Bicyclic tetrahydrothiaza derivatives

Abstract

The present invention provides novel bicyclic tetrahydrothiaza # imgabs0 # derivatives having the general formula (I), # imgabs1 # wherein R1, R2 and R4 are as defined herein; comprising the compound; methods of making the compounds; and methods of using the compounds.

Description

Technical Field

[0001] The present invention relates to bicyclic tetrahydrothiazepines that inhibit diacylglycerol kinase (DGK) α and ζ and are useful as T cell activators Compounds, their manufacture and pharmaceutical compositions containing the compounds.

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

[0003] Cancer immunity is a multi-step process that is regulated by a series of negative immune checkpoints and positive co-stimulatory receptors and related intracellular signal cascades, and anti-tumor responses can be achieved when effectively triggered (Mellman, I. et al. (2011) Cancer Immunotherapy Comes of Age, Nature 480 (7378), 480-489). In fact, PD1 / PDL1 targeted inhibitors and other immune checkpoint inhibitors have brought changes to cancer immunotherapy, but more than 70% of patients still cannot benefit from immune checkpoint inhibition. Similarly, for T cell bispecific antibodies, even in the most promising indications (non-Hodgkin's lymphoma), these T cell binders (TCBs) have only achieved complete remission in less than 50% of patients. T cell exhaustion seems to play an important role in many examples of primary or secondary resistance to cancer immunotherapy. One possible reason for the lack of efficacy is that T cell activation occurs via targeting and cross-linking CD3 (signal 1), but lacks costimulation such as via CD28 or 4-1BB (signal 2). This hypothesis has been clinically validated in CAR T-cell therapy, where it was shown that clinically relevant efficacy was only observed after incorporation of the co-stimulatory domain.

[0004] Diacylglycerol kinase (DGK) is a lipid kinase that catalyzes the conversion of diacylglycerol (DAG) to phosphatidic acid (PA), thereby limiting DAG-regulated functions and promoting PA-dependent functions (Merida, I., Avila-Flores, A. and Merino, E. 2008: Diacylglycerol kinases: at the hub of cells ignalling. Biochem. J. 409 (1), 1–18). The DGK family consists of ten isoforms, which can be divided into five subtypes based on the presence of different regulatory domains in their structures. In addition, the lack of structural data still hinders a more thorough understanding of the mode of action of DGK. In addition, information about some prokaryotic DGKs and other lipid kinases, such as sphingosine kinase and phosphatidylinositol 3-kinase (PI3K), provides only limited information on the DGK catalytic mechanism, which appears to be different from classical kinases (Arranz-Nicolás, J. and Mérida, I., 2020. Biological regulation of diacylglycerol kinases in normal and neoplastic tissues: New opportunities for cancer immunotherapy, Advances in Biological Regulation, Vol. 75; Ma, Q., Gabelli, SB, Raben, DM, 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 intensively studied in this regard. As PA generators, both enzymes have been implicated in a variety of processes that promote tumor growth and metastasis. On the other hand, as DAG-consumers, DGKα and DGKζ have been extensively 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 drugtargets for a variety of diseases: an update. Front Cell Dev Biol 4, 82; Arranz-Nicolás, J. and Merida, I., 2020. Biological regulation of diacylglycerol kinases abnormal 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 co-stimulatory receptors and the T cell receptor (TCR), and their function is to limit the amount of DAG generated and ultimately limit T cell activation (Merida, I., Andrada, E., Gharbi, SI, 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, YV, Topham, MK, Epand, RM, 2011. Regulation and functions of diacylglycerol kinases. Chem. Rev. 111(10), 6186–6208). A summary of representative DGK-regulated signaling pathways is shown in Figure 1 (Sim, JA; Kim, J.; Yang, D. Beyond Lipid Signaling: Pleiotropic Effects of Diacylglycerol Kinases in Cellular Signaling. Int. J. Mol. Sci. 2020, 21, 6861): Activated PLC1 cuts PIP2 in the plasma membrane to generate two second messengers, DAG and IP3. DAG activates PKC, Ras / MEK / ERK / AP-1, and NF-kB, while IP3 is involved in the activation of intracellular Ca2+ flux. Upregulated Ca2+ signaling then activates the transcription factor NFAT. In short, DAG production and levels determine the duration and intensity of Ras / MEK / ERK and PKC-dependent signaling pathways, and they are critical for T cell activation. Therefore, DGK is used 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) can limit tumor destruction. Experiments using CAR T cells for human mesothelioma transplanted into nude mice showed that tumor-infiltrating CAR T cells expressed elevated concentrations of surface inhibitory receptors and inhibitory enzymes SHIP-1, DGKα, and DGKζ (Moon et al., 2014). Further, high DGKα expression was also observed in TILs isolated from human renal tumors (Prinz et al., 2012). In mouse mesoCAR T cells, dual deletion of DGKα and DGKζ resulted in enhanced cytokine expression and cytotoxicity against tumor cells (Riese et al., 2013). Human CAR T cells have been reported to have similar results, in which CRISPR / Cas9 was used to silence the expression of both DGKα and DGKζ (Jung et al., 2018). All these studies support the rationale for targeting DGKα / ζ in the development of anticancer therapies (Arranz-Nicolás, J. and Mérida, I., 2020. Biological regulation of diacylglycerol kinases in normal and neoplastic tissues: New opportunities for cancer immunotherapy, Advances in Biological Regulation, Vol. 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 DGKα or DGKζ exhibit 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 anergy 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 anergy.Nat.Immunol.7(11),1174–1181).

[0008] In summary, there is a wealth of evidence that DGKα and DGKζ are high-value targets for cancer immunotherapy. At the same time, there is a lack of compounds that can effectively inhibit DGKα and DGKζ with good selectivity relative to other diacylglycerol kinases, protein kinases, and / or other lipid kinases.

[0009] The present invention describes such dual DGKα / ζ inhibitors, which have excellent selectivity in the safety / off-target group and in comparison with other protein kinases and with other lipid kinases. These compounds effectively activate suboptimally stimulated T cells, thereby acting as intracellular enhancers of the co-stimulatory signaling cascade. These DGKα / ζ inhibitors are likely to increase the proliferation, cytotoxicity, and lifespan of targeted T cells, thereby improving the anti-cancer activity of CPIs, CD3-engaged T cell bispecific antibodies, and CAR T cells. Further, by engaging the central signaling nodes of both TCR and co-stimulatory receptors, these molecules appear to enhance both signals 1 and 2, thereby achieving single agent activity, such as in inflammatory tumors.

[0010] There is a continuing need for novel compounds that are able to activate and proliferate T cells and thereby treat, prevent and / or delay cancer progression.

[0011] Therefore, the object of the present invention is to provide compounds useful as DGKα / ζ inhibitors for the treatment or prevention or alleviation of such diseases, which compounds have improved therapeutic properties, in particular improved pharmacokinetic properties. Summary of the invention

[0012] The first object of the present invention is a compound of formula (I)

[0013]

[0014] or a pharmaceutically acceptable salt thereof, wherein:

[0015] R 1 is a 6-membered heteroaryl group, wherein R 1 Optionally, one or more R 10 replace;

[0016] R 2 is hydrogen or halogen;

[0017] R 4 is selected from phenyl or pyridine, which is optionally substituted by one or more R 11 replace;

[0018] R 10 Selected from:

[0019] i)C 1-10 -alkyl, which is optionally substituted with one or more halogen, hydroxyl, -S(O)2(C 1-6 -alkyl), -N(R 10e R 10f )、-S(O)2(C 1-6 -cycloalkyl), cyano substituted;

[0020] ii) C3-10 -cycloalkyl, which is optionally substituted with one or more -S(O)2(C 1-6 -alkyl) substituted;

[0021] iii) a 3- to 10-membered heterocyclic group, which is optionally substituted with one or more halogens, -C(O)O-(R 10q )replace;

[0022] iv) phenyl, which is optionally substituted with one or more C 1-6 -alkyl, halogen or halo-C 1-6 - alkyl substitution;

[0023] v)-N(R 10e R 10f );

[0024] vi)-OR 10g ;

[0025] vii) halogen;

[0026] R 10e and R 10f are each independently selected from hydrogen and C 1-6 -alkyl;

[0027] R 10g Selected from C 1-6 -alkyl and halo-C 1-6 -alkyl;

[0028] R 10q C 1-10 -alkyl;

[0029] Or two R 10 Together with the carbon atoms to which they are attached, they form a 3- to 10-membered heterocyclic group, which is optionally substituted by one or more C 1-10 - alkyl substitution;

[0030] R 11 Selected from:

[0031] i) halogen;

[0032] ii) C 1-6 -alkyl, which is optionally substituted with one or more cyano groups;

[0033] iii) C 1-6 - alkoxy;

[0034] iv) C 3-7 - cycloalkyl;

[0035] v) a 5- to 6-membered heteroaryl group, which is optionally substituted with one or more halogenated -C 1-6 -alkyl, C 3-10 - Cycloalkyl substitution;

[0036] vi) phenyl, which is optionally substituted with one or more halogen, C 1-6 -alkoxy, halo-C 1-6 - alkyl substitution;

[0037] vii)-SO2(R 11d );

[0038] R 11d Selected from hydrogen, C 1-6 -alkyl and halo-C 1-6 -alkyl.

[0039] The second object of the present invention is a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof as described above, which comprises making a compound of formula (XI)

[0040]

[0041] Where R 1 , R 2 and R 4 As defined herein and PG is an amino protecting group, reacted with a suitable deprotecting agent to form the compound of formula (I).

[0042] The third object of the present invention is a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein, and a pharmaceutically acceptable excipient.

[0043] The 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.

[0044] 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 the invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below.

[0045] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0046] Unless otherwise indicated, the nomenclature used in this application is based on the IUPAC systematic nomenclature. DETAILED DESCRIPTION

[0047] definition

[0048] "Alkoxy" refers to an alkyl group, as defined above, attached to the parent molecular moiety through an oxygen atom. Unless otherwise specified, alkoxy groups contain 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 -alkoxy"). In some preferred embodiments, the alkoxy group contains 1 to 4 carbon atoms. In 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.

[0049] "Alkoxyalkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group has been replaced by an alkoxy group. Preferably, "alkoxyalkyl" refers to an alkyl group in which 1, 2 or 3 hydrogen atoms, and most preferably one hydrogen atom, of the alkyl group has been replaced by an alkoxy group. A preferred but non-limiting example of alkoxyalkyl is methoxymethyl and 2-methoxyethyl.

[0050] "Alkyl" refers to a group having a specified number of carbon atoms (i.e., C 1-10 alkyl groups are saturated linear (i.e., unbranched) or branched monovalent hydrocarbon chains of one to ten carbon atoms (“C”). 1-20 alkyl”), having 1 to 12 carbon atoms (“C 1-12 alkyl”), having 1 to 10 carbon atoms (“C 1-10 alkyl”), having 1 to 8 carbon atoms (“C 1-8 alkyl”), having 1 to 6 carbon atoms (“C 1-6 alkyl”), having 2 to 6 carbon atoms (“C 2-6 alkyl") or 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, tert-butyl, isobutyl, sec-butyl, homologues and isomers thereof, such as n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0051] "Alkynyl" refers to a radical having at least one site of acetylenic unsaturation (ie, having at least one moiety of the formula C≡C) and having the specified number of carbon atoms (ie, C 2-10 means an unsaturated straight chain (ie, unbranched) or branched monovalent hydrocarbon chain or combinations thereof of two to ten carbon atoms). Specific alkynyl groups are those having 2 to 20 carbon atoms (“C 2-20 Alkynyl”), having 2 to 8 carbon atoms (“C 2-8 Alkynyl”), having 2 to 6 carbon atoms (“C 2-6Alkynyl”) or a group having 2 to 4 carbon atoms (“C 2-4 Examples of alkynyl groups include, but are not limited to, groups such as ethynyl (ethynyl or acetylenyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, but-3-ynyl, homologs and isomers thereof, and the like.

[0052] "Amino" used alone or in combination with other groups refers to NH2.

[0053] "Aminoalkyl" refers to an alkyl group in which one or more of the hydrogen atoms of the alkyl group has been replaced by an amino moiety.

[0054] "Aromatic" means the conventional concept of aromatics as defined in the literature, in particular in IUPAC-Compendium of Chemical Terminology, 2nd edition, AD McNaught & A. Wilkinson (Eds.). Blackwell Scientific Publications, Oxford (1997).

[0055] "Aryl" refers to 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 bicyclic rings having two fused five-membered aryl rings (denoted 5-5), fused bicyclic rings having one five-membered aryl ring and one fused six-membered aryl ring (denoted 5-6 and 6-5), and fused bicyclic rings having two fused six-membered aryl rings (denoted 6-6). The aryl group may be optionally substituted as described above. Examples of aryl substituents include, but are not limited to, phenyl, naphthyl, phenanthrenyl, 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, each of which is optionally substituted.

[0056] "Cancer" refers to a disease characterized by the presence of a neoplasm or tumor due to abnormal, uncontrolled growth of cells (such cells are "cancer cells"). As used herein, the term cancer specifically includes, but is not limited to, hepatocellular carcinoma, malignant tumors and hyperproliferative diseases of the colon (colon cancer), lung cancer, breast cancer, prostate cancer, melanoma, and ovarian cancer.

[0057] "Cyano" used alone or in combination with other groups refers to CN (ie, nitrile).

[0058] "Cyanoalkyl" refers to an alkyl group in which one or more of the hydrogen atoms of the alkyl group has been replaced by a cyano moiety.

[0059] "Cycloalkyl" refers to a saturated or partially unsaturated carbocyclic moiety having a monocyclic, bicyclic (including bridged bicyclic and cycloalkyl spirocyclic substituents) or tricyclic ring and 3 to 10 carbon atoms (i.e., C3-C 10 )cycloalkyl). The cycloalkyl moiety may be optionally substituted with one or more substituents. In particular aspects, the cycloalkyl contains 3 to 8 carbon atoms (i.e., (C3-C8)cycloalkyl). In other particular aspects, 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 partially unsaturated (cycloalkenyl) derivatives thereof (e.g., cyclopentenyl, cyclohexenyl, and cycloheptenyl), bicyclo[3.1.0]hexane, bicyclo[3.1.0]hexenyl, bicyclo[3.1.1]heptane, bicyclo[3.1.1]heptenyl, and bicyclo[1.1.1]pentane. The cycloalkyl moiety may be attached in a "spiro-cycloalkyl" or "cycloalkyl spiro" manner, such as a "spirocyclopropyl".

[0060] "ECx" refers to the effective concentration of a particular compound (for example) in culture medium or plasma required to obtain x% of the maximum specific effect in vitro or in vivo. Examples of "ECx" are EC20, EC50 and EC100, which represent the concentrations of a particular compound in culture medium or plasma required to obtain 20%, 50% and 100% of the maximum specific effect in vitro or in vivo, respectively. "Halo" or "halogen" refers to fluorine, chlorine, bromine and / or iodine. When a residue is substituted with more than one halogen, this can be referred to by using a prefix corresponding to the number of halogen substituents attached, for example, dihaloaryl, dihaloalkyl, trihaloaryl, etc. refer to aryl and alkyl substituted with two ("di") or three ("tri") halo groups, which can be, but are not necessarily, the same halo; thus, 4-chloro-3-fluorophenyl is within the scope of dihaloaryl. An alkyl group in which one or more hydrogens are replaced by a halo group is called a "haloalkyl", for example, "C 1-6 A preferred haloalkyl group is trifluoroalkyl (-CF3).

[0061] "Haloalkoxy" refers to an alkoxy group in which at least one halogen replaces each H in the hydrocarbon constituting the alkyl portion of the alkoxy group. Examples of haloalkoxy groups are difluoromethoxy (-OCHF2), trifluoromethoxy (-OCF3).

[0062] "Haloalkyl" refers to an alkyl group as defined above containing at least one carbon atom substituted with at least one halo group, the "halo" being as defined herein. Examples of "haloalkyl" groups that can be used in the present invention include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl and n-butyl independently substituted with one or more halo groups (e.g., fluorine, chlorine, bromine and iodine).

[0063] "Haloaryl" refers to an aryl group in which at least one hydrogen has been replaced by a halogen.

[0064] "Heteroaryl" refers to an aromatic heterocyclic monocyclic, bicyclic or tricyclic ring system of 5 to 14 ring atoms, preferably 5 to 10 ring atoms, more preferably 5 to 6 ring atoms, containing 1, 2, 3 or 4 heteroatoms selected from N, O and S, the remaining ring atoms being carbon. In some aspects, the monocyclic heteroaryl ring can be 5-membered to 6-membered. Bicyclic heteroaryl ring systems include fused bicyclic rings with two fused five-membered heteroaryl rings (denoted 5-5), fused bicyclic rings with one five-membered heteroaryl ring and one fused six-membered heteroaryl ring (denoted 5-6 and 6-5), and fused bicyclic rings with two fused six-membered heteroaryl rings (denoted 6-6). The heteroaryl group can be optionally substituted as described above. Examples of heteroaryl substituents include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, isoxazolyl, benzofuranyl, isothiazolyl, benzothiophenyl, benzophenylthio, indolyl, azaindolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzo Isoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, pyrrolopiperidinyl, furopyridinyl, thienopyridinyl, pyridopyrazinyl, pyrrolopyrimidinyl, pyrrolopyrazinyl, thienopyridazinyl, thienopyrimidinyl, thienopyrazinyl, furapyridazinyl, furapyrimidinyl and furopyrazinyl. Most preferably, "5-membered heteroaryl" refers to the following groups:

[0065]

[0066] "Heteroatom" refers to atoms other than carbon or hydrogen.

[0067] "Heterocycle" or "heterocyclyl" refers to a 3, 4, 5, 6, 7, 8, 9 and 10-membered monocyclic, 7, 8, 9 and 10-membered bicyclic (including bridged bicyclic and cycloalkyl spirocyclic substituents) or 10, 11, 12, 13, 14 and 15-membered tricyclic heterocyclic moiety, which is saturated or partially unsaturated and has one or more (e.g., 1, 2, 3 or 4) heteroatoms selected from oxygen, nitrogen and sulfur located in the ring, and the remaining ring atoms are carbon. In some aspects, the heterocycle is a heterocycloalkyl. In a specific aspect, the heterocycle or heterocyclyl refers to a 4, 5, 6 or 7-membered heterocycle. When used to refer to the ring atoms of the heterocycle, nitrogen or sulfur can also be in an oxidized form, and the nitrogen can be substituted with one or more (C1-C6) alkyl or groups. The heterocycle can be attached to its side group at any heteroatom or carbon atom, resulting in a stable structure. Any ring atom of the heterocycle can be optionally substituted with one or more substituents described herein. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepine, oxepinyl, thiepineyl, morpholinyl, pyrrolidine 1-oxide, N-hydroxypiperidine, 1-methylpyrrolidine N-oxide, diaziridinyl and quinuclidinyl. The term heterocycle also includes groups in which the 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.

[0068] "Hydroxyl" used alone or in combination with other groups refers to OH.

[0069] "Hydroxyalkyl" refers to an alkyl group in which one or more of the hydrogen atoms of the alkyl group has been replaced by a hydroxy moiety. Examples include alcohols and glycols.

[0070] "Moiety" and "substituent" refer to an atom or group of chemically bonded atoms that is attached to another atom or molecule by one or more chemical bonds to form a part of a molecule.

[0071] When indicating the number of substituents, the term "one or more" refers to the range from one substituent to the highest possible number of substituents, i.e., replacement of one hydrogen up to replacement of all hydrogens by substituents, particularly where "one or more" refers to one, two or three, most particularly "one or more" refers to one or two.

[0072] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where the event or circumstance does not occur. For example, "an aryl group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the aryl group is substituted with an alkyl group and instances where the aryl group is not substituted with an alkyl group.

