide or chloride. TIFF2025526683000021.tif233170TIFF2025526683000022.tif97170

[0178] Example 5 (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (Example 1, step e) (150 mg, 0.274 mmol, equivalents: 1) in analogy to general procedure 3b, and obtained as a brown oil (141 mg, 100% yield). MS (ESI): 457,4 [M-isobutene + H] + .

[0179] Step b) tert-Butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[4-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (141 mg, 0.274 mmol, 1 equiv.) with 2-chloro-4-(trifluoromethyl)pyridine (CAS: 81565-18-6) in analogy to general procedure 4b, and obtained as a white solid (120 mg, 71% yield). MS (ESI) 558.1 [M-isobutene + H] + .

[0180] Step c) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[4-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (120 mg, 0.195 mmol, equivalents: 1) in analogy to general procedure 5b, and obtained as a white powder (34.8 mg, 0.060 mmol, 30% yield). MS (ESI): 514.5 [M + H] + .

[0181] Intermediate 1 6-tert-butyl-3-chloro-1,2,3-triazine [ka] Step a) 6-tert-butyl-3-amino-1,2,3-triazine and 5-tert-butyl-3-amino-1,2,3-triazine [ka] A solution of 1,1-dibromo-3,3-dimethyl-butan-2-one (4.67 g, 18 mmol, 1.0 equiv), aminoguanidine hydrochloride (2.0 g, 18 mmol, 1.0 equiv), and potassium acetate (5.33 g, 54 mmol, 3.0 equiv) in ethanol (30 mL) was stirred at 80 °C for 12 h. After cooling, the reaction was poured into water (100 mL) and extracted with EtAc (300 mL). The organic phase was washed with water (300 mL), brine (100 mL), dried over Na SO , and concentrated in vacuo. The crude product was purified by silica gel column chromatography (PE:EtAc = 1:0 to 1:1) to give 6-tert-butyl-1,2,4-triazin-3-amine (130 mg, 0.85 mmol, 3.3% yield) as a pale yellow solid (MS(ESI): 153.2 [M+H] + .), 6-tert-butyl-1,2,4-triazin-3-amine was obtained, which was further purified by preparative HPLC to give 5-tert-butyl-1,2,4-triazin-3-amine (1.9 g, 12 mmol, 57% yield) as a pale yellow solid. MS(ESI): 153.2 [M+H] + .

[0182] Step b) 6-tert-butyl-3-chloro-1,2,3-triazine [ka] To a solution of 6-tert-butyl-1,2,4-triazin-3-amine (130 mg, 0.85 mmol, 1.0 equiv.) and copper(II) chloride (137 mg, 1 mmol, 1.2 equiv.) in acetonitrile (2 mL) was added tert-butyl nitrite (132 mg, 1.28 mmol, 1.5 equiv.) dropwise at 20 °C. The mixture was stirred at 60 °C for 1 h. The mixture was poured into water (30 mL) and extracted with EtOAc (50 mL). The organic phase was washed with brine (50 mL), dried over Na SO , and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (PE:EtAc = 1:0 to 1:1) to give the title compound (70.0 mg, 0.41 mmol, 42% yield) as a yellow solid. MS (ESI): 172.2 [M+H] + .

[0183] Examples 9-12 in the table below were prepared similarly to Example 5 using the appropriate bromide or chloride. TIFF2025526683000030.tif200170

[0184] Example 13 (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) Methyl (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylate [ka] A solution of methyl (3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylate (CAS: 2002449-37-66) (350 mg, 0.7 mmol, 1 equiv.) was dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (18.6 mL). 4 M HCl in 1,4-dioxane (25.7 mg, 21.48 μL, 0.707 mmol, 1 equiv.) was added to the solution, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo. The remaining residue was dissolved in DCM, and the solution was poured into water. NaHCO3 and 1 M NaOH were added to the aqueous phase to adjust the pH to 13. The aqueous phase was extracted with DCM (2 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound as an orange solid (268 mg, 77% yield), which was used crude in the next step. MS (ESI) 395.1 [M+H] + .

[0185] Step b) Methyl (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylate [ka] To a solution of methyl (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylate (159 mg, 0.4 mmol, 1 equiv.) in ethyl acetate (0.415 mL) was added Na2CO3 (128 mg, 1.21 mmol, 3 equiv.) in water (0.3 mL). Allyl chloroformate (58.2 mg, 51.54 µL, 0.48 mmol, 1.2 equiv.) was added slowly while maintaining the reaction mixture temperature at 20-25 °C, and the resulting biphasic mixture was stirred vigorously at room temperature for 1 h. The mixture was diluted with water (100 mL) and EtOAc (50 mL). The layers were separated, and the aqueous layer was extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (100 mL) and dried over sodium sulfate. The crude product was purified by column chromatography on silica gel (0-30% EtOAc in heptane) to give the title compound (202 mg, 99% yield) as an orange viscous oil. MS (ESI) 479.2 [M+H] + .

[0186] Step c) (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylic acid [ka] To a solution of methyl (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylate (202 mg, 0.422 mmol, 1 equiv.) in methanol (2.52 mL), THF (2.52 mL), and water (0.619 mL) was added lithium hydroxide monohydrate (26.5 mg, 0.63 mmol, 1.5 equiv.). The mixture was stirred at 22°C for 1.5 hours. The solvent was concentrated in vacuo, and the remaining residue was dissolved in EtOAc and washed with 1N aqueous HCl and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound (190 mg, 78% yield) as an orange viscous oil. MS (ESI) 465.2 [M+H] + The product was used as received in the next step.

[0187] Step d) Allyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(hydrazinecarbonyl)-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate [ka] To a solution of (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine-7-carboxylic acid (190 mg, 0.4 mmol, 1 equiv.) in THF (1.75 mL) was added CDI (86.1 mg, 0.53 mmol, 1.3 equiv.) in one portion, and the mixture was stirred for 2 h. This solution was then added in one portion to a solution of hydrazine hydrate (95.9 mg, 93.11 μL, 1.23 mmol, 3 equiv.) in THF (0.29 mL), and the mixture was stirred at room temperature for 1 h. The reaction mixture was partitioned between EtOAc and water, and brine (5 mL) was added. The layers were separated, and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (0-100% EtOAc in heptane) to give the title compound (109 mg, 53% yield) as a red solid. MS (ESI) 479.1 [M+H] + .

[0188] Step e) Allyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate [ka] A microwave vial was charged with allyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(hydrazinecarbonyl)-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate (55 mg, 0.115 mmol, 1 equiv.), 1-phenyl-1,2-propanedione (CAS: 579-07-7) (17.0 mg, 15.47 uL, 0.11 mmol, 1 equiv.), and ammonium acetate (88.5 mg, 1.15 mmol, 10 equiv.) in acetic acid (0.574 mL). The reaction mixture was heated to 140° C. for 10 minutes under microwave irradiation and then heated again to 180° C. for 5 minutes. The reaction mixture was diluted with DCM and water was added. The layers were separated, and the aqueous layer was extracted with DCM. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound (30 mg, 42%) as an off-white solid. MS (ESI) 590.2 [M+H] + The product was used directly in the next step.

[0189] Step f) Allyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] To a solution of allyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate (29 mg, 0.049 mmol, 1 equiv.) in DCM (0.98 mL) was added 3-chloroperbenzoic acid (22.0 mg, 0.098 mmol, 2 equiv.), and the solution was stirred at room temperature for 4 h. The solution was diluted with DCM and washed with 1 M NaOH solution. The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The crude material was purified by column chromatography on silica gel (0-100% EtOAc in heptane) to give the title compound (14.2 mg, 42%) as an off-white solid. MS (ESI) 622.2 [M+H] + .

[0190] Step g) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] To a mixture of tetrakis(triphenylphosphine)palladium (1.3 mg, 0.001 mmol, 0.050 equiv.) and allyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (14 mg, 0.023 mmol, 1 equiv.) in DCM (0.13 mL) under an argon atmosphere, phenylsilane (12.1 mg, 13.87 µL, 0.113 mmol, 5 equiv.) was added, and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with water and DCM, the phases were separated, and the pH of the aqueous layer was adjusted to 13 by adding aqueous NaHCO3 and 1 M aqueous NaOH. The aqueous layer was then extracted with DCM (2 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by preparative HPLC to give the title compound (3.1 mg, 19% yield) as a white solid. MS (ESI) 538.1 [M+H] + .

[0191] Example 14 (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepin-3-yl]carbamate (600 mg, 1.53 mmol) with 1-(chloromethyl)-4-isopropoxy-benzene (CAS: 40141-12-6) in analogy to general procedure 1 and obtained as a dark red oil (800 mg, 97% yield). MS (ESI): 541.1 [M+H] + .

[0192] Step b) tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate (780 mg, 1.45 mmol, 1 equiv.) in analogy to General Procedure 2 and obtained as a pale yellow solid (550 mg, 67% yield). MS (ESI): 592.9 [M+Na] + .

[0193] Step c) tert-butyl N-[(3R)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (300 mg, 0.525 mmol, 1 equiv.) in analogy to general procedure 3a and obtained as a colorless oil (150 mg, 46% yield).

[0194] Step d) tert-butyl N-[(3R)-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (140 mg, 0.226 mmol, equivalents: 1) and 5-tert-butyl-3-chloro-1,2,4-triazine (CAS: 83413-03-0) in analogy to general procedure 4a, and obtained as a white solid (40 mg, 28% yield). MS (ESI): 628.2 [M+H] + .

[0195] Step e) (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (35 mg, 0.056 mmol, equivalents: 1) in analogy to general procedure 5b, and obtained as a white powder (25.5 mg, 0.05 mmol, 79% yield). MS (ESI): 528.1 [M + H] + .

