Compositions and methods for treating MALT1-related diseases

Abstract

This disclosure provides MALT1 inhibitors for reducing signaling mediated by activation of the Jun / Fos and / or NF-κB and / or mTor pathway (e.g., mTORc1 and / or mTORc2) in cells having gain-of-function alleles that act via MALT1. This disclosure also provides treatment for patients having these gain-of-function alleles by administering MALT1 inhibitors. In one application, the MALT1 inhibitor is administered to a patient having BENTA.

Description

[Technical Field]

[0001] background BENTA, or B-cell lymphocytosis with NF-κB and T-cell anergy, is a rare genetic disorder of the immune system caused by mutations in the CARD11 (caspase mobilization domain family member 11) gene. The disorder is characterized by elevated levels of certain B cells (B-cell lymphocytosis), splenomegaly (enlargement of the spleen), enlargement of lymph nodes (lymphadenopathy), immunodeficiency, and an increased risk of lymphoma, beginning in infancy. [Background technology]

[0002] Currently, there are very limited treatment options available to people with BENTA disease. The majority of treatment options address BENTA-associated infections and / or B-cell cancers. There is a need for novel medical treatments that address the underlying cause of BENTA as a cure, rather than simply treating the symptoms from BENTA. The purpose of this disclosure is to provide a method for addressing the underlying genetic cause of BENTA. [Overview of the project]

[0003] overview This disclosure relates to the use of MALT1 inhibitors to reduce the activation of the Jun / Fos, and / or NF-κB, and / or mTor pathways (e.g., mTORc1 and / or mTORc2) resulting from gain-of-function mutations (e.g., CARD11 or CARD14 gain-of-function variants) that act via MALT1. Such gain-of-function mutations result in diseases such as B-cell proliferation with NF-κB and T-cell anergy (BENTA), A20 haploinsufficiency, CARD14 systemic pustular psoriasis, HOIL1 hypomorphism, and NF-κB gain-of-function syndromes. Examples of MALT1 inhibitors include compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4. Other MALT1 inhibitors include compounds of formula 1:

[0004] [Chemical]

[0005] or a pharmaceutically acceptable salt thereof (wherein: R 1 is selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and 5-10 member heterocyclyl, and C1-C6 alkyl, C3-C6 cycloalkyl, and 5-10 member heterocyclyl are each independently selected from one, two, three, or more substituents selected from R 1a and may be optionally substituted on one or more available carbons by one, two, three, or more substituents, and when the 5-10 member heterocyclyl contains a ring nitrogen atom that can be substituted, the ring nitrogen atom may be optionally substituted by R 1b and when the 5-10 member heterocyclyl contains a ring sulfur atom that can be substituted, the ring sulfur atom may be optionally substituted by two O atoms; R 2 is CH3 or CF3; Is R3 hydrogen; or R3 is selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, C3-C7 cycloalkyl, 5-6 member heterocyclyl, 5-6 member heterocyclyl-C1-C3 alkyl 5-6 member heterocyclyl-O-, phenyl, and 5-6 member heteroaryl, and any of these may be optionally substituted by one, two, or three substituents each independently selected from R 3a ; R4 is C1-C6 alkyl; R 1a is independently, for each occurrence, selected from the group consisting of cyano, halogen, hydroxyl, oxo, C1-C6 alkyl, -C(O)OR A , -C(O)N(R A )2, -N(R A )2, C1-C6 alkoxy, 5-6 member heterocyclyl, and 5-6 member heteroaryl, and C1-C6 alkyl is N(R A)2 is substituted as needed, and if the 5-6 member heterocyclyl contains a substituted ring nitrogen atom, that ring nitrogen atom is R p It may be replaced as needed; R 1b These are C1-6 alkyl, -C(O)OR A -C(O)C1~6alkyl, -C(O)C3~6cycloalkyl, -C(O)N(R A )2, and selected from the group consisting of -S(O)2C1~6 alkyl groups; R 3a These are, independently, each occurrence, halogen, C1-4 alkyl, C1-4 haloalkyl, C1-4 alkoxy, C1-4 haloalkoxy, hydroxy, C1-4 alkenyl, cyano, azide, -NR C R D Selected from the group consisting of C3-6 cycloalkyl, Ci1-4 alkoxy, 5-6 member heterocyclyl-O-, 5-6 member heterocyclyl, and phenyl, where C3-6 cycloalkyl, 5-6 member heterocyclyl-O-, 5-6 member heterocyclyl, and phenyl are R p They are optionally substituted with one, two, or three substituents, each independently selected from; R p These are, independently, each occurrence, halogen, C1-4 alkyl, C1-4 haloalkyl, hydroxy, C1-4 alkoxy, C1-4 alkoxyC1-4 alkyl, NR C R D , and selected from the group consisting of amino C1-3 alkyl groups; R A Each of these is independently selected from the group consisting of hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, and -C(O)OC1-6 alkyl; R B The group is selected from the group consisting of Ci1-6 alkyl, C1-6 cycloalkyl, and -C(O)OC1-6 alkyl; R C and R D Each instance is independently selected from the group consisting of hydrogen, C1-6 alkyl, halo-C1-6 alkyl, and C3-4 cycloalkyl, or R C and R D These, together with the nitrogen atom to which they are bonded, form a 4-6 membered heterocycline or 4-6 membered heteroaryl, which may contain further nitrogen or oxygen atoms and may be substituted as needed with one or two fluorocarbons; T is either 0 or 1. [Brief explanation of the drawing]

[0006] [Figure 1] Figure 1 shows a diagram of the MALT1 signaling pathway up to NF-κB. [Figure 2] Figure 2 shows a diagram of the pathway to NF-κB related to MALT1 signaling with the CARD11 gain-of-function allele. This diagram also shows the inhibition of CARD11 stimulation by a MALT1 inhibitor. [Figure 3] Figure 3 shows a diagram of the pathway to NF-κB related to MALT1 signaling with the CARD14 gain-of-function allele. This diagram also shows the inhibition of CARD14 stimulation by a MALT1 inhibitor. [Modes for carrying out the invention]

[0007] Detailed explanation Before describing various embodiments, it should be understood that the teachings of this disclosure are not limited to the specific embodiments described and may be modified accordingly. Since the scope of these teachings is limited only by the appended claims, it should also be understood that the terminology used herein is intended solely to describe specific embodiments and is not intended to limit them.

[0008] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which this disclosure belongs. Any methods and materials similar to or equivalent to those described herein may be used in carrying out or testing this instruction, but some exemplary methods and materials are described herein.

[0009] As will be apparent to those skilled in the art when reading this disclosure, each individual embodiment described and shown herein has separate components and features that can be readily separated from or readily combined with any of several other embodiments without departing from the scope or spirit of this teaching. Any of the listed methods may be carried out in the order of the listed events or in any other logically possible order.

[0010] As used in this specification and the attached claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly indicates otherwise. Therefore, for example, a reference to "polypeptide (a polypeptide)" includes one or more polypeptides.

[0011] The headings of the sections used herein are for structural purposes only and should not be construed as limiting the subject matter of the inventions described herein.

[0012] definition Definitions of specific functional groups and chemical terms are described in more detail below. Chemical elements are identified according to the periodic table, CAS version, and the inside cover of the 75th edition of the Handbook of Chemistry and Physics, and specific functional groups are generally defined as they appear therein. Furthermore, general principles of organic chemistry, as well as specific functional parts and reactivity, are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987.

[0013] The compounds described herein may contain one or more chiral centers and therefore may exist in various isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers or geometric isomers, or in the form of a mixture of stereoisomers, including a racemic mixture and a mixture enriched with one or more stereoisomers. The isomers can be isolated from the mixture by methods known to those skilled in the art, including chiral high-performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron, Vol. 33: p. 2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions, p. 268 (Ed. E.L. Eliel, Univ. of Notre Dame Press, Notre Dame, 1972). This disclosure further encompasses the compounds described herein as individual isomers substantially free from other isomers, or as mixtures of various isomers.

[0014] As used herein, the term “pure enantiomerized compound” means that the compound is substantially free of other enantiomers or stereoisomers (i.e., enantiomerized), in other words, the “S” form of the compound is substantially free of the “R” form of the compound, and is therefore enantiomerized in the “R” form. The terms “enantiomerized pure” or “pure enantiomer” indicate that the compound contains more than 75% by weight, more than 80% by weight, more than 85% by weight, more than 90% by weight, more than 91% by weight, more than 92% by weight, more than 93% by weight, more than 94% by weight, more than 95% by weight, more than 96% by weight, more than 97% by weight, more than 98% by weight, more than 98.5% by weight, more than 99% by weight, more than 99.2% by weight, more than 99.5% by weight, more than 99.6% by weight, more than 99.7% by weight, more than 99.8% by weight, or more than 99.9% by weight of enantiomers. In one embodiment, the weight is based on the total weight of all enantiomers or stereoisomers of the compound.

[0015] In the compositions provided herein, the enantiomer pure compound may be present together with other active or inactive components. For example, a pharmaceutical composition containing an enantiomer pure R-compound may, for example, contain about 90% excipients and about 10% enantiomer pure R-compound. In certain embodiments, the enantiomer pure R-compound in such a composition may, for example, contain at least about 95% by weight of the R-compound and up to about 5% by weight of the S-compound, based on the total weight of the compound. For example, a pharmaceutical composition containing an enantiomer pure S-compound may, for example, contain about 90% excipients and about 10% enantiomer pure S-compound. In certain embodiments, the enantiomer pure S-compound in such a composition may, for example, contain at least about 95% by weight of the S-compound and up to about 5% by weight of the R-compound, based on the total weight of the compound. In certain embodiments, the active ingredient may be formulated with little or no excipients or carriers.

[0016] The compounds described herein may also contain one or more isotopic substitutions. For example, H may be 'H, H(D or deuterium), and 3 It may be any isotopic form containing H (T or tritium); C is 12 C, 13 C, and 14 It may be any isotopic form containing C; O is 16 O and 18 It may be any isotopic form containing O; F is 15 F and 19 This may include any isotopic form containing F; similar forms may also be included.

[0017] When various values ​​are listed, each value is intended to encompass a subrange within that range. For example, "Ci~6 alkyl" is intended to encompass Ci, Cu, Cs, Cr, Cs, O, Ci~e, Ci~5, Ci~4, Ci~3, Ci~2, C2~6, C2~5, C2~4, C2~3, C3~6, C3~5, C3~4, C4~6, C4~5, and C5~6 alkyl.

[0018] As used herein, the term "alkyl" refers to, for example, a linear or branched saturated hydrocarbon radical having 1 to 20 carbon atoms ("C1-20 alkyl"). In some embodiments, the alkyl group has 1 to 10 carbon atoms ("Ci-io alkyl"). In some embodiments, the alkyl group has 1 to 9 carbon atoms ("Ci-9 alkyl"). In some embodiments, the alkyl group has 1 to 8 carbon atoms ("Ci-8 alkyl"). In some embodiments, the alkyl group has 1 to 7 carbon atoms ("Ci-7 alkyl"). In some embodiments, the alkyl group has 1 to 6 carbon atoms ("Ci-6 alkyl"). In some embodiments, the alkyl group has 1 to 5 carbon atoms ("Ci-5 alkyl"). In some embodiments, the alkyl group has 1 to 4 carbon atoms ("C1-4 alkyl"). In some embodiments, the alkyl group has 1 to 3 carbon atoms ("C1-3 alkyl"). In some embodiments, the alkyl group has 1 to 2 carbon atoms ("C1-2 alkyl"). In some embodiments, alkyl groups have one carbon atom ("C1 alkyl"). Examples of Ci~6 alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, and hexyl.

[0019] As used herein, the term “alkenyl” refers to a linear or branched hydrocarbon group radical having 2 to 20 carbon atoms, one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds), and optionally one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds) ("C2-20 alkenyl"). In certain embodiments, the alkenyl does not contain any triple bonds. In some embodiments, the alkenyl group has 2 to 10 carbon atoms ("C2-10 alkenyl"). In some embodiments, the alkenyl group has 2 to 9 carbon atoms ("C2-9 alkenyl"). In some embodiments, the alkenyl group has 2 to 8 carbon atoms ("C2-8 alkenyl"). In some embodiments, the alkenyl group has 2 to 7 carbon atoms ("C2-7 alkenyl"). In some embodiments, the alkenyl group has 2 to 6 carbon atoms ("C2-6 alkenyl"). In some embodiments, the alkenyl group has 2 to 5 carbon atoms ("C2-5 alkenyl"). In some embodiments, the alkenyl group has 2 to 4 carbon atoms ("C2-4 alkenyl"). In some embodiments, the alkenyl group has 2 to 3 carbon atoms ("C2-3 alkenyl"). In some embodiments, the alkenyl group has 2 carbon atoms ("C2 alkenyl"). One or more carbon-carbon double bonds may be located internally (e.g., as 2-butenyl) or at the terminal (e.g., as 1-butenyl). Examples of C2-4 alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), and butadienyl (C4). Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkenyl groups, as well as pentenyl (C5), pentadienyl (C5), and hexenyl (Ce). Additional examples of alkenyls include heptenyl (C7), octenyl (C8), and octatrienyl (C8).

[0020] As used herein, the term "alkynyl" refers to a linear or branched hydrocarbon group radical having 2 to 20 carbon atoms, one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triple bonds), and optionally one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon double bonds) ("C 2~20 (Alkynyl). In certain embodiments, the alkynyl group contains no double bonds. In some embodiments, the alkynyl group has 2 to 10 carbon atoms ("C2-10 alkynyl"). In some embodiments, the alkynyl group has 2 to 9 carbon atoms ("C2-9 alkynyl"). In some embodiments, the alkynyl group has 2 to 8 carbon atoms ("C2-8 alkynyl"). In some embodiments, the alkynyl group has 2 to 7 carbon atoms ("C2-7 alkynyl"). In some embodiments, the alkynyl group has 2 to 6 carbon atoms ("C2-6 alkynyl"). In some embodiments, the alkynyl group has 2 to 5 carbon atoms ("C2-5 alkynyl"). In some embodiments, the alkynyl group has 2 to 4 carbon atoms ("C2-4 alkynyl"). In some embodiments, the alkynyl group has 2 to 3 carbon atoms ("C2-3 alkynyl"). In some embodiments, the alkynyl group has two carbon atoms ("C2 alkynyl"). One or more carbon-carbon triple bonds may be located internally (e.g., as 2-butynyl) or at the terminal (e.g., as 1-butynyl). Examples of C2-4 alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), and 2-butynyl (C4). Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkynyl groups, as well as pentynyl (C5) and hexynyl (C6). Additional examples of alkynyls include heptynyl (C7) and octinyl (C8).

[0021] As used herein, the terms “alkylene,” “alkenylene,” “alkylene,” “cycloalkylene,” “heterocyclylene,” “heteroarylene,” and “phenylene” refer to the divalent radicals of alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl (e.g., saturated and partially saturated), heteroaryl, and phenyl groups, respectively. When a range or number of carbon atoms is applied to a particular “alkylene,” “alkenylene,” or “alkylene” group, it should be understood that the range or number refers to the range or number of carbon atoms in the entire linear carbon chain. The “alkylene,” “alkenylene,” and “alkylene” groups may be substituted or unsubstituted with one or more substituents as described herein.

[0022] As used herein, the term “aryl” refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14π electrons shared in the cyclic arrangement) having 6 to 14 ring carbon atoms and having 0 heteroatoms in the aromatic ring system ("C6-14 aryl"). In some embodiments, the aryl group has 6 ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl group has 10 ring carbon atoms ("C10 aryl"; e.g., naphthyl, e.g., 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 ring carbon atoms ("C14 aryl"). “Aaryl” also includes ring systems in which the aryl ring as defined above is fused with one or more carbocyclic or heterocyclic groups, and the radical or bond site resides on the aryl ring, in which case the number of carbon atoms still indicates the number of carbon atoms in the aryl ring system.

[0023] Typical aryl groups, though not limited to these, include those derived from acetantrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluorantene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indan, indene, naphthalene, octacene, octafen, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiaden, pyrene, pyranthrene, rubicene, triphenylene, and trinaphthalene. Particularly noteworthy are phenyl, naphthyl, indenyl, and tetrahydronaphthyl aryl groups.