[0073] "Optionally substituted" means unsubstituted or substituted. In general, the substituents may be the same or different.

[0074] "Oxo", used alone or in combination with other groups, means =0.

[0075] "Pharmaceutically acceptable salts" refer to those salts which retain the biological effects and properties of the free bases or free acids and which are not biologically or otherwise undesirable. The salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, especially hydrochloric acid, and with 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, benzylformic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine.

[0076] Particularly preferably, the pharmaceutically acceptable salts of the compounds of formula (I) are salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid and methanesulfonic acid.

[0077] As used herein, the term "protecting group" (PG) represents a group that selectively blocks a reactive site in a multifunctional compound so that another unprotected reactive site that is usually associated therewith in synthetic chemistry selectively reacts. The protecting group can be removed at an appropriate point. Exemplary protecting groups are amino protecting groups, carboxyl protecting groups, or hydroxy protecting groups. Specific protecting groups are tert-butyloxycarbonyl (Boc), benzyloxycarbonyl (Cbz), fluorenylmethoxycarbonyl (Fmoc), and benzyl (Bn). Further specific protecting groups are tert-butyloxycarbonyl (Boc) and fluorenylmethoxycarbonyl (Fmoc). A more specific protecting group is tert-butyloxycarbonyl (Boc). Exemplary protecting groups and their use in organic synthesis are described in, for example: TW Greene and PGM Wutts, Protective Groups in Organic Chemistry, 5th edition, 2014, John Wiley & Sons, NY.

[0078] As used herein, "prevention" includes preventing or delaying the onset of clinical symptoms of a relevant state, disease or condition in a mammal, and particularly a human who may have or be susceptible to a state, disease or condition but has not yet developed or displayed clinical or subclinical symptoms of the state, disease or condition.

[0079] "Substituted" refers to that at least one hydrogen atom of a compound or part is replaced by another substituent or part. 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 group (as defined below) are replaced by one or more halogen atoms (e.g., trifluoromethyl, difluoromethyl, fluoromethyl, chloromethyl, etc.). In one aspect, substitution as used herein may refer to that at least one hydrogen atom of a compound or part as described herein is replaced by halogen or alkyl.

[0080] "Therapeutically effective amount" refers to an amount of a compound or molecule of the invention that, when administered to a subject, (i) treats or prevents a particular disease, disorder or condition, (ii) reduces, ameliorates or eliminates one or more symptoms of a particular disease, disorder or condition, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, disorder or condition as described herein. The therapeutically effective amount depends 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 medical or veterinary doctor and other factors.

[0081] "Therapeutically inert carrier" refers to any ingredient that is not therapeutically active and non-toxic and is used in formulating a drug product, such as a disintegrant, binder, filler, solvent, buffer, tonicity agent, stabilizer, antioxidant, surfactant, or lubricant.

[0082] In particular, the chemical groups whose definitions are given above are those specifically exemplified in the Examples.

[0083] The following abbreviations are used in this text:

[0084] 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, D IPEA = N,N-diisopropylethylamine, EDC = 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, ESI = electrospray ionization, EtOAc = ethyl acetate, EtOH = ethanol, h = hour, HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate, HBTU = O-benzotriazole-N,N,N', N'-Tetramethyluronium hexafluorophosphate, HFIP = hexafluoroisopropanol, HOBt = hydroxybenzotriazole, HPLC = high performance liquid chromatography, m-CPBA = meta-chloroperbenzoic acid, MeCN = acetonitrile, MeI = iodomethane, MeOH = methanol, min = minute, MS = mass spectrometry, NBS = N-bromosuccinimide, PE = petroleum ether, PyBroP = bromotris(pyrrolidino)phosphonium hexafluorophosphate, RT = Room temperature, TBAF = tetrabutylammonium fluoride, TBAOH = tetrabutylammonium hydroxide, TBDMS = tert-butyldimethylsilyl, TEA = triethylamine, TFA = trifluoroacetic acid, THF = tetrahydrofuran, TMSOTF = trimethylsilyl trifluoromethanesulfonate, TLC = thin layer chromatography, T3P = 2,4,6-tripropyl-1,3,5,2,4,6-trioxotriphosphorinane 2,4,6-trioxide.

[0085] In the present specification, if there is a discrepancy between a depicted structure and the name given to that structure, the depicted structure will control. Additionally, if the stereochemistry of a structure or portion of a structure is not indicated, for example, by a bold wedge or dashed line, that structure or portion of a structure is interpreted as encompassing all stereoisomers thereof. However, in some cases where there is more than one chiral center, the structure and name may be represented as a single enantiomer to aid in describing the relative stereochemistry.

[0086] Unless otherwise indicated, the term "the compound of formula..." or "the compound of formula..." or "the compounds of formula..." or "compounds of formula..." refers to any compound selected from the group of compounds defined by the formula (including any pharmaceutically acceptable salt of any such compound if not otherwise indicated).

[0087] 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 separate individual tautomers generally produce a mixture with chemical and physical properties consistent with the compound mixture. The position of equilibrium depends on the chemical features within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates, while in phenols, the enol form predominates. Common prototropic tautomers include keto / enol Amide / Imidic Acid and amidine The latter two are particularly common in heteroaryls and heterocycles, and the present invention encompasses all tautomeric forms of the compounds.

[0088] Furthermore, the present invention includes all optical isomers of the compounds of formula (I), ie diastereomers, diastereomeric mixtures, racemic mixtures, all corresponding enantiomers and / or tautomers thereof and solvates thereof.

[0089] Compounds of formula (I) may contain one or more asymmetric centers and may therefore occur as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. Additional asymmetric centers may be present, depending on the characteristics of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and it is intended that all possible optical isomers and diastereomers in the form of mixtures and as pure compounds or partially purified compounds are included in the present invention. The present invention is intended to cover all such isomeric forms of these compounds. The independent synthesis of these diastereomers or their chromatographic separation can be achieved by making appropriate changes to the methods disclosed herein as known in the art. The absolute stereochemistry can be determined by x-ray crystallography of crystalline products or crystalline intermediates, and if necessary, these products or intermediates can be derivatized using reagents containing asymmetric centers of known absolute configuration. If necessary, the racemic mixture of the compound can be separated so that the individual enantiomers are separated. Separation can be carried out by methods well known in the art, such as coupling a racemic mixture of the compounds to an enantiomerically pure compound to form a diastereomeric mixture followed by separation of the individual diastereomers by standard methods such as fractional crystallization or chromatography.

[0090] In embodiments where an optically pure enantiomer is provided, an optically pure enantiomer means that the compound contains >90% by weight of the desired isomer, particularly >95% by weight of the desired isomer, or more particularly >99% by weight of the desired isomer, the weight percentages being based on the total weight of the isomers of the compound. Chirally pure compounds or chirally enriched compounds can be prepared by chiral selective synthesis or by enantiomer separation. Separation of enantiomers can be performed on the final product, or alternatively on a suitable intermediate.

[0091] In some embodiments, compounds of formula (I) are isotopically labeled by replacing one or more atoms with atoms having a different atomic mass or mass number. Such isotopically labeled (i.e., radiolabeled) compounds of formula (I) are considered to be 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, such as but not limited to 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 I. Certain isotopically-labeled compounds of formula (I) (e.g., those containing a radioactive isotope) are useful in drug and / or substrate tissue distribution studies. 3 H) and carbon-14 (i.e. 14 C) are particularly useful for this purpose because they are easy to incorporate and detection means are readily available. For example, compounds of formula (I) may be enriched with 1, 2, 5, 10, 25, 50, 75, 90, 95 or 99% of a given isotope.

[0092] Using heavier isotopes (such as deuterium, 2 H) substitutions may provide certain therapeutic advantages due to greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements.

[0093] Using positron-emitting isotopes such as 11 C. 18 F. 15 O and 13N) substitution can be used to examine positron emission tomography (PET) studies of 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 methods analogous to those described in the Examples set forth below using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously used.

[0094] Compounds of the present invention

[0095] In one embodiment, provided herein is a compound of formula (I),

[0096]

[0097] or a pharmaceutically acceptable salt thereof, wherein:

[0098] R 1 is a 6-membered heteroaryl group, wherein R 1 Optionally, one or more R 10 replace;

[0099] R 2 is hydrogen or halogen;

[0100] R 4 is selected from phenyl and pyridine, which is optionally substituted by one or more R 11 replace;

[0101] R 10 Selected from:

[0102] i)C 1-10 -alkyl, which is optionally substituted with one or more halogen, hydroxyl, -S(O)2(C 1-6 -alkyl), -N(R 10e R 10f )、-S(O)2(C 1-6 -cycloalkyl), cyano substituted;

[0103] ii) C 3-10 -cycloalkyl, which is optionally substituted with one or more -S(O)2(C 1-6 -alkyl) substituted;

[0104] iii) a 3- to 10-membered heterocyclic group, which is optionally substituted with one or more halogens, -C(O)O-(R 10q )

[0105] replace;

[0106] iv) phenyl;

[0107] v)-N(R 10e R 10f );

[0108] vi)-OR 10g ;

[0109] vii) halogen;

[0110] R 10e and R 10f are each independently selected from hydrogen and C 1-6 -alkyl;

[0111] R 10g Selected from C 1-6 -alkyl and halo-C 1-6 -alkyl;

[0112] R 10q C 1-10 -alkyl;

[0113] Or two R 10 Together with the carbon atoms to which they are attached, they form a 3- to 10-membered heterocyclic group, which is optionally substituted by one or more C 1-10 - alkyl substitution;

[0114] R 11 Selected from:

[0115] i) halogen;

[0116] ii) C 1-6 -alkyl, which is optionally substituted with one or more cyano groups;

[0117] iii) C 1-6 - alkoxy;

[0118] iv) C 3-7 - cycloalkyl;

[0119] v) a 5- to 6-membered heteroaryl group, which is optionally substituted with one or more halogenated -C 1-6 -alkyl, C 3-10 - Cycloalkyl substitution;

[0120] vi) phenyl, which is optionally substituted with one or more halogen, C 1-6 -alkoxy, halo-C 1-6 - alkyl substitution;

[0121] vii)-SO2(R 11d );

[0122] R 11d Selected from hydrogen, C 1-6 -alkyl and halo-C 1-6 -alkyl.

[0123] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 1is pyridyl, pyrimidinyl, pyridazinyl, triazinyl, which is optionally substituted by one or more R 10 replace.

[0124] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 1 is pyridyl, pyrimidinyl, triazine, which is optionally substituted by one or more R 10 replace.

[0125] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 1 is a pyridyl group, a triazine group, which is optionally substituted by one or more R 10 replace.

[0126] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 2 is hydrogen or fluorine.

[0127] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 2 For fluorine.

[0128] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 4 is phenyl, which is 11 replace.

[0129] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 10 is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidinyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy;

[0130] or R 10 and R 1 Together they form trimethyl-6,8-dihydro-1,7-naphthyridinyl and dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl.

[0131] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 10 It is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile.

[0132] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 11 Selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propionitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridinyl.

[0133] In another embodiment, provided herein is a compound of formula (I) as described herein, wherein R 11 Selected from chlorine, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy.

[0134] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein, wherein:

[0135] R 1 is pyridyl, pyrimidinyl, pyridazinyl, triazinyl, which is optionally substituted by one or more R 10 replace;

[0136] R 2 is hydrogen or halogen;

[0137] R 4 is phenyl, which is 11 replace;

[0138] R 10 Selected from:

[0139] i)C 1-10 -alkyl, which is optionally substituted with one or more halogen, hydroxyl, -S(O)2(C 1-6 -alkyl

[0140] -N(R 10e R 10f )、-S(O)2(C 1-6 -cycloalkyl), cyano substituted;

[0141] ii) C 3-10 -cycloalkyl, which is optionally substituted with one or more -S(O)2(C 1-6 -alkyl) substituted;

[0142] iii) a 3- to 10-membered heterocyclic group, which is optionally substituted with one or more halogens, -C(O)O-(R 10q )

[0143] replace;

[0144] iv) phenyl;

[0145] v)-N(R 10e R 10f );

[0146] vi)-OR 10g ;

[0147] vii) halogen;

[0148] R 10e and R 10f are each independently selected from hydrogen and C 1-6 -alkyl;

[0149] R 10g Selected from C 1-6 -alkyl and halo-C 1-6 -alkyl;

[0150] R 10q C 1-10 -alkyl;

[0151] R 11 Selected from:

[0152] i) halogen;

[0153] ii) C 1-6 -alkyl, which is optionally substituted with one or more cyano groups;

[0154] iii) C 1-6 - alkoxy;

[0155] iv) C 3-7 - cycloalkyl;

[0156] v) a 5- to 6-membered heteroaryl group, which is optionally substituted with one or more halogenated -C 1-6 -alkyl, C 3-10 - Cycloalkyl substitution;

[0157] vi) phenyl, which is optionally substituted with one or more halogen, C 1-6 -alkoxy, halo-C 1-6 -alkyl

[0158] replace;

[0159] vii)-SO2(R 11d );

[0160] R 11d Selected from hydrogen, C 1-6 -alkyl and halo-C 1-6 -alkyl.

[0161] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein, wherein:

[0162] R 1is pyridyl, pyrimidinyl, triazine, which is optionally substituted by one or more R 10 replace;

[0163] R 2 is hydrogen or fluorine;

[0164] R 4 is phenyl, which is 11 replace;

[0165] R 10 is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidinyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy;

[0166] or R 10 and R 1 Together they form trimethyl-6,8-dihydro-1,7-naphthyridinyl, dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl;

[0167] R 11 Selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propionitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridinyl.

[0168] In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein, wherein:

[0169] R 1 is a pyridyl group, a triazine group, which is optionally substituted by one or more R 10 replace;

[0170] R 2 For fluorine;

[0171] R 4 is phenyl, which is 11 replace;

[0172] R 10 is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile;

[0173] R 11 Selected from chlorine, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy.

[0174] In another embodiment, provided herein is a compound of formula (I) as described herein, which is selected from

[0175] (3R)-3-Amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0176] (3R)-3-Amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0177] (3R)-3-Amino-7-(2-tert-butyl-4-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0178] (3R)-3-Amino-7-(6-tert-butylpyrimidin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0179] (3R)-3-Amino-5-(4-chlorobenzyl)-8-fluoro-1,1-diketo-7-[5-(trifluoromethoxy)-3-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0180] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0181] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-isopropoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0182] (3R)-3-Amino-7-(6-tert-butylpyridazin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0183] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0184] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[6-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0185] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)pyrimidin-2-yl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0186] (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0187] (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-benzothiazepine -4-Keto;

[0188] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazine-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0189] (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0190] (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-benzothiazepine -4-Keto;

[0191] (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-benzothiazepine -4-Keto;

[0192] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidinyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0193] 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-1,2,4-triazine-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylic acid methyl ester;

[0194] (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-benzothiazepine -4-Keto;

[0195] (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-benzothiazepine -4-Keto;

[0196] (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-benzothiazepine -4-Keto;

[0197] (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-benzothiazepine -4-Keto;

[0198] 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-trione-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-3-pyridinyl]-2-methyl-propionitrile;

[0199] (3R)-3-Amino-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-7-[5-(trifluoromethyl)-3-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0200] (3R)-3-Amino-8-fluoro-1,1-diketo-7-[5-(1-methylsulfonyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0201] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0202] (3R)-3-Amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0203] (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-benzothiazepine -4-Keto;

[0204] (3R)-3-Amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0205] (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-benzothiazepine -4-Keto;

[0206] (3R)-3-amino-8-fluoro-1,1-dioxo-7-(3-tert-butyl-1,2,4-triazine-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0207] (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-benzothiazepine -4-Keto;

[0208] (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-benzothiazepine -4-ketone; and

[0209] (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-benzothiazepine -4-Keto;

[0210] or a pharmaceutically acceptable salt thereof.

[0211] In another embodiment, provided herein is a compound of formula (I) as described herein, which is selected from

[0212] (3R)-3-Amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0213] (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0214] (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0215] (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-benzothiazepine -4-Keto;

[0216] (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-benzothiazepine -4-Keto;

[0217] 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-trione-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-3-pyridinyl]-2-methyl-propionitrile;

[0218] (3R)-3-Amino-8-fluoro-1,1-diketo-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-7-[5-(trifluoromethyl)-3-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0219] (3R)-3-Amino-8-fluoro-1,1-diketo-7-[5-(1-methylsulfonyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0220] (3R)-3-Amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto;

[0221] (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-benzothiazepine -4-Keto;

[0222] (3R)-3-Amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-ketone; and

[0223] (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-benzothiazepine -4-ketone; or a pharmaceutically acceptable salt thereof.

[0224] Production method

[0225] Processes for the manufacture of compounds of formula (I) or pharmaceutically acceptable salts thereof as described herein are also an object of the present invention.

[0226] The present invention provides a method for preparing a compound as described herein or a pharmaceutically acceptable salt thereof, the method comprising:

[0227]

[0228] Where R 1 , R 2 and R 4 As defined herein and PG is an amino protecting group, reacted with a suitable deprotecting agent to form the compound of formula (I).

[0229] The preparation of the compound of formula (I) of the present invention can be carried out according to sequential or concurrent synthetic routes. The synthesis of the present invention is shown in the following general scheme. The skills required for performing the reaction and purifying the resulting product are known to those skilled in the art. Unless otherwise indicated, the substituents and indices used in the following method descriptions have the meanings provided herein.

[0230] If one of the starting materials, intermediates or compounds of formula (I) contains one or more functional groups that are unstable or reactive under the reaction conditions of one or more reaction steps, appropriate protecting groups may be introduced before applying key steps of methods well known in the art (such as "Protective Groups in Organic Chemistry" by TW Greene and PGM Wutts, 5th edition, 2014, John Wiley & Sons, NY). Such protecting groups may be removed at a later stage of the synthesis using standard methods described in the literature.

[0231] If the starting material or intermediate contains a stereogenic center, the compound of formula (I) can be obtained in the form of a mixture of diastereomers or enantiomers, which can be separated by methods well known in the art (e.g., chiral HPLC, chiral SFC or chiral crystallization). Racemic compounds can be separated into corresponding enantiomers, for example, by diastereomeric salts, which are crystallized with optically pure acids, or by specific chromatography separations of enantiomers using chiral adsorbents or chiral eluents. Starting materials and intermediates containing stereogenic centers can also be separated to provide starting materials and intermediates enriched in diastereomers / enantiomers. Using such diastereomer / enantiomer-enriched starting materials and intermediates in the synthesis of compounds of formula (I) will generally produce diastereomers / enantiomer-enriched compounds of the corresponding formula (I).

[0232] Those skilled in the art will recognize that in the synthesis of compounds of formula (I) (if not desired), an "orthogonal protecting group strategy" will be applied, which allows multiple protecting groups to be cleaved each time without affecting other protecting groups in the molecule. The principle of orthogonal protection is well known in the art and has also been reported in the literature (e.g., Barany and RB Merrifield, J. Am. Chem. Soc. 1977, 99, 7363; H. Waldmann et al., Angew. Chem. Int. Ed. Engl. 1996, 35, 2056).

[0233] Those skilled in the art will recognize that the order of reactions may vary depending on the reactivity and nature of the intermediates.

