Use of Bicyclic Compounds for the Treatment of Diseases - Patent application

Abstract

Provided herein are compounds and compositions thereof for modulating hepatocyte growth factor. In some embodiments, the compounds and compositions are provided for the treatment of diseases, including neurological disorders.

Description

[Technical field]

[0001] The present disclosure relates generally to the use of compounds and compositions to treat diseases such as mild cognitive impairment. [Background technology]

[0002] Hepatocyte growth factor (HGF) is a pleiotropic protein factor involved in numerous biological processes, including embryonic and organ development, regeneration, and inflammation. HGF plays a key role in the development and maturation of cortical, motor, sensory, sympathetic, and parasympathetic neurons. HGF is translated and secreted as an inactive pro-HGF, but upon cleavage, the resulting α and β subunits are connected by disulfide bonds to form an active heterodimer. HGF expression occurs primarily in mesenchymal cells, such as fibroblasts, chondroblasts, adipocytes, and endothelial cells. It has also been shown to be expressed in the central nervous system (CNS), including neurons, astrocytes, and ependymal cells (Nakamura and Mizuno, 2010). All biological activities of HGF are mediated by MET, a transmembrane receptor tyrosine kinase that serves as the only known receptor for HGF. MET is known to be involved in a variety of biological processes, and roles in development, regeneration, and response to injury have been shown. Binding of HGF to the extracellular domain of MET results in homodimerization of the MET protein and autophosphorylation of the intracellular domain. Phosphorylation of the MET intracellular domain leads to the recruitment and phosphorylation of various effector proteins, including Gab1, GRB2, phospholipase C, and Stat3 (Gherardi et al., 2012; Organ and Tsao, 2011). These effector proteins then interact with downstream signaling pathways, including PI3K / Akt, Ras / Raf / MAPK, RAC1 / CDC42, and RAP / FAK, among others, to affect various cellular components, including gene regulation, cytoskeletal reorganization, cell cycle progression, cell adhesion, survival, and proliferation (Organ and Tsao, 2011).

[0003] Because HGF has been shown to play a role in development (Nakamura et al., 2011), homeostasis (Funakoshi and Nakamura, 2003), suppression of cell death, and regeneration (Matsumoto et al., 2014), stimulation of the HGF / MET signaling pathway is an ideal target for therapeutic agents in various disease states. Modulation of HGF activity has been proposed as a treatment for diseases and injuries in a wide variety of tissue types, including the liver, kidney, gastrointestinal tract, cardiovascular system, lung, skin, nervous system, and muscular system (Matsumoto et al., 2014). However, highly effective compounds useful for modulating HGF / MET signaling activity have yet to be explored or discovered.

[0004] Although progress has been made in the field, there remains a need for improved compounds and methods for the treatment of HGF-regulated diseases. Thus, in one aspect, compounds that modulate HGF are provided for use in the treatment of neurodegenerative diseases. Summary of the Invention

[0005] In certain embodiments, described herein are compounds and compositions thereof for modulating hepatocyte growth factor (HGF) for the treatment of disease. Non-limiting exemplary embodiments include the following:

[0006] Embodiment 1. A method of treating mild cognitive impairment in a subject in need thereof, comprising administering to a subject a compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof, L is a direct bond, -C(=O)-, -(CR a R b ) m -C(=O)-, -C(=O)-(CR a R b ) m - or -(CR a R b )m - and Each R a and R b are independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, or C 2 -C 6 is alkynyl, R 1a and R 1b are independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, halo, or C 6 -C 10 is arylalkyl, R 2 is H, oxo, or thioxo, R 3 is C 2 -C 6 Alkyl, C 3 -C 6 Alkenyl, C 3 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 6 Cycloalkylalkyl, C 6 -C 10 arylalkyl, 5- to 10-membered heteroarylalkyl, or 5- to 10-membered heterocyclylalkyl; wherein the 5- to 10-membered heteroarylalkyl or 5- to 10-membered heterocyclylalkyl contains 1 to 3 heteroatoms selected from nitrogen and oxygen; R 4 is C 6 -C 10 aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heterocyclyl; wherein the 5- to 10-membered heteroaryl or 5- to 10-membered heterocyclyl contains 1 to 3 heteroatoms selected from nitrogen and oxygen; Each R 5 is independently 1 -C 6 alkyl, oxo, or halo; R 6 , H, C 1 -C 6 alkyl, or oxo; R 7 is H or oxo, m is 1 or 2; n is an integer from 0 to 3, Here, each C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 12 Cycloalkylalkyl, C 6 -C 10 Aryl, C 6 -C 10 Arylalkyl, 5-10 membered heteroaryl, 5-10 membered heteroarylalkyl, 5-10 membered heterocyclyl, and 5-10 membered heterocyclylalkyl are hydroxyl, halo, amino, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, cyano, -(C=O)NH 2 , Nitro, -SO 2 (C 1 -C 6 alkyl), and -CO 2 H).