[0024] As used herein, the term “heteroaryl” refers to a radical of a 5- to 10-membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 electrons shared in the cyclic arrangement) having a ring carbon atom and providing 1 to 4 ring heteroatoms in an aromatic ring system, each heteroatom independently selected from nitrogen, oxygen, and sulfur (“5- to 10-membered heteroaryl”). In a heteroaryl group containing one or more nitrogen atoms, the bond site may be a carbon atom or a nitrogen atom, as long as the valence allows. A heteroaryl bicyclic ring system may contain one or more heteroatoms in one or both rings. “Heteroaryl” includes a ring system in which the heteroaryl ring as defined above is fused with one or more carbocyclic or heterocyclic groups, and the bond site is located on the heteroaryl ring, in which case the ring member number still indicates the ring member number in the heteroaryl ring system. A "heteroaryl" is a ring system in which the heteroaryl ring defined above is fused with one or more aryl groups, and the bond site is located on either the aryl ring or the heteroaryl ring. In such cases, the number of ring members indicates the number of ring members in the fused (aryl / heteroaryl) ring system. In the case of bicyclic heteroaryl groups in which one of the rings does not contain a heteroatom (e.g., indoyl, quinolyl, carbazolyl), the bond site may be located on either ring, i.e., on either the ring containing a heteroatom (e.g., 2-indolyl) or the ring not containing a heteroatom (e.g., 5-indolyl).

[0025] In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms provided in an aromatic ring system, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-10 membered heteroaryl"). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms provided in an aromatic ring system, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-8 membered heteroaryl"). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-6 membered heteroaryl"). In some embodiments, a 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0026] Exemplary five-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Exemplary five-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary five-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary five-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary six-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary six-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary six-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetradinyl, respectively. Examples of seven-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl. Examples of 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranil, benzoisofuranil, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolidinyl, and prinyl. Examples of 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthylidinyl, pteridinyl, quinolinyl, isoquinolinyl, cinolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Representative examples of heteroaryls include:

[0027] [ka]

[0028] (wherein each Z is selected from carbonyl, N, NR65, O, and 8; R65 is independently hydrogen, Ci-8 alkyl, C3-10 carbocyryl, 4-10 member heterocyclyl, C6-C10 aryl, and 5-10 member heteroaryl).

[0029] As used herein, the terms "carbocyclyl" or "carbocyclic" refer to the radical of a non-aromatic cyclic hydrocarbon group ("C3-10 carbocyclyl") having 3 to 10 ring carbon atoms in a non-aromatic ring system and 0 heteroatoms, wherein in some embodiments the carbocyclyl group has 3 to 8 ring carbon atoms ("C3-8 carbocyclyl"). In some embodiments the carbocyclyl group has 3 to 7 ring carbon atoms ("C3-7 carbocyclyl"). In some embodiments the carbocyclyl group has 3 to 6 ring carbon atoms ("C3-6 carbocyclyl"). In some embodiments the carbocyclyl group has 5 to 10 ring carbon atoms ("C5-10 carbocyclyl"). Examples of C3-6 carbocyrill groups include, but are not limited to, cyclopropyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (G6), and cyclohexadienyl (G6). Examples of C3-8 carbocyrill groups include, but are not limited to, the aforementioned C3-6 carbocyrill groups, as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclooctanyl (C8), and bicyclo[2.2.2]octanyl (C8). Examples of C3-10 carbocyrill groups include, but are not limited to, the aforementioned C3-8 carbocyrill groups, as well as cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-l-indenyl (C9), decahydronaphthalenyl (C10), spiro[4.5]decanyl (C10), and others. As illustrated in the examples above, in certain embodiments, the carbocyrill group may be monocyclic ("monocyclic carbocyrill"), or may include condensed, cross-linked, or spirocyclic systems, such as bicyclic systems ("bicyclic carbocyrill"), and may be saturated or partially unsaturated.The term "carbocyclic system" also includes a cyclic system in which the carbocyclyl ring defined above is fused with one or more aryl or heteroaryl groups, and the bonding site is located on the carbocyclyl ring, in which case the carbon number continues to indicate the carbon number in the carbocyclic system.

[0030] As used herein, the term "cycloalkyl" refers to a monovalent saturated cyclic, bicyclic, or bridged cyclic (e.g., adamantyl) hydrocarbon group having 3 to 12, 3 to 8, 4 to 8, or 4 to 6 carbon atoms, and is referred to herein, for example, as a "cycloalkyl" derived from cycloalkanes. Examples of cycloalkyl groups include, but are not limited to, cyclohexane, cyclopentane, cyclobutane, and cyclopropane.

[0031] As used herein, the terms "C3-6 monocyclic cycloalkyl" or "monocyclic C3-6 cycloalkyl" refer to saturated 3- to 7-membered monocyclic hydrocarbon ring systems. Examples of 3- to 7-membered monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Substituents on the cycloalkyl (e.g., in the case of optionally substituted cycloalkyls) may be located at any substituted position, including, for example, the position to which the cycloalkyl group is attached.

[0032] As used herein, the terms “heterocyclyl” or “heterocyclic” refer to a radical of a 3- to 10-membered non-aromatic ring system having a ring carbon atom and 1 to 4 ring heteroatoms, each heteroatom independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3- to 10-membered heterocyclyl”). In a heterocyclyl group containing one or more nitrogen atoms, the bond site may be a carbon atom or a nitrogen atom, as long as the valence allows. The heterocyclyl group may be monocyclic (“monocyclic heterocyclyl”), condensed, bridged, or spirocyclic, for example, a bicyclic system (“bicyclic heterocyclyl”), and may be saturated or partially unsaturated. A heterocyclyl bicyclic ring system may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which the heterocyclyl ring as defined above is fused with one or more carbocykyl groups, with the bond site located on either the carbocykyl or heterocyclyl ring, or in which the heterocyclyl ring as defined above is fused with one or more aryl or heteroaryl groups, with the bond site located on the heterocyclyl ring, in which case the number of ring members continues to express the number of ring members in the heterocyclyl ring system. The terms "heterocyclyl," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" may be used interchangeably.

[0033] In some embodiments, the heterocyclic group is a 4-7 membered non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("4-7 membered heterocyclil"). In some embodiments, the heterocyclyl group is a 5-10 membered non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, each heteroatom independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5-10 membered heterocyclil").

[0034] In some embodiments, the heterocyclyl group is a 5-8 member non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("3-8 member heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-6 member non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("5-6 member heterocyclyl"). In some embodiments, the 5-6 member heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 member heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 member heterocyclyl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0035] Examples of three-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azilidinyl, oxylanil, and thiorenyl. Examples of four-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azetidinyl, oxetanyl, and thietanyl. Examples of five-membered heterocyclyl groups containing one heteroatom include, but are not limited to, tetrahydrofuranil, dihydrofuranil, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Examples of five-membered heterocyclyl groups containing two heteroatoms include, but are not limited to, dioxolanil, oxasulfuranil, disulfuranil, and oxazolidine-2-one. Examples of five-membered heterocyclyl groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl.

[0036] Examples of six-membered heterocyclyl groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranil, dihydropyridinyl, and thianyl. Examples of six-membered heterocyclyl groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinil, dithianyl, and dioxanil. Examples of seven-membered heterocyclyl groups containing two heteroatoms include, but are not limited to, triazinyl. Examples of seven-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azepanyl, oxepanyl, and thiepanyl. Examples of eight-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azocanyl, oxecanyl, and thiocanyl. Examples of five-membered heterocyclyl groups condensed to a C5 aryl ring (also referred to herein as 5,6-bicyclic heterocyclic rings) include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, and benzoxazolinonyl. Examples of six-membered heterocyclyl groups condensed to an aryl ring (also referred to herein as 6,6-bicyclic heterocyclic rings) include, but are not limited to, tetrahydroquinolinyl and tetrahydroisoquinolinyl.

[0037] Examples of saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyridinonyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, oxetanyl, azetidinyl, and tetrahydropyrimidinyl. Substituents on the heterocyclyl (e.g., in the case of an optionally substituted heterocyclyl) may be located at any substituted position, including, for example, the position to which the heterocyclyl group is attached.

[0038] As used herein, the term "hetero" means, when used to describe a compound or a group present on a compound, that one or more carbon atoms in the compound or group are replaced by a heteroatom of nitrogen, oxygen, or sulfur. Hetero may apply to any of the above-mentioned hydrocarbyl groups, e.g., alkyl, e.g., heteroalkyl; carbocyryl, e.g., heterocyclyl; aryl, e.g., heteroaryl; and similar groups having one to five heteroatoms, particularly one to three heteroatoms.

[0039] As used herein, the term "cyano" refers to -CN. As used herein, the terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), and iodine (iod, -I). In certain embodiments, the halo group is either fluoro or chloro.

[0040] In this specification, the term As used herein, the term "alkoxy" refers to an alkyl group bonded to another part via an oxygen atom (-O(alkyl)). Non-limiting examples include, for example, methoxy, ethoxy, propoxy, and butoxy.

[0041] As used herein, the term "fluoroalkoxy" refers to a haloalkyl group bonded to another part via an oxygen atom, such as, but not limited to, -OCHCFH2 or -OCF3.

[0042] As used herein, the term "oxo" refers to -C=O. As used herein, the term “substituted” means that at least one hydrogen atom present on a group (e.g., a carbon or nitrogen atom) is replaced by an acceptable substituent, such as a substituent that results in a stable compound upon substitution, such as a substituent that is not spontaneously transformed by rearrangement, cyclization, disappearance, or other reaction. Unless otherwise indicated, a “substituted” group has substituents at one or more substitutable positions on the group, and if more than one position in any given structure is substituted, the substituents at each position are either the same or different.

[0043] The nitrogen atom may be substituted or unsubstituted as long as its valence allows, and includes primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents, but not limited to, include hydrogen, -OH, -OR, -N(R)2, -CN, -C(=O)R, -C(=O)N(R)2, CO-R N SO'R'3, -C(NR)R, -C(-NR)R, -C(-NR)N(R)2, and SO.NIR. SO.'R\SO.-OR”.-SOR, -C(=S)N(RCC)2, -C(=O)SR, C(S)SR-, P(=O)2R, -P(=O)(R)2, -P(=O)2N(R)2, -P(=O)(NR)2, C1-10 alkyl, C1-10 perhaloalkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyrill, 3-14 membered heterocyclyl, C6-11 aryl, and 5-14 membered heteroaryl, or two Rcc groups bonded to a nitrogen atom form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, carbocyrill, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5RQQ groups.

[0044] As used herein, the term "coding sequence" is defined to mean the portion of nucleic acid (e.g., a gene) that codes for the amino acid sequence of a protein.

[0045] As used herein, the term “effective dose” of a compound refers to an amount sufficient to induce a desired biological response. As will be understood by those skilled in the art, the effective dose of a compound in the present invention may be modified depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and condition of the subject. The effective dose encompasses both therapeutic and prophylactic treatments.

[0046] As used herein, the term “wild-type” is defined to mean the form that is primarily found in nature. For example, a wild-type polypeptide or polynucleotide sequence is a sequence that can be isolated from naturally occurring raw materials and is primarily found in organisms that have not been intentionally modified by human intervention.

[0047] As used herein, the terms “recombinant,” “genetically modified,” and “non-naturally occurring” are used without distinction and are defined as modified polypeptides or nucleic acids that are modified in a manner not otherwise found in nature, or produced or induced from synthetic materials and / or by manipulation using recombinant technology. Non-limiting examples include, among others, recombinant cells that express genes not found within the range of the cell’s natural (non-recombinant) form, or that express naturally occurring genes that would normally be expressed at different levels.

[0048] As used herein, the terms “percentage of sequence identity” and “percentage homology” are used interchangeably and are defined as meaning a comparison between polynucleotides or polypeptides, determined by comparing two sequences optimally aligned across a comparison region, where portions of the polynucleotide or polypeptide sequences in the comparison region may contain additions or deletions (i.e., gaps) compared to a reference sequence for optimal alignment of the two sequences. The percentage may be calculated by determining the number of positions in which identical nucleic acid bases or amino acid residues appear in both sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison region, and multiplying the result by 100 to obtain the percentage of sequence identity. Alternatively, the percentage may be calculated by determining the number of positions in which identical nucleic acid bases or amino acid residues appear in both sequences, or the number of positions in which nucleic acid bases or amino acid residues are aligned with gaps to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison region, and multiplying the result by 100 to obtain the percentage of sequence identity. Those skilled in the art will understand that there are many established algorithms available for aligning two arrays.Optimal alignment of sequences for comparison may be performed, for example, by Smith and Waterman's local homology algorithm, Adv Appl Math. vol. 2: p. 482, 1981; by Needleman and Wunsch's homology alignment algorithm, J Mol Biol. vol. 48: p. 443, 1970; by Pearson and Lipman's similarity search method, Proc Natl Acad Sci. USA vol. 85: p. 2444, 1988; by computer implementation of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the GCG Wisconsin software package) or by visual inspection (see in general Current Protocols in Molecular Biology, edited by FMAusubel et al., Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (1995 Supplement)). Examples of suitable algorithms for determining percent sequence identity and sequence similarity include the BLAST and BLAST2.0 algorithms, described by Altschul et al., J.Mol.Biol. 215: pp. 403-410, 1990; and Altschul et al., Nucleic Acids Res. 25(17): pp. 3389-3402, 1977. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information website. For nucleotide sequences, BLAST can be performed using the BLASTN program with default parameters, e.g., word length (W) 11, expected value (E) 10, M=5, N=-4, and comparison of both strands. For amino acid sequences, BLAST can be used with the BLASTP program using default parameters, such as word length (W) 3, expected value (E) 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff, Proc Natl Acad Sci. USA, Vol. 89: p. 10915, 1989).Exemplary determinations of sequence alignment and % sequence identity may also be performed using the BESTFIT or GAP programs from the GCG Wisconsin software package (Accelrys, Madison, Wisconsin), using the provided default parameters.

[0049] As used herein, the term “pharmaceutically acceptable carrier” refers to a non-toxic carrier, adjuvant, or vehicle that does not impair the pharmacological activity of the compound being formulated. Examples of pharmaceutically acceptable carriers, adjuvants, or vehicles that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, e.g., human serum albumin, buffers, e.g., phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes, e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0050] As used herein, “pharmaceutically acceptable salt” means a salt that is suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, or allergic response, within the bounds of sound medical judgment, and that has a reasonable benefit-to-risk ratio. pharmaceutically acceptable salts are well known in the art. For example, Berge et al., in J Pharmaceutical Sciences (1977), Vol. 66, pp. 1-19, describe pharmaceutically acceptable salts in detail, which are incorporated by reference in their entirety for all purposes. Suitable pharmaceutically acceptable salts of the compounds of the present invention are derived from inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid adduct salts are salts of amino groups formed using inorganic acids, e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids, e.g., acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, e.g., ion exchange. Other pharmaceutically acceptable salts include adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfonate, heptaneate, hexanoate, hydroiodide, and 2-hydroxyethanesulfone. Examples include salts, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfons, 2-naphthalenesulfons, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectins, persulfates, 3-phenylpropionates, phosphates, picrates, pivaphosphates, propions, stearates, succinates, sulfates, tartrates, thiocyans, p-toluenesulfons, undecanoates, and valersates. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N +There are (Ci~4alkyl)4 salts. Typical alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions, where appropriate, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0051] As used herein, the term “reference sequence” is defined to mean a defined sequence used as a basis for comparing sequences. A reference sequence may be a subset of a larger sequence, for example, a segment of a full-length gene or polypeptide sequence. Generally, a reference sequence is at least 20 nucleotides or amino acid residues long, at least 25 residues long, at least 50 residues long, or the full length of a nucleic acid or polypeptide. Since each of two polynucleotides or polypeptides may (1) contain sequences that are similar between the two sequences (i.e., parts of the complete sequence) and (2) contain sequences that are different between the two sequences, sequence comparisons between two (or more) polynucleotides or polypeptides are typically performed by comparing the sequences of the two polynucleotides or polypeptides across a “comparison region” to identify and compare local regions of sequence similarity. In some embodiments, the “reference sequence” may be based on a primary amino acid sequence, and the reference sequence may be a sequence that has one or more changes from the primary sequence.

[0052] As used herein, the term “substantial identity” means a polynucleotide or polypeptide sequence having at least 80 percent sequence identity, at least 85 percent identity, and 89 to 95 percent sequence identity, more commonly at least 99 percent sequence identity, compared to a reference sequence over a comparison region of at least 20 residue positions, and more frequently over a region of at least 30 to 50 residues, the percentage of sequence identity being calculated by comparing the reference sequence with a sequence containing a total of 20 percent or less of deletions or additions of the reference sequence over the comparison region. In specific embodiments applied to polypeptides, the term “substantial identity” means that two polypeptide sequences share at least 80 percent sequence identity, preferably at least 89 percent sequence identity, at least 95 percent sequence identity, or greater sequence identity (e.g., 99 percent sequence identity), if optimally aligned by, for example, a GAP or BESTFIT program using standard parameters, i.e., default parameters. Preferably, the positions of non-identical residues differ by conservative amino acid substitutions.