[0234] In more detail, the compound of formula (I) can be manufactured by the method given below, by the method given in the examples or by similar methods. The appropriate reaction conditions for each reaction step are known to those skilled in the art. Similarly, the reaction conditions affecting the reaction reported in the relevant literature are described, for example: Comprehensive Organic Transformations: A Guide to Functional Group Preparations, 2nd Edition, Richard C. Larock. John Wiley & Sons, New York, NY. 1999. It is convenient to react in the presence or absence of a solvent. There are no particular restrictions on the nature of the solvent used, as long as it has no adverse effect on the reaction or the reagents involved and can dissolve the reagents at least to a certain extent. The described reaction can occur over a wide temperature range, and the precise reaction temperature is not critical to the present invention. The above reaction can be conveniently performed in a temperature range of -78°C to reflux. The time required for the reaction can also vary over a wide range, depending on many factors, particularly the reaction temperature and the nature of the reagents. However, it usually takes 0.5 hours to several days to obtain the intermediates and compounds. The reaction sequence is not limited to the one shown in the schemes, but, depending on the starting materials and their respective reactivity, the order of the reaction steps can be freely varied.

[0235] If starting materials or intermediates were not commercially available or their synthesis was not reported in the literature, they could be prepared by analogy to existing preparations of close analogs or as outlined in the experimental section.

[0236] The compounds of formula (I) of the present invention or their pharmaceutically acceptable salts can be prepared by the method described below (Scheme 1) together with synthetic methods known in the field of organic chemistry, or modification and derivatization methods familiar to those skilled in the art.

[0237]

[0238]

[0239] Solution 1

[0240] Suitable starting materials for preparing compounds of formula (I) are nitro compounds of formula (II) wherein X 2 is F or Cl and X 1 Already R 1 or a group such as Br or -CO2 alkyl, which may then be detailed as R 1. The compound of formula (II) can be reacted with a suitably protected cysteine ​​derivative (III) in a solvent such as 1,2-dichloroethane or DMF at elevated temperature in the presence of a base such as DIPEA or potassium carbonate to obtain a compound of formula (IV). The preferred protecting group (PG) of the cysteine ​​derivative (III) is Boc. The nitro group in the compound of formula (IV) can be reduced using iron in a solvent mixture of water and ethanol at elevated temperature in the presence of hydrogen chloride or ammonium chloride to obtain a compound of formula (V). Alternatively, this conversion can be achieved by other reducing agents such as tin (II) chloride or by catalytic hydrogenation. Standard amide coupling conditions can be used to cyclize the compound of formula (V) to a compound of formula (VI). Preferably, the cyclization is carried out using 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphaninane 2,4,6-trioxide (50% solution in EtOAc) at room temperature and with a base such as DIPEA in a solvent such as DMF. 1 The reaction of a compound of formula (VII) wherein X is Cl, Br, I or a sulfonic acid group in the presence of a base (such as potassium carbonate) and (if necessary) with an additive (such as potassium iodide) in a solvent (such as DMSO or DMF) at room temperature gives a compound of formula (VIII). The sulfur atom in the compound of formula (VIII) is oxidized with an appropriate amount of an oxidant such as m-CPBA in a solvent such as DCM at room temperature to give a compound of formula (IX). For compounds of formula (VIII), wherein X is 1 is Br, which group may be detailed at this stage as a substituent R 1, as described in the following scheme, by reacting with bis(pinacol)diboron and a base (such as potassium acetate) in an aprotic solvent (such as 1,4-dioxane) at elevated temperature in the presence of a palladium(0) catalyst (e.g., 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride) to obtain a boronic acid derivative (X). Boronic acid derivative (X) is cross-coupled with a heteroaryl halide in an aprotic solvent (such as 1,4-dioxane or THF) at elevated temperature in the presence of a palladium(0) catalyst (e.g., 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride or tetrakis(triphenylphosphine)palladium(0) or Xphos Pd G4), a base (e.g., potassium carbonate, sodium carbonate, potassium phosphate or potassium acetate) to obtain a compound of formula (X1). Alternatively, compounds of formula (XI) can be prepared from the corresponding boronic acid derivatives (X) with heteroaryl methyl sulfide at elevated temperature in an aprotic solvent (e.g. THF) in the presence of a palladium (0) precursor catalyst (e.g. [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium (II), a copper salt (e.g. thiophene-2-carboxylic acid copper (I) hydrate), an additive (e.g. zinc acetate) to give compounds of formula (X1). Finally, deprotection provides compounds of formula (I). If 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. If the N-protecting group (PG) is Alloc, typical conditions are palladium (0) catalyzed saponification in an aprotic solvent in the presence of a nucleophile (e.g. morpholine) or a reducing agent (e.g. phenylsilane).

[0241] In addition, the substituent R 1 and R 4 May contain functional groups, which may be modified prior to removal of the N-protecting group (PG), or may require the use of suitable protecting groups during the synthesis. These protecting groups may be removed prior to removal of the N-protecting group (PG), or they may be removed simultaneously using suitable methods [Peter GMWuts, Greene'sprotective groups in organic synthesis, 5th edition, Hoboken, NJ: Wiley-Interscience].

[0242] Alternatively, compounds of formula (I) wherein R1 is triazole can be prepared as shown in Scheme 2.

[0243]

[0244] Solution 2

[0245] Esters of formula (XI) can be converted to acids of formula (XII) by treatment with MeOH (e.g., LiOH) in protic solvents such as MeOH, water, and solvents such as THF. Coupling of acid (XII) with hydrazine in the presence of a coupling agent such as CDI affords hydrazide (XIII), which is coupled with a 1,2-diketo compound at elevated temperature in the presence of ammonium acetate to afford an aryl heterocycle of formula (XIV). Oxidation as described in Scheme 1 affords sulfones of formula (XV), and final deprotection as described in Scheme 1 affords the title compound of formula (I).

[0246] Diketones / ketoaldehydes can be prepared by oxidation of the corresponding methyl ketones with, for example, selenodioxide at elevated temperatures.

[0247] R may contain a boc protecting group and may be chemoselectively deprotected and acylated using a chloroformate (eg methyl chloroformate) in the presence of an unprotected cyclic amine of formula (I).

[0248] Pharmaceutical compositions and administration

[0249] Another object of the present invention is to provide a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein, and a pharmaceutically acceptable excipient.

[0250] Formula (I) compound and its pharmaceutical salt can be used as medicine in the form of pharmaceutical preparation. Pharmaceutical preparation can be internally administered, such as oral (for example, in the form of tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions or suspensions), nasal (for example, in the form of nasal sprays) or rectal (for example, in the form of suppositories). However, administration can also be implemented parenterally such as intramuscularly or intravenously (for example, in the form of injections). Administration can also be implemented topically, for example, percutaneously, or in the form of eye drops or ear drops.

[0251] The compound of formula (I) and its pharmaceutically acceptable salt can be processed with a pharmaceutically inert inorganic or organic carrier to prepare a pharmaceutical preparation such as a tablet, a coated tablet, a dragee, a hard gelatin capsule, an injection or a topical preparation. For example, lactose, corn starch or its derivatives, talcum powder, stearic acid or its salts etc. can be used as such carriers for tablets, coated tablets, dragees and hard gelatin capsules.

[0252] 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, carriers are often not required in soft gelatin capsules.

[0253] Suitable carriers for the preparation of solutions and syrups are, for example, water, alcohols, polyols, sucrose, glucose, invert sugar, vegetable oils and the like.

[0254] Suitable carriers for injection solutions are, for example, water, alcohol, polyol, glycerol, vegetable oil and the like.

[0255] Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

[0256] Suitable liquids for topical ophthalmic formulations are, for example, cyclodextrins, mannitol or many other carriers and excipients known in the art.

[0257] In addition, the pharmaceutical preparations may contain preservatives, solubilizers, viscosity-increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorings, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They may also contain other therapeutically valuable substances.

[0258] Medicaments containing compounds of formula (I) or pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients and a process for their production, which comprises bringing one or more compounds of formula (I) and / or pharmaceutically acceptable salts thereof and, if desired, one or more other therapeutically valuable substances together with one or more pharmaceutically acceptable excipients into a galenical administration form, are also objects of the present invention.

[0259] Dosage is variable over a wide range, but will certainly have to be adjusted according to the individual needs of each particular case. Generally speaking, in the case of oral administration, the daily dose is about 0.1 mg to 20 mg per kg body weight, preferably 0.5 mg to 4 mg per kg body weight (e.g., about 300 mg per person), preferably divided into 1 to 3 separate doses (which may be composed of, for example, the same amount) should be suitable. In the case of topical administration, the preparation may contain 0.001% to 15% of the drug by weight, and the desired dose may be between 0.1 mg and 25 mg, and may be administered in a single dose every day or every week, or in multiple doses every day (2 to 4 times), or in multiple doses every week. However, it is apparent that if the upper or lower limit given herein is indicated, the upper or lower limit may be exceeded.

[0260] The pharmaceutical composition according to the present invention can be prepared as follows.

[0261] Preparation of pharmaceutical compositions containing the compounds of the present invention Tablet formulation (wet granulation)

[0262]

[0263] Manufacturing process:

[0264] 1. Mix ingredients 1, 2, 3 and 4 and granulate with purified water.

[0265] 2. Dry the granules at 50°C.

[0266] 3. Pass the particles through suitable grinding equipment.

[0267] 4. Add ingredient 5 and mix for three minutes; press on a suitable press.

[0268] Capsule preparation

[0269]

[0270] Manufacturing process:

[0271] 1. Mix ingredients 1, 2 and 3 in a suitable mixer for 30 minutes.

[0272] 2. Add ingredients 4 and 5 and mix for 3 minutes.

[0273] 3. Fill into suitable capsules.

[0274] Injection

[0275]

[0276]

[0277] Manufacturing process:

[0278] The compound of formula (I) is dissolved in a mixture of polyethylene glycol 400 and water for injection (part). The pH is adjusted to 5.0 by acetic acid. The volume is adjusted to 1.0 ml by adding the remainder of water. The solution is filtered, filled into a vial with an appropriate excess and sterilized.

[0279] Indications

[0280] The compounds of formula (I) may be used in an effective amount for treating a subject, particularly a human, affected by cancer.

[0281] 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.

[0282] In another 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.

[0283] In another aspect, the present invention provides a use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein for treating, preventing and / or delaying the progression of cancer.

[0284] In another aspect, the present invention provides a use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein for preparing a medicament for treating, preventing and / or delaying the progression of cancer.

[0285] In another aspect, the present invention provides a method for treating, preventing and / or delaying the progression of cancer, which method comprises administering a therapeutically effective amount of a compound of formula (I) as described herein or a pharmaceutically acceptable salt thereof.

[0286] As used herein, the terms "treatment" and "treating" and grammatical variants thereof refer to therapeutic therapy. With respect to a particular condition, treatment means: (1) ameliorating the condition or one or more of the biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that causes or contributes to the condition or (b) one or more of the 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 one or more of the biological manifestations of the condition. Prophylactic treatment using the methods and / or compositions of the present invention is also contemplated. Those skilled in the art will recognize that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to significantly reduce the likelihood or severity of a condition or its biological manifestation, or to delay the onset of such a condition or its biological manifestation. For example, when a subject is considered to be at high risk for cancer, such as when the subject has a strong family history of cancer or when the subject has been exposed to a carcinogen, prophylactic therapy is appropriate.

[0287] As an immunotherapeutic agent that acts on immune cells rather than directly on cancer cells, the present disclosure also foresees use as an anti-cancer vaccine. This also includes methods in which immune cells are cultured and manipulated ex vivo and the molecules disclosed herein are used as a means of conferring co-stimulation to the ex vivo manipulated cells.

[0288] In one embodiment, the cancer is a blood cancer, such as a lymphoma, leukemia, or myeloma. The blood cancers contemplated herein include, but are not limited to, 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); additional blood cancers or blood disorders, including, but not limited to, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse The following are some of the most common hematologic malignancies: non-Hodgkin's lymphoma, large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small-cell or large-cell follicular lymphoma, malignant lymphoproliferative disorders, mucosa-associated lymphoid tissue (MALT) lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, and "preleukemia," which is a collection of various hematologic disorders caused by ineffective production (or dysplasia) of blood cells in the bone marrow.

[0289] In a further embodiment, 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. The compound of formula (I) and Co-administration of other medications

[0290] The compound of formula (I) or its salt, or the compound disclosed herein or its pharmaceutical salt, can be used alone or in combination with other agents for treatment. For example, the second agent of the drug combination formulation or dosing regimen may have an activity complementary to the compound of formula (I) so that they do not adversely affect each other. The compounds can be administered together or separately in a single pharmaceutical composition. In one embodiment, the compound or pharmaceutical salt can be co-administered with a cytotoxic agent to treat proliferative diseases and cancer.

[0291] The term "co-administration" refers to the simultaneous administration of a compound of formula (I) or a salt thereof or a compound disclosed herein or a pharmaceutically acceptable salt thereof with one or more other active pharmaceutical ingredients (including cytotoxic agents and radiotherapy) or administered separately and sequentially in any manner. If the administration is not simultaneous, the compounds should be administered at a time close to each other. In addition, it is irrelevant whether the compounds are administered in the same dosage form, for example, one compound may be administered topically and another compound may be administered orally.

[0292] Generally, any agent with anticancer activity may be co-administered. Examples of such agents can be found in Cancer Principles and Practice of Oncology (V.T. Devita and S. Heilman, eds.), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. Based on the specific characteristics of the drugs and diseases involved, one of ordinary skill in the art will be able to discern which combination of agents will be useful.

[0293] In one aspect, the present invention provides a pharmaceutical composition as described herein, further comprising an additional therapeutic agent.

[0294] In one embodiment, the additional therapeutic agent is a chemotherapeutic agent.

[0295] In one embodiment, the additional therapeutic agent is a cytotoxic agent.

[0296] In one embodiment, the additional therapeutic agent is an immuno-oncology agent.

[0297] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (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 nucleolytic enzymes; and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof.

[0298] Exemplary cytotoxic agents can 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 cancer metabolism inhibitors.

[0299] "Chemotherapeutic agents" include chemical compounds that can be used to treat cancer. Examples of chemotherapeutic agents include erlotinib ( Genentech / OSI Pharm.), bortezomib ( Millennium Pharm.), disulfiram, epigallocatechin gallate, salisosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant ( AstraZeneca), sunitinib ( Pfizer / Sugen), letrozole ( Novartis), imatinib mesylate ( Novartis), phenaxate ( Novartis), oxaliplatin ( Sanofi), 5-FU (5-fluorouracil), folinic acid, rapamycin (sirolimus, Wyeth), Lapatinib ( GSK572016, GlaxoSmith Kline), Rhonafarmib (SCH 66336), Sorafenib ( Bayer Labs), gefitinib ( AstraZeneca), AG1478; alkylating agents such as thiotepa and Cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquinone, meturedopa, and uredopa; ethylimines and methylmelamines including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; annona lactones (especially bullatacin and bullatacinone); camptothecins (including topotecan and irinotecan); bryostatin; callystatin; CC-106 5 (including its synthetic analogs of adozelesin, carzelesin and bizelesin); cryptophycin (particularly cryptophycin I and cryptophycin 8); adrenocortical steroids (including prednisone and prednisolone); cyproterone acetate; 5a-reductase (including finasteride and dutasteride); vorinostat, romidepsin, panbicistat, valproic acid, mocetinostat, dolastatin; aldesleukin, talc, dukamycin (including synthetic analogs of KW-2189 and CBI-TM I); eleutherobin; sarcodictine; sarcodictyin; spongestatin; nitrogen mustards such as chlorambucil, chlorpheniramine, clofosamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine hydrochloride, melphalan, novembichin, phenambucil, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and lanimustine; antibiotics such as enediyne antibiotics (e.g. calicheamicin, especially calicheamicin and calicheamicin coll (Angew Chem. Inti. Ed. Engl. 1994 33:183-186); dynemicins, including dynemicin A; bisphosphonates, such as clodronate; esperamicin; and the neocarcinogen chromophores and related chromoprotein enediyne antibiotic chromophores, aclacinomycins, actinomycins, authramycins, azaserine, bleomycin, cactinomycin, carabicin, carminomicin, carzinophilin, chromomycin, dactinomycin, daunomycin, detopicin, 6-diazo-5-oxo-L-norleucine, (doxorubicin) (morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogamycin, olive mycins, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptomycin, streptozotocin, tuberculin, ubenimex, zorubicin, antimetabolites such as methotrexate and 5-fluorouracil (5 -FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calosterone, drostanolone propionate, cyclothiodine, melastane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folinic acid (frolinic acid) acid; aceglucuronide; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; colcemid; diaziquone; elfornithine; elliptonium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan Sugar; lonidainine; maytansinoids, such as maytansine and ansamitocin; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazine; procarbazine; polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizoran; germanium spiroamine; tenuazonic acid; triazoquinone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, baculocin A, and anguidine); urethane; vindesine; dacarbazine; mannitol mustard; dibromomannitol; dibromodulanol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, for example, TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), (without Cremophor), albumin-engineered paclitaxel nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, 111.) and (docetaxel, docetaxel; Sanofi-Aventis); chloranbucil; (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; (Changchunorelbine); Novantrone; Teniposide; Edatrexed; Daunorubicin; Aminopterin; Capecitabine Ibandronate; CPT-II; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; and pharmaceutically acceptable salts, acids and derivatives of any of the above.

[0300] Chemotherapeutic agents also include: (i) antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including tamoxifen (including Tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and (toremifine citrate); (ii) aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production by the adrenal glands, such as 4(5)-imidazole, aminoglutethimide, (Megestrol acetate), (exemestane; Pfizer), formestanie, fadrozole, (vorozole), (letrozole; Novartis) and (anastrozole; AstraZeneca); (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, triptorelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all trans-retinoic acid, fenretinide, and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation, such as PKC-α, Ralf, and H-Ras; (vii) ribozymes, such as inhibitors of VEGF expression (e.g. ) and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, such as and rIL-2; topoisomerase I inhibitors, e.g. and (ix) any of the pharmaceutically acceptable salts, acids and derivatives of the foregoing.

[0301] Chemotherapeutic agents also include antibodies such as alemtuzumab (Campath), bevacizumab ( Genentech; Cetuximab ( Imclone; Panitumumab ( Amgen), rituximab ( Genentech / Biogen Idee), Pertuzumab ( 2C4, Genentech), trastuzumab ( Genentech), tositumomab (Bexxar, Corixia) and antibody-drug conjugates, gemtuzumab ozogamicin ( Wyeth). Additional humanized monoclonal antibodies with therapeutic potential for use as agents in combination with the compounds of the invention include: apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumabmertansine, cantuzumab mertansine, cedilizumab, certolizumab pegol-conjugated pegol), cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin), ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab, tucotuzumabThe drugs include anti-interleukin 12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), a recombinant, fully human sequence-specific IgGiλ antibody that has been genetically modified to recognize the interleukin 12p40 protein.

[0302] Chemotherapeutic agents also include "EGFR inhibitors", which refer to compounds that bind to or directly interact with EGFR and prevent or reduce its signaling activity, and are 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 MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see, U.S. Pat. No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimeric 225 (C225 or cetuximab; ) and reshaped human 225 (H225) (see, WO 96 / 40210, Imclone Systems Inc.); IMC-11F8, a fully human antibody targeting EGFR (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 (see 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 with EGF and TGF-α for binding to EGFR (EMD / Merck); human EGFR antibody, HuMax-EGFR (GenMab); fully human antibodies, referred to as E1.1, E2.4, E2.5, E6.2, E6.4, E2.ll, E6.3 and E7.6.3, and described in US 6,235,883; MDX-447 (Medarex Inc); and mAb 806 or humanized mAb 806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)). Anti-EGFR antibodies can be conjugated to cytotoxic agents to generate immunoconjugates (see, e.g., EP 659,439 A2, Merck Patent GmbH). EGFR antagonists include small molecules, e.g., U.S. Pat. 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, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008 and 5,747,498, and the following PCT publications: compounds described in WO98 / 14451, WO98 / 50038, WO99 / 09016 and WO99 / 24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, Genentech / OSI Pharmaceuticals); PD 183805 (Cl 1033, 2-acrylamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer); ZD1839, gefitinib 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 (6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(l-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-butenamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); dual EGFR / HER2 tyrosine kinase inhibitors such as lapatinib ( GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[(2-methylsulfonyl)ethyl]amino]methyl]-2-furyl]-4-quinazolinamine).