[0007] Embodiment 2. L is -C(=O)- or -(CR a Rb ) m The method of embodiment 1,

[0008] Embodiment 3. The method of embodiment 1 or 2, wherein L is -C(=O)-.

[0009] Embodiment 4. L is -(CR a R b ) m The method of embodiment 1 or 2, wherein

[0010] Embodiment 5.R a and R b The method of embodiment 4, wherein each is H and m is 1.

[0011] Embodiment 6.R 1a and R 1b each independently represents H; halo, -CO 2 H and -C(=O)NH 2 C optionally substituted with 1 to 3 substituents selected from 1 -C 6 Alkyl;C 1 -C 6 C optionally substituted with 1 to 3 substituents selected from alkoxy; halo; or halo and amino. 6 -C 10 The method of any one of embodiments 1 to 5, wherein the is arylalkyl.

[0012] Embodiment 7.R 1a and R 1b are each independently H, methyl, fluoro, 2-methylbutyl, -CH 2 F, methoxy, -CH 2 CO 2 H, -CH 2 C(=O)NH 2 7. The method of embodiment 6, wherein the aryl group is 4-aminobenzyl, benzyl, or 4-aminobenzyl.

[0013] Embodiment 8.R 1a and R 1b are each independently H or C 1 -C 3The method of embodiment 6, wherein the alkyl group is alkyl.

[0014] Embodiment 9.R 1a is methyl and R 1b The method of embodiment 8, wherein

[0015] Embodiment 10.R 1a and R 1b The method of embodiment 8, wherein each is H.

[0016] Embodiment 11.R 2 The method of any one of embodiments 1-10, wherein

[0017] Embodiment 12.R 2 The method of any one of embodiments 1-10, wherein is thioxo.

[0018] Embodiment 13.R 2 The method of any one of embodiments 1-10, wherein is oxo.

[0019] Embodiment 14.R 3 But, C 3 -C 6 Alkyl, C 3 -C 6 Alkenyl, C 3 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 6 Cycloalkylalkyl, C 6 -C 10 arylalkyl, 5-10 membered heteroarylalkyl, or 5-10 membered heterocyclylalkyl, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, arylalkyl, heteroarylalkyl, or heterocyclylalkyl is selected from the group consisting of hydroxyl, halo, amino, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6Haloalkoxy, cyano, -(C=O)NH 2 , Nitro, -SO 2 (C 1 -C 6 alkyl), and -CO 2 The method of any one of embodiments 1-13, wherein the compound is optionally substituted with 1 to 5 substituents selected from H.

[0020] Embodiment 15.R 3 But, Halo, C 1 -C 3 Alkoxy, hydroxy, -NH 2 , -SO 2 (C 1 -C 3 alkyl), and -C(=O)NH 2 C optionally substituted with 1 to 3 substituents selected from 2 -C 6 Alkyl;C 2 -C 6 Alkenyl; C 3 -C 6 Cycloalkylalkyl; 5-6 membered heteroarylalkyl; 5-6 membered heterocyclylalkyl; or C 6 The method of any one of embodiments 1-13, wherein the is arylalkyl.

[0021] Embodiment 16.R 3 But, C 1 -C 3 Alkoxy, hydroxy, -NH 2 , and -SO 2 (C 1 -C 3 C substituted by 1 to 3 substituents selected from alkyl 2 The method of embodiment 15, wherein the alkyl group is alkyl.