[0196] Example 15 (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka]

[0197] Step a) tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazin-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (100 mg, 0.19 mmol, formula: 1) and 3-chloro-N,N-diethyl-1,2,4-triazin-5-amine (CAS: 1247362-85-1) in analogy to general procedure 4c, and obtained as a yellow solid (6 mg, 5% yield). MS (ESI): 619.2, [M+H] + .

[0198] Step b) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazin-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (6 mg, 0.01 mmol, Formula 1) in analogy to general procedure 5b, and obtained as a light brown solid (3.2 mg, 0.01 mmol, 58% yield). MS (ESI): 519.0 [M+H] + .

[0199] Example 16 in the table below was prepared similarly to Example 15 using the appropriate boronic ester. TIFF2025526683000048.tif104170

[0200] Example 17 (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) (5E)-3-Methylsulfanyl-1,2,4-triazine-5-carbaldehyde oxime [ka] To a stirred suspension of powdered potassium hydroxide (3088 mg, 55.0 mmol, 7.0 equiv.) in dry DMSO (20 mL) was added a solution of 3-methylthio-1,2,4-triazine (1000 mg, 7.86 mmol, 1.0 equiv.) and nitromethane (1439 mg, 23.59 mmol, 3.0 equiv.) in DMSO (2 mL) in one portion at 25 °C, and the mixture was then stirred at 25 °C under N for 16 h. The reaction was poured into water (200 mL) and extracted with EtOAc (100 mL × 2). The aqueous phase was acidified to pH 2 with 1 N HCl and extracted with EtOAc (100 mL × 3). The combined organic phase was washed with brine (200 mL × 2), dried over anhydrous NaSO, and concentrated to give the crude product, which was purified by column chromatography on silica gel (PE / EtAc = 5 / 1 to 3 / 1). The eluent was concentrated in vacuo to give the title compound (350 mg, 2.06 mmol, 24% yield) as a yellow solid. MS (ESI): 171.1 [M+H] + .

[0201] Step b) 3-methylsulfanyl-1,2,4-triazine-5-carbonitrile [ka] To a solution of (5E)-3-methylsulfanyl-1,2,4-triazine-5-carbaldehyde oxime (200 mg, 1.18 mmol, 1.0 equiv.) and triethylamine (0.02 mL, 0.12 mmol, 0.1 equiv.) in toluene (5 mL) was added isocyanatobenzene (0.25 mL, 2.35 mmol, 2.0 equiv.) at 25 °C. The mixture was then degassed three times with N, heated to reflux, and stirred for 2 h. The mixture was cooled to room temperature and then filtrated. The cake was filtered and washed with EtOAc (3 mL). The filtrate was diluted with EtOAc (5 mL), quenched with brine (5 mL × 2), dried over anhydrous NaSO, and concentrated to give the crude product, which was purified by column chromatography on silica gel (PE / EtAc = 8 / 1 to 4 / 1). The eluent was concentrated in vacuo to give the title compound (140 mg, 0.92 mmol, 76% yield) as a dark red oil. MS (ESI): 153.1 [M+H] +.

[0202] Step c) 1,1-difluoro-4-(3-methylsulfanyl-1,2,4-triazin-5-yl)-1,4-thiazinane [ka] To 4,4-difluoropiperidine (47.7 mg, 0.39 mmol, 1.0 equiv.) was added 3-methylsulfanyl-1,2,4-triazine-5-carbonitrile (60.0 mg, 0.39 mmol, 1.0 equiv.), and the mixture was stirred at 25° C. for 1 hour. The mixture was diluted with EtOAc (5 mL) and concentrated in vacuo onto silica gel (500 mg). The residue was purified by column chromatography on silica gel (PE / EtAc=2 / 1 to 1 / 1), and the eluent was concentrated in vacuo to give the title compound (80.0 mg, 0.32 mmol, 80% yield) as a pale solid. MS (ESI): 247.0 [M+H] + .

[0203] Step d) tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (124.9 mg, 0.24 mmol, 1.2 equiv.) and 5-(4,4-difluoro-1-piperidyl)-3-methylsulfanyl-1,2,4-triazine in analogy to general procedure 4c, and obtained as a white solid (50 mg, 35% yield). MS (ESI) 667.1 [M+H] + .

[0204] Step e) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (50 mg, 0.075 mmol, equivalents: 1) in analogy to general procedure 5b, and obtained as a light brown solid (19.4 mg, 0.030 mmol, 43% yield). MS (ESI): 566.9 [M+H] + .

[0205] Example 18 Methyl 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylate [ka]

[0206] Step a) tert-butyl 3-ethyl-5-[methoxy(methyl)carbamoyl]piperidine-1-carboxylate [ka] To a solution of 3-tert-butoxycarbonyl-3-azabicyclo[3.1.1]heptane-1-carboxylic acid (3.2 g, 13.2 mmol, 1.0 equiv.), N,N-diisopropylethylamine (5.78 mL, 33.1 mmol, 2.5 equiv.), 1-hydroxybenzotriazole (2.69 g, 19.8 mmol, 1.5 equiv.), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.81 g, 19.8 mmol, 1.5 equiv.) in DMF (35 mL) was added O,N-dimethylhydroxylamine hydrochloride (1.68 g, 17.2 mmol, 1.3 equiv.) at 25 °C. The mixture was stirred at 25 °C for 16 h and poured into water (40 mL). The aqueous phase was extracted with EtOAc (40 mL × 3). The combined organic phase was washed with brine (90 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (PE: EtOAc 20-60%) to give the title compound (3.61 g, 12.7 mmol, 95% yield) as a white solid. MS (ESI): 229.1 [M-isobutene + H] + .

[0207] Step b) tert-butyl 1-acetyl-3-azabicyclo[3.1.1]heptane-3-carboxylate [ka] To a solution of tert-butyl 3-ethyl-5-[methoxy(methyl)carbamoyl]piperidine-1-carboxylate (3.61 g, 12.0 mmol, 1.0 equiv.) in THF (50 mL) was added methylmagnesium bromide (6.01 mL, 18 mmol, 1.5 equiv.) at -20 °C, and the mixture was stirred at 25 °C for 3 h. The reaction was quenched by adding saturated aqueous NH4Cl (50 mL) at 0 °C. The aqueous phase was extracted with EtOAc (50 mL × 3). The combined organic phases were washed with brine (120 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (PE / EtOAc 20-100%) to give the title compound (2.74 g, 10.7 mmol, 89% yield) as a colorless oil. MS (ESI): 184.0 [M+H] + .

[0208] Step c) tert-Butyl 1-oxyaldehyde-3-azabicyclo[3.1.1]heptane-3-carboxylate [ka] To a solution of tert-butyl 1-acetyl-3-azabicyclo[3.1.1]heptane-3-carboxylate (400 mg, 1.67 mmol, 1.0 equiv.) in 1,4-dioxane (4 mL) and water (1 mL) was added selenium dioxide (380 mg, 3.42 mmol, 2.05 equiv.) at 25° C. The mixture was heated to 80° C. and stirred for 16 hours. The reaction was filtered and washed with EtOAc (20 mL×3). The filtrate was concentrated in vacuo, purified by preparative HPLC, and lyophilized to give the title compound (344 mg, 1.36 mmol, 81% yield) as a pale yellow solid. MS (ESI): 198.2 [M-isobutene + H] + .

[0209] Step d) tert-butyl 1-(3-methylsulfanyl-1,2,4-triazin-5-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate and tert-butyl 1-(3-methylsulfanyl-1,2,4-triazin-6-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate [ka] To a solution of tert-butyl 1-oxyaldehyde-3-azabicyclo[3.1.1]heptane-3-carboxylate (172 mg, 0.68 mmol, 1.0 equiv.) and sodium bicarbonate (85.5 mg, 1.02 mmol, 1.5 equiv.) in ethanol (2 mL) was added 1-amino-2-methyl-isothiourea hydroiodide (158 mg, 0.68 mmol, 1.0 equiv.) in water (2 mL) at 25 °C, and the mixture was stirred at 25 °C for 4 hours. The reaction mixture was poured into water (4 mL). The aqueous phase was extracted with EtOAc (4 mL × 3). The combined organic phase was washed with brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product, which was purified by preparative TLC (PE: EtOAc = 3:1) to give tert-butyl 1-(3-methylsulfanyl-1,2,4-triazin-5-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate (244 mg, 0.76 mmol, 111% yield) as a pale yellow oil and tert-butyl 1-(3-methylsulfanyl-1,2,4-triazin-6-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate (30.0 mg, 0.09 mmol, 13% yield) as a pale red oil. MS (ESI) 323.1 [M+H] + .

[0210] Step e) tert-butyl 1-[3-[(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylate [ka] The title compound was prepared from (3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (232 mg, 0.45 mmol, 1.2 equivalents) and tert-butyl 1-(3-methylsulfanyl-1,2,4-triazin-5-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate in analogy to general procedure 4c, and obtained as an orange oil (162 mg, 58% yield). MS (ESI): 687.0 [M-Boc-isobutene + H] + .

[0211] Step f) (3R)-3-amino-7-[5-(3-azabicyclo[3.1.1]heptan-1-yl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared by the procedure of general procedure 5b, similar to that of tert-butyl 1-[3-[(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ 6[1,5-benzothiazepin-7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylate (152 mg, 0.205 mmol, equiv.: 1) and obtained as an orange solid (145 mg, 100% yield). MS (ESI): 543.1 [M+H] + .

[0212] Step g) Methyl 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylate [ka] To a solution of (3R)-3-amino-7-[5-(3-azabicyclo[3.1.1]heptan-1-yl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one (30.0 mg, 0.050 mmol, 1 equiv.) in DCM (1 mL) was added dimethyl dicarbonate (0.01 mL, 0.050 mmol, 1 equiv.) at 0° C. The mixture was stirred at 25° C. for 30 minutes. The solvent was concentrated in vacuo, and the remaining residue was purified by preparative HPLC to give the title compound (20.4 mg, 0.030 mmol, 66% yield) as a light brown solid. MS (ESI) 600.9 [M+H] + .