[0303] Chemotherapeutic agents also include "tyrosine kinase inhibitors", including the EGFR targeted drugs described in the previous paragraph; small molecule FIER2 tyrosine kinase inhibitors, such as TAK165 available from Takeda Pharmaceuticals; 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 simultaneously inhibits cells overexpressing HER2 and EGFR; lapatinib (GSK572016; available from GlaxoSmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033; Pharmacia); Raf-I inhibitors, such as ISIS-5132, an antisense agent that inhibits Raf-I signaling, available from ISIS Pharmaceuticals; non-HER targeted TK inhibitors, such as imatinib mesylate ( available from GlaxoSmithKline); multitargeted tyrosine kinase inhibitors such as sunitinib ( VEGF receptor tyrosine kinase inhibitors such as vatalanib (PTK787 / ZK222584, available from Novartis / Schering AG); MAPK extracellular regulated kinase I inhibitor Cl-1040 (available from Pharmacia); quinazolines such as PD153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines such as CGP59326, CGP 60261 and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (difluoroformylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrosine containing a nitrothiophene moiety; PD-0183805 (Wamer-Lamber); antisense molecules (e.g., molecules that bind to HER encoding nucleic acids); quinoxalines (U.S. Pat. No. 5,804,396); tyrosine phosphorylation inhibitors (U.S. Pat. No. 5,804,396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); pan-HER inhibitors, such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); imatinib mesylate PKI 166 (Novartis); GW2016 (GlaxoSmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); seramatinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering); INC-ICl I (Imclone), rapamycin (sirolimus, ); or any of the following patent publications: U.S. Pat. No. 5,804,396, WO 1999 / 09016 (American Cyanamid), WO 1998 / 43960 (American Cyanamid), WO 1997 / 38983 (Warner Lambert), WO 1999 / 06378 (Warner Lambert), WO 1999 / 06396 (Warner Lambert), WO 1996 / 30347 (Pfizer, Inc), WO 1996 / 33978 (Zeneca), WO 1996 / 3397 (Zeneca), and WO 1996 / 33980 (Zeneca).

[0304] Chemotherapeutic agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, live BCG, bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, alfa), erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, interferon alfa-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium sodium), quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, valrubicin, zoledronate and zoledronic acid and pharmaceutically acceptable salts thereof.

[0305] Chemotherapeutic agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, thiocortisone pivalate, triamcinolone acetonide, triamcinolone acetonide alcohol, mometasone, amcinonide, budesonide, desonide, fluocinolone acetate, fluocinolone, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone-17-butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, and phenoxyethanol. dipropionate), betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, flucortolone hexanoate, flucortolone valerate, and fluprednidine acetate; immunoselective anti-inflammatory peptides (ImSAIDs), such as phenylalanine-glutamine-glycine (PEG) and its D-isomer form (feG) (IMULAN Biotherapeutics Ltd. BioTherapeutics, LLC); antirheumatic drugs such as azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomide, minocycline, sulfasalazine; tumor necrosis factor alpha (TNFa) blockers such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), golimumab (Simponi); interleukin I (IL-I) blockers such as anakinra (Kineret); T-cell costimulation blockers such as abatacept (Orencia); interleukin 6 (IL-6) blockers such as tocilizumab Interleukin 13 (IL-13) blockers, such as lebrikizumab; interferon alpha (IFN) blockers, such as lonavirin; β7 integrin blockers, such as rhuMAb β7; IgE pathway blockers, such as anti-M1 primers; secreted homotrimeric LTa3 and membrane-bound heterotrimeric LTa I / β2 blockers, such as anti-lymphotoxin α (LTa); radioisotopes (e.g., At 211 ,I 131 ,I 125 , Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 , P 32 , Pb 212and radioisotopes of Lu); various experimental agents such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3 or farnesyl transferase inhibitors (L-739749, L-744832); polyphenols such as quercetin, resveratrol, picrophenol, epigallocatechin gallate, theaflavins, flavanols, proanthocyanidins, betulinic acid and its derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, ); β-lapachone; lapaol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin); podophyllotoxin; tegafur Bexarotene Bisphosphonates, such as clodronate (e.g. or ), etidronate NE-58095, Zoledronic Acid / Zoledronate Alendronate Pamidronate Tirodronate Risedronate and epidermal growth factor receptor (EGF-R); vaccines such as Vaccines; 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 Pixantrone; farnesyl transferase inhibitors such as lonafamib (SCH 6636, SARASAR TM ); and pharmaceutically acceptable salts, acids or derivatives of any of the above; and combinations of two or more of the above, such as CHOP (an abbreviation for the combination therapy of cyclophosphamide, doxorubicin, vincristine and prednisolone); and FOLFOX (oxaliplatin (ELOXATIN TM ) is an abbreviation for the combination therapy with 5-FU and folinate).

[0306] In another embodiment, the compound of formula (I) can be co-formulated with an immuno-oncology agent. Immuno-oncology agents include, for example, small molecule drugs, antibodies or other biological or small molecules. Examples of biological immuno-oncology 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.

[0307] In one aspect, the immuno-oncology agent is an agonist of (i) stimulatory (including co-stimulatory) receptors or (ii) an antagonist of inhibitory (including co-inhibitory) signals on T cells, both of which result in amplification of antigen-specific T cell responses (commonly referred to as immune checkpoint modulators).

[0308] Certain stimulatory and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). An important family of membrane-bound ligands that bind to co-stimulatory 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 co-stimulatory or co-inhibitory receptors is the family of TNF molecules that bind to cognate TNF receptor family members, which include 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 / Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTfiR, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin alpha / TNP beta, TNFR2, TNF alpha, LTR, lymphotoxin alpha 1β2, FAS, FASL, RELT, DR6, TROY, NGFR.

[0309] In one aspect, T cell responses can be stimulated by a combination of a compound of Formula (I) and one or more of: (i) antagonists of proteins that inhibit T cell activation (e.g., immune checkpoint inhibitors), 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) agonists of proteins that stimulate T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0310] Other agents that can be combined with compounds of formula (I) for the treatment of cancer include antagonists of inhibitory receptors on NK cells or agonists of activating receptors on NK cells. For example, compounds of formula (I) can be combined with KIR antagonists such as lirilumab.

[0311] Other agents for combination therapy include agents that inhibit or deplete macrophages or monocytes, including but not limited to CSF-1R antagonists, such as CSF-1R antagonist antibodies, including RG7155 or FPA-008.

[0312] In another aspect, the compounds of formula (I) may be used with one or more of the following agents: agonists that bind to positive co-stimulatory receptors; blockers 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 engagement (e.g., PD-L1 / PD-1 interaction), 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 agents that trigger innate immune activation and / or inflammation at the tumor site.

[0313] In some embodiments, the immuno-oncology agent is a CTLA-4 antagonist, such as an antagonist CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, Ivo (ipilimumab) or tremelimumab. On the other hand, the immuno-oncology agent is a PD-1 antagonist, such as an antagonist PD-1 antibody. Suitable PD-1 antibodies include, for example, Opdivo (nivolumab), Keyruida (pembrolizumab) or MEDI-0680 (AMP-514; WO2012 / 145493). Immuno-oncology agents may also include pidilizumab (CT-011), although its specificity for binding to PD-1 is questioned. Another method of targeting the PD-1 receptor is a recombinant protein consisting of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, called AMP-224.

[0314] In another aspect, the immuno-oncology agent is a PD-L1 antagonist, such as an antagonist PD-L1 antibody. Suitable PD-L1 antibodies include, for example, Tecentriq (atezolizumab) (RG7446; WO2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO2007 / 005874), and MSB0010718C (WO2013 / 79174).

[0315] In another aspect, the immuno-oncology agent is a LAG-3 antagonist, such as an antagonist LAG-3 antibody. Suitable LAG3 antibodies include, for example, BMS-986016 (WO2010 / 19570, WO2014 / 08218), or IMP-731 or IMP-321 (WO2008 / 132601, WO2009 / 44273).

[0316] In another aspect, the immuno-oncology agent is a CD137 (4-1BB) agonist, such as an agonistic CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566 (WO2012 / 32433).

[0317] In another aspect, the immuno-oncology agent is a GITR agonist, such as 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).

[0318] In another aspect, the immuno-oncology agent is an IDO antagonist. Suitable IDO antagonists include, for example, INCB-024360 (WO2006 / 122150, WO2007 / 75598, WO2008 / 36653, WO2008 / 36642), indoximod or NLG-919 (WO2009 / 73620, WO2009 / 1156652, WO2011 / 56652, WO2012 / 142237).

[0319] In another aspect, the immuno-oncology agent is an OX40 agonist, such as an agonistic OX40 antibody. Suitable OX40 antibodies include, for example, MEDI-6383 or MEDI-6469. In another aspect, the immuno-oncology agent is an OX40L antagonist, such as an antagonist OX40 antibody. Suitable OX40L antagonists include, for example, RG-7888 (WO06 / 029879).

[0320] In another aspect, the immuno-oncology agent is a CD40 agonist, such as an agonistic CD40 antibody. In yet another embodiment, the immuno-oncology agent is a CD40 antagonist, such as an antagonistic CD40 antibody. Suitable CD40 antibodies include, for example, lucatumumab or dacetuzumab.

[0321] In another aspect, the immuno-oncology agent is a CD27 agonist, such as an agonistic CD27 antibody. Suitable CD27 antibodies include, for example, varlilumab.

[0322] In another aspect, the immuno-oncology agent is MGA271 (to B7H3) (WO2011 / 109400).

[0323] Examples

[0324] The present invention will be more fully understood by reference to the following examples. However, the claims should not be interpreted as being limited to the scope of the examples.

[0325] 1) Preparation Example

[0326] If not stated otherwise, all reaction examples and intermediates were prepared under argon atmosphere.

[0327] 1.1) General process:

[0328] · Alkylation: General Procedure 1

[0329] Potassium carbonate (1.14 g, 8.23 ​​mmol), potassium iodide (228 mg, 1.37 mmol) and a reagent (3.29 mmol) of formula (VII) were added to a solution of intermediate (2.74 mmol) of formula (VI) in DMSO (10 mL) at RT. The reaction was stirred for 2 h at RT, quenched with water and extracted twice with EtOAc. The combined organic layer was washed with water and saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. Required product (VIII) was used in the next step as a crude product, or purified by silica gel flash column chromatography or by reverse phase preparative HPLC.

[0330] · Oxidation: General Procedure 2

[0331] The solution of formula (VIII) intermediate (2.74mmol) and m-CPBA (1.18g, 6.85mmol) in DCM (10mL) was stirred at RT for 1 day. The reaction was diluted with ethyl acetate and THF, washed with 2N sodium hydroxide aqueous solution, 1N HCl aqueous solution and saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and the solvent was removed under reduced pressure. Required product (IX) is used in the next step as a crude product, or purified by silica gel flash column chromatography or by reverse phase preparative HPLC.

[0332] · Boropinacol ester formation: General procedure 3a

[0333] Argon is bubbled through a mixture of compound of formula (IX) (1 eq), bis(pinacol) diboron (CAS: 73183-34-3) (2 eq) and potassium acetate (3 eq) in 1,4-dioxane (0.2 M) for 5 minutes. 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (II) (0.1 eq) is added at RT, and the reaction mixture is heated to 80 ° C for 6 h. The reaction mixture is cooled to RT, diluted with EtOAc and THF, washed with saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and the solvent is removed under reduced pressure. The desired product (X) is used as a crude product in the next step, or purified by silica gel flash column chromatography.

[0334] · Boronic acid formation: General procedure 3b

[0335] Argon is bubbled through a mixture of formula (IX) compound (1eq), bis(pinacol) diboron (CAS:73183-34-3) (2.5eq) and potassium acetate (3.1eq) in 1,4-dioxane (0.05M) for 5 minutes. 1,1'-bis(diphenylphosphine) ferrocene palladium dichloride (II) (0.12eq) is added at RT, and the reaction mixture is heated to reflux for 3h. The reaction mixture is cooled to RT and filtered through diatomite, and the solvent is removed under reduced pressure. The desired product (X) is purified by preparative HPLC.

[0336] · Cross-coupling reactions: General procedure 4a

[0337] Argon is bubbled through a mixture of compound (X) (1 eq), heteroaryl halide (1.3 eq) and K2CO3 or KOAc (3 eq) in 1,4-dioxane and water (0.1 M) for 5 minutes. 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (II) (0.2 eq) is added, and the reaction mixture is heated to 80 ° C and stirred for 5 h. The reaction mixture is cooled to RT and diluted with EtOAc. The catalyst and solid are removed by filtration through diatomaceous earth. The filtrate is washed with water, saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and the solvent is evaporated under reduced pressure. The desired product (XI) is used as a crude product in the next step and purified by silica gel flash column chromatography or by reverse phase preparative HPLC.

[0338] · Cross-coupling reactions: General procedure 4b

[0339] Argon is bubbled through a mixture of formula (X) compound (1eq), heteroaryl halide (1.3eq) and Na2CO3 (2.5eq) in 1,4-dioxane and water (0.1M) for 5 minutes. Tetrakis (triphenylphosphine) palladium (0) (CAS: 14221-01-3) (0.1eq) is added, and the reaction mixture is heated to 90 ° C and stirred for 2h. The reaction mixture is cooled to RT and diluted with EtOAc. The catalyst and solid are removed by filtering through diatomaceous earth. The filtrate is washed with water, saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and the solvent is evaporated under reduced pressure. The desired product (XI) is used as a crude product in the next step, purified by silica gel flash column chromatography, preparative TLC or by reverse phase preparative HPLC.

[0340] · Cross-coupling reactions: General procedure 4c

[0341] Argon is bubbled through a mixture of compound of formula (X) (1.2 eq), heteroaryl (methyl sulfide) (1 eq), copper (I) thiophene-2-carboxylate hydrate (CAS: 1292766-17-6) (2.25 eq) and zinc acetate (1.2 eq) in THF (0.5 M) for 5 minutes. [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium (II) (0.1 eq) is added, and the reaction mixture is heated to 70 ° C for 6 h. After cooling, the mixture is filtered through celite, the filtrate is diluted with EtOAc, then washed with saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and the solvent is evaporated under reduced pressure. The desired product (XI) is used as a crude product in the next step, or purified by silica gel flash column chromatography, preparative TLC or by reverse phase preparative HPLC.

[0342] · Boc deprotection: General procedure 5a

[0343] To the solution of formula (XI) intermediate (38.9 μmol) in DCM (1 mL), dioxane (200 μl, 800 μmol, 20.6 eq) containing 4M HCl was added, and the reaction was stirred overnight at RT. The solvent was evaporated, and the residue was suspended in DCM and ether. The solid was filtered off, washed with ether and dried in vacuo to obtain the desired product (I).

[0344] · Boc deprotection: General procedure 5b

[0345] 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 eq) 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 obtain the desired product (I).

[0346] · Boc deprotection: General procedure 5c

[0347] A solution of the intermediate of formula (XI) (22.7 μmol) in 1,1,1,3,3,3-hexafluoropropane-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).

[0348] · Boc deprotection: General procedure 5d

[0349] To a solution of the intermediate of formula (XI) (0.25 mmol) in 1,1,1,3,3,3-hexafluoropropane-2-ol (4 mL) was added HCl / dioxane or HCl / Et2O (0.5 mmol, 2 eq) at 0°C. The reaction mixture was stirred at 20°C for 2 hours. The solvent was evaporated, and the resulting solid was absorbed in DCM and concentrated again to remove traces of 1,1,1,3,3,3-hexafluoropropane-2-ol. The process was repeated twice and then dried under high vacuum to obtain the desired product (I).

[0350] · Boc deprotection: general procedure 5e

[0351] A solution of the intermediate of formula (XI) (0.2mmol) in DCM (1.5mL) and TFA (0.5mL, 0.02mmol, 1.0eq) was stirred at 20°C for 0.3h. The reaction was carefully alkalized to pH=8 with saturated NaHCO3 solution at 0°C to 5°C. The mixture was extracted with DCM (10mL×3). The organic layer was washed with brine (10mL), dried over Na2SO4, and concentrated under vacuum at 35°C or less. The residue was dissolved in EtAOAc (30mL), and 4M HCl (0.1mL) was added dropwise to the mixture at 0°C to 5°C with stirring. The mixture was concentrated under vacuum at 35°C or less, dissolved in deionized water (20mL) and lyophilized to obtain the desired product (I).

[0352] 1.2) Synthesis of Examples

[0353] Example 1

[0354] (3R)-3-amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-

[0355] Dihydro-1λ6,5-benzothiazepine -4-Keto

[0356]

[0357] Step a) (2R)-3-(4-bromo-5-fluoro-2-nitro-phenyl)sulfanyl-2-(tert-butoxycarbonylamino)propanoic acid

[0358]

[0359] To a solution of N-Boc-L-cysteine ​​(CAS: 20887-95-0) (23.3 g, 105 mmol, 1.0 eq) and 1-bromo-2,4-difluoro-5-nitrobenzene (CAS: 345-24-4) (25.0 g, 105 mmol, 1.0 eq) in THF (150 mL) was added NaCO (22.3 g, 210.3 mmol, 2.0 eq). The mixture was stirred at 50 ° C for 12 h and then filtered. The filtrate was diluted with H O (200 mL) and extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (300 mL), dried over Na SO, filtered, and concentrated under reduced pressure. The crude title compound (32.6 g, 74.2 mmol, 71% yield) was obtained as a yellow oil and used in the next step without further purification. MS (ESI): 340.8 [M-isobutylene-CO2+H]+.

[0360] Step b) (2R)-3-(2-amino-4-bromo-5-fluoro-phenyl)sulfanyl-2-(tert-butoxycarbonylamino)propanoic acid

[0361]

[0362] 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 eq) in THF (150 mL) and H2O (100 mL) was added Fe (12.43 g, 222.6 mmol, 3.0 eq) and NH4Cl (11.91 g, 222.6 mmol, 3.0 eq), 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 Na2SO4, filtered, and concentrated under reduced pressure, and the remaining crude title compound (28.1 g, 68.66 mmol, 93% yield) was obtained as a brown oil and used in the next step without further purification. MS (ESI): 353.0 [M-isobutylene + H] +

[0363] Step c) N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0364]

[0365] To a solution of (2R)-3-(2-amino-4-bromo-5-fluoro-phenyl)sulfanyl-2-(tert-butoxycarbonylamino)propanoic acid (28.1 g, 68.6 mmol, 1.0 eq) in THF (150 mL) was added T3P (52.4 g, 82.3 mmol, 49.00 mL, 50% in EtOAc, 1.2 eq) and DPIEA (17.7 g, 137.3 mmol, 23.92 mL, 2.0 eq), and the mixture was stirred at 65° C. for 2 h. The reaction mixture was quenched by the addition of H2O (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, yield 20%) as a white solid. MS (ESI): 334.9 [M-isobutylene + H] + .

[0366] Step d) N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0367]

[0368] Analogously to General Procedure 1, the title compound was prepared from N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepine] -3-yl]carbamic acid tert-butyl ester (2.7 g, 6.9 mmol) and 1-(bromomethyl)-4-chloro-benzene (CAS: 622-95-7) were prepared and obtained as a white solid (5.4 g, yield 76%). MS (ESI): 517.1 [M+H] + .

[0369] Step e) N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0370]

[0371] Analogously to General Procedure 2, the title compound was prepared from N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (1.2 g, 2.33 mmol, 1 eq) was prepared and obtained as a white solid (1.26 g, yield 97.8%). MS (ESI): 491.1 / 493.1 [M-isobutylene + H] + .