[0022] Embodiment 17.R 3 17. The method of any one of embodiments 14-16, wherein [ka]

[0023] Embodiment 18.R 3 18. The method of embodiment 17, wherein: [ka]

[0024] Embodiment 19.R 4 But halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 C optionally substituted with 1 to 3 substituents selected from haloalkoxy 6 -C 10 The method of any one of embodiments 1-18, wherein is aryl.

[0025] Embodiment 20.R 4 But -CF 3 , -OCHF 2 20. The method of embodiment 19, wherein the aryl group is phenyl substituted with 1 to 3 substituents selected from: -OH, fluoro, and chloro.

[0026] Embodiment 21.R 4 21. The method of embodiment 20, wherein: [ka]

[0027] Embodiment 22.R 4 22. The method of embodiment 21, wherein: [ka]

[0028] Embodiment 23.R 4 But halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 The method of any one of embodiments 1-18, wherein the heteroaryl is 5-10 membered optionally substituted with 1-3 substituents selected from haloalkoxy.

[0029] Embodiment 24.R 4 But halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 The method of embodiment 23, wherein the pyridyl or indolyl is optionally substituted with 1 to 3 substituents selected from haloalkoxy.

[0030] Embodiment 25.R 4 but, [ka] 25. The method of embodiment 24, wherein

[0031] Embodiment 26.R 4 but, [ka] 26. The method of embodiment 25, wherein

[0032] Embodiment 27.R 4 But halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 The method of any one of embodiments 1-18, wherein the heterocyclyl is 5-10 membered optionally substituted with 1-3 substituents selected from haloalkoxy.

[0033] Embodiment 28.R 4 The method of embodiment 27, wherein is indolinyl.

[0034] Embodiment 29.R 4 but, [ka] 29. The method of embodiment 28, wherein

[0035] Embodiment 30.-LR 4 27. The method of any one of embodiments 1-26, wherein [ka]

[0036] Embodiment 31. The method of any one of embodiments 1 to 30, wherein n is 0.

[0037] Embodiment 32. The method of any one of embodiments 1 to 30, wherein n is 1.

[0038] Embodiment 33.R 5 The method of embodiment 32, wherein is oxo or halo.

[0039] Embodiment 34.R 5 The method of embodiment 33, wherein is oxo or fluoro.

[0040] Embodiment 35.R 2 The method of any one of embodiments 1-34, wherein

[0041] Embodiment 36.R 2 The method of any one of embodiments 1-35, wherein is oxo.

[0042] Embodiment 37. The method of any one of embodiments 1-10, 13-31, 35, and 36, wherein the compound has formula (V): [ka]

[0043] EMBODIMENT 38 L is -C(=O)- or -CH 2 - and R 1a and R 1b are independently H, or -CO 2 C optionally substituted with H 1 -C 3 is alkyl, R 3 But, C 4 -C 5 Alkyl, C4 -C 5 Alkenyl, or C 3 -C 5 Cycloalkyl-substituted C 1 -C 3 is alkyl, R 4 But -CF 3 , -OCHF 2 38. The method of embodiment 37, wherein the aryl group is phenyl or pyridyl substituted with 1 to 3 substituents selected from -OH, fluoro, and chloro.

[0044] Embodiment 39. A method of treating mild cognitive impairment in a subject in need thereof comprising administering an effective amount of a compound selected from the compounds of Table 1A and pharma- ceutically acceptable salts thereof.

[0045] Embodiment 40 The method of any one of the preceding embodiments, wherein the method slows the progression of dementia in the subject.

[0046] Embodiment 41 The method of any one of the preceding embodiments, wherein the method improves cognitive function and / or slows the progression of cognitive impairment in the subject. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0047] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the following description, certain specific details are set forth to provide a thorough understanding of various embodiments of the present disclosure. It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not intended to limit any of the claimed subject matter. In the event that any material incorporated herein by reference contradicts the explicit content of the present disclosure, the explicit content shall control. In this application, the use of the singular includes the plural unless expressly stated. It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. In this application, the use of "or" means "and / or" unless otherwise stated. Furthermore, the use of the term "including," as well as other forms such as "include," "includes," and "included," is not limiting.

[0048] Unless the context requires otherwise, throughout this specification and claims, the word "comprise" and variations thereof, such as "comprises" and "comprising," are intended to be interpreted in their open and inclusive sense, i.e., "including, but not limited to."