[0053] As used herein, the term “therapeutic dose” of a compound means an amount sufficient to provide therapeutic benefit in the treatment of a disease, disorder, or condition, or to delay or minimize one or more signs associated with a disease, disorder, or condition. The therapeutic dose of a compound means the amount of the therapeutic agent, either alone or in combination with other therapies, that provides therapeutic benefit in the treatment of a disease, disorder, or condition. The term “therapeutic dose” may also include amounts that improve the overall treatment, reduce or avoid the signs or causes of a disease or condition, or enhance the therapeutic effect of another therapeutic agent.

[0054] As used herein, the terms “amino acid substitution” or “amino acid difference” are defined to mean a change in an amino acid residue at a position in a polypeptide sequence compared to an amino acid residue at a corresponding position in a reference sequence, which is the primary translation product beginning with a methionine start codon. The position of the amino acid difference is generally referred to herein as “Xn,” where n is the corresponding position in the reference sequence based on the residue difference. For example, “residue difference at position X compared to the primary translation product beginning with a methionine start codon” refers to a change in an amino acid residue at the polypeptide position corresponding to position X in the wild-type protein. Therefore, if the reference polypeptide of the primary translation beginning with a methionine start codon has valine at position X with respect to the wild-type gene, then “amino acid substitution” or “residue difference at position X compared to the reference sequence” refers to an amino acid substitution of any residue other than valine at the polypeptide position corresponding to position X in the reference sequence. In most cases, in this specification, a specific amino acid substitution or amino acid residue difference at a given position is indicated as "XnY," where "Xn" indicates the corresponding position as described above, and "Y" is a single-letter identifier of the amino acid found in the genetically engineered polypeptide (i.e., a residue different from that in the reference polypeptide). In some embodiments, if more than one amino acid can appear at a given residue position, the alternative amino acids may be listed in the form XnY / Z, where Y and Z represent the alternative amino acid residues. In some cases, this disclosure also provides specific amino acid differences indicated by the conventional notation "AnB," where A is a single-letter identifier of a residue in the reference sequence, "n" is the number of residues located in the reference sequence, and B is a single-letter identifier of a residue substitution in the sequence of the genetically engineered polypeptide. Furthermore, in some cases, the polypeptides of this disclosure may contain one or more amino acid residue differences with respect to a reference sequence, indicated by a list of designated positions where modifications have been made with respect to the reference sequence.

[0055] As used herein, the terms “conservative amino acid substitution” or “conservative amino acid difference” are defined as meaning a change in an amino acid at a residue position to a different residue having a similar side chain, and therefore typically involve amino acid substitutions in a polypeptide of amino acids within a range of similar or similar defined classes of amino acids. For example, but not limited to, amino acids having aliphatic side chains may be substituted with other aliphatic amino acids, e.g., alanine, valine, leucine, and isoleucine; amino acids having hydroxyl side chains may be substituted with other amino acids having hydroxyl side chains, e.g., serine and threonine; amino acids having aromatic side chains may be substituted with other amino acids having aromatic side chains, e.g., phenylalanine, tyrosine, tryptophan, and histidine; amino acids having basic side chains may be substituted with other amino acids having basic side chains, e.g., lysine and arginine; amino acids having acidic side chains may be substituted with other amino acids having acidic side chains, e.g., aspartic acid or glutamic acid; and hydrophobic or hydrophilic amino acids may be replaced with other hydrophobic or hydrophilic amino acids, respectively. Exemplary conservative substitutions are provided in Table 1 below.

[0056] [Table 1]

[0057] As used herein, the terms “unconservative substitution” or “unconservative amino acid difference” are defined as changes in an amino acid at a residue position to a different residue having significantly different side-chain properties. Unconservative substitutions may involve amino acids between defined groups rather than within the defined groups and affect (a) the structure of the peptide backbone in the substitution region (e.g., substitution of glycine with proline), (b) the charge or hydrophobicity, or (c) the bulkiness of the side chain. Examples of unconservative substitutions, though not limited to examples, may include acidic amino acids substituted with basic or aliphatic amino acids; aromatic amino acids substituted with small amino acids; and hydrophilic amino acids substituted with hydrophobic amino acids.

[0058] As used herein, the term “deletion” is defined as a modification of a polypeptide by removing one or more amino acids from a reference polypeptide, or a modification of a nucleic acid by removing one or more nucleotides from a reference nucleic acid. For example, a deletion may include the removal of one or more amino acids, two or more amino acids, five or more amino acids, ten or more amino acids, fifteen or more amino acids, or twenty or more amino acids, up to 10% of the total number of amino acids constituting the reference polypeptide, or up to 20% of the total number of amino acids. A deletion may target an internal and / or terminal portion of the polypeptide. In various embodiments, a deletion may include a continuous segment or be discontinuous.

[0059] As used herein, the term “insertion” is defined as modifying a reference polypeptide into a polypeptide by adding one or more amino acids, or modifying a nucleic acid by adding one or more nucleic acids. Insertions can be made in the interior of a polypeptide, at the carboxyl terminus, or at the amino terminus. Examples of insertions as used herein include fusion proteins known in the art. Insertions may be in adjacent amino acid segments or separated by one or more amino acids in the reference polypeptide.

[0060] As used herein, the term "specificity" is defined, when used in reference to a biocatalyst or enzyme, as meaning the identification of a biocatalyst with respect to a substrate compound.

[0061] As used herein, the term “relative specificity” is defined as meaning that the specificity of a biocatalyst or enzyme for one substrate compound is greater than that for another or other substrate compounds.

[0062] As used herein, the term “stringent hybridization conditions” is defined as hybridizing in 50% formamide in 5×SSC at a temperature of 42°C and washing the filter in 0.2×SSC at 60°C (1×SSC is 0.15M NaCl, 0.015M sodium citrate). Stringent hybridization conditions also include low ionic strength and high temperature for washing, e.g., 0.015M sodium chloride / 0.0015M sodium citrate / 0.1% sodium dodecyl sulfate, 50°C; hybridization at 42°C using a denaturing agent, e.g., formamide, e.g., 50% (v / v) formamide containing 0.1% bovine serum albumin / 0.1% Ficoll / 0.1% polyvinylpyrrolidone / 50 mM sodium phosphate buffer at pH 6.5, along with 750 mM sodium chloride and 75 mM sodium citrate; or 50% formamide, 5×SSC(0.75M Hybridization was performed at 42°C using NaCl (0.075M sodium citrate), 50mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5× Denhardt's solution, sonicated salmon sperm DNA (50 μg / ml), 0.1% SDS, and 10% dextran sulfate. The samples were then washed at 42°C in 0.2× SSC (sodium chloride / sodium citrate) with 50% formamide at 55°C, followed by a high-stringency washing solution at 55°C consisting of 0.1× SSC containing EDTA.

[0063] As defined herein, the term “heterogeneous” polynucleotide or polypeptide means any polynucleotide or polypeptide not found in nature in a host cell. Such a term includes polynucleotides that have been removed from a host cell, subjected to laboratory manipulation, and then reintroduced into the host cell. In some embodiments, the introduced polynucleotide expresses a heterogeneous polypeptide.

[0064] As used herein, the term “control sequence” is defined to include all components necessary or advantageous for the expression of the polynucleotides and / or polypeptides of this disclosure. Each control sequence may be native or foreign to the nucleic acid sequence encoding the polypeptide. Such control sequences include, but are not limited to, leaders, polyadenylated sequences, propeptide sequences, promoters, signal peptide sequences, and transcriptional terminators. A control sequence includes at least a promoter, a transcriptional stop signal, and, where appropriate, a translational stop signal. A control sequence may have a linker for the purpose of introducing a specific restriction site to facilitate ligation between the control sequence and the coding region of the nucleic acid sequence encoding the polypeptide.

[0065] As used herein, the term “operable linked” is defined as a configuration in which the control sequence is appropriately positioned relative to the polynucleotide of interest (i.e., a functional relationship exists) so that the control sequence directs or regulates the expression of the polynucleotide and / or polypeptide of interest.

[0066] As used herein, the term “promoter sequence” is defined as a nucleic acid sequence recognized by a host cell for the expression of the polynucleotide of interest, e.g., a coding sequence or gene. The promoter sequence includes a transcriptional regulatory sequence that mediates the expression of the polynucleotide of interest. The promoter may be any nucleic acid sequence that exhibits transcriptional activity in a selected host cell, including mutant promoters, truncated promoters, and hybrid promoters, and may be obtained from a gene encoding an extracellular or intracellular polypeptide, either homogeneous or heterogeneous, to the host cell.

[0067] MALT1 inhibitors Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1). MALT1 is a caspase-like protease that plays a role in BCL10-induced activation of NF-κB, and / or activation of mTOR pathway signaling (e.g., mTORc1 and / or mTORc2), and / or activation of Jun / Fos. The protein may be a component of the CARMA1-BCL10-MALT1 (CBM) signalosome and other signalosomes (e.g., those involving CARD11 or CARD14) that result in lymphocyte activation after antigen-receptor stimulation that induces NF-κB signaling, mTOR pathway signaling (e.g., mTORc1 and / or mTORc2), and Jun / Fos signaling. Bi-allelic loss-of-function mutations in this gene lead to immunodeficiency 12 (IMD12).

[0068] MALT1 inhibitors are compounds of formula I:

[0069] [ka]

[0070] or a pharmaceutically acceptable salt thereof (in the formula: R 1 The group is selected from the group consisting of C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, and 5-10 membered heterocyclines, and C1-6 alkyl, C3-6 cycloalkyl, and 5-10 membered heterocyclines are R 1a One or more available carbons may be optionally substituted by one, two, three, or more substituents independently selected from each of the following, and if the 5- to 10-membered heterocyclyl contains a substituted ring nitrogen atom, the ring nitrogen atom is R 1b If a 5- to 10-membered heterocyclyl contains a substituteable ring sulfur atom, that ring sulfur atom may be substituted with two oxygen atoms as needed; R 2 It is either CH3 or CF3; Is R3 hydrogen; or R3 is selected from the group consisting of C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heterocyclyl-C1-3 alkyl5-6 membered heterocyclyl-O-, phenyl, and 5-6 membered heteroaryl, and any of these is R 3a They may be substituted as needed with one, two, or three substituents, each independently selected from them; R4 is a C1-6 alkyl group; R 1a These are independently, with each appearance being cyano, halogen, hydroxyl, oxo, C1-6 alkyl, -C(O)OR A ,-C(O)N(R A )2, -N(R A )2, selected from the group consisting of C1-6 alkoxy, 5-6 membered heterocyclyl, and 5-6 membered heteroaryl, where C1-6 alkyl is N(R A )2 is substituted as needed, and if the 5-6 member heterocyclyl contains a substituted ring nitrogen atom, that ring nitrogen atom is R p It may be replaced as needed; R 1b These are C1-6 alkyl, -C(O)OR A -C(O)C1~6alkyl, -C(O)C3~6cycloalkyl, -C(O)N(R A )2, and selected from the group consisting of -S(O)2C1~6 alkyl groups; R 3a These are, independently, each occurrence, halogen, C1-4 alkyl, C1-4 haloalkyl, C1-4 alkoxy, C1-4 haloalkoxy, hydroxy, C1-4 alkenyl, cyano, azide, -NR C R D Selected from the group consisting of C3-6 cycloalkyl, Ci1-4 alkoxy, 5-6 member heterocyclyl-O-, 5-6 member heterocyclyl, and phenyl, where C3-6 cycloalkyl, 5-6 member heterocyclyl-O-, 5-6 member heterocyclyl, and phenyl are R p They are optionally substituted with one, two, or three substituents, each independently selected from; Rp These are, independently, each occurrence, halogen, C1-4 alkyl, C1-4 haloalkyl, hydroxy, C1-4 alkoxy, C1-4 alkoxyC1-4 alkyl, NR C R D , and selected from the group consisting of amino C1-3 alkyl groups; R A Each of these is independently selected from the group consisting of hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, and -C(O)OC1-6 alkyl; R B The group is selected from the group consisting of Ci1-6 alkyl, C1-6 cycloalkyl, and -C(O)OC1-6 alkyl; R C and R D Each instance is independently selected from the group consisting of hydrogen, C1-6 alkyl, halo-C1-6 alkyl, and C3-4 cycloalkyl, or R C and R D These, together with the nitrogen atom to which they are bonded, form a 4-6 membered heterocycline or 4-6 membered heteroaryl, which may contain further nitrogen or oxygen atoms and may be substituted as needed with one or two fluorocarbons; T is either 0 or 1.

[0071] MALT1 inhibitors also include, for example,

[0072] [ka]

[0073] Includes. Other compounds used as MALT1 inhibitors are recognized in WO2022 / 081967 (PCT / US21 / 55173), which shall be incorporated by reference in their entirety for all purposes.

[0074] Furthermore, other compounds used as MALT1 inhibitors are recognized in WO2023 / 192506 (PCT / US2023 / 016941), which shall be incorporated by reference in their entirety for all purposes. Other MALT1 inhibitors include:

[0075] [ka]

[0076] things, or there are stereoisomers and / or pharmaceutically acceptable salts thereof (in the formula: R 1 C 1~6 Alkyl or C 1~3 It is a haloalkyl, C 1~6 Alkyl is -OC 1~3 It may be substituted with alkyl as needed; R 2 This is an aryl or a 5-6 member heteroaryl, where the aryl may be substituted with a cyano as needed; R 4 It is -C(O)OH or a 5-6 member heteroaryl; m is either 0 or 1; n is either 0 or 1.

[0077] Other MALT1 inhibitors include, for example:

[0078] [ka]

[0079] There is. MALT1 inhibitors include pharmaceutically acceptable salts, pharmaceutically acceptable esters, tautomers, polymorphs, and prodrugs of such compounds. In some embodiments, MALT1 inhibitors include pharmaceutically acceptable additive salts, pharmaceutically acceptable esters, solvates (e.g., hydrates) of additive salts, tautomers, polymorphs, enantiomers, enantiomer mixtures, stereoisomers or stereoisomer mixtures (as pure stereoisomers or racemic or non-racemic mixtures), compounds described herein, e.g., pure or highly concentrated stereoisomers of compounds of formula I; e.g., compounds of formulas named herein.

[0080] Synthesis of MALT1 inhibitors A mixture of tert-butyl N-{2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-yl}carbamate [INT 1-e] (420 mg, 1.28 mmol) in 4N HCl / dioxane (5 mL) was stirred at 2.5 °C for 2 hours. LC-MS indicated completion of the reaction, and one novel peak with the desired MS was detected (Rt = 0.611 min, m / z: 227.8 [M + H]+). The mixture was concentrated under reduced pressure to obtain 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine hydrochloride [INT 1.1] (400 mg, crude) as a solid.

[0081] (R)-2-chloro-7-(1-methoxyethyl)-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine [INT 1.3] may be prepared using (2R)-2-methoxypropanoic acid as a starting material by the same synthetic route outlined for INT 1.1, with m / z:[M+H]+ C8H11C1N50 calculated value 228.1; experimental value 228.1.

[0082] (R)-2-methylpropane-2-sulfinamide (72.1 g, 595 mmol, 1.5 eq) was added at 25°C to a solution of 4-bromobenzaldehyde [INT 2-a] (100 g, 541 mmol, 1.0 eq) in toluene (500 mL). The mixture was stirred at 25°C for 15 minutes. Then, NaOH (21.6 g, 541 mmol, 1.0 eq) was added to the reaction product, and the mixture was stirred at 2.5°C for 12 hours. Na2SO4 (50 g) was added to the mixture and stirred for 20 minutes. The four reaction mixtures were combined, filtered by Celite to obtain the filtrate, and concentrated under vacuum to obtain the crude product as an oily substance. The crude product was dissolved in petroleum ether (1.0 L), stirred at -50°C for 1.0 hour, and filtered to obtain (R,E)-N-(4-bromobenzylidene)-2-methylpropane-2-sulfinamide [INT 2-b] (620 g, 2.15 rnol, yield 99.5%) as a solid.