[0213] Example 19 (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) 5-(bromomethyl)-3-methylsulfanyl-1,2,4 triazine [ka] To a solution of 5-methyl-3-methylsulfanyl-1,2,4-triazine (20.0 g, 141.6 mmol, 1.0 equiv.) and N-bromosuccinimide (22.69 g, 127.4 mmol, 0.9 equiv.) in MeCN (400 mL) was added AIBN (6.79 g, 41.33 mmol, 0.29 equiv.). The mixture was stirred at 25 °C for 3 h. The reaction mixture was poured into brine (100 mL) and extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (120 mL × 2), dried over Na2SO4, concentrated in vacuo, and purified by silica gel column chromatography (PE / EtOAc = 2-10%) to give the title compound (2500 mg, 11.36 mmol, 5% yield) as a light brown solid.

[0214] Step b) 3-methylsulfanyl-5-(methylsulfonylmethyl)-1,2,4-triazine [ka] To a mixture of sodium methanesulfinate (2899 mg, 28.4 mmol, 2.5 equiv.) and potassium iodide (0.6 mL, 11.36 mmol, 1.0 equiv.) in DMF (52 mL) was added 5-(bromomethyl)-3-methylsulfanyl-1,2,4-triazine (2500 mg, 11.3 mmol, 1.0 equiv.) at 25 °C, and the mixture was stirred at 50 °C for 16 h. The solution was diluted with EtOAc (100 mL), washed with brine (50 mL × 3), dried over anhydrous Na2OS4, concentrated in vacuo, and purified by silica gel column chromatography (PE / EtAc = 10 / 1 to 1 / 1). The eluent was concentrated in vacuo to give the title compound (220 mg, 1.0 mmol, 8% yield) as a brown oil. MS (ESI): 220.2 [M+H] + .

[0215] Step c) 3-methylsulfanyl-5-(1-methylsulfonylcyclopropyl)-1,2,4-triazine [ka] A solution of 3-methylsulfanyl-5-(methylsulfonylmethyl)-1,2,4-triazine (330 mg, 1.5 mmol, 1.0 equiv.), 1-bromo-2-chloroethane (0.17 mL, 2.11 mmol, 1.4 equiv.), potassium iodide (0.02 mL, 0.3 mmol, 0.2 equiv.), and K2CO3 (519.2 mg, 3.76 mmol, 2.5 equiv.) in DMF (15 mL) was heated to 60 °C and stirred for 16 h under N2. Cs2CO3 (560 mg, 1.72 mmol, 1.14 equiv.) and potassium iodide (0.02 mL, 0.3 mmol, 0.2 equiv.) were added to the mixture, which was then stirred at 60 °C for an additional 16 h. The mixture was diluted with EtOAc (20 mL), washed with brine (20 mL x 2), dried over anhydrous NaSO, and concentrated to give the crude material, which was purified by preparative TLC (EtOAc / PE = 1 / 1) to give the title compound (30 mg, 3% yield) as a brown gum. MS (ESI): 282.1 [M+H] + .

[0216] Step d) tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic (62.7 mg, 0.122 mmol, 1.2 equivalents) and 3-methylsulfanyl-5-(1-methylsulfonylcyclopropyl)-1,2,4-triazine in analogy to general procedure 4c, and obtained as a light brown solid (18 mg, 0.030 mmol, 26% yield). MS (ESI): 610.2 [M-isobutene + H] +.

[0217] Step e) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (9 mg, 0.014 mmol, equivalents: 1) in analogy to general procedure 5c, and obtained as a brown solid (4.3 mg, 0.010 mmol, 48% yield). MS (ESI): 566.2 [M+H] + .

[0218] Example 20 (3R)-3-amino-7-[5-(3-chloro-1-methylsulfonyl-propyl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) (3R)-3-amino-7-[5-(3-chloro-1-methylsulfonyl-propyl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (9.0 mg, 0.01 mmol, 1 equiv.) in analogy to general procedure 5b, and obtained as a brown solid (6.8 mg, 0.01 mmol, 74% yield). MS (ESI): 602.2 [M+H] +

[0219] Example 21 (3R)-3-Amino-8-fluoro-1,1-diketo-7-(5-morpholino-3-pyridyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) N'-hydroxy-4-methyl-benzamidine [ka] To a solution of 4-methylbenzonitrile (5000 mg, 42.6 mmol, 1.0 equiv.) in ethanol (143 mL) was added hydroxylamine hydrochloride (3.71 g, 53.3 mmol, 1.25 equiv.), followed by NaHCO3 (17.9 g, 213 mmol, 5 equiv.). The reaction mixture was heated to reflux for 6 h, cooled to room temperature, and concentrated. The resulting white solid was suspended in water and sonicated for 10 min. The mixture was filtered and washed with water. The filter cake was then washed with heptane and dried in vacuo to afford the title compound (5.16 g, 74%) as a white solid. MS (ESI): 151.1 [M+H] +

[0220] Step b) 3-(p-tolyl)-5-(trifluoromethyl)-1,2,4-oxadiazole [ka] N'-Hydroxy-4-methyl-benzamidine (5.61 g, 37.3 mmol, 1 equiv.) was dissolved in THF (superhydrous) (100 mL). The reaction mixture was cooled to 0 °C. TFAA (12.5 g, 8.44 mL, 59.7 mmol, 1.6 equiv.) was then added dropwise. The ice bath was removed, and the mixture was stirred at room temperature for 4 h. The reaction mixture was poured into brine and basified with saturated aqueous NaHCO3 (200 mL) to pH = 7-8. The mixture was then extracted with ethyl acetate (3 × 100 mL), and the combined organic phases were dried over MgSO4, filtered, and concentrated to give the title compound (7.98 g, 88%) as a colorless oil. MS (ESI): 226.9 [MH] -

[0221] Step c) 3-[4-(bromomethyl)phenyl]-5-(trifluoromethyl)-1,2,4-oxadiazole [ka] To a solution of 3-(p-tolyl)-5-(trifluoromethyl)-1,2,4-oxadiazole (7.98 g, 34.9 mmol, 1 equiv.) in acetonitrile (140 mL) was added N-bromosuccinimide (7.47 g, 41.9 mmol, 1.2 equiv.) and 2,2-azobis(2-methylpropionitrile) (1.72 g, 10.49 mmol, 0.3 equiv.). The reaction mixture was stirred at 80 °C for 3.5 h, poured into brine (100 mL), and extracted with ethyl acetate (3 × 150 mL). The combined organic phases were dried over MgSO4, filtered, concentrated, and purified by column chromatography on silica gel (0–10% ethyl acetate in heptane) to give the title compound (4.69 g, 41.49%) as a white solid.

[0222] Step d) N-[(3R)-7-bromo-8-fluoro-4-keto-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester [ka] To a solution of N-[(3R)-7-bromo-8-fluoro-4-keto-3,5-dihydro-2H-1,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester (500 mg, 1.15 mmol, 1 equiv.) in N,N-dimethylformamide (superhydrous) (8.85 mL), 3-[4-(bromomethyl)phenyl]-5-(trifluoromethyl)-1,2,4-oxadiazole (557 mg, 1.73 mmol, 1.5 equiv.), potassium carbonate (476 mg, 3.45 mmol, 3 equiv.), and potassium iodide (95 mg, 0.57 mmol, 0.5 equiv.) were added at room temperature. The reaction mixture was stirred for 2 h. The reaction was quenched with 10 mL of water. The crude material was extracted with AcOEt (3 × 20 mL). The combined organic layers were washed with water (2 x 10 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude material was purified by silica gel column chromatography (6-60% DCM in heptane) to give the title compound (407 mg, 55%) as a yellow solid. MS(ESI): 615.1 [MH] - .

[0223] Step e) N-[(3R)-7-bromo-8-fluoro-1,1,4-triketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester [ka] The title compound was prepared from N-[(3R)-7-bromo-8-fluoro-4-keto-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester (402.7 mg, 0.652 mmol, 1 equiv.) in analogy to general procedure 2 and obtained as a white solid (282 mg, 66% yield). MS (ESI): 685.1 [M+HCOO] - .

[0224] Step f) N-[(3R)-5-(4-chlorobenzyl)-8-fluoro-1,1,4-triketo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester [ka] The title compound was prepared from tert-N-[(3R)-7-bromo-8-fluoro-1,1,4-triketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester (140 mg, 0.216 mmol, 1 eq.) in analogy to general procedure 3a and obtained as a yellow solid (129 mg, 85% yield). MS (ESI): 559.0 [M-2 isobutene + H] + .

[0225] Step g) N-[(3R)-7-(5-tert-butyl-3-pyridyl)-5-(4-chlorobenzyl)-8-fluoro-1,1,4-triketo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester [ka] The title compound was prepared from N-[(3R)-8-fluoro-1,1,4-triketo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester (58 mg, 0.083 mmol, 1 eq.) and 4-(5-bromo-3-pyridyl)morpholine (26.32 mg, 0.108 mmol, 1.3 eq.) in analogy to general procedure 4a, and obtained as a white solid (15.9 mg, 26% yield). MS (ESI): 733.2 [M+H] + .

[0226] Step h) (3R)-3-amino-8-fluoro-1,1-diketo-7-(5-morpholino-3-pyridyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from N-[(3R)-8-fluoro-1,1,4-triketo-7-(5-morpholino-3-pyridyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamic acid tert-butyl ester (15.9 mg, 0.022 mmol, 1 equiv.) in analogy to general procedure 5d and obtained as the hydrochloride salt as a yellow solid (11.5 mg, 79% yield). MS (ESI): 677.3 [M+HCOO] - .