[0049] In the present description, any concentration range, percentage range, ratio range, or integer range is understood to include any integer value within the recited range, and, where appropriate, fractions thereof (such as 1 / 10 and 1 / 100 of an integer), unless otherwise specified. Also, any numerical range recited herein for any physical characteristic, such as polymer subunits, size, or thickness, is understood to include any integer within the recited range, unless otherwise specified. As used herein, the terms "about" and "approximately" mean ±20%, ±10%, ±5%, or ±1% of the indicated range, value, or structure, unless otherwise specified.

[0050] References throughout this specification to "one embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrase "one embodiment" or "embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0051] "Amino" is -NH 2 Refers to radicals.

[0052] "Carboxy" or "carboxyl" is -CO 2 H radical.

[0053] "Cyano" refers to the -CN radical.

[0054] "Hydroxy" or "hydroxyl" refers to the --OH radical.

[0055] "Nitro" is -NO 2 Refers to radicals.

[0056] "Oxo" refers to the =O substituent.

[0057] "Thioxo" refers to the =S substituent.

[0058] "Thiol" refers to a -SH substituent.

[0059] "Alkyl" means an unbranched or branched saturated hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, having from 1 to 12 carbon atoms (C 1 -C 12 alkyl), preferably having 1 to 8 carbon atoms (C 1 -C 8 alkyl), 1 to 6 carbon atoms (C 1 -C 6 alkyl), or 1 to 3 carbon atoms (C 1 -C 3 alkyl) and is attached to the remainder of the molecule by a single bond, for example, methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless stated otherwise in the specification, alkyl groups are optionally substituted.

[0060] "Alkenyl" means an unbranched or branched unsaturated hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, containing one or more carbon-carbon double bonds and having 2 to 12 carbon atoms (C 2 -C 12 alkenyl), preferably having 2 to 8 carbon atoms (C 2 -C 8 alkenyl) or 2 to 6 carbon atoms (C 2 -C 6 alkenyl) which is attached to the remainder of the molecule by a single bond, e.g., ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, etc. Unless stated otherwise in the specification, alkenyl groups are optionally substituted.

[0061] "Alkynyl" means an unbranched or branched unsaturated hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, containing one or more carbon-carbon triple bonds and having 2 to 12 carbon atoms (C 2 -C 12 alkynyl), preferably having 2 to 8 carbon atoms (C 2 -C 8 alkynyl) or 2 to 6 carbon atoms (C 2 -C 6 alkynyl) which is attached to the remainder of the molecule by a single bond, e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc. Unless stated otherwise in the specification, alkynyl groups are optionally substituted.

[0062] "Alkoxy" means a group of the formula -OR a where R a is an alkyl radical as defined above containing 1 to 12 carbon atoms. Preferred alkoxy groups are those containing 1 to 6 carbon atoms (i.e., C 1 -C 6 alkoxy) or 1 to 3 carbon atoms (i.e., C 1 -C 3 Unless stated otherwise in the specification, an alkoxy group is optionally substituted.

[0063] "Aromatic ring" refers to a cyclic planar portion (i.e., radical) of a molecule having a resonance-bonded ring that exhibits enhanced stability compared to other connection arrangements by the same atomic group. In general, aromatic rings contain covalently bonded coplanar atoms and contain an even number of pi electrons (e.g., alternating double and single bonds) that is not a multiple of four (i.e., 4n+2 pi electrons, where n=0, 1, 2, 3, etc.). Aromatic rings include, but are not limited to, phenyl, naphthenyl, imidazolyl, pyrrolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridonyl, pyridazinyl, and pyrimidonyl. Unless otherwise specified herein, aromatic rings include all radicals that are optionally substituted.

[0064] "Aryl" refers to an alkyl group having 6 to 18 carbon atoms and at least one aromatic ring (i.e., C 6 -C 18 aryl), preferably containing 6 to 10 carbon atoms (i.e., C 6 -C 10 For purposes of the present disclosure, aryl radicals are monocyclic, bicyclic, tricyclic or tetracyclic ring systems, which may include fused or bridged ring systems. Aryl radicals include, but are not limited to, aryl radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, phenyl, pleiadene, pyrene, and triphenylene. Unless otherwise stated in the specification, aryl groups are optionally substituted.