[0372] Step f) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0373]

[0374] The title compound was prepared in analogy to General Procedure 3a from N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl] tert-butyl carbamate (1.2 g, 2.33 mmol, 1 eq) was prepared and obtained as a light brown solid (699 mg, 43% yield). MS (ESI): boronate ester cleavage was observed in MS: 457.2 ([M-(2,3-dimethyl-butadiene)-isobutylene+H] +).

[0375] Step g) N-[(3R)-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0376]

[0377] The title compound was prepared in analogy to General Procedure 4a from N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (60 mg, 0.1 mmol, Eq: 1) was prepared with 2-bromo-4-tert-butyl-pyridine (CAS: 50488-34-1) and obtained as a white solid (21.8 mg, yield 35%). MS (ESI): 602.4 [M+H] + .

[0378] 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-benzothiazepine -4-Keto

[0379]

[0380] The title compound was prepared in analogy to General Procedure 5a from N-[(3R)-7-(4-tert-butyl-2-pyridinyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (21.8 mg, 0.036 mmol, Eq: 1) was prepared and obtained as a light brown powder (16.8 mg, yield 85%). MS (ESI): 502.3 [M+H] + .

[0381] Examples 2 to 7 and 31 in the following table were prepared analogously to Example 1 using the appropriate bromide or chloride.

[0382]

[0383]

[0384] *As hydrochloride

[0385] **As formate

[0386] Example 5

[0387] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0388]

[0389] Step a) [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid

[0390]

[0391] The title compound was prepared in analogy to General Procedure 3b from N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl] tert-butyl carbamate (Example 1, step e) (150 mg, 0.274 mmol, Eq: 1) was prepared and obtained as a light brown oil (141 mg, yield 100%). MS (ESI): 457.4 [M-isobutylene + H] + .

[0392] Step b) N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[4-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0393]

[0394] The title compound was prepared in analogy to General Procedure 4b from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid (141 mg, 0.274 mmol, 1 eq) was prepared with 2-chloro-4-(trifluoromethyl)pyridine (CAS: 81565-18-6) and obtained as a white solid (120 mg, yield 71%). MS (ESI) 558.1 [M-isobutylene + H] + .

[0395] Step c) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0396]

[0397] The title compound was prepared in analogy to General Procedure 5b from N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-7-[4-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (120 mg, 0.195 mmol, Eq: 1) was prepared and obtained as a white powder (34.8 mg, 0.060 mmol, yield 30%). MS (ESI): 514.5 [M+H] + .

[0398] Intermediate 1

[0399] 6-tert-Butyl-3-chloro-1,2,3-triazine

[0400]

[0401] Step a) 6-tert-butyl-3-amino-1,2,3-triazine and 5-tert-butyl-3-amino-1,2,3-triazine

[0402]

[0403] A solution of 1,1-dibromo-3,3-dimethyl-butane-2-one (4.67 g, 18 mmol, 1.0 eq), aminoguanidine hydrochloride (2.0 g, 18 mmol, 1.0 eq) and potassium acetate (5.33 g, 54 mmol, 3.0 eq) 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 Na2SO4, and concentrated under vacuum. 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-triazine-3-amine (130 mg, 0.85 mmol, 3.3% yield) as a light yellow solid (MS (ESI): 153.2 [M + H] +), and 6-tert-butyl-1,2,4-triazine-3-amine, which were further purified by preparative HPLC to give 5-tert-butyl-1,2,4-triazine-3-amine (1.9 g, 12 mmol, 57% yield) as a light yellow solid. MS (ESI): 153.2 [M+H] + .

[0404] Step b) 6-tert-butyl-3-chloro-1,2,3-triazine

[0405]

[0406] To a solution of 6-tert-butyl-1,2,4-triazine-3-amine (130 mg, 0.85 mmol, 1.0 eq) and copper (II) chloride (137 mg, 1 mmol, 1.2 eq) in acetonitrile (2 mL) was added tert-butyl nitrite (132 mg, 1.28 mmol, 1.5 eq) dropwise at 20 °C. The mixture was stirred at 60 °C for 1 h. The mixture was poured into water (30 mL), extracted with EtOAc (50 mL), the organic phase was washed with brine (50 mL), dried over Na2SO4, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (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] + .

[0407] Examples 9 to 12 in the table below were prepared analogously to Example 5 using the appropriate bromide or chloride.

[0408]

[0409] *As hydrochloride

[0410] Example 13

[0411] (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazine-3-yl)-1,1-

[0412] Dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0413]

[0414] Step a) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine 7-Methyl formate

[0415]

[0416] (3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -7-Methyl formate (CAS: 2002449-37-6 6) (350mg, 0.7mmol, 1eq) was dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (18.6mL). 1,4-dioxane (25.7mg, 21.48uL, 0.707mmol, 1eq) containing 4M HCl was added to the solution, and the reaction mixture was stirred at RT for 1h. The reaction mixture was concentrated in vacuo. The remaining residue was dissolved in DCM, and the solution was poured on water. NaHCO3 and 1M NaOH were added to the aqueous phase to adjust the pH to 13. The aqueous phase was extracted with DCM (2×50mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the title compound as an orange solid (268mg, 77% yield), which was used in the next step as a crude product. MS (ESI) 395.1 [M+H] + .

[0417] Step b) (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -7-Methyl formate

[0418]

[0419] To (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -7-Methyl formate (159mg, 0.4mmol, 1eq) in ethyl acetate (0.415mL) was added with water (0.3mL) containing Na2CO3 (128mg, 1.21mmol, 3eq). Allyl chloroformate (58.2mg, 51.54uL, 0.48mmol, 1.2eq) was slowly added, the temperature of the reaction mixture was maintained at 20°C to 25°C, and the resulting biphasic mixture was vigorously stirred at RT for 1h. The mixture was diluted with water (100mL) and EtOAc (50mL). The layers were separated, and the aqueous layer was extracted with EtOAc (3×30mL). The combined organic layers were washed with brine (100mL) and dried over sodium sulfate. The crude product was purified by silica gel column chromatography (heptane containing 0% to 30% EtOAc) to obtain the title compound (202mg, yield 99%), which was an orange viscous oil. MS (ESI) 479.2 [M+H] + .

[0420] Step c) (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine 7-Formic acid

[0421]

[0422] To (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine To a solution of methyl-7-formate (202 mg, 0.422 mmol, 1 eq) 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 eq). The mixture was stirred at 22 ° C for 1.5 h. The solvent was concentrated in vacuo and the remaining residue was dissolved in EtOAc and washed with 1N HCl aqueous solution and brine. The organic layer was dried over sodium sulfate and concentrated in vacuo to give the title compound (190 mg, yield 78%) as an orange viscous oil. MS (ESI) 465.2 [M + H] + The product was used in the next step as received.

[0423] Step d) N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(hydrazinecarbonyl)-4-oxo-2,3-dihydro-1,5-benzothiazepine 3-aminophenyl]allyl carbamate

[0424]

[0425] To (3R)-3-(allyloxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -7-formic acid (190mg, 0.4mmol, 1eq) was added to a solution of THF (1.75mL) with CDI (86.1mg, 0.53mmol, 1.3eq) at one time, and the mixture was stirred for 2h. The solution was then added to a solution of hydrazine hydrate (95.9mg, 93.11uL, 1.23mmol, 3eq) in THF (0.29mL) at one time, and the mixture was stirred at room temperature for 1h. The reaction mixture was distributed between EtOAc and water, and brine (5mL) was added. The layers were separated, and the aqueous layer was extracted with EtOAc (2×50mL). The combined organic layers were washed with brine and dried over sodium sulfate, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (heptane containing 0% to 100% EtOAc) to give the title compound (109mg, yield 53%) as a red solid. MS (ESI) 479.1 [M+H] + .

[0426] Step e) 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-benzothiazepine -3-yl] allyl carbamate

[0427]

[0428] A microwave vial was charged with N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(hydrazinecarbonyl)-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl] allyl carbamate (55mg, 0.115mmol, 1eq), 1-phenyl-1,2-propanedione (CAS: 579-07-7) (17.0mg, 15.47uL, 0.11mmol, 1eq) and ammonium acetate (88.5mg, 1.15mmol, 10eq) and acetic acid (0.574mL). Under microwave irradiation, the reaction mixture was heated to 140°C for 10min and then heated to 180°C for 5min. 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 sodium sulfate and concentrated in vacuo to give the title compound (30mg, 42%) as an off-white solid. MS (ESI) 590.2 [M + H] +The product was used as is in the next step.

[0429] Step f) N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazine-3-yl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl] allyl carbamate

[0430]

[0431] 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-benzothiazepine To a solution of allyl]carbamate (29 mg, 0.049 mmol, 1 eq) in DCM (0.98 mL) was added 3-chloroperoxybenzoic acid (22.0 mg, 0.098 mmol, 2 eq), and the solution was stirred at RT for 4 h. The solution was diluted with DCM and washed with 1 M NaOH solution. The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude residue was purified by silica gel column chromatography (heptane containing 0% to 100% EtOAc) to give the title compound (14.2 mg, 42%) as an off-white solid. MS (ESI) 622.2 [M+H] + .

[0432] Step g) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazine-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0433]

[0434] Tetrakis(triphenylphosphine)palladium (1.3 mg, 0.001 mmol, 0.050 eq) and N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazine-3-yl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine were added to the mixture under argon atmosphere. -3-yl] allyl carbamate (14 mg, 0.023 mmol, 1 eq) was added to a mixture of DCM (0.13 mL) with phenylsilane (12.1 mg, 13.87 uL, 0.113 mmol, 5 eq), and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with water and DCM, and the phases were separated, and an aqueous NaHCO3 solution and a 1M NaOH aqueous solution were added to adjust the pH of the aqueous layer to 13. The aqueous layer was then extracted with DCM (2×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product 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] + .

[0435] Example 14

[0436] (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0437]

[0438] Step a) N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0439]

[0440] Analogously to General Procedure 1, the title compound was prepared from N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepine] -3-yl]carbamic acid tert-butyl ester (600 mg, 1.53 mmol) and 1-(chloromethyl)-4-isopropoxy-benzene (CAS: 40141-12-6) were prepared and obtained as a dark red oil (800 mg, yield 97%). MS (ESI): 541.1 [M+H] + .

[0441] Step b) N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0442]

[0443] Analogously to General Procedure 2, the title compound was prepared from N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (780 mg, 1.45 mmol, 1 eq) was prepared and obtained as a light yellow solid (550 mg, yield 67%). MS (ESI): 592.9 [M+Na] + .

[0444] Step c) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0445]

[0446] The title compound was prepared in analogy to General Procedure 3a from N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (300 mg, 0.525 mmol, 1 eq) was prepared and obtained as a colorless oil (150 mg, yield 46%).

[0447] Step d) N-[(3R)-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0448]

[0449] The title compound was prepared in analogy to General Procedure 4a from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (140 mg, 0.226 mmol, Eq: 1) and 5-tert-butyl-3-chloro-1,2,4-triazine (CAS: 83413-03-0) were prepared and obtained as a white solid (40 mg, yield 28%). MS (ESI): 628.2 [M+H] + .

[0450] Step e) (3R)-3-amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0451]

[0452] The title compound was prepared in analogy to General Procedure 5b from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (35 mg, 0.056 mmol, Eq: 1) was prepared and obtained in the form of a white powder (25.5 mg, 0.05 mmol, yield 79%). MS (ESI): 528.1 [M+H] + .

[0453] Example 15

[0454] (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-benzothiazepine -4-Keto

[0455]

[0456] Step a) N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(diethylamino)-1,2,4-triazine-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0457]

[0458] The title compound was prepared in analogy to General Procedure 4c from N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl] tert-butyl carbamate (100 mg, 0.19 mmol, Eq: 1) and 3-chloro-N,N-diethyl-1,2,4-triazine-5-amine (CAS: 1247362-85-1) were prepared and obtained as a yellow solid (6 mg, yield 5%). MS (ESI): 619.2, [M+H] + .

[0459] 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-benzothiazepine -4-Keto

[0460]

[0461] The title compound was prepared in analogy to General Procedure 5b from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (6 mg, 0.01 mmol, Eq: 1) was prepared and obtained as a light brown solid (3.2 mg, 0.01 mmol, yield 58%). MS (ESI): 519.0 [M+H] + .

[0462] Example 16 in the table below was prepared similarly to Example 15 using the appropriate boronate ester.

[0463]

[0464] *As hydrochloride

[0465] Example 17

[0466] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidinyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0467]

[0468] Step a) (5E)-3-Methylthio-1,2,4-triazine-5-aldehyde oxime

[0469]

[0470] To a stirred suspension of powdered potassium hydroxide (3088 mg, 55.0 mmol, 7.0 eq) in anhydrous DMSO (20 mL) was added 3-methylthio-1,2,4-triazine (1000 mg, 7.86 mmol, 1.0 eq) and nitromethane (1439 mg, 23.59 mmol, 3.0 eq) in DMSO (2 mL) at 25 ° C. The mixture was stirred at 25 ° C. for 16 h under N2. The reaction was poured into water (200 mL) and extracted with EtOAc (100 mL×2). The aqueous phase was acidified to pH 2 with 1N HCl and extracted with EtOAc (100 mL×3). The combined organic phase was washed with brine (200 mL x 2), dried over anhydrous Na2SO4, and concentrated to give a crude product, which was purified by silica gel column chromatography (PE / EtAc=5 / 1 to 3 / 1), and the eluent was concentrated under vacuum to give the title compound as a yellow solid (350 mg, 2.06 mmol, yield 24%). MS (ESI): 171.1 [M+H] + .

[0471] Step b) 3-Methylthio-1,2,4-triazine-5-carbonitrile

[0472]

[0473] To a solution of (5E)-3-methylthio-1,2,4-triazine-5-aldehyde oxime (200 mg, 1.18 mmol, 1.0 eq) and triethylamine (0.02 mL, 0.12 mmol, 0.1 eq) in toluene (5 mL) was added isocyanatobenzene (0.25 mL, 2.35 mmol, 2.0 eq) at 25° C. The mixture was then degassed three times with N2, heated to reflux, and stirred for 2 h. The mixture was cooled to room temperature and then filtered, the filter cake was washed with EtOAc (3 mL), the filtrate was diluted with EtOAc (5 mL), and quenched with brine (5 mL×2), dried over anhydrous Na2SO4, and concentrated to give a crude product, which was purified by silica gel column chromatography (PE / EtAc=8 / 1 to 4 / 1), and the eluent was concentrated under vacuum to give the title compound (140 mg, 0.92 mmol, yield 76%) as a dark red oil. MS (ESI): 153.1 [M+H] + .

[0474] Step c) 1,1-difluoro-4-(3-methylthio-1,2,4-triazine-5-yl)-1,4-thiazinane

[0475]

[0476] 3-Methylsulfanyl-1,2,4-triazine-5-carbonitrile (60.0 mg, 0.39 mmol, 1.0 eq) was added to 4,4-difluoro-piperidine (47.7 mg, 0.39 mmol, 1.0 eq), and the mixture was stirred at 25 ° C for 1 h. The mixture was diluted with EtOAc (5 mL) and concentrated under vacuum with silica gel (500 mg). The residue was purified by silica gel column chromatography (PE / EtAc=2 / 1 to 1 / 1), and the eluent was concentrated under vacuum to give the title compound (80.0 mg, 0.32 mmol, 80% yield) as a light solid. MS (ESI): 247.0 [M+H] + .

[0477] Step d) N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidinyl)-1,2,4-triazine-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0478]

[0479] The title compound was prepared in analogy to General Procedure 4c from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid (124.9 mg, 0.24 mmol, 1.2 eq) and 5-(4,4-difluoro-1-piperidinyl)-3-methylsulfanyl-1,2,4-triazine were prepared and obtained as a white solid (50 mg, yield 35%). MS (ESI) 667.1 [M+H] + .

[0480] Step e) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidinyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine 4-Keto

[0481]

[0482] The title compound was prepared in analogy to General Procedure 5b from N-[(3R)-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidinyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (50 mg, 0.075 mmol, Eq: 1) was prepared and obtained as a light brown solid (19.4 mg, 0.030 mmol, yield 43%). MS (ESI): 566.9 [M+H] + .

[0483] Example 18

[0484] 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylic acid methyl ester

[0485]

[0486] Step a) tert-Butyl 3-ethyl-5-[methoxy(methyl)carbamoyl]piperidine-1-carboxylate

[0487]

[0488] To a solution of 3-tert-butoxycarbonyl-3-azabicyclo[3.1.1]heptane-1-carboxylic acid (3.2 g, 13.2 mmol, 1.0 eq), N,N-diisopropylethylamine (5.78 mL, 33.1 mmol, 2.5 eq), 1-hydroxybenzotriazole (2.69 g, 19.8 mmol, 1.5 eq) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.81 g, 19.8 mmol, 1.5 eq) in DMF (35 mL) at 25°C was added O,N-dimethylhydroxylamine hydrochloride (1.68 g, 17.2 mmol, 1.3 eq). The mixture was stirred at 25°C for 16 hours and poured into water (40 mL). The aqueous phase was extracted with EtOAc (40 mL×3). The combined organic phases were washed with brine (90 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude product was purified by silica gel column chromatography (PE: EtOAc 20% to 60%) to give the title compound (3.61 g, 12.7 mmol, 95% yield) as a white solid. MS (ESI): 229.1 [M-isobutylene + H] + .

[0489] Step b) tert-Butyl 1-acetyl-3-azabicyclo[3.1.1]heptane-3-carboxylate

[0490]

[0491] Methylmagnesium bromide (6.01 mL, 18 mmol, 1.5 eq) was added to a solution of tert-butyl 3-ethyl-5-[methoxy(methyl)carbamoyl]piperidine-1-carboxylate (3.61 g, 12.0 mmol, 1.0 eq) in -THF (50 mL) at -20 ° C, and the mixture was stirred at 25 ° C for 3 hours. The reaction was quenched by adding saturated NH4Cl aqueous solution (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 under vacuum. The residue was purified by silica gel column chromatography (PE / EtOAc 20% to 100%) to give the title compound (2.74 g, 10.7 mmol, yield 89%) as a colorless oil. MS (ESI): 184.0 [M+H] + .

[0492] Step c) tert-Butyl 1-oxalyl-3-azabicyclo[3.1.1]heptane-3-carboxylate

[0493]

[0494] To a solution of tert-butyl 1-acetyl-3-azabicyclo[3.1.1]heptane-3-carboxylate (400 mg, 1.67 mmol, 1.0 eq) in 1,4-dioxane (4 mL) and water (1 mL) was added selenodioxide (380 mg, 3.42 mmol, 2.05 eq) 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 and purified by preparative HPLC and lyophilized to give the title compound (344 mg, 1.36 mmol, 81% yield) as a light yellow solid. MS (ESI): 198.2 [M-isobutylene + H] + .

[0495] Step d) tert-Butyl 1-(3-methylthio-1,2,4-triazine-5-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate and tert-butyl 1-(3-methylthio-1,2,4-triazine-6-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate

[0496]

[0497] To a solution of 1-oxalyl-3-azabicyclo[3.1.1]heptane-3-carboxylic acid tert-butyl ester (172 mg, 0.68 mmol, 1.0 eq) and sodium bicarbonate (85.5 mg, 1.02 mmol, 1.5 eq) in ethanol (2 mL) was added water (2 mL) containing 1-amino-2-methyl-isothiourea hydroiodide (158 mg, 0.68 mmol, 1.0 eq) at 25 ° C, and then the mixture was stirred at 25 ° C for 4 hours. The reaction 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 x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a crude product, which was purified by preparative TLC (PE: EtOAc = 3: 1) to give tert-butyl 1-(3-methylsulfanyl-1,2,4-triazine-5-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate (244 mg, 0.76 mmol, yield 111%) as a pale yellow oil and tert-butyl 1-(3-methylsulfanyl-1,2,4-triazine-6-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylate (30.0 mg, 0.09 mmol, yield 13%) as a pale red oil. MS (ESI) 323.1 [M+H] + .