[0227] Intermediate 2 3-Bromo-5-(1-mesyl-1-methyl-ethyl)pyridine [ka] Step a) 3-Bromo-5-(mesylmethyl)pyridine [ka] Sodium methanesulfinate (61 mg, 0.59 mmol, 1.5 eq) and 3-bromo-5-(bromomethyl)pyridine (100 mg, 0.39 mmol, 1 eq) were dissolved in DMSO (0.996 mL) and heated to 50° C. for 2 hours and 30 minutes.

[0228] The reaction is diluted with 100 mL of water, extracted with (3 x 20 mL) AcOEt, and washed with (2 x 10 ml) water and (1 x 20 mL) brine. The organic layers are combined, dried over Na2SO4, and concentrated to give the title compound (80 mg, 67%) as an orange powder, which is used without further purification. MS (ESI): 249.7 [M+H] +

[0229] Step b) 3-Bromo-5-(1-mesyl-1-methyl-ethyl)pyridine [ka] 3-Bromo-5-(mesylmethyl)pyridine (80 mg, 0.26 mmol, 1 equiv.) was dissolved in N,N-dimethylformamide (superhydrous) (2.69 mL). Cesium carbonate (262 mg, 0.8 mmol, 3 equiv.) was added and the reaction was stirred at room temperature under Ar for 30 min. Methyl iodide (152 mg, 4 equiv.) was then added and the reaction was stirred overnight. Cesium carbonate (87.5 mg, 0.269 mmol, 1 equiv.) and iodomethane (76 mg, 33.6 μL, 2 equiv.) were added and the reaction was left stirring for an additional 24 h. The mixture was extracted (3 × 40 mL), and the AcOEt was washed with distilled water (2 × 50 mL) and once with brine (1 × 40 mL). The organic phase was then dried over Na2SO4, concentrated, and purified by preparative HPLC to give the title compound (20 mg, 26% yield) as a pale red solid. MS (ESI): 278.9 [M+H] +

[0230] Examples 22-25 in the table below were prepared similarly to steps g)-h) of Example 21 using the appropriate bromide or chloride. TIFF2025526683000083.tif233170

[0231] Example 26 (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) tert-Butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] A mixture of tert-butyl N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (100 mg, 0.18 mmol, 1.0 equiv), 5-methoxypyridine-3-boronic acid (36.2 mg, 0.24 mmol, 1.3 equiv), K2CO3 (75.6 mg, 0.55 mmol, 3.0 equiv), Pd(dppf)Cl2*DCM (14.9 mg, 0.02 mmol, 0.1 equiv) in 1,4-dioxane (10 mL) and water (1 mL) was stirred at 80 °C under N2 for 2 h. The solution was poured into water (30 mL) and extracted with EtAc (50 mL), and the organic phase was washed with brine (50 mL), dried over Na2SO4, concentrated in vacuo, and purified by preparative HPLC to give the title compound (50 mg, 0.09 mmol, 47% yield) as a white solid. MS (ESI): 576.1 [M+H] + .

[0232] Step b) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (40 mg, 0.07 mmol, 1 equiv.) in analogy to general procedure 5b and obtained as the dihydrochloride salt as a white solid after preparative HPLC (33.1 mg, 0.06 mmol, 92% yield). MS (ESI): 476.2 [M+H] + .

[0233] Example 27 (3R)-3-Amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) 1-(chloromethyl)-4-(4-methoxyphenyl)benzene [ka] To a solution of [4-(4-methoxyphenyl)phenyl]methanol (1900 mg, 8.87 mmol, 1.0 equiv) in DCM (38 mL) was added thionyl chloride (3.86 mL, 53.2 mmol, 6.0 equiv), and the mixture was stirred at 25° C. for 6 h and concentrated in vacuo to give the crude product, which was evaporated twice with THF (2 mL) to give the title compound (2000 mg, 8.59 mmol, 92% yield) as a brown solid, which was used directly in the next step. MS (ESI): 197.3 [M-Cl] - .

[0234] Step b) tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from 1-(chloromethyl)-4-(4-methoxyphenyl)benzene (654 mg, 2.8 mmol, 1.1 equiv.) and tert-butyl N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepin-3-yl]carbamate (1000 mg, 2.56 mmol, 1.0 equiv.) in DMF in analogy to general procedure 1 and obtained as a white solid after silica gel column chromatography (1270 mg, 2.16 mmol, 84% yield). MS (ESI): 533.2 [M-isobutene + H] + .

[0235] Step c) tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate (1270.0 mg, 2.16 mmol) in analogy to general procedure 2 and obtained as a light brown solid (1000 mg, 1.61 mmol, 69% yield). MS (ESI): 643.2 [M+2+Na] + .

[0236] Step d) tert-butyl N-[(3R)-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared in the same manner as in step a) of Example 26 from tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (100 mg, 0.16 mmol, 1 equivalent) and 5-chloropyridine-3-boronic acid (40.6 mg, 0.26 mmol, 1.6 equivalent), and obtained as a white solid (70 mg, 0.11 mmol, 63% yield) after silica gel column chromatography. MS (ESI): 652.2 [M + H] + .

[0237] Step e) (3R)-3-amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; dihydrochloride [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (70 mg, 0.11 mmol, 1 equiv.) in analogy to general procedure 5b and obtained as the dihydrochloride salt as a yellow solid after preparative HPLC (28.5 mg, 0.05 mmol, 47% yield). MS (ESI): 552.2 [M+H] + .

[0238] Examples 28-30 in the table below were prepared similarly to steps d)-e) of Example 27 using the appropriate bromide or chloride. TIFF2025526683000093.tif169170

[0239] Example 37 (3R)-3-Amino-8-fluoro-1,1-dioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) tert-butyl N-[rac-(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from 4-(bromomethyl)benzonitrile (275.58 mg, 1.41 mmol, 1.1 equiv.) and tert-butyl N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepin-3-yl]carbamate (500 mg, 1.28 mmol, 1.0 equiv.) in DMF in analogy to general procedure 1 and obtained as a pale yellow oil after silica gel column chromatography (450 mg, 68% yield). MS (ESI): 450.1 [M-isobutene + H] + .

[0240] Step b) tert-Butyl N-[rac-(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[rac-(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepin-3-yl]carbamate (2000.0 mg, 3.95 mmol, 1.0 equiv.) in analogy to General Procedure 2 and obtained as an off-white solid (1900.0 mg, 3.53 mmol, 87.48% yield). MS (ESI): 484.0 [M-isobutene + H] + .

[0241] Step c) [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (1000.0 mg, 1.86 mmol, 1.0 equiv.) in analogy to general procedure 3a and obtained as a yellow solid (900 mg, 1.79 mmol, 96% yield). MS (ESI): 448.0 [M-isobutene + H] + .

[0242] Step d) 3-[1-(trifluoromethyl)vinyl]pyridazine [ka] To a solution of 1-(trifluoromethyl)vinylboronic acid hexylene glycol ester (6.28 g, 28.3 mmol, 1.0 equiv.), 3-bromopyridazine (4.5 g, 28.3 mmol, 1.0 equiv.), and cesium carbonate (18.4 g, 56.6 mmol, 2.0 equiv.) in toluene (100 mL) and water (10 mL), the mixture was stirred at 90 °C for 12 h, and then Pd(dppf)Cl2CH2Cl2 (1.16 g, 1.42 mmol, 0.05 equiv.) was added at 20 °C. EtOAc (100 mL) and water (300 mL) were added, and the mixture was filtered. The filtrate was extracted with EtOAc (100 mL × 2). The combined extracts were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (PE / EA = 5 / 1 to 3 / 1) to give the title compound (4250 mg, 24.4 mmol, 86% yield) as a pale yellow oil. MS (ESI): 175.3 [M+H] +

[0243] Step e) 3-[1-(trifluoromethyl)cyclopropyl]pyridazine [ka] To a solution of 3-[1-(trifluoromethyl)vinyl]pyridazine (300 mg, 1.7 mmol, 1.0 equiv.) and methyl(diphenyl)sulfonium tetrafluoroborate (496 mg, 1.72 mmol, 1.0 equiv.) in THF (2 mL) was added NaHMDS in THF (3.45 mL, 3.45 mmol, 2.0 equiv.) at 0-5 °C. The solution was stirred at 20 °C for 2.5 h. The reaction mixture was quenched by the slow addition of saturated aqueous NH4Cl (30 mL) under stirring. EtOAc (30 mL) and water (30 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (30 mL × 2). The combined extracts were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by silica gel chromatography (PE / EA = 10 / 1 to 2 / 1) to give the title compound (180 mg, 0.96 mmol, 53% yield) as a pale yellow oil. MS (ESI): 189.3 [M+H] +

[0244] Step f) Trimethyl-[methyl-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-trimethylsilyloxy-silyl]oxy-silane [ka] To a solution of 3-[1-(trifluoromethyl)cyclopropyl]pyridazine (100 mg, 0.53 mmol, 1.0 equiv), trimethyl-[methyl(trimethylsilyloxy)silyl]oxy-silane (124 mg, 0.56 mmol, 1.05 equiv), and 2,9-dimethyl-1,10-phenanthroline (22.14 mg, 0.11 mmol, 0.2 equiv) in THF (5 mL) was added [Ir(COD)OMe] (38.43 mg, 0.05 mmol, 0.1 equiv) under N at 20° C. The mixture was stirred in a sealed tube at 100° C. for 12 h, after which the solvent was removed under reduced pressure. The residue was purified by silica gel chromatography (PE / EA = 20 / 1 to 10 / 1; 3 / 1 to 1 / 1) to give the title compound (88 mg, 0.21 mmol, 35% yield) as a yellow oil. MS (ESI): 409.2 [M+H] +.