[0065] An "arylalkyl" is a group of the formula -R b -R c where R b is an alkylene chain, and R c is one or more aryl radicals as defined above, e.g., benzyl, diphenylmethyl, etc. An arylalkyl group is 6 -C 10 C connected to an aryl radical 1 -C 10 Alkylene chain (i.e., C 6 -C 10 Unless stated otherwise in the specification, an arylalkyl group may be optionally substituted.

[0066] "Cycloalkyl" means a stable non-aromatic monocyclic or polycyclic carbocyclic radical, consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from 3 to 15 carbon atoms (i.e., C 3 -C 15 cycloalkyl), preferably having 3 to 10 carbon atoms (i.e., C 3 -C10 cycloalkyl) or 3 to 6 carbon atoms (i.e., C 3 -C 6 Cycloalkyl refers to groups having a cyclic or cyclic ring structure, which may be saturated or unsaturated and connected to the remainder of the molecule by a single bond. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cycloalkyl also includes "spirocycloalkyl" when there are two substitution positions on the same carbon atom. Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless stated otherwise in the specification, cycloalkyl groups are optionally substituted.

[0067] "Cycloalkylalkyl" refers to a group of the formula -R b -R c where R b is an alkylene chain, and R c is one or more cycloalkyl radicals as defined above, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. A cycloalkylalkyl group is one or more of the following: 3 -C 12 C attached to a cycloalkyl radical 1 -C 10 Alkylene chain (i.e., C 3 -C 12 Cycloalkylalkyl) or C 3 -C 6 C attached to a cycloalkyl radical 1 -C 10 Alkylene chain (i.e., C 3 -C 6 Unless stated otherwise in the specification, a cycloalkylalkyl group may be optionally substituted.

[0068] "Fused" refers to any ring structure described herein that is fused to an existing ring structure in a compound of the present disclosure. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom on the existing ring structure that becomes part of the fused heterocyclyl ring or fused heteroaryl ring is replaced with a nitrogen atom.

[0069] "Halo" or "halogen" refers to bromo, chloro, fluoro, or iodo.

[0070] "Haloalkyl" refers to an alkyl radical, as defined above, substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. Preferred haloalkyl groups include alkyl groups having 1 to 6 carbon atoms and substituted by one or more halo radicals (i.e., C 1 -C 6 The halo radicals can all be the same or the halo radicals can be different. Unless stated otherwise in the specification, a haloalkyl group is optionally substituted.

[0071] "Haloalkoxy" refers to a group of the formula -OR a where R a is a haloalkyl radical, as defined herein, containing 1 to 12 carbon atoms. Preferred haloalkoxy groups include those containing 1 to 6 carbon atoms (i.e., C 1 -C 6 haloalkoxy) or 1 to 3 carbon atoms (C 1 -C 3 haloalkoxy) and includes alkoxy groups substituted with one or more halo radicals. The halo radicals can all be the same or all different. Unless stated otherwise in the specification, a haloalkoxy group is optionally substituted.

[0072] "Heteroaryl" refers to an aromatic group having a monocyclic, polycyclic, or multiple fused rings (e.g., a 5-14 membered ring system) containing one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. As used herein, heteroaryl contains 1-10 ring carbon atoms and 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur in the ring. Preferred heteroaryl groups have 5-10 membered ring systems containing 1-4 heteroatoms selected from nitrogen, oxygen, and sulfur (i.e., 5-10 membered heteroaryl) and 5-6 membered ring systems containing 1-4 heteroatoms selected from nitrogen, oxygen, and sulfur (i.e., 5-6 membered heteroaryl). For purposes of embodiments of the present disclosure, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems. Examples of heteroaryl groups include pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Heteroaryl can include one or more N-oxide (NO-) moieties, such as pyridine-N-oxide. Unless otherwise stated herein, heteroaryl groups are optionally substituted.