[0083] To a solution of (R,E)-N-(4-bromobenzylidene)-2-methylpropane-2-sulfinamide [INT 2-b] (2.06 g, 715 mmol, 1.0 eq) and tetrabutylammonium acetate (216 g, 715 mmol, 21.8 mL, 1.0 eq) in DMF (1.4 L), TMSCF3 (259 g, 1.82 mol, 2.5 eq) was added at 0°C. The mixture was stirred at 5°C for 1.5 hours. This process was repeated twice, and the three reaction mixtures were combined. The mixture was poured into a saturated NH4Cl solution (1.3 L) and stirred for 10 minutes to obtain a suspension. The suspension was filtered to obtain a filtrate, which was eluted with water (5 L). The filtered mass was ground with METBE / petroleum ether (v / v=1:4, 2.0 L) to obtain the product as a solid, the mother liquor was concentrated under vacuum to obtain the crude product as an oily substance, and this was purified by silica gel column chromatography using petroleum ether / ethyl acetate (10 / 1~1 / 1) to obtain (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 2.1] (389 g, 1.09 mol, yield 50.6%) as a solid. 1HNMR (400 MHz, CDCb) δ = 1.25 (s, 9H), 3.64 (d, J = 6.40 Hz, 1H), 4.79-4.83 (m, 1H), 7.32 (d, I = 8.40 Hz, 2H), 7.56 (d, I = 6.40 Hz, 2H). (S)-N-((R)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 14-a] was prepared using (S)-(-)-2-methyl-2-propanesulfinamide by the same synthetic route outlined for (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 2.1].

[0084] A mixture of (S)-N-((R)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 14-a] (8 g, 22.3 mmol) in MEOH (60 mL) was mixed with 4 M HCl in dioxane (20 mL). The mixture was stirred at 20°C for 1.5 hours. The mixture was concentrated under reduced pressure to obtain the crude product. The mixture was diluted with water (50 mL) and extracted with ELISA (50 mL × 2). The combined organic layers were washed with 1 M HCl (50 mL × 2). The aqueous phase was basicized to pH=9-10 using 2N NaOH and extracted with CH₂Cl₂ (50 mL × 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product (R)-1-(4-bromophenyl)-2,2,2-trifluoroethane-1-amine [INT 14-b] (3.50 g, 13.7 mmol, yield 61.8%) as a yellow solid. Calculated m / z values ​​for [M+H]+ C8H8BrF3N were 254.0 and 256.0; measured value was 254.1.

[0085] (R)-1-(4-bromophenyl)-2,2,2-trifluoroethane-1-amine [INT 14-b] (2g, 7.87 mmol) was added to a mixture of tetrahydro-2H-thiopyran-4-carboxylic acid 1,1-dioxide [INT 4-a] (1.68 g, 9.44 mmol), EDCl (2.26 g, 11.8 mmol), and HOBt (1.59 g, 11.8 mmol) in CH2Cl2 (20 ml). The mixture was stirred at 25°C for 16 hours. The reaction was quenched by adding water (50 mL) and extracted with SiO2 (50 mL x 3). The combined organic layers were washed with saline solution (50 mL x 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain the crude product. This was purified by silica gel flash chromatography (siRNA / PE = 0 / 1 to 1 / 5) to obtain (R)-N-(1-(4-bromophenyl)-2,2,2-trifluoroethyl)tetrahydro-2H-thiopyran-4-carboxamide 1,1-dioxide [INT 14-c] (2.20 g, 5.31 mmol, yield 67.4%) as a white solid. Calculated m / z values ​​for [M+H]+ C14H16BrF3N03S were 414.0 and 416.0; measured value was 416.2.

[0086] To a solution of (R)-N-(1-(4-bromophenyl)-2,2,2-trifluoroethyl)tetrahydro-2H-thiopyran-4-carboxamide 1,1-dioxide [INT 4-c] (1 g, 2.41 mmol) in DMF (10 mL), Cs2CO3 (1.57 g, 4.82 mmol) was added, and the reaction mixture was stirred at 25°C for 1 hour. Then, methyl iodide (1.02 g, 7.23 mmol) at 0°C was added, and the reaction mixture was stirred at 25°C for 2 hours. The reaction was quenched by adding water (50 mL), and then extracted with ELISA (50 mL x 3). The combined organic layers were washed with saline solution (50 mL x 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain the crude product. This crude product was purified by silica gel flash chromatography (PE / siRNA = 1 / 0 to 1 / 1) to obtain (R)-N-(1-(4-bromophenyl)-2,2,2-trifluoroethyl)-N-methyltetrahydro-2H-thiopyran-4-carboxamide 1,1-dioxide [INT 14.1] (700 mg, 1.63 mmol, yield 67.9%) as a colorless oil. m / z: Calculated values ​​for [M+H]+ C15H18BrF3NO3S are 428.0 and 430.0; measured value is 430.1.

[0087] (R)-2-methylpropane-2-sulfinamide (12.1 g, 100 mmol) and 4-bromo-2-methylbenzaldehyde [INT 15-a] (10 g, 50.2 mmol) were dissolved in tetrahydrofuran (50 mL), and titanium ethoxide (34.2 g, 150 mmol) was added. The mixture was stirred at 25°C for 10 hours. The reaction mixture was then poured into water (500 mL) and extracted with ethyl acetate (3 × 300 mL). The organic extracts were combined, dried over Na₂SO₄, and evaporated under vacuum. The residue was purified by flash chromatography (hexane / MTBE = 1 / 0 to 0 / 1) to obtain (R,E)-N-(4-bromo-2-methylbenzylidene)-2-methylpropane-2-sulfinamide [INT 15-b] (10.4 g, 34.5 mmol, yield 68.8%) as a yellow solid. II NMR (400 MHz, CDCh) δ 8.76 is. 1H), 7.75 (d, J=8.7 Hz, 1H), 7.46 - 7.37 (m, 2H), 2.56 (s, 3H), 1.24 (s, 9H). (R)-2-methylpropane-2-sulfinamide (18.1 g, 150 mmol) and 4-bromo-3-methylbenzaldehyde [INT 15-e] (15 g, 75.3 mmol) were dissolved in tetrahydrofuran (100 mL), and titanium ethoxide (51.3 g, 225 mmol) was added. The mixture was stirred at 60°C for 10 hours. The reaction mixture was then poured into water (500 mL) and extracted with METBE (3 × 300 mL). The organic extract was re-extracted with water (3 × 200 mL), dried over Na₂SO₄, and evaporated under vacuum to obtain (R,E)-N-(4-bromo-3-methylbenzylidene)-2-methylpropane-2-sulfinamide [INT 15-f] (15.2 g, 50.2 mmol, yield 66.9%) as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.49 (s, 1H). 7.67 (d, J=2.2 Hz, 1H), 7.65 - 7.52 (m, 1H), 7.51 - 7.43 (m, III). 2.43 (s, 3H), 1.23 (s, 9H). (R,E)-N-(4-bromo-3-methylbenzylidene)-2-methylpropane-2-sulfinamide [INT 15-f] (21 g, 69.4 mmol) and tetrabutylammonium triphenyl difluorosilicate (56.1 g, 104 mmol) were dissolved in THF (500 ml). Trifluoromethyltrimethylsilane (49.3 g, 347 mmol) was added dropwise at -80°C. The mixture was stirred at -30°C for 30 minutes, and then NH4Cl aqueous solution (300 mL) was added. The mixture was extracted with ELISA (2 × 300 ml). The organic phase was dried over sodium sulfate and evaporated under vacuum at 45°C. The residue was purified by flash chromatography to obtain (R)-N-((S)-1-(4-bromo-3~methylphenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 15-g] (18.5g, 49.7 mmol, yield 71.7%) as a white solid, with a calculated m / z of 372.0 for [M+H]+C13H18BrF3NOS; an experimental value of 372.0. 1 H NMR (500 MHz, CDCl3) δ 7.56 (d, J=8.2 Hz, 1H), 7.30 - 7.26 (m, 1H). 7.14 - 7.08 (m, 1H), 4.75 (p, J=7.1 Hz, 1H), 3.58 (d, J=6.4 Hz, 1H), 2.41 (s, 3H), 1.25 (s, 9H). (R)-N-((S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 15-g] (10g, 26.8 mmol) was dissolved in THF (200 mL). Lithium (1+) bis(trimethylsilyl) azanide (74.3 mL, 80.3 mmol) was added at 0°C. The mixture was stirred at 0°C for 20 minutes. Methyl iodide (22.7 g, 160 mmol) was added. The mixture was stirred at 20°C for 10 hours, and then aqueous NH4Cl solution (200 mL) was added. The mixture was extracted with ELISA (2 × 200 mL). The organic phase was dried with sodium sulfate and evaporated in a vacuum at 45°C to obtain crude (R)-N-((S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoroethyl)-N,2-dimethylpropane-2-sulfinamide [INT 15-h] (9.09 g, 23.5 mmol, yield 88.2%) as a brown oily substance. m / z: [M+H]+ C14H20BrF3NOS Calculated value 386.0; Measured value 386.0. 1 H NMR (400 MHz, CDCl3) δ 7.57 (d, J=8.3 Hz, 1H). 7.34 (s, 1H), 7.17 (d, J=8.6 Hz, 1H), 5.03 (q, J=8.5 Hz, 1H), 2.50 - 2.41 (m, 6H), 1.27 (s, 9H). (R)-N-((S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoroethyl)-N,2-dimethylpropane-2-sulfinamide [INT 15-h] (10.7 g, 27.7 mmol) was dissolved in methanol (20 mL), and then hydrochloric acid (4 M in 1,4-dioxane, 100 mL, 2.54 mol) was added. The mixture was stirred at 20°C for 10 hours, and then evaporated under vacuum at 50°C. MTE (100 mL) was added. The formed solid was filtered and washed with MTBE (50 mL) to obtain (S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoro-N-methylethane-1-amine hydrochloride [INT 15.2] (5.41 g, 16.9 mmol, yield 61.3%) as a beige solid, with calculated m / z values ​​of 282.0 and 284.0 for [M+H]+C10H12BrF3N; measured value 284.0. 1 H NMR(500 MHz, DMSO-d6) δ 10.51 (s, 2H), 7.76 (d, J=8.3 Hz, 1H), 7.63 (d, J=2.2 Hz, 1H), 7.41 (dd, J=8.3, 2.2 Hz, 1H), 5.42 (s, 1H), 2.43 (s, 3H), 2.37 (s, 3H). Phosphorus oxychloride (788 mg, 5.14 mmol) is mixed with [(1S)-1-(4-bromo-2-methylphenyl)-2;2,2-trifluoroethyl](methyl)amine hydrochloride [INT 15.1] (500 mg, 1.56 mmol) and 1,1-dioxo-1λ in pyridine (3 mL). 6 The mixture was added at 0°C to a solution of -thian-4-carboxylic acid [INT 4-a] (833 mg, 4.68 mmol). The reaction mixture was stirred overnight. 3 mL of aqueous sodium bicarbonate solution was added, the mixture was extracted with RINKAN (3 × 10 mL), and washed with NaHSOr (3 × 10 mL). The combined organic layer was dried over anhydrous NaiSCb and evaporated under reduced pressure. The crude product was purified by HPLC (see conditions below) and then N-[(1S)-1-(4-bromo-2-methylphenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6-Chiang-4-carboxamide [INT 16.3] (132 mg, 0.2985 mmol, yield 19.1%) was obtained as a pink solid, with calculated values ​​of m / z [M+H]+ C16H20BrF3NO3S 442.0, 444.0; measured value 444.0.

[0088] [(1S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoroethyl](methyl)amine hydrochloride [INT 15.2] (0.5g, 1.56 mmol) and 1,1-dioxo-1λ 6 -Thian-4-carboxylic acid [INT 4-a] (833 mg, 4.68 mmol) was mixed in pyridine (2 mL). Phosphorus oxychloride (788 mg, 5.14 mmol) was added. The mixture was stirred at 90°C for 10 hours. Ether (20 mL) was added, and the mixture was washed with NaHSOr aqueous solution (3 × 5 mL). The organic phase was dried over sodium sulfate and evaporated in vacuum at 45°C to obtain crude N-[(1S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -Chiang-4-carboxamide [INT 17.3] (656 mg, 1.48 mmol) was obtained as a yellow solid, with a calculated m / z value of 442.0 for [M+H]+ C16H20BrF3NO3S and an actual measured value of 442.0.

[0089] In dioxane (2 mL), N-[(1R)-1-(4-bromophenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6To a solution of -thian-4-carboxamide [INT 14.1] (50 mg, 116 μmol) and 2-chloro-7-[(1S)H-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine hydrochloride [INT 1.1] (31.6 mg, 139 μmol), Pd2(dba)3 (10.6 mg, 11.6 μmol), Cs2CO3 (113 mg, 348 μmol), and xanthophos (13.4 mg, 23.2 μmol) were added. The reaction mixture was stirred at 100°C under N2 conditions for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified by preparative HPLC (column: Boston Green ODS 150*30mm*5pm, table: 24~64%B (A=water (0.05% ammonia hydroxide)), B=acetonitrile), flow rate: 30 mL / min, UV detector 220 nm) and N-[(1R)-1-[4-({2-chloro-7-[(1S)-1-methoxyethyl]-[L2,4]triazolo[1,5-a]pyrimidine-6-yl}amino)phenyl]-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -Chiane-4-carboxamide [Compound 1.26] (16.4 mg, 28.5 pmol, yield 24.6%) was obtained as a yellow dry powder. Calculated value for m / z: [M+H]+ C23H27ClF3N6O4S: 575.1; measured value: 575.3. 1 H NMR (400MHZ, DMSO-d6) δ = 8.83 (s, 1H), 8.07 - 7.97 (m, 1H), 7.29 - 7.17 (m, 2H), 7.03 - 6.92 (m, 2H), 6.49 - 6.05 (m, 1H), 5.16 (q, J=6.8 Hz, 1H), 3,26 - 3.19 (m, 2H), 3.16 (s, 3H), 3.13 - 3.08 (m, 2H), 2.90 (s, 3H), 2.65 (s, 1H), 2.10 - 1.95 (m, 4H), 1.59 (d, J=6.8 Hz, 3H). Dioxane (2 ml) contains 2-chloro-7-[(1R)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine hydrochloride|INT1.3] (60 mg, 227 μmol), N-[(1R)-1-(4-bromophenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 A mixture of -thian-4-carboxamide [INT 14.1] (97.2 mg, 227 μmol), Pd2(dba)3 (20.7 mg, 22.7 μmol), xanthophos (26.2 mg, 45.4 μmol), and Cs2CO3 (221 mg, 681 μmol) was stirred at 100°C for 3 hours under a Ni2 atmosphere. The mixture was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel flash chromatography (methanol / dichloromethane = 0 / 1 to 1 / 20). The obtained product was purified by preparative HPLC (column: YMC Triart C18 250*50mm*7um, table: 26~66%B (A=water (0.05% ammonia hydroxide v / v)), B=acetonitrile), flow rate: 60 mL / min, UV detector 220 nm) and preparative TLC (SiO2, dichloromethane:methanol {=20:1) and then N-[(1R)-1-[4-({2-chloro-7-[(1R)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-yl}amino)phenyl]-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -Chiane-4-carboxamide [Compound 1.27] (5.20 mg, 9.04 μmol, yield 4.0%) was obtained as a white, dry powder. Calculated m / z value for m / z: [M+H.]+ C23H27ClF3N6O4S was 575.1; measured value was 575.3. 11H NMR (400 MHz, CD3OD) δ = 8.86 (s, 1H), 7.30 (d, J=8.4 Hz, 2H), 7.06 (d, J=8.4 Hz, 2H), 6.52 (q, J=9.2 Hz, 1H), 5.36 (q, J=6.8 Hz, 1H), 3.36 (s, 3H), 3.25 - 3.10 (m, 5H), 3.02 - 2.75 (m, 3H), 2.35 - 2.12 (m, 4H), 1.64 (d, J=6.8 Hz, 3H). N-[(1S)-1-(4-Bromo-2-methylphenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -thia-4-carboxamide [INT 16.3] (100 mg, 0.2260 mmol), 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-6-amine [free base of INT 1.I] (51.4 mg, 226 μmol), C S2 CO3 (220 mg, 678 μmol), and dioxane (3 mL) mixture was purged with argon. Then, Xantphos (26.1 mg, 45.2 μmol) and Pd2(dba)3 (20.6 mg, 22.6 μmol) were added and the reaction mixture was stirred at 100 °C for 10 hours. After cooling, the reaction mixture was diluted with EtOAc (30 mL) and concentrated under reduced pressure. The obtained residue was purified by HPLC to give N-[(1S)-1-[4-({2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl}amino)-2-methylphenyl]-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -thia-4-carboxamide [Compound 1.30] (12.5 mg, 0.02128 mmol, yield 9.4%) was obtained as a yellow solid, m / z: calculated value for [M+H]+ C24H29ClF3N6O4S 589.2; measured value 589.0. 1H NMR (500 MHz, DMSO-d6) δ = 8,80 (s, 1H), 7.90 (s, 1H), 7.29 (d, J=8.3 Hz, 1H). 6.87 - 6.75 (m, 2H), 6.43 (q, J=9.1 Hz, 1H), 5.13 (q, 2.01 (s, 3H), 1.99 - 1.94 (m, 2H), 1.57 (d, J=6.7 Hz, 3H). N-[(1S)-1-(4-bromo-3-methylphenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -Chiane-4-carboxamide [INT 17.3] (0.1g, 0.2260 mmol), 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine [free base of INT 1.1] (51.4mg, 226pmol), C s2 CO3 (220 mg, 678 μmol) and xanthophos (13.0 mg, 22.6 μmol) were mixed in dioxane (2 mL), and the reaction mixture was degassed with argon for 5 minutes. Pd2(dba)3 (10.3 mg, 11.3 μmol) was added. The reaction mixture was then degassed with argon for 5 minutes and stirred at 100 °C for 10 hours. The reaction mixture was cooled to room temperature, and solid matter was removed by filtration. The filtrate was purified by HPLC (see conditions below) to obtain N-[(1S)-1-[4-({2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5~a]pyrimidine-6-yl}amino)~3-methylphenyl]-2,2,2-trifluoroethyl]-N-methyl1,1-dioxo-1λ 6 -Chiane-4-carboxamide [Compound 1.33] (20.7 mg, 0.03515 mmol, yield 15.5%) was obtained as a yellow solid. Calculated value for m / z: [M+H]+ C24H29C1F3N6)4S was 589.2; measured value was 589.0. 1H NMR (600 MHz, DMSO-d6) δ = 8,74 (s, 1H), 7.2.4 (s, 1H), 7.15 (s, 1H), 7.03 (d, J=8,6 Hz, 1H), 6.84 (d, J=8.4 Hz, 1H), 6.41 (q, J=9.4 Hz, 1H), 5.16 (q, J=6.7 Hz, 1H), 3.21 (d, J=13.9 Hz, 3H), 3.18-3.05 (m, 5H), 2.89 (s, 3H), 2.34 - 2.28 (m, 3H), 2.11 - 1.95 (m, 4H), 1.54 (d, J=6.7 Hz, 3H). Synthesis of 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine hydrochloride (intermediate 1.1)