[0245] Step g) 5-Bromo-3-[1-(trifluoromethyl)cyclopropyl]pyridazine [ka] To a solution of trimethyl-[methyl-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-trimethylsilyloxy-silyl]oxy-silane (30 mg, 0.07 mmol, 1.0 equiv.), N-bromosuccinimide (13 mg, 0.07 mmol, 1.0 equiv.) in ACN (2 mL) was added silver(I) fluoride (13.9 mg, 0.11 mmol, 1.5 equiv.) at 20° C., and the mixture was then stirred at 20° C. for 1 hour. The reaction mixture was concentrated in vacuo to give a residue below 40° C. The residue was triturated in TBME (10 mL) and collected by filtration. The filtrate was concentrated in vacuo to give the title compound (25 mg, 0.09 mmol, 126% yield) as a pale yellow oil, which was used directly in the next step. (ESI): 269.0 [M+H] + .

[0246] Step h) tert-butyl N-[(3R)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] To a solution of [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (20.0 mg, 0.04 mmol, 1.0 equiv.), 5-bromo-3-[1-(trifluoromethyl)cyclopropyl]pyridazine (21.2 mg, 0.08 mmol, 2.0 equiv.) pyridazine, and K3PO4 (25.3 mg, 0.12 mmol, 3.0 equiv.) in 1,4-dioxane (1 mL) and water (0.1 mL), the mixture was stirred at 80 °C under N2 for 2 h, and then the resulting mixture was treated with cataCXiumA Pd G3 (2.89 mg, 0.0 mmol, 0.1 equiv, CAS: 1651823-59-4) was added under N at 20 °C. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was dissolved in EtOAc (30 mL) and water (30 mL), and then extracted with EtOAc (30 mL x 2). The combined extracts were washed with brine (20 mL), dried over NaSO, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative TLC (PE / EA = 2 / 1, Rf = 0.2) to give the title compound (9.0 mg, 0.015 mmol, 31% yield) as a pale yellow solid. MS (ESI): 646.2 [M+H] + .

[0247] Step i) tert-butyl N-[(3R)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] To a solution of tert-butyl N-[(3R)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (15.0 mg, 0.02 mmol, 1.0 equiv.) and NaOAc (3.81 mg, 0.05 mmol, 2.0 equiv.) in ethanol (1 mL), NH2OH.HCl (1.94 mg, 0.03 mmol, 1.2 equiv.) was added at 20 °C. The mixture was stirred at 50 °C for 18 h, then poured into water (20 mL), and the layers were separated. The aqueous phase was extracted with EtOAc (10 mL × 3). The combined extracts were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated in vacuo to give the title compound (20.0 mg, 0.03 mmol, 109% yield) as a pale yellow solid. MS (ESI): 679.2 [M+H] + .

[0248] Step j) tert-Butyl N-[(3R)-8-fluoro-1,1,4-trioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] To a solution of tert-butyl N-[(3R)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (140 mg, 0.21 mmol, 1.0 equiv) in THF (4 mL) was added TFAA (86.6 mg, 0.41 mmol, 2.0 equiv) at 20 °C after stirring the mixture for 1 h at 20 °C. The reaction mixture was carefully basified with a saturated solution of NaHCO to pH = 8 and extracted with EtOAc (20 mL × 3). The organic layers were washed with brine (10 mL), combined, dried over NaSO, and concentrated in vacuo to give a residue, which was purified by preparative TLC (PE / EA=1 / 1, Rf=0.5) to give the title compound (75.0 mg, 0.1 mmol, 46% yield) as a pale yellow solid. MS (ESI): 757.2 [M+H] + .

[0249] Step k) (3R)-3-amino-8-fluoro-1,1-dioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; hydrochloride [ka] A solution of tert-butyl N-[(3R)-8-fluoro-1,1,4-trioxo-7-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (71.0 mg, 0.09 mmol, 1.0 equiv.) in HCl in EtOAc (3.0 mL, 12.0 mmol, 127.88 equiv.) was stirred at 20° C. for 0.5 h. The mixture was concentrated under vacuum below 35° C. to give a residue. The residue was dissolved in deionized water (20 mL) and lyophilized to give the title compound (49.6 mg, 0.07 mmol, 74% yield) as a white solid. MS (ESI): 657.1 [M+H] + .

[0250] Example 36 (3R)-3-amino-8-fluoro-1,1-dioxo-7(3R)-3-amino-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) 3-tert-butyl-1,2,4-triazin-5-ol [ka] To a solution of 2,2-dimethylpropanamidine (5000 mg, 49.9 mmol, 1.0 equiv., CAS 59950-56-0) and triethylamine (27.8 mL, 199 mmol, 4.0 equiv.) in ethanol (50 mL), hydrazine (4780 mg, 149.1 mmol, 2.99 equiv.) was added at 25 °C for 1 h, and then ethyl glyoxalate (30578 mg, 149 mmol, 3.0 equiv.) was added at 25 °C. The mixture was then stirred at 60 °C for 11 h. The reaction was concentrated in vacuo to give a residue, which was purified by column chromatography (PE:EA = 10:1 to 1:1) to give the title compound (4500 mg, 29.3 mmol, 585% yield) as a yellow solid. MS (ESI): 154.3 [M+H] + .

[0251] Step b) 3-tert-butyl-1,2,4 triazine-5-thiol [ka] To a solution of 3-tert-butyl-1,2,4-triazin-5-ol (400 mg, 2.1 mmol, 1.0 equiv.) in 1,4-dioxane (16 mL) was added Lawesson's reagent (528 mg, 1.31 mmol, 0.5 equiv.) at 25° C., and the mixture was then stirred at 100° C. for 2 h. The reaction was concentrated in vacuo to give a residue, which was purified by column chromatography (PE:EA=0:1 to 1:1) to give the title compound (180 mg, 1.06 mmol, 40% yield) as an orange solid. MS (ESI): 170.3 [M+H] + .

[0252] Step c) 3-tert-butyl-5-methylsulfanyl-1,2,4 triazine [ka] To a solution of 3-tert-butyl-1,2,4-triazin-5-ol (400 mg, 2.1 mmol, 1.0 equiv.) in 1,4-dioxane (16 mL) was added Lawesson's reagent (528 mg, 1.31 mmol, 0.5 equiv.) at 25° C., and the mixture was then stirred at 100° C. for 2 h. The reaction was concentrated in vacuo to give a residue, which was purified by column chromatography (PE:EA=0:1 to 1:1) to give the title compound (180 mg, 1.06 mmol, 40% yield) as an orange solid. MS (ESI): 170.3 [M+H] + .

[0253] Step d) tert-butyl N-[(3R)-7-(3-tert-butyl-1,2,4-triazin-5-yl)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (180 mg, 0.36 mmol, 1.0 equiv.) and 3-tert-butyl-5-methylsulfanyl-1,2,4-triazine (98.3 mg, 0.54 mmol, 1.5 equiv.) analogously to general procedure 4c and obtained as a yellow solid (80.0 mg, 0.13 mmol, 33% yield). MS (ESI): 595.2 [M+H] + .

[0254] Step e) tert-butyl N-[(3R)-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(3-tert-butyl-1,2,4-triazin-5-yl)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (80.0 mg, 0.13 mmol, 1.0 equiv.) in analogy to step i) of Example 37, and obtained as a yellow solid (80 mg, 0.13 mmol, 67% yield). MS (ESI): 628.2 [M + H] + .

[0255] Step f) tert-butyl N-[(3R)-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (80.0 mg, 0.13 mmol, 1.0 equiv.) in analogy to step j) of Example 37, and obtained as a yellow solid (50 mg, 0.07 mmol, 54% yield). MS (ESI): 706.1 [M+H] + .

[0256] Step g) (3R)-3-amino-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (50.0 mg, 0.07 mmol, 1.0 equiv.) in analogy to step k) of Example 37, and obtained as a yellow solid (26.3 mg, 0.04 mmol, 57% yield). MS (ESI): 606.1 [M+H] + .

[0257] 5-Bromo-3-(cyclopropoxy)pyridazine [ka] To a solution of cyclopropanol (150 mg, 2.58 mmol, 1.0 equiv) in THF (5 mL) was added sodium hydride (113 mg, 2.84 mmol, 1.1 equiv) in portions at 0 °C. After 0.5 h, the reaction mixture was added to a solution of 5-bromo-3-chloropyridazine (500 mg, 2.58 mmol, 1.0 equiv) in THF (5 mL) at 0 °C and stirred at 0 °C for 1.5 h. The reaction mixture was poured into water (30 mL). The aqueous phase was extracted with ethyl acetate (20 mL × 3). The combined organic phase was washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated in vacuo to give a residue that was purified by column chromatography (PE:EA = 1:0 to 5:1) to give the title compound (280 mg, 1.3 mmol, 50% yield) as a yellow oil, which was used as crude in the next step. MS(ESI):217.1[M+H] + .

[0258] 5-Bromo-3-tert-butyl-pyridazine [ka] Step a) (6-tert-butylpyridazin-4-yl)-methyl-bis(trimethylsilyloxy)silane WUX003530-273 [ka] The title compound was prepared from 3-tert-butylpyridazine (300 mg, 2.2 mmol, 1.0 equiv., CAS 215452-12-3) in analogy to step f) of Example 37 and obtained as a pale yellow oil (666 mg, 1.86 mmol, 84% yield) as a yellow solid after silica gel chromatography (PE / EA=10 / 1). MS (ESI): 357.2 [M+H] + .

[0259] Step b) 5-Bromo-3-tert-butyl-pyridazine [ka] The title compound was prepared from (6-tert-butylpyridazin-4-yl)-methyl-bis(trimethylsilyloxy)silane (40 mg, 0.11 mmol) in analogy to step h) of Example 37 to give the crude material as a light brown oil (40 mg, 0.19 mmol). MS (ESI): 217.0 [M+H] + .