[0073] "Heteroarylalkyl" refers to a group of the formula -R b -R c where R b is an alkylene chain, and R c is one or more heteroaryl radicals as defined above. A heteroarylalkyl group is a C alkyl group attached to a 5-10 membered heteroaryl group. 1 -C 10 C attached to an alkylene chain (i.e., 5- to 10-membered heteroarylalkyl) or a 5- to 6-membered heteroaryl group 1 -C 10 It may contain an alkylene chain (i.e., a 5-6 membered heteroarylalkyl).Unless stated otherwise in the specification, a heteroarylalkyl group is optionally substituted.

[0074] "Heterocyclyl" refers to a saturated or unsaturated cyclic alkyl group containing one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. The term "heterocyclyl" includes heterocycloalkenyl groups (i.e., heterocyclyl groups having at least one double bond), bridged heterocyclyl groups, fused heterocyclyl groups, and spiroheterocyclyl groups. Heterocyclyls can be monocyclic or polycyclic, where the polycyclic rings can be fused, bridged, or spiro, and can contain one or more oxo (C=O) or N-oxide (NO-) moieties. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of the bond (i.e., it can be bonded through a carbon atom or a heteroatom). Additionally, the term heterocyclyl is intended to encompass any non-aromatic ring containing at least one heteroatom that can be fused to an aryl ring or a heteroaryl ring, regardless of the bond to the rest of the molecule. As used herein, a heterocyclyl has 1 to 10 ring carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms and 1 to 5 ring heteroatoms, 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 to 2 heteroatoms independently selected from nitrogen, sulfur, and oxygen. Preferred heterocyclyls have 5 to 10 members in a ring system containing 1 to 4 heteroatoms selected from nitrogen and oxygen (i.e., 5-10 membered heterocyclyl) or 5 to 8 members in a ring system containing 1 to 4 heteroatoms selected from nitrogen and oxygen (i.e., 5-8 membered heterocyclyl).Examples of heterocyclyl groups include dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise in the specification, heterocyclyl groups are optionally substituted.

[0075] "Heterocyclylalkyl" refers to a group of the formula -R b -R c where R b is an alkylene chain, and R c is one or more heterocyclyl radicals as defined above. A heterocyclylalkyl group is a C 1 -C 10 C attached to an alkylene chain (i.e., 5- to 10-membered heterocyclylalkyl) or a 5- to 8-membered heterocyclyl radical 1 -C 10 It may contain an alkylene chain (i.e., a 5- to 8-membered heterocyclylalkyl).Unless stated otherwise in the specification, a heterocyclylalkyl group is optionally substituted.

[0076] In some embodiments, the term "substituted" as used herein refers to any of the above groups or other substituents (e.g., C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 12In any of the cycloalkyl, alkyl, aryl, and heteroaryl, at least one hydrogen atom (e.g., one, two, three, or all hydrogen atoms) may be replaced with any of the following: halogen atoms (i.e., "halo"), such as F, Cl, Br, and I; oxygen atoms in groups such as hydroxyl or alkoxy groups (e.g., alkoxy or haloalkoxy); amines (e.g., -NH 2 ), amides (e.g., -(C=O)NH 2 ), and a nitrogen atom in the group, such as nitro; alkyl groups that contain one or more halogens, such as F, Cl, Br, and I (e.g., haloalkyl); and cyano.

[0077] L, R 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 It is understood that each selection for is optionally substituted as above, unless otherwise specified, provided that all valences are satisfied by substitution. Specifically, L, R 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 Each of the options for is optionally substituted, unless otherwise specified, provided that such substitution results in a stable molecule (e.g., groups such as H and halo are not optionally substituted).

[0078] As used herein, "mild cognitive impairment" and "MCI" refer to a decline in cognitive function that is intermediate between age-related cognitive changes and early dementia. Individuals exhibiting MCI show evidence of cognitive impairment that indicates a decline from a previous period, but are able to function substantially independently in daily activities. Mild cognitive impairment includes cognitive impairment that may be a precursor to dementia, but is not severe enough to be classified as dementia. MCI may affect one or more cognitive functions, including, but not limited to, learning and memory, language, visuospatial skills, executive function, and psychomotor function. See, e.g., Knopman et al., 2014, Mayo Clin Proc, 89(10):1452-1459.