[0090] [ka]

[0091] Synthesis of tert-butyl(S)-4-methoxy-3-oxopentanoate (INT1-b): A solution of (S)-2-methoxypropanoic acid [INT 1-a] (20 g, 192 mmol) in anhydrous tetrahydrofuran (342 mL) was cooled to 0°C. Carbonyl diimidazole (30.6 g, 189 mmol) was added in several portions at 0°C, and the mixture was stirred at this temperature for 1.25 hours. In a separate flask, magnesium (1+)1-methylethyl chloride (249 mL, 499 mmol, 2 M in THF) was added to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (46.1 g, 288 mmol) in anhydrous tetrahydrofuran (342 mL) at 0°C, and the mixture was stirred at room temperature for 1.25 hours. This solution was then added to the acylimidazole solution via a cannula at 0°C, and the resulting mixture was stirred overnight at room temperature. The reaction mixture was cooled to 0°C, quenched by adding a 10% aqueous citric acid solution, extracted with ethyl acetate, washed with saturated aqueous NaHCO3 solution, dried over anhydrous Na2SO4, concentrated under reduced pressure to obtain the crude product, which was purified by silica gel flash chromatography (acetone / hexane = 0 / 1 to 1 / 9) to obtain tert-butyl(S)-4-methoxy-3-oxopentanoate [INT 1-b] (30.0 g, 148 mmol, 55.6%) as an oily substance.

[0092] Synthesis of tert-butyl(S)-2-amino-7-(1-methoxyethyl)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate(INT1-c): A solution of tert-butyl(S)-4-methoxy-3-oxopentanoate [INT 1-b] (25 g, 123 mmol) and (dimethoxymethyl)dimethylamine (11.1 mL, 83.6 mmol) was heated at 120°C for 1.5 hours. The mixture was cooled to room temperature, and 4H-1,2,4-triazole-3,5-diamine (12.1 g, 123 mmol), followed by ethanol (123 mL), was added. The mixture was then heated at 85°C for 1 hour. After completion, the mixture was concentrated under reduced pressure, recrystallized from EtOH / water (1:1, 600 mL), filtered, and the filtrate was washed with 30% EtOH / water, followed by MTBE to obtain tert-butyl 2-amino-7-(1-methoxyethyl)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate (12.7 g, 43.2 mmol, 52%) as a solid. The filtrate was concentrated under reduced pressure to remove MTBE, the solid was filtered, and washed with hexane to further obtain tert-butyl(S)-2-amino-7-(1-methoxyethyl)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate [INT 1-c] (5.3 g, 18.0 mmol, 23%) as a solid. Total 18.0 g, yield 75%. Chiral HPLC showed 96.9% ee. 1 H NMR (400 MHz, CDCl3) δ = 8.75 (s, 1H), 5.40 (q, J=6.8 Hz, 1H), 4.95 (br s, 2H), 3.30 (s, 3H), 1.75 (d, J=6.8 Hz, 3H), 1.62 (s, 9H). Synthesis of 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid (INT1-d): A mixture of tert-butyl 2-amino-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate[INT 1-c] (1.2 g, 4.09 mmol) and copper(II) chloride dihydrate (173 mg, 1.02 mmol) in concentrated HCl (20 mL) was mixed with sodium nitrite solution (338 mg, 4.90 mmol) in H2O (5 mL) at 5°C in an ice bath, and the mixture was stirred at 5°C for 30 minutes. The mixture was then heated to 25°C and stirred for 16 hours. Water (100 mL) was added, and the pH was adjusted to 3-4 by adding 1N NaOH aqueous solution. The mixture was extracted with CHCl3:i-PrOH=3:1 (100 mL x 3), the combined organic layer was dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid[INT 1-d] (962 mg, yield 92.4%) as a solid. m / z:Calculated value for [M+H]+ C9H10ClN4O3 was 257.0; measured value was 256.9. 1 H NMR (400 MHz, DMSO-d6) δ = 14.00 (br s, 1H), 9.07 (s, 1H), 5.39 (q, J=6.4 Hz, 1H), 3.21 (s, 3H), 1.63 (d, J=6.4 Hz, 3H). Synthesis of tert-butyl N-{2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-yl}carbamate (INT1-e): A solution of 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid [INT 1-d] (1.3 g, 5.06 mmol) in t-BuOH (10 mL) was added with {[azido(phenoxy)phosphoryl]oxy}benzene (2.08 g, 7.58 mmol) and triethylamine (1.02 g, 10.1 mmol), and the mixture was stirred at 100 °C for 2 h under a N2 atmosphere. The mixture was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel flash chromatography (EtOAc / petroleum ether = 1 / 10 to 1 / 5) to give tert-butyl N-{2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl}carbamate [INT 1-e] (420 mg, yield 25.4%) as a solid. m / z: calculated value for [M+H]+ C13H19ClN5O3 is 328.1; measured value is 328.0. 1 H NMR (400 MHz, CDCl3) δ = 9.62 (br s, 1H), 8.05 (s, 1H), 5.45 (q, J=6.8 Hz, 1H), 3.48 (s, 3H), 1.63 (d, J=6.8 Hz, 3H), 1.56 (s, 9H). Synthesis of 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-6-amine hydrochloride (INT 1.1): A mixture of tert-butyl N-{2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl}carbamate [INT 1-e] (420 mg, 1.28 mmol) in 4N HCl / dioxane (5 mL) was stirred at 25 °C for 2 h. The completion of the reaction was indicated by LCMS, and one new peak with the desired MS was detected (Rt = 0.611 min, m / z: 227.8 [M+H]+). The mixture was concentrated under reduced pressure to obtain 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-6-amine hydrochloride [INT 1.1] (400 mg, crude) as a solid.

[0093] Synthesis of tetrahydro-2H-thiopyran-4-carbonyl chloride 1,1-dioxide (intermediate 4.1):

[0094] [ka]

[0095] To a solution of tetrahydro-2H-thiopyran-4-carboxylic acid 1,1-dioxide [INT 4-a] (41.0 g, 230 mmol, 1.0 eq) in DCM (410 mL), (COCl)2 (58.4 g, 460 mmol, 40.3 mL, 2.0 eq) and DMF (168 mg, 2.30 mmol, 177 μL, 0.01 eq) were added under N2 at 0°C. The mixture was heated to 20°C and stirred at 20°C for 2 hours. The suspension turned clear, indicating that most of the starting material had been consumed. The reaction mixture was concentrated under vacuum to obtain the crude product as a solid, which was concentrated by an oil pump to remove solvent residue and obtain tetrahydro-2H-thiopyran-4-carbonyl chloride 1,1-dioxide [INT 4.1] (46.5 g, crude) as a solid.

[0096] Synthesis of (S)-N-(1-(4-bromophenyl)-2,2,2-trifluoroethyl)-N-methyltetrahydro-2H-thiopyran-4-carboxamide 1,1-dioxide (intermediate 5.1)

[0097] [ka]

[0098] To a solution of (S)-1-(4-bromophenyl)-2,2,2-trifluoro-N-methylethane-1-amine hydrochloride [INT 3.1] (39.0 g, 128 mmol, 1.0 eq, HCl) and TEA (45.7 g, 451 mmol, 62.8 mL, 3.5 eq) in DCM (200 mL), tetrahydro-2H-thiopyran-4-carbonyl chloride 1,1-dioxide [INT 4.1] (45.3 g, 231 mmol, 1.8 eq) was added at 0-10°C. The mixture was stirred at 20°C for 12 hours. The mixture was divided to obtain the organic layer, and the aqueous layer was extracted with DCM (100 mL). The combined organic layers were concentrated under vacuum to obtain the crude product as an oily substance. The crude product was purified by silica gel column chromatography using petroleum ether / ethyl acetate (15 / 1~3 / 1) to obtain (S)-N-(1-(4-bromophenyl)-2,2,2-trifluoroethyl)-N-methyltetrahydro-2H-thiopyran-4-carboxamide 1,1-dioxide [INT 5.1] (26.0 g, 60.7 mmol, yield 47.4%, purity 100%) as a solid. 1 H NMR (400 MHz, CDCl3) δ 2.25-2.37 (m, 1H), 2.38-2.40 (m, 3H), 2.88-3.00 (m, 6H), 3.30-3.31 (m, 1H), 3.22-3.45 (m, 1H), 6.56-6.63 (m, 1H), 7.23 (d, J = 8.00 Hz, 2H), 7.55 (d, J = 8.40 Hz, 2H). SFC: Rt=1.21 min, 100.0%ee; Column: Chiralpak AD-3, 50 x 4.6 mm i.d., 3 um; Mobile phase: A: CO2, B: MeOH (0.05% IPAm, v / v); Flow rate: 3.4 mL / min; Column temperature: 35 °C.

[0099] LC-MS: Rt = 2.431 min, purity 100%, m / z = 428.0, 430.0 (M+1)+. The gradient was maintained for 0.40 min at 5%B, 0.4–3.0 min at 5–95%B, 1.00 min at 95%B, and then 0.01 min at 95–5%B, with a flow rate of 1.0 ml / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50*2.1 mm column (5 μm particles). Detection methods were diode array (DAD) and positive electrospray ionization. The MS range was 100–1000.

[0100] N-[(1S)-1-[4-({2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-yl}amino)phenyl]-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 Synthesis of -thian-4-carboxamide (also known as N-((S)-1-(4-((2-chloro-7-((S)-1-methoxyethyl)-[1,2,4]triazolo[1,5-a]pyrimidine-6-yl)amino)phenyl)-2,2,2-trifluoroethyl)-N-methyltetrahydro-2H-thiopyran-4-carboxamide 1,1-dioxide) (compound 1.1)

[0101] [ka]

[0102] Dioxane (6 mL) contains 2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-amine hydrochloride [INT 1.1] (460 mg, 1.74 mmol), N-[(1S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6A mixture of -thian-4-carboxamide [INT 5.1] (779 mg, 1.82 mmol), Pd2(dba)3 (159 mg, 174 μmol), xanthophos (201 mg, 348 μmol), and Cs2CO3 (1.70 g, 5.22 mmol) was stirred at 100°C for 4 hours. The mixture was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel flash chromatography (methanol / dichloromethane = 0 / 1 to 1 / 10) and preparative HPLC (column: YMC Triart C18 250*50mm*7μm, table: 29~58%B (A = water (0.05% ammonia hydroxide v / v)), B = acetonitrile), flow rate: 60 mL / min, UV detector 220 nm), and then N-[(1S)-1-[4-({2-chloro-7-[(1S)-1-methoxyethyl]-[1,2,4]triazolo[1,5-a]pyrimidine-6-yl}amino)phenyl]-2,2,2-trifluoroethyl]-N-methyl-1,1-dioxo-1λ 6 -Thiane-4-carboxamide [Compound 1.1] (211 mg, 368 μmol) was obtained as a dry powder. m / z: [M+H]+ C23H27ClF3N6O4S Calculated value 575.2; Measured value 575.3. 1 ¹H NMR (400 MHz, DMSO-d6) δ = 8.80 (s, 1H), 8.02 - 7.94 (m, 1H), 7.26 - 7.12 (m, 2H), 6.99 - 6.89 (m, 2H), 6.45 - 6.02 (m, 1H), 5.12 (q, J=6.8 Hz, 1H), 3.25 - 3.14 (m, 3H), 3.12 (s, 3H), 3.10 - 3.05 (m, 2H), 2.86 (s, 3H), 2.08 - 1.94 (m, 4H), 1.55 (d, J=6.8 Hz, 3H). The chiral purity of compound 1.1 was determined to be at least 89%.

[0103] The synthesis of other MALT1 inhibitors is described in WO2022 / 081967 (PCT / US21 / 55173), which is incorporated by reference in its entirety for all purposes.

[0104] Synthesis of MALT1 inhibitors of formula Ia Scheme 2

[0105] [ka]

[0106] The starting material G-2a is treated with a base (LiOH, H2O, or NaOH) to provide the compound of formula (B). R2 is phenyl, pyridyl, or pyridazine; R1 is CF3 or (S)-methoxyethane; m and n are independently 0 or 1. R3' is methyl or benzyl. Synthesis of 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-amine

[0107] [ka]

[0108] Triethylamine (2.10 g, 20.8 mmol) was added to a solution of ethyl 2-(ethoxymethylidene)-4,4,4-trifluoro-3-oxobutanoate (5 g, 20.8 mmol) and 2-hydrazinylpyridine [INT 1-iii] (2.26 g, 20.8 mmol) in EtOH (30 mL). The mixture was stirred at 80°C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel flash chromatography (siRNA / PE = 0 / 1 to 10 / 1) to obtain ethyl 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate [INT 1-jjj] (5.00 g, 17.5 mmol, yield 84.3%) as a yellow oil. m / z: Calculated value for [M+H]+ C12H11F3N3O2: 286.1; Measured value: 285.9. 1H NMR (400 MHz, DMSO-d6) δ = 8.64 - 8.59 (m, 1H), 8.34 (s, 1H), 8.19 - 8.12 (m, 1H), 7.80 (dd, J = 0.8, 8.0 Hz, 1H), 7.68 - 7.63 (m, 1H), 4.33 (q, J = 7.2 Hz, 2H), 1.31 (t, J = 7.2 Hz, 3H). Lithium hydroxide monohydrate (440 mg, 10.5 mmol) was added to a mixture of ethyl 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate [INT 1-jjj] (1 g, 3.50 mmol) in THF (6 mL) and H2O (2 mL). The reaction mixture was stirred at 15°C for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was acidified with 1 M HCl to pH=4. The reaction mixture was quenched by adding H2O (20 mL) and extracted with ELISA (20 mL x 2). The combined organic layers were washed with saline solution (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid [INT 1-kkk] (800 mg, 3.11 mmol, yield 88.8%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ = 13.43 (br s, 1H), 8.60 (td, J = 0.8, 4.8 Hz, 1H), 8.28 (s, 1H), 8.14 (dt, J = 2.0, 7.6 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.64 (ddd, J = 0.8, 4.8, 7.6 Hz, 1H). A mixture of 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid [INT 1-kkk] (400 mg, 1.55 mmol), triethylamine (784 mg, 7.75 mmol), and diphenyl phosphoryl azide (564 mg, 2.32 mmol) in t-BuOH (3 mL) and dioxane (3 mL) was stirred at 100°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by silica gel flash chromatography (petroleum ether / ethyl ammonium = 1 / 0 to 1 / 1) to obtain tert-butyl N-[1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-yl]carbamate [INT 1-lll] (470 mg, 1.43 mmol, yield 92.5%) as a white solid. m / z:[M+H]+ C14H16F3N4O2 Calculated value 329.1; Measured value 328.9. 1 H NMR (400 MHz, DMSO-d6) δ = 9.14 (br s, 1H), 8.56 - 8.51 (m, 1H), 8.06 (dt, J = 1.8, 7.6 Hz, 1H), 7.95 (s, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.55 - 7.46 (m, 1H), 1.46 (s, 9H). A solution of tert-butyl N-[1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-yl]carbamate [INT 1-III] (470 mg, 1.43 mmol) in 4 M HCl / dioxane (4 mL) was stirred at 15°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-amine [INT 1.15] (260 mg, 1.13 mmol, yield 79.7%) as a white solid. m / z: Calculated value for [M+H]+ C9H8F3N4: 229.1; Measured value: 228.9. 1H NMR (400 MHz, DMSO-d6) δ = 8.48 (dd, J = 1.2, 4.8 Hz, 1H), 8.01 (dt, J = 1.6, 7.6 Hz, 1H), 7.76 - 7.72 (m, 2H), 7.65 (br s, 2H), 7.43 (dd, J = 4.8, 6.8 Hz, 1H). Synthesis of (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide

[0109] [ka]

[0110] To a solution of 4-bromobenzaldehyde [INT 2-a] (100 g, 541 mmol, 1.0 eq) in toluene (500 mL), (R)-2-methylpropane-2-sulfinamide (72.1 g, 595 mmol, 1.1 eq) was added at 25°C. The mixture was stirred at 25°C for 15 minutes. Then, NaOH (21.6 g, 541 mmol, 1.0 eq) was added to the reaction product, and the mixture was stirred at 25°C for 12 hours. Na2SO4 (50 g) was added to the mixture and stirred for 20 minutes. The four reaction mixtures were combined, filtered by Celite to obtain the filtrate, and concentrated under vacuum to obtain the crude product as an oily substance. The crude product was dissolved in petroleum ether (1.0 L), stirred at -50°C for 1.0 hour, and filtered to obtain (R,E)-N-(4-bromobenzylidene)-2-methylpropane-2-sulfinamide [INT 2-b] (620 g, 2.15 mol, yield 99.5%) as a solid.