[0260] Examples 34-35 in the table below were prepared analogously to steps h)-k) of Example 37 using the appropriate boronic ester and aryl bromide. TIFF2025526683000118.tif82170

[0261] Example 33 (3R)-3-Amino-8-fluoro-1,1-dioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) 5-bromo-3-(2,2,2 trifluoroethoxy)pyridazine [ka] To a solution of 2,2,2-trifluoroethanol (0.19 mL, 2.58 mmol, 1.0 equiv) in THF (5 mL) was added sodium hydride (113 mg, 2.84 mmol, 1.1 equiv) in portions at 0° C. After 0.5 h, the reaction mixture was added to a solution of 5-bromo-3-chloropyridazine (500 mg, 2.58 mmol, 1.0 equiv) in THF (5 mL) at 0° C. The mixture was stirred at 0° C. for 1.5 h. The reaction mixture was poured into water (30 mL). The aqueous phase was extracted with ethyl acetate (20 mL × 3). The combined organic phase was washed with brine (50 mL), dried over anhydrous NaSO, filtered, concentrated in vacuo, and purified by silica gel column chromatography (PE:EA=1:0 to 5:1) to give the title compound (350 mg, 1.36 mmol, 50% yield) as a yellow oil. MS(ESI): 257.1 [M+H] + .

[0262] Step b) tert-butyl N-[(3R)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] To a solution of [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (140 mg, 0.28 mmol, 1.0 equiv), 5-bromo-3-(2,2,2 trifluoroethoxy)pyridazine (107 mg, 0.42 mmol, 1.5 equiv), potassium carbonate (115 mg, 0.83 mmol, 3.0 equiv) in 1,4-dioxane (4 mL) and water (0.5 mL) was added Brettphos Pd g3 (37.8 mg, 0.04 mmol, 0.15 equiv) under N2 at 25 °C. The reaction was stirred at 90°C for 2 hours, quenched with water (15 mL), and extracted with EtOAc (10 mL x 3). The organic phase was washed with brine (30 mL x 2), dried over NaSO, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 1:0 to 3:1) to give the title compound (140 mg, 0.22 mmol, 79% yield) as a yellow solid. MS (ESI): 636.1 [M+H] + .

[0263] Step c) tert-butyl N-[(3R)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (130 mg, 0.2 mmol, 1.0 equiv.) in analogy to step i) of Example 37, and obtained as a yellow solid (158.0 mg, 0.24 mmol, 81% yield). MS (ESI): 669.2 [M + H] + .

[0264] Step d) tert-butyl N-[(3R)-8-fluoro-1,1,4-trioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (148.0 mg, 0.22 mmol, 1.0 equiv.) in analogy to step j) of Example 37, and obtained as a yellow solid after preparative HPLC (50 mg, 0.07 mmol, 30% yield). MS (ESI): 747.1 [M+H] + .

[0265] Step e) (3R)-3-amino-8-fluoro-1,1-dioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-8-fluoro-1,1,4-trioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (45.0 mg, 0.06 mmol, 1.0 equiv.) in analogy to general procedure 5b, and obtained as a yellow solid (48.1 mg, 0.07 mmol, 120% yield). MS (ESI): 647.0 [M+H] + .

[0266] Example 32 (3R)-3-Amino-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] Step a) tert-butyl N-[(3R)-7-bromo-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-1,1,4-trioxo-3,5-dihydro-2H-1λ6,5-benzothiazepin-3-yl]carbamate (250 mg, 0.59 mmol) with 2-[4-(chloromethyl)phenyl]-5-(trifluoromethyl)pyridine; hydrochloride (CAS: 613239-76-2) in analogy to general procedure 1, and obtained as a pale yellow solid (720 mg, 1.09 mmol, 153.65% yield). MS (ESI): 660.0 [M+H] + .

[0267] Step b) [(3R)-3-(tert-butoxycarbonylamino)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid [ka] The title compound was prepared from tert-butyl N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (300 mg, 0.525 mmol, 1 equiv.) in analogy to general procedure 3a and obtained as a colorless oil (150 mg, 46% yield).

[0268] Step c) tert-butyl N-[(3R)-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate [ka] The title compound was prepared from [(3R)-3-(tert-butoxycarbonylamino)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]boronic acid (20 mg, 0.03 mmol, 1.0 equiv.) and 5-bromo-3-tert-butyl-pyridazine (13.8 mg, 0.06 mmol, 2.0 equiv.) in analogy to general procedure 4a, and obtained as a white solid (2.5 mg, 0.004 mmol, 10% yield). MS (ESI): 714.2 [M + H] + .

[0269] Step d) (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one [ka] The title compound was prepared from tert-butyl N-[(3R)-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-3-yl]carbamate (37.0 mg, 0.05 mmol) in analogy to general procedure 5b and obtained as a white solid after preparative HPLC and SFC (12.2 mg, 0.02 mmol, 35.88% yield). MS (ESI): 614.1 [M+H] + .

[0270] 2) Biological Examples 2.1) In vitro DGK inhibition assay DGKα and ζ kinases use ATP to phosphorylate the substrate 1,2-dilauroyl-sn-glycerol (DLG incorporated into liposomes), resulting in the conversion of ATP to ADP.

[0271] After the kinase reaction, an ATP depletion reagent is added to stop the kinase reaction and deplete any remaining ATP, leaving only ADP. Second, a detection reagent is added to simultaneously convert ADP to ATP and convert the newly synthesized ATP to light using a coupled luciferase / luciferin reaction.

[0272] Reagents and materials TIFF2025526683000130.tif55170

[0273] TIFF2025526683000131.tif80170

[0274] Full-length DGK α and ζ were expressed in Sf21 insect cells by infecting the cells with a baculovirus stock at an MOI of 2. Purification of both enzymes was performed as previously described by Takahashi et al., PeerJ, 2018 (Takahashi, D.; Sakane, F. Expression and purification of human diacylglycerol kinase alpha from baculovirus-infected insect cells for structural studies. PeerJ 2018, 6, No. e5449). TIFF2025526683000132.tif47170

[0275] TIFF2025526683000133.tif82170

[0276] Assay procedure Concentrated liposome solutions were prepared in assay buffer without DTT and BSA: 2 mM DLG in 21 mM total liposomes (2 mM DLG / 8 mM PS / 11 mM PC). The reaction mixtures contained assay buffer with a final DLG concentration of 25 μM (for the DGKA assay) or 50 μM (for the DGKZ assay) at 125 μM ATP concentration. The reaction was initiated by the addition of DGKα and ζ kinases at final concentrations of 4 nM and 2 nM, respectively. After 1 h of reaction, the amount of ADP formed was detected using the ADP-Glo kinase assay (Promega) according to the manufacturer's instructions. Compounds were added in an 11-point dose response, starting at 10 mM, 1:3 dilution, with a final DMSO concentration of 2%. A Multidrop Combi was used as the liquid handler, and luminescence was read in 0.5 s using a vision reader (PE).

[0277] result TIFF2025526683000134.tif250170

[0278] In vitro DGK inhibition assay: ADP Glo

[0279] DGKα and ζ kinases use ATP to phosphorylate the substrate 1,2-dilauroyl-sn-glycerol (DLG), resulting in the conversion of ATP to ADP.

[0280] After the kinase reaction, an ATP depletion reagent is added to stop the kinase reaction and deplete any remaining ATP, leaving only ADP. Second, a detection reagent is added to simultaneously convert ADP to ATP and convert the newly synthesized ATP to light using a coupled luciferase / luciferin reaction.

[0281] Experimental procedures, reagents and materials The DGKα and ζ kinase ADP Glo assay was performed by Reaction Biology Corp. (1 Great Valley Parkway, Suite 2, Malvern, PA, 19355, USA). The information provided by the service provider is as follows: DGKα and ζ kinase were used at a final concentration of 2 nM. Reactions were performed with 50 μM ATP. 500 μM of the substrate DLG (dilauroyl-sn-glycerol) was used. Compounds were received as 10 mM DMSO stock solutions and tested at 10 doses of IC50 replicates using 3-fold serial dilutions starting at 1 μM. The control compound, calphostin C, was tested at 10 doses of IC50 replicates using 3-fold serial dilutions starting at 100 μM.

[0282] result TIFF2025526683000135.tif50170

[0283] 2.2) IL2 secretion measurement As a readout of T cell activation, IL2 secretion after 24 hours and proliferation after 5 days were measured. The increase in IL2 secretion and proliferation upon compound treatment was evaluated as % of the maximum value of reference compound A1. WO 2016 / 139181 discloses reference compound A1 as Example 70. As a counterscreen and to ensure that undesired TCR-independent activation was not caused, PBS conditions were performed for all compounds.

[0284] Reagents and materials TIFF2025526683000136.tif244170TIFF2025526683000137.tif141170

[0285] cell culture Expanded primary human T cells were thawed and cultured at a density of 2 μl / ml in RPMI 1640 (Gibco, #61870-010) + 5% human serum (HS, Sigma, #H 3667) + 1 mM sodium pyruvate (Gibco, #11360-039) + 50 μM 2-mercaptoethanol (Gibco, #31350-010) and 1× Pen-Strep (Life Technologies, #15140122) medium at 5% CO2, 37°C, and 95% humidity for 3 hours. For plate coating, 100 μl / well of PBS++ and PBS-- or PBS++ and CD3 antibody (concentration donor-dependent and determined by CD3 titration) was added to a poly-D lysine-coated 96-well plate. The plate was sealed and incubated at room temperature for 3 hours on a tabletop rocking platform. After incubation, plates were washed once with PBS and filled with 40 μl / well of culture medium alone. Compounds were then added to the medium-only plates (see next section). After 3 hours of T cell culture, cells were filtered through a cell strainer (Miltenyi Biotech, #130-041-407), counted again, and the concentration was adjusted to 1.25 Mio / ml.