[0079] An "effective amount" or "therapeutically effective amount" of a compound or composition refers to the amount of the compound or composition that produces the intended result desired based on the disclosure herein. An effective amount can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, ED 50 (the dose therapeutically effective in 50% of the population) and LD 50 (the dose that is lethal to 50% of the population). In some embodiments, an effective amount of a compound results in a reduction or inhibition of symptoms or a prolongation of survival in a subject (i.e., a human patient). Multiple administrations of a compound may be required for results.

[0080] "Treating" or "treatment" of a disease of interest refers to 1) preventing the occurrence of the disease in a patient who is predisposed to or does not yet exhibit symptoms of the disease, 2) inhibiting or arresting the development of the disease, or 3) alleviating or causing remission of the disease. As used herein, "treatment" or "treating" is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of this disclosure, beneficial or desired results include, but are not limited to, one or more of the following: relieving one or more symptoms caused by a disease or disorder of a subject, reducing the extent of a disease or disorder, stabilizing a disease or disorder (e.g., preventing or delaying the worsening of a disease or disorder), delaying the onset or recurrence of a disease or disorder, delaying or slowing the progression of a disease or disorder, improving the condition of a disease or disorder, providing remission (whether partial or total) of a disease or disorder, reducing the dosage of one or more other drugs required to treat a disease or disorder, enhancing the effect of another drug used to treat a disease or disorder, delaying the progression of a disease or disorder, improving quality of life, and / or prolonging survival. Relief of the pathological consequences of a disease or disorder is also encompassed by "treatment". The method of the present invention contemplates any one or more of these modes of treatment.

[0081] As used herein, the terms "individual(s)," "subject(s)," and "patient(s)" refer to any mammal. Examples include, but are not limited to, mice, rats, hamsters, guinea pigs, pigs, rabbits, cats, dogs, goats, sheep, cows, and humans. In some embodiments, the mammal is a human.

[0082] The term "therapeutic benefit" as used herein encompasses therapeutic and / or prophylactic benefits as described herein. A therapeutic benefit includes delaying or eliminating the appearance of a disease or condition; delaying or eliminating the onset of symptoms of a disease or condition; slowing, halting, or reversing the progression of a disease or condition; causing partial or complete remission of a disease or condition; or any combination thereof.

[0083] The terms "co-administration," "administered in combination with," and their grammatical equivalents, as used herein, encompass administration of two or more agents to an animal, including a human, such that both agents and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in one composition in which both agents are present.

[0084] "Pharmaceutically acceptable" refers to compounds, salts, compositions, dosage forms and other materials useful in the preparation of pharmaceutical compositions suitable for animal or human pharmaceutical use.

[0085] "Pharmaceutically acceptable salt" includes both acid and base addition salts.

[0086] "Pharmaceutically acceptable acid addition salts" refers to salts which retain the biological effectiveness and properties of the free base and which are not biologically or otherwise undesirable, for example, but not limited to, those formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, as well as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, geranilic acid, glycerin, glycerol ... It is formed with organic acids such as antiseptic, glucoheptonic, gluconic, glucuronic, glutamic, glutaric, 2-oxo-glutaric, glycerophosphoric, glycolic, hippuric, isobutyric, lactic, lactobionic, lauric, maleic, malic, malonic, mandelic, methanesulfonic, mucic, naphthalene-1,5-disulfonic, naphthalene-2-sulfonic, 1-hydroxy-2-naphthoic, nicotinic, oleic, orotic, oxalic, palmitic, pamoic, propionic, pyroglutamic, pyruvic, salicylic, 4-aminosalicylic, sebacic, stearic, succinic, tartaric, thiocyanic, p-toluenesulfonic, trifluoroacetic, and undecylenic acids.

[0087] "Pharmaceutically acceptable base addition salt" refers to a salt that retains the biological effectiveness and properties of its free acid and is not biologically or otherwise undesirable. These salts are prepared from the addition of an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0088] In some embodiments, pharma- ceutically acceptable salts include quaternary ammonium, such as a quaternary amine alkyl halide salt (eg, methyl bromide).