[0498] Step e) 1-[3-[(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-1,2,4-triazin-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylic acid tert-butyl ester

[0499]

[0500] The title compound was prepared in analogy to General Procedure 4c from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid (232 mg, 0.45 mmol, 1.2 eq) and 1-(3-methylsulfanyl-1,2,4-triazine-5-yl)-3-azabicyclo[3.1.1]heptane-3-carboxylic acid tert-butyl ester were prepared and obtained as an orange oil (162 mg, yield 58%). MS (ESI): 687.0 [M-Boc-isobutylene + H] + .

[0501] Step f) (3R)-3-amino-7-[5-(3-azabicyclo[3.1.1]heptane-1-yl)-1,2,4-triazine-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine 4-Keto

[0502]

[0503] The title compound was prepared in analogy to General Procedure 5b from 1-[3-[(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-1,2,4-triazine-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylic acid tert-butyl ester (152 mg, 0.205 mmol, Eq: 1) was prepared and obtained as an orange solid (145 mg, yield 100%). MS (ESI): 543.1 [M+H] + .

[0504] Step g) 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine 7-yl]-1,2,4-triazine-5-yl]-3-azabicyclo[3.1.1]heptane-3-carboxylic acid methyl ester

[0505]

[0506] To (3R)-3-amino-7-[5-(3-azabicyclo[3.1.1]heptane-1-yl)-1,2,4-triazin-3-yl]-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine was added at 0°C. To a solution of -4-ketone (30.0 mg, 0.050 mmol, 1 eq) in DCM (1 mL) was added dimethyl dicarbonate (0.01 mL, 0.050 mmol, 1 eq). The mixture was stirred at 25 °C for 30 min. 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] + .

[0507] Example 19

[0508] (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-benzothiazepine -4-Keto

[0509]

[0510] Step a) 5-(bromomethyl)-3-methylthio-1,2,4-triazine

[0511]

[0512] To a solution of 5-methyl-3-methylthio-1,2,4-triazine (20.0 g, 141.6 mmol, 1.0 eq) and N-bromosuccinimide (22.69 g, 127.4 mmol, 0.9 eq) in MeCN (400 mL) was added AIBN (6.79 g, 41.33 mmol, 0.29 eq). The mixture was stirred at 25 ° C for 3 h. The reaction 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% to 10%) to give the title compound (2500 mg, 11.36 mmol, 5% yield) as a light brown solid.

[0513] Step b) 3-methylthio-5-(methylsulfonylmethyl)-1,2,4-triazine

[0514]

[0515] To a mixture of sodium methanesulfinate (2899 mg, 28.4 mmol, 2.5 eq) and potassium iodide (0.6 mL, 11.36 mmol, 1.0 eq) in DMF (52 mL) was added 5-(bromomethyl)-3-methylthio-1,2,4-triazine (2500 mg, 11.3 mmol, 1.0 eq) at 25 ° C, and the mixture was stirred at 50 ° C for 16 h. The solution was diluted with EtOAc (100 mL), then washed with brine (50 mL×3), dried over anhydrous Na2OS4, concentrated under vacuum, and purified by silica gel column chromatography (PE / EtAc=10 / 1 to 1 / 1), and the eluent was concentrated under vacuum to obtain the confirmed title compound (220 mg, 1.0 mmol, yield 8%) as a brown oil. MS (ESI): 220.2 [M+H] +Step c) 3-methylthio-5-(1-methylsulfonylcyclopropyl)-1,2,4-triazine

[0516]

[0517] A solution of 3-methylthio-5-(methylsulfonylmethyl)-1,2,4-triazine (330 mg, 1.5 mmol, 1.0 eq), 1-bromo-2-chloroethane (0.17 mL, 2.11 mmol, 1.4 eq), potassium iodide (0.02 mL, 0.3 mmol, 0.2 eq) and K2CO3 (519.2 mg, 3.76 mmol, 2.5 eq) in DMF (15 mL) was heated to 60° C. and stirred for 16 h under N2. Cs2CO3 (560 mg, 1.72 mmol, 1.14 eq) and potassium iodide (0.02 mL, 0.3 mmol, 0.2 eq) were added to the mixture, and then stirred at 60° C. for another 16 h. The mixture was diluted with EtOAc (20 mL), washed with brine (20 mL×2), dried over anhydrous Na 2 SO 4 , and concentrated to give a crude product, which was purified by preparative TLC (EtOAc / PE=1 / 1) to give the title compound (30 mg, yield 3%) as a brown gum. MS (ESI): 282.1 [M+H] + .

[0518] Step d) 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-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0519]

[0520] The title compound was prepared in analogy to General Procedure 4c from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid (62.7 mg, 0.122 mmol, 1.2 eq) and 3-methylthio-5-(1-methylsulfonylcyclopropyl)-1,2,4-triazine were prepared and obtained as a light brown solid (18 mg, 0.030 mmol, yield 26%) MS (ESI): 610.2 [M-isobutylene + H] + .

[0521] 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-benzothiazepine -4-Keto

[0522]

[0523] The title compound was prepared in analogy to General Procedure 5c from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (9 mg, 0.014 mmol, Eq: 1) was prepared and obtained as a brown solid (4.3 mg, 0.010 mmol, yield 48%). MS (ESI): 566.2 [M+H] + .

[0524] Example 20

[0525] (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-benzothiazepine -4-Keto

[0526]

[0527] 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-benzothiazepine -4-Keto

[0528]

[0529] The title compound was prepared in analogy to General Procedure 5b from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (9.0 mg, 0.01 mmol, 1 eq) was prepared and obtained as a brown solid (6.8 mg, 0.01 mmol, yield 74%). MS (ESI): 602.2 [M+H] +

[0530] Example 21

[0531] (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-benzothiazepine -4-Keto

[0532]

[0533] Step a) N'-Hydroxy-4-methyl-benzamidine

[0534]

[0535] To a solution of 4-methylbenzonitrile (5000 mg, 42.6 mmol, 1.0 eq) in ethanol (143 mL) was added hydroxylamine hydrochloride (3.71 g, 53.3 mmol, 1.25 eq) followed by NaHCO3 (17.9 g, 213 mmol, 5 eq). The reaction mixture was heated to reflux for 6 hours, cooled to room temperature and concentrated. The resulting white solid was suspended in water and sonicated for 10 minutes. The mixture was filtered and washed with water. The filter cake was then washed with heptane and dried in vacuo to give the title compound (5.16 g, 74%) as a white solid. MS (ESI): 151.1 [M+H] +

[0536] Step b) 3-(p-Tolyl)-5-(trifluoromethyl)-1,2,4-oxadiazole

[0537]

[0538] N'-Hydroxy-4-methyl-benzamidine (5.61 g, 37.3 mmol, 1 eq) was dissolved in ultra-dry THF (100 mL). The reaction mixture was cooled to 0 ° C. TFAA (12.5 g, 8.44 mL, 59.7 mmol, 1.6 eq) was then added dropwise. The ice-water bath was removed and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into brine and basified to pH = 7 to 8 with saturated NaHCO3 aqueous solution (200 ml). 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] -

[0539] Step c) 3-[4-(bromomethyl)phenyl]-5-(trifluoromethyl)-1,2,4-oxadiazole

[0540]

[0541] To a solution of 3-(p-tolyl)-5-(trifluoromethyl)-1,2,4-oxadiazole (7.98 g, 34.9 mmol, 1 eq) in acetonitrile (140 mL) were added N-bromosuccinimide (7.47 g, 41.9 mmol, 1.2 eq) and 2,2-azobis(2-methylpropionitrile (1.72 g, 10.49 mmol, 0.3 eq). The reaction mixture was stirred at 80° C. for 3.5 hours, poured into brine (100 mL), and extracted with ethyl acetate (3×150 mL). The combined organic phases were dried over MgSO , filtered, and concentrated, and purified by silica gel column chromatography (heptane containing 0% to 10% ethyl acetate) to give the title compound (4.69 g, 41.49%) as a white solid.

[0542] 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0543]

[0544] N-[(3R)-7-bromo-8-fluoro-4-keto-3,5-dihydro-2H-1,5-benzothiazepine] -3-yl] tert-butyl carbamate (500mg, 1.15mmol, 1eq) was added to a solution of ultra-dry N,N-dimethylformamide (8.85mL) with 3-[4-(bromomethyl)phenyl]-5-(trifluoromethyl)-1,2,4-oxadiazole (557mg, 1.73mmol, 1.5eq), potassium carbonate (476mg, 3.45mmol, 3eq) and potassium iodide (95mg, 0.57mmol, 0.5eq). The reaction mixture was stirred for 2h. The reaction was quenched with 10mL of water. The crude product was extracted with AcOEt (3×20mL). The combined organic layers were washed with water (2×10mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (heptane containing 6% to 60% DCM) to give the title compound (407mg, 55%) as a yellow solid. MS (ESI): 615.1 [MH] - .

[0545] Step e) N-[(3R)-7-bromo-8-fluoro-1,1,4-trione-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0546]

[0547] Analogously to General Procedure 2, 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-benzothiazepine -3-yl] tert-butyl carbamate (402.7 mg, 0.652 mmol, 1 eq) was prepared and obtained as a white solid (282 mg, yield 66.%). MS (ESI): 685.1 [M+HCOO] - .

[0548] Step f) N-[(3R)-5-(4-chlorobenzyl)-8-fluoro-1,1,4-trione-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0549]

[0550] The title compound was prepared in analogy to General Procedure 3a from N-[(3R)-7-bromo-8-fluoro-1,1,4-trione-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -3-yl] tert-butyl carbamate (140 mg, 0.216 mmol, 1 eq) was prepared and obtained as a yellow solid (129 mg, yield 85%). MS (ESI): 559.0 [M-2 isobutylene + H] + .

[0551] Step g) N-[(3R)-7-(5-tert-butyl-3-pyridyl)-5-(4-chlorobenzyl)-8-fluoro-1,1,4-trione-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0552]

[0553] The title compound was prepared in analogy to General Procedure 4a from N-[(3R)-8-fluoro-1,1,4-trione-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-benzothiazepine -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) were prepared and obtained as a white solid (15.9 mg, yield 26%). MS (ESI): 733.2 [M+H] + .

[0554] Step h) (3R)-3-amino-8-fluoro-1,1-diketo-7-(5-morpholino-3-pyridinyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0555]

[0556] The title compound was prepared in analogy to General Procedure 5d from N-[(3R)-8-fluoro-1,1,4-trione-7-(5-morpholino-3-pyridinyl)-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (15.9 mg, 0.022 mmol, 1 eq) was prepared and obtained as a yellow solid (11.5 mg, yield 79%) as a hydrochloride salt. MS (ESI): 677.3 [M+HCOO] - .

[0557] Intermediate 2

[0558] 3-Bromo-5-(1-methylsulfonyl-1-methyl-ethyl)pyridine

[0559]

[0560] Step a) 3-bromo-5-(methylsulfonylmethyl)pyridine

[0561]

[0562] 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 maintained at 50° C. for 2 h30.

[0563] The reaction was diluted in 100 mL of water and extracted with AcOEt (3×20 mL), washed with water (2×10 ml) and brine (1×20 mL). The organic layers were combined, dried over Na2SO4, and concentrated to give the title compound (80 mg, 67%) as an orange powder and used without further purification. MS (ESI): 249.7 [M+H] +

[0564] Step b) 3-bromo-5-(1-methylsulfonyl-1-methyl-ethyl)pyridine

[0565]

[0566] 3-bromo-5-(methylsulfonylmethyl)pyridine (80mg, 0.26mmol, 1eq) was dissolved in ultra-dry N,N-dimethylformamide (2.69mL). Cesium carbonate (262mg, 0.8mmol, 3eq) was added, and the reaction was stirred at room temperature for 30min under Ar. Methyl iodide (152mg, 4eq) was then added, and the reaction was stirred overnight. Cesium carbonate (87.5mg, 0.269mmol, 1eq) and methyl iodide (76mg, 33.6uL, 2eq) were added, and the reaction was stirred for another 24h. The mixture was extracted with AcOEt (3×40ml), washed with distilled water (2×50ml), and washed once with brine (1×40mL). The organic phase was then dried over Na2SO4, concentrated, and purified by preparative HPLC to obtain the title compound (20mg, yield 26.%) as a light red solid. MS (ESI): 278.9 [M+H] +

[0567] Examples 22 to 25 in the table below were prepared analogously to Example 21 steps g) to h) using the appropriate bromide or chloride.

[0568]

[0569]

[0570] *As hydrochloride

[0571] Example 26

[0572] (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0573]

[0574] Step a) N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0575]

[0576] N-[(3R)-7-bromo-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine A mixture of tert-butyl 2-(4-(4-(4-methoxy-3-yl)-1-carbamate (100 mg, 0.18 mmol, 1.0 eq), 5-methoxypyridine-3-boronic acid (36.2 mg, 0.24 mmol, 1.3 eq), K2CO3 (75.6 mg, 0.55 mmol, 3.0 eq), Pd(dppf)Cl2*DCM (14.9 mg, 0.02 mmol, 0.1 eq) in 1,4-dioxane (10 mL) and water (1 mL) was stirred at 80°C for 2 h under N2. The solution was poured into water (30 mL), extracted with EtAc (50 mL), the organic phase was washed with brine (50 mL), dried over Na2SO4, concentrated in vacuo, and purified by preparative HPLC to provide the title compound (50 mg, 0.09 mmol, 47% yield) as a white solid. MS (ESI): 576.1 [M+H] + .

[0577] Step b) (3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0578]

[0579] The title compound was prepared in analogy to General Procedure 5b from N-[(3R)-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridinyl)-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (40 mg, 0.07 mmol, 1 eq) was prepared and obtained as a white solid after preparative HPLC (33.1 mg, 0.06 mmol, yield 92%) as the dihydrochloride salt. MS (ESI): 476.2 [M+H] + .

[0580] Example 27

[0581] (3R)-3-Amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0582]

[0583] Step a) 1-(Chloromethyl)-4-(4-methoxyphenyl)benzene

[0584]

[0585] To a solution of [4-(4-methoxyphenyl)phenyl]methanol (1900 mg, 8.87 mmol, 1.0 eq) in DCM (38 mL) was added thionyl chloride (3.86 mL, 53.2 mmol, 6.0 eq) and the mixture was stirred at 25 °C for 6 h, concentrated under vacuum to give a 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] - Step b) N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0586]

[0587] Analogously to General Procedure 1, the title compound was prepared from a mixture containing 1-(chloromethyl)-4-(4-methoxyphenyl)benzene (654 mg, 2.8 mmol, 1.1 eq) and N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepine] -3-yl] tert-butyl carbamate (1000 mg, 2.56 mmol, 1.0 eq) was prepared in DMF and obtained as a white solid after silica gel column chromatography (1270 mg, 2.16 mmol, yield 84%). MS (ESI): 533.2 [M-isobutylene + H] + .

[0588] Step c) N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0589]

[0590] Analogously to General Procedure 2, the title compound was prepared from N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl] tert-butyl carbamate (1270.0 mg, 2.16 mmol), and obtained as a light brown solid (1000 mg, 1.61 mmol, yield 69%). MS (ESI): 643.2 [M+2+Na] + .

[0591] Step d) N-[(3R)-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0592]

[0593] Analogously to step a) of Example 26, the title compound was prepared from N-[(3R)-7-bromo-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (100 mg, 0.16 mmol, 1 eq) and 5-chloropyridine-3-boronic acid (40.6 mg, 0.26 mmol, 1.6 eq) were prepared and obtained as a white solid after silica gel column chromatography (70 mg, 0.11 mmol, yield 63%). MS (ESI): 652.2 [M+H] + .

[0594] 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-benzothiazepine 4-Keto; dihydrochloride

[0595]

[0596] The title compound was prepared by analogy to General Procedure 5b from N-[(3R)-7-(5-chloro-3-pyridinyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (70 mg, 0.11 mmol, 1 eq) was prepared and obtained as a yellow solid after preparative HPLC (28.5 mg, 0.05 mmol, yield 47%) as the dihydrochloride salt. MS (ESI): 552.2 [M+H] + .

[0597] Examples 28 to 30 in the table below were prepared analogously to Example 27 steps d) to e) using the appropriate bromide or chloride.

[0598]

[0599]

[0600] *As dihydrochloride

[0601] Example 37

[0602] (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-benzothiazepine -4-Keto

[0603]

[0604] Step a) N-[rac-(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0605]

[0606] Analogously to General Procedure 1, the title compound was prepared from a mixture containing 4-(bromomethyl)benzonitrile (275.58 mg, 1.41 mmol, 1.1 eq) and N-[(3R)-7-bromo-8-fluoro-4-oxo-3,5-dihydro-2H-1,5-benzothiazepine] -3-yl] tert-butyl carbamate (500 mg, 1.28 mmol, 1.0 eq) was prepared in DMF and obtained as a light yellow oil after silica gel column chromatography (450 mg, yield 68%). MS (ESI): 450.1 [M-isobutylene + H] + .

[0607] Step b) N-[rac-(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0608]

[0609] Analogously to General Procedure 2, the title compound was prepared from N-[rac-(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-4-oxo-2,3-dihydro-1,5-benzothiazepine -3-yl] tert-butyl carbamate (2000.0 mg, 3.95 mmol, 1.0 eq) was prepared and obtained as an off-white solid (1900.0 mg, 3.53 mmol, yield 87.48%). MS (ESI): 484.0 [M-isobutylene + H] + .

[0610] Step c) [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid

[0611]

[0612] The title compound was prepared in analogy to General Procedure 3a from N-[(3R)-7-bromo-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl] tert-butyl carbamate (1000.0 mg, 1.86 mmol, 1.0 eq) was prepared and obtained as a yellow solid (900 mg, 1.79 mmol, yield 96%). MS (ESI): 448.0 [M-isobutylene + H] + .

[0613] Step d) 3-[1-(trifluoromethyl)vinyl]pyridazine

[0614]

[0615] After the mixture was stirred at 90°C for 12 h, Pd(dppf)Cl2 CH2Cl2 (1.16 g, 1.42 mmol, 0.05 eq) was added to a solution of 1-(trifluoromethyl)vinylboronic acid hexanediol ester (6.28 g, 28.3 mmol, 1.0 eq), 3-bromopyridazine (4.5 g, 28.3 mmol, 1.0 eq) and cesium carbonate (18.4 g, 56.6 mmol, 2.0 eq) in toluene (100 mL) and water (10 mL) 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, yield 86%) as a light yellow oil. MS (ESI): 175.3 [M+H] +

[0616] Step e) 3-[1-(trifluoromethyl)cyclopropyl]pyridazine

[0617]

[0618] To a solution of 3-[1-(trifluoromethyl)vinyl]pyridazine (300 mg, 1.7 mmol, 1.0 eq) and methyl(diphenyl)sulfonium tetrafluoroborate (496 mg, 1.72 mmol, 1.0 eq) in THF (2 mL) was added THF (3.45 mL, 3.45 mmol, 2.0 eq) at 0 to 5 ° C. The solution was stirred at 20 ° C for 2.5 h. The reaction mixture was quenched with a slowly added saturated NH4Cl aqueous solution (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 under vacuum to obtain 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, yield 53%) as a light yellow oil. MS (ESI): 189.3 [M+H] +

[0619] Step f) Trimethyl-[methyl-[6-[1-(trifluoromethyl)cyclopropyl]pyridazin-4-yl]-trimethylsilyloxy-silyl]oxy-silane

[0620]

[0621] Under N2, to 3-[1-(trifluoromethyl)cyclopropyl]pyridazine (100 mg, 0.53 mmol, 1.0 eq), trimethyl-[methyl(trimethylsilyloxy)silyl]oxy-silane (124 mg, 0.56 mmol, 1.05 eq) and 2,9-dimethyl-1,10-phenanthroline (22.14 mg, 0.11 mmol, 0.2 eq) in THF (5 mL) solution was added [Ir(COD)OMe]2 (38.43 mg, 0.05 mmol, 0.1 eq) at 20 ° C. The mixture was stirred at 100 ° C for 12 h in a sealed tube, and 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 obtain the title compound (88 mg, 0.21 mmol, 35% yield) as a yellow oil. MS (ESI): 409.2 [M+H] + .