[0286] Cells were then seeded with 80 μl / well to 40 μl / well of compound, distributed according to the plate layout. Compounds were further diluted 1:3 by adding cells, resulting in 100k cells / 120 μl / well. After 24 hours, 40 μl of supernatant was carefully collected from the top without disturbing the cells and transferred to a round-bottom 96-well plate. The collected and frozen supernatant was used for IL-2 detection using the IL-2 Human ProQuantum Immunoassay Kit (Invitrogen) or the Human IL-2 ELISA Kit (Thermo Fisher).

[0287] Compound treatment Compounds were added in a 5- or 6-point dose response using a Tecan D300e digital dispenser. All conditions were concentrated 3x the final concentration prior to subsequent cell addition (80 μl of treated cells to 40 μl of conditioned medium). DRs started at a final top concentration of 20 μM or 10 μM and a dilution factor of 3.333. The positive control was reference compound A1, added in a similar dose response, plus three wells at only 20 μM to represent a positive stimulator control. All wells were normalized to a final concentration of 0.6% with DMSO (0.2% final concentration).

[0288] IL2 ProQuantum Immunoassay The immunoassay is performed according to the manufacturer's manual (Invitrogen, #A35603).

[0289] Additional Information: For immunoassays, use MicroAmp™ EnduraPlate™ Optical 384-well plates. Thaw the frozen supernatant and centrifuge at 1000 x g for 5 minutes, both steps performed at 4°C. After centrifugation, collect the required sample volume from the top and dilute it in a separate LightCycler V-bottom plate (working plate) with assay dilution buffer, PBS, or CD3, depending on the condition, at a dilution factor of at least 1:3. Prepare IL-2 standards and blanks in the same V-bottom plate (extended version) with standards ranging from 0.0128 to 5000 pg / ml. After preparation, transfer 5 μl of sample dilutions or standards / blanks to the optical 384-well plate (assay plate) and follow the 10 μl reaction protocol. Use a QuantStudio 12K Flex system for measurements. Extract raw data and calculate IL-2 concentrations using the Thermo Fisher online app (apps.thermofisher.com / apps / proquantum).

[0290] IL2 Elisa The ELISA is performed according to the manufacturer's manual (Thermo Fisher Scientific, #88-7025-88).

[0291] Additional information: For ELISA, Nunc MaxiSorp 96-well plates are used. Frozen supernatants are thawed and centrifuged at 1000 x g for 5 minutes, both steps being carried out at 4°C. The required sample volume is then collected from the top and placed in a separate V-bottom plate, diluted with ELISA diluent, PBS, or a dilution factor appropriate for the CD3 condition. IL-2 standards and blanks are prepared in the same V-bottom plate. After preparation, 50 μl of sample dilution and 100 μl of standard or blank are transferred to the Nunc plate.

[0292] Calculations and Data Reporting CD3 and PBS plates were analyzed separately in Genedata Screener using Roche Normalization PCT_POS_CTRL, DMSO was set as neutral control and 20 μM of reference compound A1 was set as stimulator control / 100%.

[0293] For CD3 conditions, the EC50 and Emax of the fitted sigmoidal curve were reported. If a curve could not be fitted, the EC50 was reported as a blank field, and Emax was based on individual data points. Emax did not necessarily correspond to the highest concentration tested. Compounds that activated unstimulated cells or adversely affected viability (see proliferation assays) were flagged.

[0294] result TIFF2025526683000138.tif255170TIFF2025526683000139.tif66170

[0295] 2.3) Proliferation assay Reagents and materials TIFF2025526683000140.tif244170TIFF2025526683000141.tif37170

[0296] Thaw expanded primary human T cells and culture them at a density of 2 μl / ml in RPMI 1640 (Gibco, #61870-010) + 5% human serum (HS, Sigma, #H3667) + 1 mM sodium pyruvate (Gibco, #11360-039) + 50 μM 2-mercaptoethanol (Gibco, #31350-010) and 1x Pen-Strep (Life Technologies, #15140122) medium at 5% CO2, 37°C, and 95% humidity for 3 hours. For plate coating, add 100 μl / well of PBS++ alone or PBS++ with CD3 antibody (concentration donor-dependent and determined by CD3 titration) to a 96-well plate coated with Poly-D Lysine. Seal the plate and incubate it on a tabletop rocking platform at room temperature for 3 hours. After incubation, the plates are washed once with PBS and filled with 40 μl / well of culture medium alone. Compounds are then added to the medium-only plates (see next section). After 3 hours of culturing the T cells, the cells are filtered through a cell strainer (Miltenyi Biotech, #130-041-407), counted again, and the concentration is adjusted to 1.25 Mio / ml.

[0297] Cells are then seeded in 80 μl / well to 40 μl / well containing compounds distributed according to the plate layout. Compounds are further diluted 1:3 by adding cells to obtain 100k cells / 120 μl / well. After 48 hours, 40 μl of supernatant is carefully collected from the top without disturbing the cells. Cells are assessed for proliferation after 5 days by measuring ATP consumption using CellTiterGlo (Promega).

[0298] Compound treatment Compounds were added in a 5- or 6-point dose response using a Tecan D300e digital dispenser. All conditions were concentrated 3x the final concentration prior to subsequent cell addition (80 μl of treated cells to 40 μl of conditioned medium). DRs started at a final top concentration of 20 μM or 10 μM and a dilution factor of 3.333. The positive control was reference compound A1, added in a similar dose response, plus three wells at only 20 μM to represent a positive stimulator control. All wells were normalized to a final concentration of 0.6% with DMSO (0.2% final concentration).

[0299] Cell titer Glo measurement After 5 days, CellTiter-Glo® 2.0 Reagent is used to detect ATP, which is directly proportional to the number of cells present per well. After visually controlling for toxicity or precipitation of the tested compounds, the plate is equilibrated to room temperature for 45 minutes. The CellTiter-Glo® 2.0 Reagent is also equilibrated to room temperature. After equilibration, an equal volume of CellTiter-Glo Reagent is added to the cells (80 μl / well) using an electronic multichannel pipette. The plate is placed on a rocking platform for 15 minutes at room temperature. After incubation, the bottom of the plate is sealed with backing tape. Luminescence is measured on a PHERAstar FSX (interval time 0.5 seconds, gain 3000, focal height 15 mm) and exported as a CSV file for analysis on the Genedata screener.

[0300] Calculations and Data Reporting CD3 and PBS plates were analyzed separately in Genedata Screener using Roche Normalization PCT_POS_CTRL, DMSO was set as neutral control and 20 μM of reference compound A1 was set as stimulator control / 100%.

[0301] For CD3 conditions, the EC50 and Emax of the fitted sigmoidal curve were reported. If a curve could not be fitted, the EC50 was reported as a blank field, and Emax was based on individual data points. Emax did not necessarily correspond to the highest concentration tested. Compounds that activated unstimulated cells (see IL2 measurements) or had a negative effect on viability were flagged.

[0302] result TIFF2025526683000142.tif255170TIFF2025526683000143.tif83170

[0303] 2.5) T cell-TCB-MV3 killing assay Reagents and materials TIFF2025526683000144.tif245170TIFF2025526683000145.tif85170

[0304] cell culture All culture steps are carried out at 5% CO2, 37°C and 95% humidity.

[0305] MV-3 RFP cells were cultured in MV-3 medium (DMEM + 10% FBS, 1x PenStrep, and 0.5 μg / mL puromycin) for at least 3 weeks. 80% confluent cultured MV-3 cells were washed once with PBS and trypsinized until detached. Cells were then counted and resuspended at 1x105 cells / mL in T cell medium (RPMI 1640 + 5% human serum + 1 mM sodium pyruvate + 50 μM 2-mercaptoethanol and 1x PenStrep). Cells were seeded at 100 μL / well into a 96-well plate (TTP, #92696) and left undisturbed at room temperature for 40 minutes to achieve uniformly distributed cell attachment. The plate was then incubated until further use.

[0306] The next day, thaw the expanded primary human T cells and resuspend them in T cell medium to 4 x 106 cells / mL. Culture them in a 6-well plate for 3 hours, up to 6 mL per well. After culturing the T cells, filter them through a cell strainer (Miltenyi Biotech, #130-041-407), count them again, and adjust the cell concentration to 2 x 106 cells / mL.

[0307] Compound treatment MCSP-TCB or PBS is prediluted in T cell medium (concentration depends on the T cell donor) at 4x the final concentration. 60 μL / well of the predilution is then dispensed into a round-bottom plate (Costar, #3799) according to the plate layout. Compounds are added in a 9-point dose response using a Tecan D300e digital dispenser, also at 4x the final concentration. The DMSO concentration in all wells is adjusted to 0.8% to obtain a final concentration of 0.2%.

[0308] Add 60 μL / well of the T cell suspension to the prepared round-bottom plate and resuspend using a manual multichannel filter. Then, carefully transfer 100 μL / well of the resuspended T cell suspension containing the treatment to the overnight-cultured MV-3 cells according to the plate layout. Add 100 μL of T cell medium only to the outer MV-3 wells only. The final compound DR starts at 20 μM, with a dilution factor of 3.333. The final TCB concentration ranged from 1.5 pM to 5 pM and was determined individually for each T cell donor by performing a TCB titration. For each donor, a TCB concentration corresponding to 10-20% of the MV3 baseline cell death in the absence of compound treatment was selected. Positive controls were reference compound A1 added to the DR as well as additional wells containing only 20 μM. 20 μM reference compound A1 represented a positive stimulator control, TCB alone (DMSO wells), and a neutral control.