[0111] To a solution of (R,E)-N-(4-bromobenzylidene)-2-methylpropane-2-sulfinamide [INT 2-b] (206 g, 715 mmol, 1.0 eq) and tetrabutylammonium acetate (216 g, 715 mmol, 218 mL, 1.0 eq) in DMF (1.4 L), TMSCF3 (259 g, 1.82 mol, 2.5 eq) was added at 0°C. The mixture was stirred at 5°C for 1.5 hours. This process was repeated twice, and the three reaction mixtures were combined. The mixture was poured into a saturated NH4Cl solution (13.0 L) and stirred for 10 minutes to obtain a suspension. The suspension was filtered, and the filtrate was washed with water (5.0 L). The filtered mass was ground with MTBE / petroleum ether (v / v=1:4, 2.0 L) to obtain the product as a solid, the mother liquor was concentrated under vacuum to obtain the crude product as an oily substance, and this was purified by silica gel column chromatography using petroleum ether / ethyl acetate (10 / 1~1 / 1) to obtain (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 2.1] (389 g, 1.09 mol, yield 50.6%) as a solid. 1 H NMR (400 MHz, CDCl3) δ = 1.25 (s, 9H), 3.64 (d, J = 6.40 Hz, 1H), 4.79-4.83 (m, 1H), 7.32 (d, J = 8.40 Hz, 2H), 7.56 (d, J = 6.40 Hz, 2H). Synthesis of (S)-1-(4-bromophenyl)-2,2,2-trifluoro-N-methylethane-1-amine

[0112] [ka]

[0113] (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-2-methylpropane-2-sulfinamide [INT 2.1] (100 g, 279 mmol, 1.0 eq) was added to a solution of LiHMDS (1.0 M, 838 mL, 3.0 eq) at 0-10°C, and the resulting mixture was stirred at 0-10°C for 0.5 hours. MeI (119 g, 838 mmol, 52.1 mL, 3.0 eq) was added to the above mixture at 0-10°C, and the reaction was stirred at 25°C for 1 hour. The process was repeated twice, and the three combined reaction mixtures were poured into saturated aqueous NH4Cl (3.0 L) and diluted with HCl (1.0 L). The mixture was divided to obtain the organic layer, and the aqueous layer was extracted with HCl (500 mL). The combined organic layers were washed with saturated NaCl (1.0 L), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product as an oil. The crude product was purified by silica gel column chromatography using petroleum ether / ethyl acetate (15 / 1~1 / 1) to obtain (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-N,2-dimethylpropane-2-sulfinamide [INT 3-b] (161 g, 432.5 mmol, yield 51.6%) as an oil.

[0114] A mixture of (R)-N-((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)-N,2-dimethylpropane-2-sulfinamide [INT 3-b] (202 g, 543 mmol, 1.0 eq) in HCl / siRNA (4.0 M, 2.02 L, 14.9 eq) was gradually added. The mixture was stirred at 20°C for 1 hour. The reaction mixture was filtered to obtain a solid, which was eluted with siRNA (200 mL), and the mother liquor was concentrated under vacuum to obtain a solid. The solid was purified by silica gel column chromatography using petroleum ether / ethyl acetate (10 / 1~1 / 0), and combined with the filtrate. The mixture was concentrated at 45°C for 1 hour using an oil pump, and the solvent residue was removed to obtain (S)-1-(4-bromophenyl)-2,2,2-trifluoro-N-methylethane-1-amine hydrochloride [INT 3.1] (115 g, 378 mmol, yield 69.6%, purity 100%, HCl) as the solid.1 H NMR (400 MHz, DMSO-d6) δ = 2.45 (s, 3H), 5.51 (s, 1H), 7.62 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 8.40 Hz, 2H), 10.59 (s, 2H). SFC: Rt=0.776 min, 99.98%ee; Column: Chiralpak AD-3, 100 × 4.6 mm, inner diameter, 3 μm; Mobile phase: A: CO2, B: MeOH (0.05% IPAm); Gradient: A: B = 97:3; Flow rate: 3 mL / min; Column temperature: 35 °C.

[0115] LC-MS: Rt = 1.755 min, purity 100.0%, m / z = 268.0, 270.0 (M+1)+. The gradient was maintained for 0.40 min at 5%B, 0.4–3.0 min at 5–95%B, 1.00 min at 95%B, and then 0.01 min at 95–5%B, with a flow rate of 1.0 ml / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50*2.1 mm column (5 μm particles). Detection methods were diode array (DAD) and positive electrospray ionization. The MS range was 100–1000.

[0116] Synthesis of methyl(1r,4r)-4-(chlorocarbonyl)cyclohexane-1-carboxylate

[0117] [ka]

[0118] To a mixture of (1r,4r)-4-(methoxycarbonyl)cyclohexane-1-carboxylic acid [INT 4-d] (1.45 g, 7.78 mmol) in dichloromethane (10 mL), oxalyl dichloride (2.93 g, 23.3 mmol) and DMF (56.8 mg, 778 μmol) were gradually added, and the mixture was stirred at 40°C for 2 hours. The mixture was concentrated under reduced pressure to obtain crude methyl (1r,4r)-4-(chlorocarbonyl)cyclohexane-1-carboxylate [INT 4.4] (1.59 g, 7.76 mmol) as a yellow, rubbery substance.

[0119] Synthesis of methyl(1S,4r)-4-(((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)(methyl)carbamoyl)cyclohexane-1-carboxylate

[0120] [ka]

[0121] A mixture of methyl(1r,4r)-4-(carbonochloridoyl)cyclohexane-1-carboxylate [INT 4.4] (1.59 g, 7.76 mmol) and Et3N (2.65 g, 26.2 mmol) in dichloromethane (6 mL) was mixed with a solution of [(1S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl](methyl)amine hydrochloride [INT 3.1] (1.6 g, 5.25 mmol) in dichloromethane (6 mL), and the mixture was stirred at 25°C for 16 hours. Water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL x 2). The combined organic layers were washed with saline solution (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. This crude product was purified by silica gel flash chromatography (siRNA / petroleum ether = 1 / 10 to 1 / 5) to obtain methyl (1S,4r)-4-(((S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl)(methyl)carbamoyl)cyclohexane-1-carboxylate [INT 5.4] (1.10 g, 2.52 mmol, yield 32.5%) as a yellow oily substance. m / z: Calculated values ​​for [M+H]+ C18H22BrF3NO3 were 436.1 and 438.1; measured value was 438.0.

[0122] Synthesis of methyl(1r,4r)-4-{methyl[(1S)-2,2,2-trifluoro-1-(4-{[1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-yl]amino}phenyl)ethyl]carbamoyl}cyclohexane-1-carboxylate

[0123] [ka]

[0124] To a solution of 1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-amine [INT 1.15] (100 mg, 438 μmol), methyl (1r,4r)-4-{[(1S)-1-(4-bromophenyl)-2,2,2-trifluoroethyl](methyl)carbamoyl}cyclohexane-1-carboxylate [INT 5.4] (191 mg, 438 μmol), Cs2CO3 (426 mg, 1.31 mmol), and xanthophos (50.6 mg, 87.6 μmol) in dioxane (3 mL), Pd2(dba)3 (40.1 mg, 43.8 μmol) was added, and the reaction mixture was stirred at 100°C under N2 for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel flash chromatography (siRNA / PE = 0 / 1 to 1 / 3) to obtain methyl(1r,4r)-4-{methyl[(1S)-2,2,2-trifluoro-1-(4-{[1-(pyridine-2-yl)-5-(trifluoromethyl)-1H-pyrazole-4-yl]amino}phenyl)ethyl]carbamoyl}cyclohexane-1-carboxylate[INT 6.4] (170 mg, 291 μmol, yield 66.6%) as a yellow oil. m / z: [M+H]+ C27H28F6N5O3 Calculated value 584.2; measured value 584.1.

[0125] The synthesis of MALT1 inhibitors is also described in WO2023 / 192506 (PCT / US23 / 16941), which is incorporated in its entirety by reference for all purposes.

[0126] Pharmaceutical composition and administration The compounds provided in accordance with this disclosure are typically administered in the form of pharmaceutical compositions. This disclosure provides pharmaceutical compositions comprising one or more of the compounds described as active ingredients or pharmaceutically acceptable salts or esters thereof, and a carrier comprising one or more pharmaceutically acceptable excipients, inert solid diluents and fillers, diluents comprising sterile aqueous solutions and various organic solvents, penetration enhancers, solubilizers and adjuvants. Such compositions may also comprise buffers, e.g., neutral buffered saline, phosphate-buffered saline; carbohydrates, e.g., glucose, mannose, sucrose or dextran, mannitol; proteins; polypeptides or amino acids, e.g., glycine; antioxidants; chelating agents, e.g., EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives. The pharmaceutical compositions may be administered alone or in combination with other therapeutic agents. Such compositions are prepared in a manner well known in the pharmaceutical field (e.g., Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa., 17th edition (1985); and Modem Pharmaceutics, Marcel Dekker, Inc., 3rd edition (edited by GS Banker and C.T. Rhodes, which is incorporated in its entirety by reference for all purposes)). Suitable pharmaceutically acceptable excipients are well known to those skilled in the art. Examples of pharmaceutically acceptable excipients include phosphate-buffered saline (e.g., 0.01 M phosphate, 0.138 M NaCl, 0.0027 M KCl, pH 7.4), aqueous solutions of inorganic salts, e.g., hydrochloride, hydrobromide, phosphate, or sulfate, saline, glycol, or ethanol, and organic salts, e.g., acetate, propionate, malonate, or benzoate. Adjuvants, e.g., wetting agents or emulsifiers, and pH buffers may also be used. Pharmacochemically acceptable excipients listed in Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991) (which are incorporated herein by reference in their entirety for all purposes) can also be appropriately used. The compositions may be formulated into known forms suitable for oral administration or parenteral administration, e.g., injection or infusion. The composition may contain pharmaceutical additives, such as suspending agents, preservatives, stabilizers and / or dispersants, and preservatives to extend the shelf life during storage.

[0127] The pharmaceutical composition may be administered in a manner appropriate to the disease to be treated (or prevented). The dosage and frequency of administration will be determined by factors such as the patient's condition, the type and severity of the patient's disease, but the appropriate dosage may be determined by clinical trials.

[0128] The pharmaceutical composition may be administered in single or multiple doses by any of the acceptable modes of administration of drugs having similar utility as described in the patents and patent applications incorporated by reference, including, for example, rectal, buccal, sublingual, intranasal, and transdermal routes, by intra-arterial injection, intravenous, intraperitoneal, parenteral, intramuscular, subcutaneous, oral, topical, as an inhalant, or via an impregnated or coated device, such as a stent or cylindrical polymer for arterial insertion.

[0129] One mode of administration is parenteral, particularly by injection. Forms into which the novel compositions of this disclosure can be incorporated for injection include aqueous or oily suspensions or emulsions using sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol solutions, dextrose solutions, or sterile aqueous solutions, and similar medicinal vehicles. Aqueous solutions in physiological saline have also been conventionally used for injection, but are less preferred in the context of this invention. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, etc. (and preferred mixtures thereof), cyclodextrin derivatives, and vegetable oils may also be used. Appropriate fluidity can be maintained, for example, by using coatings, such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants. Microbial activity can be inhibited by various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc.

[0130] Sterile injectable solutions are prepared by incorporating the compounds according to this disclosure in the required amounts in a suitable solvent, along with various other components listed above as needed, followed by filtration and sterilization. Generally, dispersants are prepared by incorporating various sterile active ingredients into a sterile vehicle containing a basic dispersion medium and other components as needed from those listed above. For sterile powders for preparing sterile injectable solutions, preferred preparation methods are vacuum drying and freeze-drying techniques, which yield a powder of the active ingredient and any additional desired components from a pre-sterilized filtered solution of the powder.

[0131] Oral administration is another route for administering the compounds according to this disclosure. Administration may be carried out via capsules or enteric-coated tablets, etc. In the preparation of a pharmaceutical composition comprising at least one of the compounds described herein, the active ingredient is usually diluted with an excipient and / or encapsulated in a carrier, which may be in the form of capsules, sachets, paper or other containers. If the excipient acts as a diluent, it may be in the form of a solid, semi-solid, or liquid material (as described above) and acts as a vehicle, carrier or medium for the active ingredient. Thus, the composition may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or in a liquid medium), for example, ointments, soft gelatin and hard gelatin capsules, sterile injection solutions, and sterile packaged powders containing up to 10% by weight of the active compound.

[0132] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulation may further contain lubricants, such as talc, magnesium stearate, and mineral oil; humectants, emulsifiers and suspending agents; preservatives, such as methyl and propyl hydroxybenzoates; sweeteners; and flavoring agents.

[0133] The compositions of the present invention may be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient using procedures known in the art. Controlled-release drug delivery systems for oral administration include osmotic pump systems and dissolution systems containing polymer-coated storage units, or drug polymer matrix formulations. Examples of controlled-release systems are given in U.S. Patents 3,845,770; 4,326,525; 4,902,514; and 5,616,345. Another formulation for use in the methods of the present invention is a transdermal delivery device ("patch"). Such a transdermal patch may be used to provide a controlled amount of continuous or discontinuous infusion of the compounds of the present invention. The construction and use of transdermal patches for drug delivery are well known in the art. See, for example, U.S. Patents 5,023,252, 4,992,445, and 5,001,139. Such patches may be constructed to deliver pharmaceuticals continuously, in pulses, or on demand.

[0134] The composition is preferably formulated in unit dosage forms. The term “unit dosage form” refers to physically distinct units suitable as unit doses for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in conjunction with a suitable pharmaceutical excipient (e.g., tablets, capsules, ampoules). The compound is generally administered in pharmaceutically effective amounts. Preferably, with respect to oral administration, each dosage unit contains 1 mg to 2 g of the compound described herein, and with respect to parenteral administration, preferably 0.1 to 700 mg of the compound described herein. However, it will be understood that the actual amount of compound administered will usually be determined by the physician in terms of the condition to be treated, the selected route of administration, the actual compound administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient’s symptoms, and other relevant environmental factors.