[0622] Step g) 5-bromo-3-[1-(trifluoromethyl)cyclopropyl]pyridazine

[0623]

[0624] To a solution of trimethyl-[methyl-[6-[1-(trifluoromethyl)cyclopropyl]pyridazine-4-yl]-trimethylsilyloxy-silyl]oxy-silane (30 mg, 0.07 mmol, 1.0 eq), N-bromosuccinimide (13 mg, 0.07 mmol, 1.0 eq) in ACN (2 mL) was added silver (I) fluoride (13.9 mg, 0.11 mmol, 1.5 eq) at 20 ° C, and the mixture was stirred at 20 ° C for 1 h. The reaction mixture was concentrated under vacuum to obtain a residue below 40 ° C. The residue was ground in TBME (10 mL) and collected by filtration. The filtrate was concentrated under vacuum to obtain the title compound (25 mg, 0.09 mmol, yield 126%), which was a light yellow oil and was used directly in the next step. (ESI): 269.0 [M + H] + .

[0625] Step h) 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-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0626]

[0627] After the mixture was stirred at 80° C. for 2 h under N2, [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine was added to the mixture at 20° C. under N2. To the solution of 1,4-dioxane (1mL) and water (0.1mL) was added cataCXiumA Pd G3 (2.89mg, 0.0mmol, 0.1eq, CAS: 1651823-59-4). The reaction mixture was filtered and the filtrate was concentrated under vacuum to obtain a residue. The residue was dissolved in EtOAc (30mL) and water (30mL), then extracted with EtOAc (30mL x 2). The combined extracts were washed with brine (20mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain 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, yield 31%) as a pale yellow solid. MS (ESI): 646.2 [M+H] + .

[0628] Step i) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0629]

[0630] At 20°C, 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-benzothiazepine was added. To a solution of tert-butyl]carbamate (15.0 mg, 0.02 mmol, 1.0 eq) and NaOAc (3.81 mg, 0.05 mmol, 2.0 eq) in ethanol (1 mL) was added NH2OH.HCl (1.94 mg, 0.03 mmol, 1.2 eq), and the mixture was stirred at 50 ° C for 18 h, 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 under vacuum to give the title compound (20.0 mg, 0.03 mmol, yield 109%) as a light yellow solid. MS (ESI): 679.2 [M+H] + .

[0631] Step j) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0632]

[0633] After the mixture was stirred at 20° C. for 1 h, 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-benzothiazepine was added at 20° C. To a solution of tert-butyl]carbamate (140 mg, 0.21 mmol, 1.0 eq) in THF (4 mL) was added TFAA (86.6 mg, 0.41 mmol, 2.0 eq). The reaction mixture was carefully basified to pH = 8 with saturated NaHCO3 solution and extracted with EtOAc (20 mL×3). The organic layer was washed with brine (10 mL), combined, dried over Na2SO4, and concentrated under vacuum 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, yield 46%) as a light yellow solid. MS (ESI): 757.2 [M+H] + .

[0634] 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-benzothiazepine -4-Keto; hydrochloride

[0635]

[0636] 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-benzothiazepine A solution of tert-butyl]-3-yl]carbamate (71.0 mg, 0.09 mmol, 1.0 eq) in EtOAc (3.0 mL, 12.0 mmol, 127.88 eq) containing HCl was stirred at 20 ° C for 0.5 h. The mixture was concentrated under vacuum at 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, yield 74.%) as a white solid. MS (ESI): 657.1 [M+H] + .

[0637] Example 36

[0638] (3R)-3-amino-8-fluoro-1,1-dioxo-7-(3-tert-butyl-1,2,4-triazine-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0639]

[0640] Step a) 3-tert-butyl-1,2,4-triazine-5-ol

[0641]

[0642] To a solution of 2,2-dimethylpropionamidine (5000 mg, 49.9 mmol, 1.0 eq, CAS 59950-56-0), triethylamine (27.8 mL, 199 mmol, 4.0 eq) in ethanol (50 mL) was added hydrazine (4780 mg, 149.1 mmol, 2.99 eq) at 25°C, and after 1 h, ethyl glyoxylate (30578 mg, 149 mmol, 3.0 eq) was added at 25°C, and the mixture was stirred at 60°C for 11 h. The reaction was concentrated in vacuo to give a residue, which was purified by column (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] + .

[0643] Step b) 3-tert-butyl-1,2,4-triazine-5-thiol

[0644]

[0645] To a solution of 3-tert-butyl-1,2,4-triazine-5-ol (400 mg, 2.1 mmol, 1.0 eq) in 1,4-dioxane (16 mL) was added Lawessons reagent (528 mg, 1.31 mmol, 0.5 eq) at 25°C, and the mixture was stirred at 100°C for 2 h. The reaction was concentrated in vacuo to give a residue, which was purified by column (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] + .

[0646] Step c) 3-tert-butyl-5-methylthio-1,2,4-triazine

[0647]

[0648] To a solution of 3-tert-butyl-1,2,4-triazine-5-ol (400 mg, 2.1 mmol, 1.0 eq) in 1,4-dioxane (16 mL) was added Lawessons reagent (528 mg, 1.31 mmol, 0.5 eq) at 25°C, and the mixture was stirred at 100°C for 2 h. The reaction was concentrated in vacuo to give a residue, which was purified by column (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] + .

[0649] Step d) N-[(3R)-7-(3-tert-butyl-1,2,4-triazine-5-yl)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0650]

[0651] The title compound was prepared in analogy to General Procedure 4c from [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid (180 mg, 0.36 mmol, 1.0 eq) and 3-tert-butyl-5-methylthio-1,2,4-triazine (98.3 mg, 0.54 mmol, 1.5 eq) were prepared and obtained as a yellow solid (80.0 mg, 0.13 mmol, yield 33%). MS (ESI): 595.2 [M+H] + .

[0652] Step e) N-[(3R)-7-(3-tert-butyl-1,2,4-triazine-5-yl)-8-fluoro-5-[[4-[(Z)-N'-hydroxycarbamimidoyl]phenyl]methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0653]

[0654] Analogously to step i) of Example 37, the title compound was prepared from N-[(3R)-7-(3-tert-butyl-1,2,4-triazine-5-yl)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (80.0 mg, 0.13 mmol, 1.0 eq) was prepared and obtained as a yellow solid (80 mg, 0.13 mmol, yield 67%). MS (ESI): 628.2 [M+H] + Step f) N-[(3R)-7-(3-tert-butyl-1,2,4-triazine-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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0655]

[0656] Analogously to step j) of Example 37, the title compound was prepared from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (80.0 mg, 0.13 mmol, 1.0 eq) was prepared and obtained as a yellow solid (50 mg, 0.07 mmol, yield 54%). MS (ESI): 706.1 [M+H] + .

[0657] Step g) (3R)-3-amino-7-(3-tert-butyl-1,2,4-triazine-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto

[0658]

[0659] Analogously to Example 37, step k), the title compound was prepared from N-[(3R)-7-(3-tert-butyl-1,2,4-triazine-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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (50.0 mg, 0.07 mmol, 1.0 eq) was prepared and obtained as a yellow solid (26.3 mg, 0.04 mmol, yield 57%). MS (ESI): 606.1 [M+H] + .

[0660] 5-Bromo-3-(cyclopropyloxy)pyridazine

[0661]

[0662] To a solution of cyclopropanol (150 mg, 2.58 mmol, 1.0 eq) in THF (5 mL) was added sodium hydride (113 mg, 2.84 mmol, 1.1 eq) in portions at 0°C. After 0.5 h, the reaction was added to THF (5 mL) containing 5-bromo-3-chloropyridazine (500 mg, 2.58 mmol, 1.0 eq) at 0°C and stirred at 0°C for 1.5 h. The reaction was poured into water (30 mL). The aqueous phase was extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue, which was purified by column (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 in the next step as a crude product. MS (ESI): 217.1 [M+H] + .

[0663] 5-Bromo-3-tert-butyl-pyridazine

[0664]

[0665] Step a) (6-tert-butylpyridazin-4-yl)-methyl-bis(trimethylsilyloxy)silane WUX003530-273

[0666]

[0667] Analogously to Example 37 step f), the title compound was prepared from 3-tert-butylpyridazine (300 mg, 2.2 mmol, 1.0 eq, CAS 215452-12-3) and obtained as a yellow solid after silica gel chromatography (PE / EA=10 / 1) (666 mg, 1.86 mmol, yield 84%), which was a light yellow oil. MS (ESI): 357.2 [M+H] + .

[0668] Step b) 5-Bromo-3-tert-butyl-pyridazine

[0669]

[0670] Analogously to step h) of Example 37, the title compound was prepared from (6-tert-butylpyridazin-4-yl)-methyl-bis(trimethylsilyloxy)silane (40 mg, 0.11 mmol) and obtained as a crude product in the form of a light brown oil (40 mg, 0.19 mmol). MS (ESI): 217.0 [M+H] + .

[0671] Examples 34-35 in the table below were prepared in analogy to Example 37 steps h) to k) using the appropriate boronate and aryl bromide.

[0672]

[0673]

[0674] *As hydrochloride

[0675] Example 33

[0676] (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-benzothiazepine -4-Keto

[0677]

[0678] Step a) 5-bromo-3-(2,2,2-trifluoroethoxy)pyridazine

[0679]

[0680] To a solution of 2,2,2-trifluoroethanol (0.19 mL, 2.58 mmol, 1.0 eq) in THF (5 mL) was added sodium hydride (113 mg, 2.84 mmol, 1.1 eq) in batches at 0 ° C. After 0.5 h, the reaction was added to a solution of 5-bromo-3-chloropyridazine (500 mg, 2.58 mmol, 1.0 eq) in THF (5 mL) at 0 ° C. The mixture was stirred at 0 ° C for 1.5 h. The reaction was poured into water (30 mL). The aqueous phase was extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and 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, yield 50%) as a yellow oil. MS (ESI): 257.1 [M+H] + .

[0681] Step b) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0682]

[0683] Under N2, at 25°C, add [(3R)-3-(tert-butoxycarbonylamino)-5-[(4-cyanophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine To a solution of 1,4-dioxane (4 mL) and water (0.5 mL) was added brettphos Pd g3 (37.8 mg, 0.04 mmol, 0.15 eq) with 5-bromo-3-(2,2,2-trifluoroethoxy)pyridazine (107 mg, 0.42 mmol, 1.5 eq), potassium carbonate (115 mg, 0.83 mmol, 3.0 eq). The reaction was stirred at 90 ° C for 2 h, quenched with water (15 mL), and extracted with EtOAc (10 mL×3). The organic phase was washed with brine (30 mL×2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel 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] + .

[0684] Step c) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0685]

[0686] Analogously to step i) of Example 37, the title compound was prepared from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (130 mg, 0.2 mmol, 1.0 eq) was prepared and obtained as a yellow solid (158.0 mg, 0.24 mmol, yield 81%). MS (ESI): 669.2 [M+H] + .

[0687] Step d) 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0688]

[0689] Analogously to step j) of Example 37, the title compound was prepared from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (148.0 mg, 0.22 mmol, 1.0 eq) was prepared and obtained as a yellow solid after preparative HPLC (50 mg, 0.07 mmol, yield 30%). MS (ESI): 747.1 [M+H] + .

[0690] 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-benzothiazepine 4-Keto

[0691]

[0692] The title compound was prepared in analogy to General Procedure 5b from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (45.0 mg, 0.06 mmol, 1.0 eq) was prepared and obtained as a yellow solid (48.1 mg, 0.07 mmol, yield 120). MS (ESI): 647.0 [M+H] + .

[0693] Example 32

[0694] (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-benzothiazepine -4-Keto

[0695]

[0696] Step a) N-[(3R)-7-bromo-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester

[0697]

[0698] Analogously to General Procedure 1, the title compound was prepared from N-[(3R)-7-bromo-8-fluoro-1,1,4-trioxo-3,5-dihydro-2H-1λ6,5-benzothiazepine] -3-yl]carbamic acid tert-butyl ester (250 mg, 0.59 mmol) and 2-[4-(chloromethyl)phenyl]-5-(trifluoromethyl)pyridine; hydrochloride (CAS: 613239-76-2) were prepared and obtained as a light yellow solid (720 mg, 1.09 mmol, yield 153.65%). MS (ESI): 660.0 [M+H] + .

[0699] 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-benzothiazepine -7-yl]boronic acid

[0700]

[0701] The title compound was prepared in analogy to General Procedure 3a from N-[(3R)-7-bromo-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -3-yl]carbamic acid tert-butyl ester (300 mg, 0.525 mmol, 1 eq) was prepared and obtained as a colorless oil (150 mg, yield 46%).

[0702] Step c) 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-benzothiazepine tert-butyl 3-amino-4-nitropropane

[0703]

[0704] The title compound was prepared in analogy to General Procedure 4a from [(3R)-3-(tert-butoxycarbonylamino)-8-fluoro-1,1,4-trioxo-5-[[4-[5-(trifluoromethyl)-2-pyridinyl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]boronic acid (20 mg, 0.03 mmol, 1.0 eq) and 5-bromo-3-tert-butyl-pyridazine (13.8 mg, 0.06 mmol, 2.0 eq) were prepared and obtained as a white solid (2.5 mg, 0.004 mmol, yield 10%). MS (ESI): 714.2 [M+H] + .

[0705] 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-benzothiazepine -4-Keto

[0706]

[0707] The title compound was prepared in analogy to General Procedure 5b from 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-benzothiazepine -3-yl]carbamic acid tert-butyl ester (37.0 mg, 0.05 mmol) was prepared and obtained as a white solid after preparative HPLC and SFC (12.2 mg, 0.02 mmol, yield 35.88%). MS (ESI): 614.1 [M+H] + .

[0708] 2) Biological Examples

[0709] 2.1) In vitro DGK inhibition assay

[0710] DGKα and ζ kinases use ATP to phosphorylate the substrate 1,2-dilauroyl-sn-glycerol (DLG, incorporated into liposomes). As a result of this enzymatic reaction, ATP is converted into ADP.

[0711] After the kinase reaction, an ATP depleting reagent is added to terminate the kinase reaction and deplete any remaining ATP, leaving only ADP. Next, a detection reagent is added to simultaneously convert ADP to ATP, and the newly synthesized ATP is converted into light using a coupled luciferase / luciferin reaction.

[0712] Reagents and Materials

[0713] Buffer components (solvents and salts)

[0714] Chemicals supplier Catalog Number: DTT AppliChem Catalog Number A3668 BSA Sigma-Aldrich Catalog Number A2153-10G MOPS Sigma-Aldrich Catalog No. M1254 Sodium chloride (NaCl) Sigma-Aldrich Catalog No. S7663-1KG <![CDATA[Magnesium chloride (MgCl2)]]> MERCK Catalog number 1.05833.0250 <![CDATA[Calcium chloride (CaCl2)]]> Sigma-Aldrich Catalog Number C4901-500G

[0715] Protein / Substrate / Tracer

[0716]

[0717]

[0718] Full-length DGKα and ζ were expressed in Sf21 insect cells by infecting them with baculovirus stocks at an MOI of 2. Purification of both enzymes was performed as previously described by Takahashi et al. Peer J, 2018 (Takahashi, D.; Sakane, F. Expression and purification of human diacylglycerol kinase alpha from baculovirus-infected insect cells for structural studies. Peer J 2018, 6, No. e5449).

[0719] hardware

[0720] Material supplier Catalog Number / Volume Range 384 multiwell plate, white Greiner Catalog No. 781904 384 multiwell plate, white Corning Catalog Number CLS3574 Matrix Multichannel Pipette Thermo Fisher SCIENTIFIC 2μl to 125μl Multidrop Combi Thermo Fisher SCIENTIFIC 0.5μl / well to 50μl / well Envision Plate Reader PerkinElmer

[0721] Assay buffer (30 ml)

[0722]

[0723]

[0724] Determination procedure

[0725] Concentrated liposome solutions were prepared in assay buffer without DTT and BSA: 21 mM total liposomes (2 mM DLG / 8 mM PS / 11 mM PC) containing 2 mM DLG. The reaction mixture contained assay buffer with a final DLG concentration of 125 uM and an ATP concentration of 25 μM (for DGKA assay) or 50 μM (for DGKZ assay). The reaction was initiated by adding DGKα and ζ kinases at final concentrations of 4 nM and 2 nM, respectively. After 1 hour of reaction, the amount of ADP formed was measured 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, diluted 1:3, and a final DMSO concentration of 2%. A Multidrop Combi was used as a liquid handler, and luminescence was read at 0.5 s using an Envision plate reader (PE).

[0726] result

[0727]

[0728]

[0729]

[0730] In vitro DGK inhibition assay: ADPGlo

[0731] DGKα and ζ kinases use ATP to phosphorylate the substrate 1,2-dilauroyl-sn-glycerol (DLG). As a result of this enzymatic reaction, ATP is converted into ADP.

[0732] After the kinase reaction, an ATP depleting reagent is added to terminate the kinase reaction and deplete any remaining ATP, leaving only ADP. Next, a detection reagent is added to simultaneously convert ADP to ATP, and the newly synthesized ATP is converted into light using a coupled luciferase / luciferin reaction.

[0733] Experimental procedures, reagents, and materials

[0734] DGKα and ζ kinase ADP Glo assays were performed by Reaction Biology Corp. (1 Great Valley Parkway, Suite 2, Malvern, 19355, PA, USA). Information provided by the service provider is as follows: DGKα and ζ kinases were used at a final concentration of 2 nM. Reactions were performed at 50 μM ATP. 500 uM substrate DLG (dilauroyl-sn-glycerol) was used. Compounds were received as 10 mM DMSO stock solutions and tested in 10 doses of IC50 using 3-fold serial dilutions starting from 1 μM. The control compound Calphostin C was tested in 10 doses of IC50 using 3-fold serial dilutions starting from 100 μM.

[0735] result

[0736]

[0737] 2.2) IL2 secretion measurement

[0738] 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 after compound treatment was assessed as a percentage of the maximum value of reference compound A1. WO 2016 / 139181 discloses reference compound A1 as Example 70. As a counter-screen and to ensure that no unwanted TCR-independent activation was triggered, all compounds were run under PBS conditions.

[0739] Reagents and Materials

[0740]

[0741]

[0742]

[0743]

[0744] Cell culture

[0745] The amplified primary human T cells were thawed and cultured at a density of 2 Mio / ml for 3 hours 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 1xPen-Strep (Life Technologies, #15140122) culture medium at 5% CO2, 37°C and 95% humidity. For the coating of the plate, PBS++ and PBS--, or PBS++ and CD3 antibody (concentration depends on the donor and is determined by CD3 titration) are added to a 96-well plate coated with poly-D lysine at 100 μl / well. The plate is sealed and incubated at room temperature for 3 hours on a benchtop rocking platform. After incubation, the plate was washed once with PBS and filled with only 40 μl / well culture medium. The compound was then added to the plate containing only culture medium (see next section). After the T cells were cultured for 3 hours, the cells were filtered through a cell strainer (Miltenyi Biotech, #130-041-407), counted again, and the concentration was adjusted to 1.25 Mio / ml.