[0309] calculation After transfer of T cells with pretreatment diluent, MV-3 cells were imaged by time-lapse microscopy using IncucyteZOOM™ (Essen BioScience, Michigan, USA). Images were acquired every 3 hours for a total of 120 hours (10x objective, phase and red imaging channels, 400 ms acquisition time, green / red 4614 optical module). RFP object counts per well were analyzed using IncucyteZOOM™ software (version 2019B Rev2) using a mask previously created and optimized for MV-3 cells. Raw data were exported as object counts / well, and values were normalized as %TCL compared to MV-3-only wells, with 100% proliferation and therefore 0% TCL.

[0310] RFP measurement The calculated %TCL values are analyzed in Genedata Screener using Roche Normalization PCT_POS_CTRL, with MCSP-TCB alone set as neutral control and 20 μM reference compound A1 set as stimulation control / 100%.

[0311] The EC50 and Emax values are shown in the table below.

[0312] TCLs induced by compounds without TCB treatment or toxicity (observed in PBS conditions) were flagged.

[0313] result TIFF2025526683000146.tif52170

Claims

1. Formula (I) 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein R １ is a 6-membered heteroaryl, and R １ represents one or more R １０ is a 6-membered heteroaryl optionally substituted with R ２ is hydrogen or halogen, R ４ represents one or more R １１ selected from phenyl and pyridinyl, optionally substituted by R １０ teeth, i) one or more of halogen, hydroxy, —S(O) ２ (C １～６ -alkyl), -N(R １０ｅ R １０ｆ ), -S(O) ２ (C １～６ -cycloalkyl), optionally substituted with cyano, C １～１０ - alkyl; ii) one or more —S(O) ２ (C １～６ -alkyl), halo-C １～６ - optionally substituted with alkyl, C ３～１０ -cycloalkyl; iii) one or more halogens, —C(O)O—(R １０ｑ 3- to 10-membered heterocyclyl optionally substituted with iv) one or more C １～６ -alkyl, halogen, or halo-C １～６ -phenyl optionally substituted with alkyl; v)-N(R １０ｅ R １０ｆ )、 vi)-OR １０ｇ 、 vii) halogens is selected from R １０ｅ and R １０ｆ are each independently hydrogen and C １～６ - alkyl, R １０ｇ is C １～６ -Alkyl, halo-C １～６ -Alkyl and C ３～１０ -cycloalkyl, R １０ｑ is C １～１０ - alkyl or or two R's １０ together with the carbon atoms to which they are attached, form one or more C １～１０ - forms a 3- to 10-membered heterocyclyl optionally substituted by alkyl, R １１ teeth, i) halogen; ii) optionally substituted with one or more cyanos, C １～６ - alkyl; iii) C １～６ -alkoxy, iv) C ３～７ -cycloalkyl, v) one or more halo-C １～６ -Alkyl, C ３～１０ -5-6 membered heteroaryl optionally substituted by cycloalkyl; vi) one or more halogens, C １～６ -alkoxy, halo-C １～６ -phenyl optionally substituted with alkyl; v))) p. ２ () １１ｄ ) is selected from R １１ｄ is hydrogen, C １～６ -Alkyl and halo-C １～６ -alkyl, or a pharmaceutically acceptable salt thereof.

2. R １ is one or more R １０ The compound according to claim 1, wherein the compound is pyridyl, pyrimidinyl, pyridazinyl, or triazinyl, optionally substituted with

3. R １ is one or more R １０ The compound according to claim 1 or 2, wherein the compound is pyridyl, pyrimidinyl, or triazinyl, optionally substituted with

4. R １ is one or more R １０ The compound according to any one of claims 1 to 3, wherein the compound is pyridyl or triazinyl, optionally substituted by:

5. R ２ The compound according to any one of claims 1 to 4, wherein is hydrogen or fluorine.

6. R ２ The compound according to any one of claims 1 to 5, wherein is fluorine.

7. R ４ But one R １１ The compound of any one of claims 1 to 6, wherein the compound is phenyl substituted with

8. R １０ is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy, or Or, R １０ and R １ The compound according to any one of claims 1 to 7, wherein

9. R １０ The compound according to any one of claims 1 to 8, wherein is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile.

10. R １１ is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propanenitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridonyl.

11. R １１ The compound according to any one of claims 1 to 10, wherein is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy.

12. R １ is one or more R １０ pyridyl, pyrimidinyl, pyridazinyl, triazinyl, optionally substituted by R ２ is hydrogen or halogen, R ４ But one R １１ is a phenyl substituted with R １０ but, i) one or more of halogen, hydroxy, —S(O) ２ (C １～６ -alkyl), -N(R １０ｅ R １０ｆ ), -S(O) ２ (C １～６ -cycloalkyl), optionally substituted with cyano, C １～１０ - alkyl; ii) one or more —S(O) ２ (C １～６ -alkyl), C ３～１０ -cycloalkyl, iii) one or more halogens, —C(O)O—(R １０ｑ 3- to 10-membered heterocyclyl optionally substituted with iv) phenyl; v)-N(R １０ｅ R １０ｆ )、 vi)-OR １０ｇ 、 vii) halogens is selected from R １０ｅ and R １０ｆ are each independently hydrogen and C １～６ - alkyl, R １０ｇ But C １～６ -Alkyl and halo-C １～６ - alkyl, R １０ｑ But C １～１０ - alkyl, R １１ but, i) halogen; ii) optionally substituted with one or more cyanos, C １～６ - alkyl; iii) C １～６ -alkoxy, iv) C ３～７ -cycloalkyl, v) one or more halo-C １～６ -Alkyl, C ３～１０ -5-6 membered heteroaryl optionally substituted by cycloalkyl; vi) one or more halogens, C １～６ -alkoxy, halo-C １～６ -phenyl optionally substituted with alkyl; v))) p. ２ () １１ｄ ) is selected from R １１ｄ But hydrogen, C １～６ -Alkyl and halo-C １～６ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the aryl group is selected from the group consisting of -alkyl, ...

13. R １ is one or more R １０ pyridyl, pyrimidinyl, triazinyl, optionally substituted by R ２ is hydrogen or fluorine, R ４ But one R １１ is a phenyl substituted with R １０ is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy, or Or, R １０ and R １ together to form trimethyl-6,8-dihydro-1,7-naphthyridinyl, dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl, R １１ is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propanenitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridonyl, or a pharmaceutically acceptable salt thereof.

14. R １ is one or more R １０ pyridyl, triazinyl, optionally substituted by R ２ is fluorine, R ４ But one R １１ is a phenyl substituted with R １０ is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propanenitrile, R １１ 2. The compound of claim 1, wherein is selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, or a pharmaceutically acceptable salt thereof.

15. (3R)-3-amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(2-tert-butyl-4-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butylpyrimidin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-(4-chlorobenzyl)-8-fluoro-1,1-diketo-7-[5-(trifluoromethoxy)-3-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-isopropoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[6-(trifluoromethyl)-2-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)pyrimidin-2-yl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butyl-1,2,4-triazin-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazin-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-6-methyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; Methyl 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylate; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-[5-(1-methylsulfonylcyclopropyl)-1,2,4-triazin-3-yl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-[5-(3-chloro-1-methylsulfonyl-propyl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-diketo-7-(5-morpholino-3-pyridyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-triketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-3-pyridyl]-2-methyl-propionitrile; (3R)-3-amino-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-7-[5-(trifluoromethyl)-3-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-diketo-7-[5-(1-mesyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5,6-dimethyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-dioxo-7(3R)-3-amino-7-(3-tert-butyl-1,2,4-triazin-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-dioxo-7-[6-(2,2,2-trifluoroethoxy)pyridazin-4-yl]-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; and (3R)-3-amino-7-(6-tert-butylpyridazin-4-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; 15. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, selected from:

16. (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazin-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-6-methyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-triketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-7-yl]-3-pyridyl]-2-methyl-propionitrile; (3R)-3-amino-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-7-[5-(trifluoromethyl)-3-pyridyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-1,1-diketo-7-[5-(1-mesyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-7-(5,6-dimethyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; (3R)-3-amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; and (3R)-3-amino-7-(5-tert-butyl-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepin-4-one; 16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, selected from:

17. A process for the preparation of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, comprising the step of: 【Chemistry 2】 (In the formula, R １ , R ２ and R ４ is as defined in any one of claims 1 to 14, and PG is an amino protecting group), with a suitable deprotecting agent to form said compound of formula (I).

18. 18. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, when produced according to the process of claim 17.

19. 17. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for use as a therapeutically active substance.

20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

21. 21. The pharmaceutical composition of claim 20, further comprising an additional therapeutic agent.

22. 17. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for use in the treatment, prevention and / or delay of progression of cancer.

23. 23. The compound for use according to claim 22, wherein the cancer is associated with aberrant diacylglycerol kinase signaling, and the diacylglycerol kinase is selected from DGKα and / or DGKζ.

24. The cancer is selected from the group consisting of B-cell acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin's lymphoma, 24. The compound for use according to claim 22 or 23, wherein the compound is selected from the group consisting of leukemia, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, preleukemia, sarcoma, carcinoma, melanoma, neuroblastoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, gastric cancer, brain cancer, lung cancer (e.g., NSCLC), pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, uterine cancer, adrenal cancer, and head and neck cancer.

25. Use of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for the treatment, prevention and / or delay of progression of cancer.

26. 20. Use of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment, prevention and / or delay of progression of cancer.

27. 17. A method for treating, preventing and / or delaying the progression of cancer, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof.

28. 17. Use of the compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for inhibiting the activity of at least one diacylglycerol kinase selected from DGKα and DGKζ.

29. 17. A method for inhibiting the activity of at least one diacylglycerol kinase selected from DGKα and DGKζ, comprising administering to a subject in need thereof a therapeutically effective amount of at least one compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof.

30. The invention as hereinbefore described.

Images