[0135] With regard to the preparation of solid compositions, such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid pre-formulation composition comprising a homogeneous mixture of the compounds of this disclosure. When these pre-formulation compositions are referred to as homogeneous, it means that the active ingredient is uniformly dispersed throughout the composition, and as a result, the composition can be easily subdivided into uniformly effective unit dosage forms, such as tablets, pills, and capsules.

[0136] The tablets or pills of this disclosure may be coated or compounded to provide a dosage form that offers the benefit of long-term action or to protect from acidic conditions in the stomach. For example, the tablets or pills may contain internal and external dosage components, the latter in the form of an envelope covering the former. The two components may be separated by an enteric coating that serves to resist disintegration in the stomach and allows the internal component to pass intact into the duodenum or delays its release. Various materials may be used for such enteric coatings or coatings, such as several polymer acids and mixtures of polymer acids with materials, e.g., shellac, ethyl alcohol, and cellulose acetate.

[0137] Compositions for inhalation or inhalation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain the above-mentioned preferred pharmaceutically acceptable excipients. Preferably, compositions are administered by oral or nasal respiration for localized or systemic effects. Preferably, compositions in pharmaceutically acceptable solvents may be sprayed using an inert gas. The sprayed solution may be inhaled directly from a spraying device, or the spraying device may be attached to a face mask tent or an intermittent positive pressure respirator. The solution, suspension, or powder composition may be preferably administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0138] MALT1-related diseases Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1). MALT1 is a caspase-like protease that plays a role in BCL10-induced activation of NF-κB, and / or activation of the mTor pathway (e.g., mTORc1 and / or mTORc2), and / or activation of Jun / Fos. The protein is a component of the CARMA1-BCL10-MALT1 (CBM) signalosome that induces lymphocyte activation after antigen-receptor stimulation by inducing NF-κB signaling, and / or mTor pathway signaling (e.g., mTORc1 and / or mTORc2), and / or Jun / Fos signaling. Some MALT1-related genetic disorders result in constitutive or hyperactivation of NF-κB, and / or hyperactivation of the mTor pathway (e.g., mTORc1 and / or mTORc2), and / or hyperactivation of Jun / Fos, and / or hyperactivation of other signaling pathways, by increasing activating signals via MALT1 or decreasing downregulatory signals via MALT1.

[0139] B-cell proliferation with NF-κB and T-cell anergy (BENTA) is a rare genetic disorder of the immune system caused by mutations in the CARD11 (caspase mobilization domain family member 11) gene. The disorder can be characterized by elevated levels of certain B cells (B-cell lymphocytosis), splenomegaly (enlargement of the spleen), enlargement of lymph nodes (lymphadenopathy), immunodeficiency, and / or an increased risk of lymphoma, beginning in infancy.

[0140] CARD11 encodes membrane-associated guanylate kinase (MAGUK), a type of protein that functions as a molecular scaffold for the assembly of multiprotein complexes in specific regions of the cell membrane. This protein is also a member of the CARD protein family, which is defined by possessing a characteristic caspase-associated recruitment domain (CARD). The CARD domain of the protein specifically interacts with BCL10, a protein known to function as a positive regulator of cellular apoptosis, NF-κB activation, and mTOR pathway (e.g., mTORc1 and / or mTORc2) activation. When expressed in cells, this protein activated NF-κB, the mTOR pathway (e.g., mTORc1 and / or mTORc2), and induced phosphorylation of BCL10.

[0141] BENTA disease can be inherited in an autosomal dominant manner or it can develop de novo from spontaneous mutations in the CARD11 gene. BENTA disease is caused by "gain-of-function" mutations in the CARD11 gene, which instruct the gene to produce the CARD11 protein. These gain-of-function mutations result in overactivity of the CARD11 protein. The CARD11 protein is required in both B cells and T cells for activation of the NF-κB and / or mTor pathway (e.g., mTORc1 and / or mTORc2), and / or Jun / Fos, and is essential for a healthy immune response. B cell development and differentiation may also be partially impaired in BENTA disease. BENTA disease is inherited in an autosomal dominant manner. For a person to have BENTA disease, only one of the two copies of CARD11 must be abnormal.

[0142] In the CARD11 gain-of-function allele, the MALT1 signaling pathway leads to constant activation of the Jun / Fos and / or NF-κB and / or mTor pathways (e.g., mTORc1 and / or mTORc2) in immune cells. This results in a lymphocytic proliferative syndrome called BENTA. Patients with BENTA have elevated levels of inflammation and are highly susceptible to recurrent infections, autoimmune diseases, lymphoma, and hemophagocytic lymphohistiocytosis (HLH). HLH is a rare but potentially fatal condition in which certain white blood cells (histiocytes and lymphocytes) accumulate and damage organs, including the bone marrow, liver, and spleen, destroying other blood cells. BENTA-associated mutations may reside within the N-terminal region of CARD11, including the CARD, LATCH, and coiled-coil domains. These domains are responsible for the formation of CARD11 oligomers and the recruitment of BCL10 and MALT1, making this region a hotspot for CARD11 mutations. CARD11 mutants can spontaneously aggregate to form an active signaling cluster with BCL10, MALT1, and active IKK, inducing constitutive NF-κB activation and / or mTor pathway (e.g., mTORc1 and / or mTORc2) activation and / or Jun / Fos activation without stimulating antigen receptors. Examples of BENTA-associated mutants include, for example, the following CARD11 mutants: R30G, Q, or W, C49Y or S, E, F, or N, F115I, T117P, G123S or D, G126R, T128M, F130I, or C, and E134G.

[0143] Patients with BENTA often have mild immunodeficiency and are highly susceptible to recurrent sinusitis and pulmonary infections, as well as viral infections, such as molluscum contagiosum virus, Epstein-Barr virus, or BK virus. Patients with BENTA are also at high risk of developing lymphoma (e.g., large B-cell lymphoma).

[0144] The majority of B cells in the blood of BENTA patients are naive mature B cells with elevated levels of a B cell subtype called transitional B cells. Laboratory studies also show poor B cell differentiation and immunoglobulin or antibody secretion. Serum IgM is low in most patients, and total IgG and IgA are typically at the lower end of normal. Some patients have an inadequate immune response to certain vaccines. While T cell counts in people with BENTA are within the normal range or slightly above, these T cells may be inadequately responsive to certain exogenous pathogens, thus encompassing the word "anergy," which is part of the acronym BENTA.

[0145] Other genetic disorders, such as A20 haploinsufficiency, HOIL1 hypomorphism, CARD14 systemic pustular psoriasis, and NF-κB gain-of-function syndrome, are associated with increased signaling via the MALT1 pathway.

[0146] A20 haploinsufficiency is an autosomal dominant inherited disorder caused by pathological mutations in the tumor necrosis factor (TNF)-α-induced protein 3 gene. Consequently, there is insufficient production of the nuclear factor (NF)-κB regulatory protein A20, encoded by the TNFAIP3 gene. Protein A20, also known as TNAP3, is encoded by TNFAIP3 and plays a crucial role in the negative regulation of inflammation and immunity.

[0147] HOIL1 or HOIP deficiency is associated with autoinflammatory, immunodeficiency, and immunocompromised immune disorders with inflammatory bowel disease (IBD)-like symptoms. MALT1 paracaspase is a novel negative regulator of LUBAC by cleaving HOIL1 protein. The linear ubiquitin chain assembly complex (LUBAC), consisting of HOIL1, HOIP, and SHARPIN, catalyzes linear ubiquitination of target proteins, a post-translational modification essential for NF-κB activation. HOIL1 or HOIP deficiency is caused by mutations in HOIL1 that result in loss of function.

[0148] Other conditions associated with MALT1 include, for example, autoinflammatory conditions (severe, recurrent, or persistent inflammation that is superficially non-provocative), hyperinflammatory conditions (severe or persistent inflammation that is disproportionately high compared to the triggering stimulus), autoimmune, autoinflammatory, or hyperinflammatory conditions resulting from genetic mutations that lead to a "gain-of-function" (including partial gain-of-function) situation in proteins in the pathway that normally activate or promote the activation of cellular signaling pathways targeted by MALT1 proteases, and autoimmune, autoinflammatory, or hyperinflammatory conditions resulting from genetic mutations that lead to a "loss-of-function" (including partial loss-of-function) situation in proteins in the inhibitory pathway that normally inhibit or promote the inhibition of cellular signaling pathways targeted by MALT1 proteases.

[0149] Usage The compounds and compositions described herein are generally useful for modulating MALT1 and are useful in treating diseases or disorders, particularly those that are highly sensitive to the modulation of MALT1's proteolytic and / or autoproteolytic activity. In some embodiments, the compounds and compositions described herein are useful for inhibiting MALT1. In some embodiments, the compounds and compositions disclosed herein may be useful in treating diseases, disorders, or conditions characterized by NF-κB dysregulation and / or mTOR pathway (e.g., mTORc1 and / or mTORc2) and / or Jun / Fos activation, such as genetic disorders, autoimmune or immunological disorders and inflammatory disorders, allergic disorders, respiratory disorders, and oncological disorders. These conditions or conditions, including those described above, may be treated by administering the MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) to patients suffering from the disease, disorder, or condition.

[0150] Other autoimmune and inflammatory disorders that may be treated with MALT1 inhibitors include, for example, arthritis, ankylosing spondylitis, inflammatory bowel disease, ulcerative colitis, gastritis, pancreatitis, Crohn's disease, celiac disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, rheumatoid arthritis, rheumatic fever, gout, organ or graft rejection, acute or chronic graft-versus-host disease, chronic allograft rejection, Behçet's disease, uveitis, psoriasis, psoriatic arthritis, BENA disease, A20 haploinsufficiency, HOIL1 or HOIP deficiency, polymyositis, and skin disease. These include inflammation, atopic dermatitis, dermatomyositis, acne vulgaris, myasthenia gravis, hidradenitis suppurativa, Graves' disease, Hashimoto's thyroiditis, Sjögren's syndrome, and vesicular disorders (e.g., pemphigus vulgaris), ANCA-associated vasculitis, Henoch-Schönlein purpura, IKBKG gain-of-function (also known as NEMO), IKB gain-of-function, immune complex-driven diseases (lupus, rheumatoid arthritis, IgA nephritis, IgA vasculitis, IgG vasculitis), and antibody-mediated vasculitis syndromes, including immune complex vasculitis (primary or secondary to infection or cancer).

[0151] As shown in Figure 1, MALT1 is a key component in the signaling pathway for NF-κB activation and / or mTOR pathway (e.g., mTORc1 and / or mTORc2) activation and / or Jun / Fos activation, resulting in the activation of immune cells (e.g., B cells, T cells, macrophages and / or neutrophils). As shown in Figure 2, the CARD11 mutation may result in gain-of-function in patients, where MALT1 signals to activate NF-κB and / or mTOR pathway (e.g., mTORc1 and / or mTORc2) and / or Jun / Fos, thereby producing BENTA.

[0152] NF-κB activation by the CARD11 allele that generates gain-of-function and BENTA can be mitigated or prevented using MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). When MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4) are applied to cells (e.g., immune cells, e.g., B cells, T cells, macrophages, and / or neutrophils) with NF-κB activation from the CARD11 allele that generates gain-of-function and BENTA via the MALT1 signalosome, NF-κB activation can be reduced, thereby decreasing the cellular activation status. In some cases, MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) can return cells to a quiescent state.

[0153] Patients with a CARD11 gain-of-function mutation (e.g., BENTA) may be treated by administering a MALT1 inhibitor as disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). The MALT1 inhibitor (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) may be administered in a preferred formulation (e.g., oral dosage form) at a preferred dose (e.g., 25–1000 mg / day). The MALT1 inhibitor (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) produce a therapeutic effect by reversing some or all of the effects of the CARD11 gain-of-function allele in the patient (e.g., a BENTA patient). CARD11 gain-of-function mutations can also reduce the killing activity of natural killer cells. This effect on NK cells can be reversed by MALT1 inhibitors disclosed herein for treating viral infections (e.g., EBV or CMV, members of the herpes family). These infections can be very serious or life-threatening for patients with BENTA.

[0154] As shown in Figure 3, the CARD14 gain-of-function mutant in CARD14 systemic pustular psoriasis interacts with MALT1 in a manner similar to that of the CARD11 mutant in BENTA.

[0155] The activation of the NF-κB and / or mTor pathway (e.g., mTORc1 and / or mTORc2) and / or Jun / Fos by the CARD14 allele, which generates gain-of-function and CARD14 systemic pustular psoriasis, can be mitigated or prevented using MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4). When MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4) are administered to cells (e.g., immune cells, e.g., B cells, T cells, macrophages, and / or neutrophils) that undergo NF-κB activation from the CARD14 allele to generate gain-of-function and CARD14 systemic pustular psoriasis via the MALT1 signalosome, it is possible to reduce NF-κB activation and / or mTOR pathway activation (e.g., mTORc1 and / or mTORc2) and / or Jun / Fos activation, thereby reducing the activating state of the cells. In some cases, MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) can return cells to a quiescent state.

[0156] Patients suffering from CARD14 dermatitis (e.g., generalized pustular psoriasis, pityriasis rubra pilaris) may be treated by administering the MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). The MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) may be administered in a preferred formulation (e.g., in an oral dosage form) at a preferred dose (e.g., 25 to 1000 mg / day). MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) produce therapeutic effects by reversing some or all of the effects of the CARD14 gain-of-function allele in patients with CARD14 systemic pustular psoriasis.

[0157] Activation of the Jun / Fos and / or NF-κB and / or mTor pathways (e.g., mTORc1 and / or mTORc2) by three tumor necrosis factor (TNF)-α-induced protein alleles that generate gain-of-function and A20 haploinsufficiency can be mitigated or prevented using MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). When MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are administered to cells (e.g., immune cells, e.g., B cells, T cells, macrophages, and / or neutrophils) with activation of the NF-κB and / or mTor pathway (e.g., mTORc1 and / or mTORc2) and / or Jun / Fos via three tumor necrosis factor (TNF)-α-induced protein alleles that generate gain-of-function and A20 haploinsufficiency via the MALT1 signalosome, the activation of the NF-κB and / or mTor pathway (e.g., mTORc1 and / or mTORc2) and / or Jun / Fos can be reduced, thereby decreasing the activation status of the cells. In some cases, MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) can return cells to a quiescent state.

[0158] Patients suffering from A20 haploinsufficiency may be treated by administering MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) may be administered in a preferred formulation (e.g., oral dosage form) at a preferred dose (e.g., 25 to 1000 mg / day). MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) produce a therapeutic effect by reversing some or all of the effects of the tumor necrosis factor (TNF)-α-induced protein 3 gain-of-function alleles in patients suffering from A20 haploinsufficiency.

[0159] Activation of the Jun / Fos and / or NF-κB and / or mTor pathway (e.g., mTORc1 and / or mTORc2) by the HOIL1 allele, which generates loss of function and HOIL1 or HOIP deficiency, can be mitigated or prevented using MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). When MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are applied to cells (e.g., immune cells, e.g., B cells, T cells, macrophages, and / or neutrophils) with NF-κB activation from the CARD14 allele, which generates gain of function and HOIL1 or HOIP deficiency via the MALT1 signalosome, NF-κB activation can be reduced, thereby decreasing the cellular activation status. In some cases, MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) can return cells to a quiescent state.

[0160] In general, undesirable activation of the Jun / Fos and / or NF-κB and / or mTor pathways (e.g., mTORc1 and / or mTORc2) via the MALT1 signalosome from any source (e.g., genetic and / or receptor stimulation) can be mitigated or prevented using MALT1 inhibitors disclosed herein (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4). When MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are administered to cells (e.g., immune cells, e.g., B cells, T cells, macrophages, and / or neutrophils) exhibiting undesirable activation of the Jun / Fos and / or NF-κB and / or mTor pathways (e.g., mTORc1 and / or mTORc2) via the MALT1 signalosome, NF-κB activation can be reduced, thereby decreasing the cellular activation status. In some cases, MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) can return cells to a quiescent state.

[0161] The compounds of the compositions described herein may be administered in combination with other agents or treatments. The subjects to whom the compounds disclosed herein will be administered may have diseases, disorders, or conditions, or symptoms thereof, to which benefits would be obtained from treatment with other agents or treatments. The compounds of the compositions described herein may be administered simultaneously with or before or after one or more other therapeutic agents. The compounds of the compositions described herein may be administered individually or together in the same pharmaceutical composition by the same or different routes of administration as the other agents. The compounds described herein may be administered as a single active ingredient, or together with other drugs, such as immunosuppressants or immunomodulators or other anti-inflammatory agents, such as those for treating or preventing acute or chronic rejection of allografts or inflammatory or autoimmune disorders, or chemotherapeutic agents, such as malignant cell antiproliferative agents, for example, as adjuvants to these. For example, the compounds of the present invention may be used in combination with calcineurin inhibitors, such as cyclosporine A or FK506; mTOR inhibitors, such as rapamycin, 40-0-(2-hydroxyethyl)-rapamycin, biolimus-7 or biolimus-9; ascomycin having immunosuppressive properties, such as ABT-281 or ASM981; corticosteroids; cyclophosphamide; azathioprine; methotrexate; leflunomide; mizoribine; mycophenolic acid or salt; mycophenolate mofetil; or IL-1 beta inhibitors.