[0746] The cells were then inoculated at 80 μl / well to 40 μl / well (including compounds assigned according to the plate layout). By adding cells, the compound was further diluted by 1:3, and 100k cells / 120 μl / well were obtained. 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 supernatant collected and frozen was used to detect IL2 using an IL-2 human ProQuantum immunoassay kit (Invitrogen) or using a human IL-2 ELISA kit (Thermo Fisher).

[0747] Compound treatment

[0748] Compounds were added using a Tecan D300e digital dispenser in a 5-point or 6-point dose response, with concentrations of 3 times the final concentration for all conditions, since cells were added later (80 μl of cells were added to 40 μl of prepared culture medium after treatment). The DR starting concentration was 20 μM or 10 μM final maximum concentration, and the dilution factor was 3.333. The positive control was the reference compound A1, which was also added to the dose response, and was also added to 3 wells at only 20 μM representing the positive stimulant control. All wells were normalized to a final concentration of 0.6% (0.2% final concentration) with DMSO.

[0749] IL2 ProQuantum Immunoassay

[0750] The immunoassay was performed according to the manufacturer's manual (Invitrogen, #A35603).

[0751] Additional Information: For immunoassays, use MicroAmp TM EnduraPlate TM Optical 384-well plates. Frozen supernatants were thawed and centrifuged at 1000×g for 5 minutes, both steps were performed at 4°C. After centrifugation, the required sample volume was removed from the top and placed in a separate LightCycler V-bottom plate (working plate) and diluted with assay dilution buffer, the dilution factor depending on the PBS or CD3 conditions, but at least 1:3. IL-2 standards and blanks were prepared in the same V-bottom plate, and the standards ranged from 0.0128 pg / ml to 5000 pg / ml (extended version). After preparation, 5 μl of sample dilution or standards / blanks were transferred to an optical 384-well plate (assay plate) and a 10 μl reaction protocol was followed. Measurements were performed using the QuantStudio 12K Flex system. Raw data were extracted and IL-2 concentrations were calculated using the Thermo Fisher online application (apps.thermofisher.com / apps / proquantum).

[0752] IL2 Elisa

[0753] ELISA was performed according to the manufacturer's manual (Thermo Fisher Scientific, #88-7025-88).

[0754] Additional Information: For ELISA, Nunc MaxiSorp 96-well plates were used. The frozen supernatant was thawed and centrifuged at 1000 x g for 5 min, both steps were performed at 4°C. Then, the required sample volume was taken from the top and placed in a separate V-bottom plate and diluted with ELISA diluent, the dilution factor depending on the PBS or CD3 conditions. IL-2 standards and blanks were prepared in the same V-bottom plate. After preparation, 50 μl of sample dilution and 100 μl of standard or blank were transferred to the Nunc plate.

[0755] Calculations and data reporting

[0756] CD3 and PBS plates were analyzed separately in Genedata Screener using Roche normalized PCT_POS_CTRL, with DMSO set as neutral control and 20 μM reference compound A1 set as stimulator control / 100%.

[0757] For CD3 conditions, EC50 and Emax are reported for the fitted sigmoidal curve. If a curve could not be fitted, EC50 is reported as a blank field and Emax is based on a single data point. Emax does not always correspond to the highest concentration tested. Compounds that activate unstimulated cells or that adversely affect viability (see proliferation assay) are flagged.

[0758] result

[0759]

[0760]

[0761] 2.3) Proliferation assay

[0762] Reagents and Materials

[0763]

[0764]

[0765] The expanded primary human T cells were thawed and cultured at a density of 2 Mio / ml for 3 hours 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) culture medium at 5% CO2, 37°C and 95% humidity. For the coating of the plate, PBS++ alone or PBS++ with CD3 antibody (concentration depends on the donor and is determined by CD3 titration) was added to a 96-well plate coated with poly-D lysine at 100 μl / well. The plate was sealed and incubated at room temperature for 3 hours on a benchtop rocking platform. After incubation, the plate was washed once with PBS and filled with only 40 μl / well culture medium. The compound was then added to the plate containing only culture medium (see next section). After the T cells were cultured for 3 hours, the cells were filtered through a cell strainer (Miltenyi Biotech, #130-041-407), counted again, and the concentration was adjusted to 1.25 Mio / ml.

[0766] The cells were then inoculated at 80 μl / well to 40 μl / well (including the compounds assigned according to the plate layout). The compounds were further diluted 1:3 by adding cells and 100k cells / 120 μl / well were obtained. After 48 hours, 40 μl of supernatant was carefully collected from the top without disturbing the cells. After 5 days, the proliferation of the cells was assessed by measuring ATP consumption using CellTiterGlo (Promega).

[0767] Compound treatment

[0768] Compounds were added using a Tecan D300e digital dispenser in a 5-point or 6-point dose response, with concentrations of 3 times the final concentration for all conditions, since cells were added later (80 μl of cells were added to 40 μl of prepared culture medium after treatment). The DR starting concentration was 20 μM or 10 μM final maximum concentration, and the dilution factor was 3.333. The positive control was the reference compound A1, which was also added to the dose response, and was also added to 3 wells at only 20 μM representing the positive stimulant control. All wells were normalized to a final concentration of 0.6% (0.2% final concentration) with DMSO.

[0769] Cell Titer Glo Measurement

[0770] After 5 days, use 2.0 reagent detects ATP proportional to the number of cells present in each well. After visual control of the toxicity or precipitation of the test compounds, the plates were equilibrated to room temperature for 45 minutes. 2.0 reagent is also balanced to room temperature.After balance, equal amount of CellTiter-Glo reagent is added to cells (80 μ l / well) with electronic multichannel pipette.Plate is placed on rocking platform at room temperature for 15 minutes.After incubation, the bottom of plate is sealed with backing tape.PHERAstar FSX is used to measure luminescence (interval 0.5 second, gain 3000, focal height 15mm), and exported as CSV file, so that in Genedata Screener, analyze.

[0771] Calculations and data reporting

[0772] CD3 and PBS plates were analyzed separately in Genedata Screener using Roche normalized PCT_POS_CTRL, with DMSO set as neutral control and 20 μM reference compound A1 set as stimulator control / 100%.

[0773] For CD3 conditions, EC50 and Emax are reported for the fitted sigmoidal curve. If a curve could not be fitted, EC50 is reported as a blank field and Emax is based on a single data point. Emax does not always correspond to the highest concentration tested. Compounds that activate unstimulated cells (see IL2 measurements) or that adversely affect viability are flagged.

[0774] result

[0775]

[0776]

[0777]

[0778] 2.5) T cell-TCB-MV3 killing assay Reagents and Materials

[0779]

[0780]

[0781]

[0782] Cell culture

[0783] All incubation steps were performed under 5% CO2, 37°C and 95% humidity.

[0784] MV-3RFP cells were cultured in MV-3 ​​medium (DMEM+10% FBS, 1x PenStrep and 0.5μg / mL puromycin) for at least 3 weeks. The MV-3 ​​cells of 80% confluence of the culture were washed once with PBS and trypsinized until separated. The cells were then counted and resuspended to 1×105 cells / mL in T cell culture medium (RPMI 1640+5% human serum+1mm sodium pyruvate+50μM 2-mercaptoethanol and 1x Pen-Strep). The cells were inoculated into 96-well plates (TTP, #92696) at 100μL / well and left motionless at room temperature for 40 minutes to achieve uniform distribution of cells. The plate was then incubated until further use.

[0785] The next day, the expanded primary human T cells were thawed and resuspended in T cell culture medium to 4×106 cells / mL. In a 6-well plate (up to 6 mL per well), they were cultured for 3 hours. After the T cells were cultured, they were filtered through a cell strainer (Miltenyi Biotech, #130-041-407), counted again, and the cell concentration was adjusted to 2×106 cells / mL.

[0786] Compound treatment

[0787] MCSP-TCB or PBS was pre-diluted in T cell culture medium (concentration depends on T cell donor) at 4 times the final concentration. Then 60 μL / well of the pre-dilution was dispensed into a round bottom plate (Costar, #3799) according to the plate layout. Compounds were added in a 9-point dose response using a Tecan D300e digital dispenser, also at 4 times the final concentration. The DMSO concentration of all wells was adjusted to 0.8% for a final concentration of 0.2%.

[0788] The T cell suspension of 60 μL / well is added to the prepared round bottom plate and resuspended using a manual multichannel. Then, according to the plate layout, the resuspended T cell suspension of 100 μL / well (including treatment) is carefully transferred to the MV-3 ​​cells cultured overnight. Only 100 μL T cell culture medium is added to the external MV-3 ​​well. The starting concentration of the final compound DR is 20 μM, and the dilution factor is 3.333. The final TCB concentration is between 1.5 pM and 5 pM, and the concentration of each T cell donor is determined separately by running TCB titration. For each donor, a TCB concentration corresponding to 10% to 20% of the killing of MV3 baseline cells in the absence of compound treatment is selected. The positive control is the reference compound A1 added to DR, and only the other wells of 20 μM are added. 20 μM reference compound A1 represents a positive stimulant control, and only TCB (DMSO well) represents a neutral control.

[0789] calculate

[0790] After transferring T cells with pre-treatment dilutions, use IncucyteZOOM TM MV-3 cells were imaged by time-lapse microscopy (Essen BioScience, MI, USA). Images were taken every 3 h for a total of 120 h (10X objective, phase and red image channels, acquisition time 400 ms, green / red 4614 optical module). Using a mask previously created and optimized for MV-3 ​​cells, the images were acquired in the IncucyteZOOM TM RFP object counts per well were analyzed in the ANALYZER® software (version 2019B Rev 2). Raw data were exported as object counts / well and values ​​were normalized to % TCL compared to wells with MV-3 ​​alone (representing 100% growth, therefore 0% TCL).

[0791] RFP Measurement

[0792] %TCL values ​​calculated in Genedata Screener using Roche normalized PCT_POS_CTRL analysis, where MCSP-TCB alone was set as neutral control and 20 μM reference compound A1 was set as stimulator control / 100%.

[0793] EC50 and Emax values ​​are provided in the table below.

[0794] TCLs induced by compounds without TCB treatment or without toxicity (observed under PBS conditions) are marked.

[0795] result

[0796] #imgpt410#

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 1 is a 6-membered heteroaryl group, wherein R 1 Optionally, one or more R 10 replace; R 2 is hydrogen or halogen; R 4 is selected from phenyl and pyridyl, optionally substituted by one or more R which may be the same or different 11 replace; R 10 Selected from: i)C 1-10 -alkyl, which is optionally substituted with one or more halogen, hydroxyl, -S(O)2(C 1-6 -alkyl), -N(R 10e R 10f )、-S(O)2(C 1-6 -cycloalkyl), cyano substituted; ii) C 3-10 -cycloalkyl, which is optionally substituted with one or more -S(O)2(C 1-6 -alkyl), Halogenated-C 1-6 - alkyl substitution; iii) a 3- to 10-membered heterocyclic group, which is optionally substituted with one or more halogens, -C(O)O-(R 10q )replace; iv) phenyl, which is optionally substituted with one or more C 1-6 -alkyl, halogen or halo-C 1-6 - alkyl substitution; v)-N(R 10e R 10f ); vi)-OR 10g ; vii) halogen; R 10e and R 10f are each independently selected from hydrogen and C 1-6 -alkyl; R 10g Selected from C 1-6 -alkyl, halo-C 1-6 -alkyl and C 3-10 - cycloalkyl; R 10q C 1-10 -alkyl; Or two R 10 Together with the carbon atoms to which they are attached, they form a 3- to 10-membered heterocyclic group, It is optionally treated with one or more C 1-10 - alkyl substitution; R 11 Selected from: i) halogen; ii) C 1-6 -alkyl, which is optionally substituted with one or more cyano groups; iii) C 1-6 - alkoxy; iv) C 3-7 - cycloalkyl; v) a 5- to 6-membered heteroaryl group, which is optionally substituted with one or more halogenated -C 1-6 -alkyl, C 3-10 - Cycloalkyl substitution; vi) phenyl, which is optionally substituted with one or more halogen, C 1-6 -alkoxy, halo-C 1-6 - alkyl substitution; vii)–SO2(R 11d ): R 11d Selected from hydrogen, C 1-6 -alkyl and halo-C 1-6 -alkyl.

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

3. The compound according to claim 1 or 2, wherein R 1 is pyridyl, pyrimidinyl, triazine, which is optionally substituted by one or more R 10 replace.

4. A compound according to any one of claims 1 to 3, wherein R 1 is a pyridyl group, a triazine group, which is optionally substituted by one or more R 10 replace.

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

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

7. A compound according to any one of claims 1 to 6, wherein R 4 is phenyl, which is 11 replace.

8. A compound according to any one of claims 1 to 7, wherein R 10 is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidinyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy; or R 10 and R 1 Together they form trimethyl-6,8-dihydro-1,7-naphthyridinyl and dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl.

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

10. A compound according to any one of claims 1 to 9, wherein R 11 Selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propionitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridinyl.

11. A compound according to any one of claims 1 to 10, wherein R 11 Selected from chlorine, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy.

12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 is pyridyl, pyrimidinyl, pyridazinyl, triazinyl, which is optionally substituted by one or more R 10 replace; R 2 is hydrogen or halogen; R 4 is phenyl, which is 11 replace; R 10 Selected from: (i)C 1-10 -alkyl, which is optionally substituted with one or more halogen, hydroxyl, -S(O)2(C 1-6 -alkyl), -N(R 10e R 10f )、-S(O)2(C 1-6 -cycloalkyl), cyano substituted; ii) C 3-10 -cycloalkyl, which is optionally substituted with one or more -S(O)2(C 1-6 -alkyl) substituted; iii) a 3- to 10-membered heterocyclic group, which is optionally substituted with one or more halogens, -C(O)O-(R 10q )replace; iv) phenyl; v)-N(R 10e R 10f ); vi)-OR 10g ; vii) halogen; R 10e and R 10f are each independently selected from hydrogen and C 1-6 -alkyl; R 10g Selected from C 1-6 -alkyl and halo-C 1-6 -alkyl; R 10q C 1-10 -alkyl; R 11 Selected from: i) halogen; ii) C 1-6 -alkyl, which is optionally substituted with one or more cyano groups; iii) C 1-6 - alkoxy; iv) C 3-7 - cycloalkyl; v) a 5- to 6-membered heteroaryl group, which is optionally substituted with one or more halogenated -C 1-6 -alkyl, C 3-10 - Cycloalkyl substitution; vi) phenyl, which is optionally substituted with one or more halogen, C 1-6 -alkoxy, halo-C 1-6 - alkyl substitution; vii)–SO2(R 11d ): R 11d Selected from hydrogen, C 1-6 -alkyl and halo-C 1-6 -alkyl.

13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 is pyridyl, pyrimidinyl, triazine, which is optionally substituted by one or more R 10 replace; R 2 is hydrogen or fluorine; R 4 is phenyl, which is 11 replace; R 10 is trifluoromethoxy, tert-butyl, isopropyl, methyl, chloro, methoxy, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile, morpholino, methylsulfonylcyclopropyl, chloro-methylsulfonyl-propyl, azabicyclo[3.1.1]heptane-carboxylate, difluoro-piperidinyl, diethylamino, phenyl, aminoethyl, hydroxy-methyl-ethyl, isopropyl, isopropoxy, difluoromorpholine, (dimethylamino)ethyl, (dimethylamino)methyl, trifluoroethoxy; or R 10 and R 1 Together they form trimethyl-6,8-dihydro-1,7-naphthyridinyl, dimethyl-7,8-dihydro-6H-1,8-naphthyridinyl; R 11 Selected from chloro, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy, difluoromethylsulfonyl, methylsulfonyl, methyl-propionitrile, (trifluoromethyl)phenyl, cyclopropyl-oxadiazolyl, (trifluoromethyl)pyridinyl.

14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 is a pyridyl group, a triazine group, which is optionally substituted by one or more R 10 replace; R 2 For fluorine; R 4 is phenyl, which is 11 replace; R 10 is tert-butyl, methyl, chloro, methyl-methylsulfonyl-ethyl, trifluoromethyl, methyl-propionitrile; R 11 Selected from chlorine, methoxyphenyl, (trifluoromethyl)-oxadiazolyl, isopropoxy.

15. A compound according to any one of claims 1 to 14, selected from (3R)-3-Amino-7-(4-tert-butyl-2-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(2-tert-butyl-4-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(6-tert-butylpyrimidin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-(4-chlorobenzyl)-8-fluoro-1,1-diketo-7-[5-(trifluoromethoxy)-3-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-isopropoxypyridazin-4-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(6-tert-butylpyridazin-4-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[6-(trifluoromethyl)-2-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-7-[4-(trifluoromethyl)pyrimidin-2-yl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(6-methyl-5-phenyl-1,2,4-triazine-3-yl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -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-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-7-[5-(4,4-difluoro-1-piperidinyl)-1,2,4-triazin-3-yl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; 1-[3-[(3R)-3-amino-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1,4-trioxo-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-1,2,4-triazine-5-yl]-3-azabicyclo[ 3.1.1] Methyl 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-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-trione-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-3-pyridinyl]-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-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-8-fluoro-1,1-diketo-7-[5-(1-methylsulfonyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-5-[(4-chlorophenyl)methyl]-8-fluoro-7-(5-methoxy-3-pyridyl)-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (3R)-3-Amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (3R)-3-amino-8-fluoro-1,1-dioxo-7-(3-tert-butyl-1,2,4-triazine-5-yl)-8-fluoro-1,1-dioxo-5-[[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]phenyl]methyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (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-benzothiazepine -4-ketone; 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-benzothiazepine -4-Keto; or a pharmaceutically acceptable salt thereof.

16. A compound according to any one of claims 1 to 15, selected from (3R)-3-Amino-7-(5-tert-butyl-3-pyridyl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-5-[(4-chlorophenyl)methyl]-8-fluoro-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-tert-butyl-1,2,4-triazine-3-yl)-8-fluoro-5-[(4-isopropoxyphenyl)methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; 2-[5-[(3R)-3-amino-8-fluoro-1,1,4-trione-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -7-yl]-3-pyridinyl]-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-pyridinyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-8-fluoro-1,1-diketo-7-[5-(1-methylsulfonyl-1-methyl-ethyl)-3-pyridyl]-5-[4-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzyl]-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (3R)-3-Amino-7-(5-chloro-3-pyridyl)-8-fluoro-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-Keto; (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-benzothiazepine -4-Keto; (3R)-3-Amino-8-fluoro-7-(6-isopropyl-3-pyridyl)-5-[[4-(4-methoxyphenyl)phenyl]methyl]-1,1-dioxo-2,3-dihydro-1λ6,5-benzothiazepine -4-ketone; 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-benzothiazepine -4-Keto; or a pharmaceutically acceptable salt thereof.

17. A method for preparing a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, the method comprising making a compound of formula (XI), Where R 1 , R 2 and R 4 As defined in any one of claims 1 to 14 and PG is an amino protecting group, reacted with a suitable deprotecting agent to form the compound of formula (I).

18. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, which is prepared according to the process of claim 17.

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

22. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for use in treating, preventing and / or delaying the progression of cancer.

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

24. The compound for use according to claim 22 or 23, wherein the cancer is selected from the group consisting of: B-cell acute lymphoid leukemia, T-cell acute lymphoid leukemia, acute lymphoid 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 disorders, MALT lymphoma, mantle cell Lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma, 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 treating, preventing and / or delaying the progression of cancer.

26. 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 treating, preventing and / or delaying the progression of cancer.

27. A method for treating, preventing and / or delaying the progression of cancer, the method 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. 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. A method of inhibiting the activity of at least one diacylglycerol kinase selected from DGKα and DGKζ, the method 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 herein described.

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