[0162] Suitable genetic testing of CARD11 gain-of-function mutants (alleles) that produce BENTA can be performed using well-known methods. Such methods include, for example, high-throughput genome sequencing, exome sequencing, FISH techniques, microarrays, other hybridization techniques, PCR-related diagnostics, other nucleic acid amplification techniques, CRISPR diagnostics, denaturing HPLC, gene expression profiling, pharmacokinetic studies of CARD11, and / or CARD11 / MALT1 / NF-κB and associated metabolite levels.

[0163] Various features and embodiments of this disclosure are shown in the following representative examples, which are intended to be illustrative and not limiting. However, those skilled in the art will readily understand that the specific methods and results discussed are merely illustrative of the invention, which will be described more fully in the subsequent claims. Unless otherwise indicated, this disclosure is not limited to specific procedures, materials, etc., and may be modified as such. It should also be understood that the technical terms used herein are for the sole purpose of describing specific embodiments and are not intended to limit them. [Examples]

[0164] Examples Example 1: MALT1 inhibitor reverses the gain-of-function effect induced by the CARD11 variant. MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are incubated with T cells that lack endogenous CARD11 and have transiently transfected CARD11 cDNA containing a gain-of-function mutation. A variation as needed is the co-transfer of 50:50 copies of the wild-type CARD11 gene and the CARD11 gain-of-function mutant. The T cells contain a reporter gene (κB-GFP) that acts as a readout for NFκB signaling. NF-κB-GFP expression is measured at baseline and after anti-CD3 or PMA / ionomycin stimulation.

[0165] T cells possessing the CARD11 GOF variant exhibit high baseline and post-stimulation NF-κB-GFP activation, which is assessed by flow cytometry. MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) suppress the effects of CARD11 gain-of-function mutants, resulting in decreased NF-κB-GFP expression.

[0166] Example 2: Clinical trial of the MALT1 inhibitor BENTA MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are used to treat patients with BENTA. The MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are formulated in oral form and administered to the patient once daily at an initial dose of 25 mg / day. Alternatively, the daily dose may be gradually increased from 25 mg / day to 1000 mg / day.

[0167] Patients treated with MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) show reduced activation of Jun / Fos and / or NF-κB and / or mTOR pathways (e.g., mTORc1 and / or mTORc2) and improvement of BENTA symptoms.

[0168] Example 3: MALT1 inhibitors reverse the activation of the mTor pathway (e.g., mTORc1 and / or mTORc2) by gain-of-function CARD11 variants. Primary T cells are stimulated and activated and cultured in the presence of IL-2. For stimulation, cells are pre-incubated on ice with anti-CD3ε and anti-CD28 and washed. Cells are stimulated by crosslinking anti-CD3 antibody with a secondary antibody in complete T cell medium (e.g., anti-mouse IgG, 10 minutes) and then fixed with pre-warmed fixation buffer. Cells are then washed with FACS buffer, permeabilized with permeabilization buffer pre-cooled to -20°C, and incubated on ice for 30 minutes. Cells are washed and then stained with the following antibodies or appropriate isotype controls: anti-pAkt (Ser473) and anti-pS6 (Ser240 / 244).

[0169] MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are incubated with activated primary T cells. MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) suppress the activation of the mTor pathway (e.g., mTORc1 and / or mTORc2).

[0170] Example 4: MALT1 inhibitor reverses the gain-of-function effect of the CARD14 variant. MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are incubated with epithelial cells (e.g., HEK293 cells) that lack endogenous CARD14 and have transiently transfected CARD14 cDNA with gain-of-function mutations. The epithelial cells contain a reporter gene (κB-GFP) that acts as a readout for NFκB signaling. NF-κB-GFP expression is measured at baseline and after anti-CD3 or PMA / ionomycin stimulation.

[0171] Epithelial cells with gain-of-function variants of CARD14 exhibit high baseline and post-stimulation NF-κB-GFP activation, which is assessed by flow cytometry. MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) suppress the effects of CARD14 gain-of-function mutants, resulting in decreased NF-κB-GFP expression.

[0172] Example 5: MALT1 cleavage of HOIL1 is reduced by a MALT1 inhibitor. Recombinant full-length human MALT1 protein was expressed and purified. C-terminal Myc-FLAG-labeled full-length human HOIL1 was obtained from Origene. HOIL1 protein (0.05 μg / μL) was incubated with different concentrations of MALT1 in assay buffer (200 mM Tris-HCl, 0.8 M sodium citrate, 0.1 mM EGTA, 0.05% CHAPS, 1 mM DTT, pH 7.4) at 37°C for 2 hours. HOIL1 cleavage was analyzed by gel electrophoresis using a 4–12% bis-tris-SDS-polyacrylamide (PAGE) gradient gel (Life Technologies). The presence of small fragments was confirmed by immunoblotting using an N-terminal antibody (anti-N-terminal HOIL1; HPA024185; Sigma). Total HOIL1 was observed using a C-terminal recognition antibody (anti-FLAG, clone M2, Sigma). These assays are performed using both cleavable HOIL1 and non-cleavable HOIL1 (with Lys replaced by Arg165) as a negative control.

[0173] MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, 2.1, and / or 2.4) are added to the reaction mix for HOIL1 cleavage to reduce HOIL1 cleavage.

[0174] Example 6: MALT1 inhibitors reduce NF-κB activity in CARD11 gain-of-function mutants of Jurcut cells. Jurcut cells were genetically engineered using E134G, C49Y, or G123D CARD11 gain-of-function (GOF) mutants. The Jurcut cells contained a reporter gene (κB-GFP) that acts as a readout for NFκB signaling.

[0175] Jurcut cells possessing GOF E134G, C49Y, or G123D CARD11 showed elevated baseline NF-κB-GFP activation. MALT1 inhibitors (e.g., compound 1.1 or 2.4) were incubated with the genetically modified Jurcut cells. The MALT1 inhibitors (e.g., compound 1.1 or 2.4) suppressed the effects of the CARD11 gain-of-function mutants, resulting in decreased NF-κB-GFP expression in all three GOF CARD11 mutants. These MALT1 inhibitors reduced NF-κB-GFP expression in a dose-dependent manner from approximately 100 nM to 5 μM for both compound 1.1 and compound 2.4.

[0176] Jurcut cells possessing GOF E134G, C49Y, or G123D CARD11 were also evaluated for CYLD cleavage as a measure of MALT1 proteolytic activity. In the absence of inhibitors, Jurcut cells with GOF CARD11 mutants cleaved CYLD. MALT1 inhibitors (e.g., compound 1.1 or 2.4) suppressed this CYLD cleavage in a dose-dependent manner in the range of approximately 100 nM to 10 μM.

[0177] Example 7: MALT1 inhibitors reduce NF-κB activity in CARD11 gain-of-function mutants of B cells. B cells are obtained from patients with BENTA (B cell proliferation with NF-κB and T cell anergy). Both resting B cells and activated (CD40L+IL-4) B cells are treated with MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4).

[0178] Changes in steady-state CARD11-induced NF-κB signaling are measured at various time points over approximately 24 hours by monitoring CARD11 aggregation and p65 nuclear accumulation, or by directly quantifying the active NF-κB complex in nuclear lysates (TransAm assays). MALT1 substrate cleavage will also be monitored over time.

[0179] Changes in the expression of NF-κB-dependent genes known to be elevated in BENTA B cells (e.g., BCL2, NFKB2, c-FLIP, cyclin D1) are evaluated by quantitative PCR and / or immunoblotting of whole cell lysates with / + MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4). Alternatively, bulk RNA-seq can be performed on BENTA B cells with / + MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4) to evaluate changes in the overall B cell transcriptome.

[0180] Acquisition of CARD11 function is known to promote B cell survival and proliferation, which can lead to an excess of B cells, as in BENTA or B-cell lymphoma. MALT1 inhibitors reduce this survival. We evaluate the survival and proliferation of healthy B cells or BENTA B cells in response to combinations of stimuli (e.g., anti-IgM / IgD, rCD40L / IL-4 / IL-21, porkweed mitogen, etc.)- / + MALT1 inhibitors (e.g., compounds 1.1, 1.26, 1.27, 1.30, 1.33, and / or 2.4).

[0181] All publications, patents, patent applications, and other documents referenced herein are incorporated herein by reference in whole for all purposes to the same extent that each individual publication, patent, patent application, or other document is individually indicated as being incorporated by reference for all purposes.

[0182] While various specific embodiments have been shown and described, it will be understood that various modifications may be made without departing from the scope of the invention in this disclosure.

Claims

1. A method for reducing NF-κB activation caused by the CARD11 gain-of-function allele, comprising the steps of: obtaining cells having the CARD11 gain-of-function allele; and exposing the cells to a MALT1 inhibitor to reduce the NF-κB activation.

2. A method for treating a patient having a CARD11 gain-of-function mutation, comprising the steps of: obtaining a patient having BENTA; and administering a therapeutically effective dose of a MALT1 inhibitor to the patient, thereby reducing NF-κB activation in the patient.

3. The method according to claim 3, wherein the patient is a BENTA patient.

4. A method for reducing NF-κB activation caused by the CARD14 gain-of-function allele, comprising the steps of: obtaining cells having the CARD14 gain-of-function allele; and exposing the cells to a MALT1 inhibitor to reduce the NF-κB activation.

5. A method for treating a patient having CARD14 dermatitis, comprising the steps of: obtaining a patient having CARD14 systemic pustular psoriasis; and administering a therapeutically effective amount of a MALT1 inhibitor to the patient, thereby reducing NF-κB activation in the patient with CARD14 systemic pustular psoriasis.

6. The method according to claim 4, wherein the CARD14 dermatitis is pustular psoriasis or pityriasis rubra pilaris.

7. A method for reducing the activation of NF-κB caused by tumor necrosis factor (TNF)-α-induced protein 3 gain-of-function alleles, comprising the steps of: obtaining cells having the tumor necrosis factor (TNF)-α-induced protein 3 gain-of-function alleles; and exposing the cells to a MALT1 inhibitor to reduce the activation of NF-κB.

8. A method for treating a patient having A20 haploinsufficiency, comprising the steps of: obtaining the patient having A20 haploinsufficiency; and administering a therapeutically effective amount of a MALT1 inhibitor to the patient, thereby reducing NF-κB activation in the patient having A20 haploinsufficiency.

9. A method for reducing NF-κB activation caused by a gain-of-function allele acting on MALT1, comprising the steps of: obtaining cells having the gain-of-function allele; and exposing the cells to a MALT1 inhibitor to reduce the NF-κB activation.

10. A method for reducing NF-κB activation caused by the CARD11 gain-of-function allele, comprising the steps of: obtaining cells having the CARD11 gain-of-function allele; and exposing the cells to a MALT1 inhibitor to reduce the activation of mTORC1 or mTORc2.

11. A method for reducing NF-κB activation caused by the CARD11 gain-of-function allele, comprising the steps of: obtaining cells having the CARD11 gain-of-function allele; and exposing the cells to a MALT1 inhibitor to increase Jun / Fos inactivation.

12. The method according to any one of claims 1 to 11, wherein the cells are immune cells.

13. The method according to claim 12, wherein the immune cells are B cells, T cells, macrophages, or neutrophils.

14. The method according to claim 13, wherein the immune cells are B cells.

15. The MALT1 inhibitor is a compound of formula 1. 【Chemistry 1】 The method according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof (wherein: R1 is selected from the group consisting of C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, and 5-10 membered heterocyclils, wherein the C1-6 alkyl, C3-6 cycloalkyl, and 5-10 membered heterocyclils may be optionally substituted on one or more available carbons by one, two, three, or more substituents independently selected from R1a, and if the 5-10 membered heterocyclil contains a substituted ring nitrogen atom, that ring nitrogen atom may be optionally substituted by R1b, and if the 5-10 membered heterocyclil contains a substituted ring sulfur atom, that ring sulfur atom may be optionally substituted by two oxygen atoms; R2 is either CH3 or CF3; Is R3 hydrogen; or R3 is selected from the group consisting of C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heterocyclyl-C1-3 alkyl5-6 membered heterocyclyl-O-, phenyl, and 5-6 membered heteroaryl, and any of these may be optionally substituted with one, two, or three substituents independently selected from R3a; R4 is a C1-6 alkyl group; R1a is independently selected, for each occurrence, from the group consisting of cyano, halogen, hydroxyl, oxo, C1-6 alkyl, -C(O)ORA, -C(O)N(RA)2, -N(RA)2, C1-6 alkoxy, 5-6 membered heterocyclyl, and 5-6 membered heteroaryl, wherein the C1-6 alkyl is optionally substituted with N(RA)2, and if the 5-6 membered heterocyclyl contains a substituteable ring nitrogen atom, that ring nitrogen atom may optionally be substituted with Rp; R1b is selected from the group consisting of C1-6 alkyl, -C(O)ORA, -C(O)C1-6 alkyl, -C(O)C3-6 cycloalkyl, -C(O)N(RA)2, and -S(O)2C1-6 alkyl; R3a is independently selected, for each occurrence, from the group consisting of halogens, C1-4 alkyls, C1-4 haloalkyls, C1-4 alkoxys, C1-4 haloalkoxys, hydroxy, C1-4 alkenyls, cyanos, azides, -NRCRDs, C3-6 cycloalkyls, Ci1-4 alkoxys, 5-6 membered heterocyclyl-O-saturates, 5-6 membered heterocyclyls, and phenyls, where the C3-6 cycloalkyls, 5-6 membered heterocyclyl-O-saturates, 5-6 membered heterocyclyls, and phenyls are optionally substituted with one, two, or three substituents independently selected from Rp; Rp is independently selected, for each occurrence, from the group consisting of halogens, C1-4 alkyls, C1-4 haloalkyls, hydroxy, C1-4 alkoxys, C1-4 alkoxyC1-4 alkyls, NRCRDs, and aminoC1-3 alkyls; RA is independently selected, for each occurrence, from the group consisting of hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, and -C(O)OC1-6 alkyl; RB is selected from the group consisting of Ci1-6 alkyl, C1-6 cycloalkyl, and -C(O)OC1-6 alkyl; RC and RD are independently selected, for each occurrence, from the group consisting of hydrogen, C1-6 alkyl, halo-C1-6 alkyl, and C3-4 cycloalkyl, or RC and RD, together with the nitrogen atom to which they are bonded, form a 4-6 membered heterocycline or 4-6 membered heteroaryl, which may contain further nitrogen or oxygen atoms and may be optionally substituted with one or two fluorocarbons; T is either 0 or 1.

16. The method according to claim 15, wherein the MALT1 inhibitor comprises compound 1.1, compound 1.26, compound 1.27, compound 1.30, or compound 1.33, or a combination thereof.

17. The method according to claim 16, wherein the MALT1 inhibitor comprises compound 1.

1.

18. The method according to claim 16, wherein the MALT1 inhibitor comprises compound 1.

26.

19. The method according to claim 16, wherein the MALT1 inhibitor comprises compound 1.

27.

20. The method according to claim 16, wherein the MALT1 inhibitor comprises compound 1.

30.

21. The method according to claim 16, wherein the MALT1 inhibitor comprises compound 1.

33.

22. The MALT1 inhibitor is a compound of formula Ia. 【Chemistry 2】 The method according to any one of claims 1 to 11, or a stereoisomer thereof and / or a pharmaceutically acceptable salt thereof (wherein: R1 is a C1-6 alkyl or C1-3 haloalkyl, and the C1-6 alkyl may be substituted with an -O-C1-3 alkyl as needed; R2 is an aryl or a 5- to 6-membered heteroaryl, wherein the aryl may be substituted with a cyano as needed; R4 is -C(O)OH or a 5- to 6-membered heteroaryl; m is either 0 or 1; n is either 0 or 1.

23. The method according to claim 22, wherein the MALT1 inhibitor is compound 2.1 or compound 2.

4.

24. The method according to claim 23, wherein the MALT1 inhibitor is compound 2.

1.

25. The method according to claim 23, wherein the MALT1 inhibitor is compound 2.4.

Images