[0089] As used herein, a "therapeutic agent" refers to a biological, pharmaceutical, or chemical compound or other moiety. Non-limiting examples include simple or complex organic or inorganic molecules, peptides, proteins, oligonucleotides, antibodies, antibody derivatives, antibody fragments, vitamin derivatives, carbohydrates, toxins, or chemotherapeutic compounds. A variety of compounds can be synthesized, including small molecules and oligomers (e.g., oligopeptides and oligonucleotides), as well as synthetic organic compounds based on a variety of core structures. In addition, compounds for screening can be obtained from a variety of natural sources, such as plant or animal extracts.

[0090] The term "in vivo" refers to an event that takes place inside a subject's body.

[0091] Embodiments of the present disclosure are also intended to encompass all pharma- ceutically acceptable compounds of formula (I) that are isotopically labeled by having one or more atoms replaced by an atom having a different atomic mass or mass number (i.e., "isotopic forms" of compounds of formula (I). Examples of isotopes that can be introduced into compounds of formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, e.g., 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125I. These radiolabeled compounds may be useful to aid in determining or measuring the effectiveness of a compound, for example, by characterizing the site or mode of action, or binding affinity to a pharmacologically important site of action. Certain isotopically labeled compounds of formula (I), for example those incorporating a radioisotope, are useful in drug and / or substrate tissue distribution studies. The radioisotope tritium, i.e. 3 H, and carbon-14, i.e. 14 C is particularly useful for this purpose because of its ease of incorporation and ready means of detection.

[0092] Deuterium, i.e. 2 Substitution with heavier isotopes, such as H, may be preferable in some circumstances as they may confer certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements.

[0093] 11 C. 18 F, 15 O and 13 Substitution with positron emitting isotopes, such as N, can be useful in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described in the Examples set forth below, using appropriate isotopically labeled reagents in place of previously employed non-labeled reagents.

[0094] Certain embodiments are also intended to encompass in vivo metabolic products of the disclosed compounds. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound, primarily due to enzymatic processes. Thus, embodiments include compounds produced by a process comprising administering a compound of the present disclosure to a mammal for a period of time sufficient to produce its metabolic products. Such products are typically identified by administering a detectable dose of a radiolabeled compound of the present disclosure to an animal, such as a rat, mouse, guinea pig, monkey, or human, allowing sufficient time for metabolism to occur, and isolating the conversion products from urine, blood, or other biological samples.

[0095] "Stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture and formulation into an efficacious therapeutic agent.

[0096] In many cases, crystallization produces a solvate of the disclosed compound. As used herein, the term "solvate" refers to an aggregate that includes one or more molecules of the compound of formula (I) together with one or more solvent molecules. In some embodiments, the solvent is water, in which case the solvate is a hydrate. Alternatively, in other embodiments, the solvent is an organic solvent. Thus, the compound of formula (I) can exist as a hydrate, including monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate, etc., as well as the corresponding solvates. In some aspects, the compound of formula (I) is a true solvate, and in other cases, the compound of the disclosed compound simply retains adventitious water or is a mixture of water and some adventitious solvent.

[0097] "Optional" or "optionally" means that the subsequently described circumstance event may or may not occur, and the description includes both cases where the event or circumstance occurs and does not occur. For example, "optionally substituted aryl" means that the aryl radical may or may not be substituted, and the description includes both substituted and unsubstituted aryl radicals. It is not intended to include herein polymers or similar infinite structures that are reached by defining a substituent with an unlimited number of further substituents added (e.g., a substituted aryl with a substituted alkyl, which is itself substituted with a substituted aryl group, which is further substituted with a substituted heteroalkyl group, etc.). Similarly, the above definition is not intended to include impermissible substitution patterns (e.g., a methyl substituted with five fluorines, or a heteroaryl group with two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to those skilled in the art.

[0098] A "pharmaceutical composition" or "pharmaceutical acceptable composition" refers to a formulation of a compound of the present disclosure with a vehicle generally accepted in the art for delivering a biologically active compound to a mammal, e.g., a human. Such a vehicle includes any pharmaceutically acceptable carrier, diluent, or excipient therefor.

[0099] A "pharmaceutically acceptable carrier, diluent, or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, flow agent, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonicity agent, solvent, or emulsifier approved by the U.S. Food and Drug Administration as acceptable for use in humans or domestic animals.

[0100] The compounds of formula (I), or pharma- ceutically acceptable salts or isotopic forms thereof, may contain one or more centers resulting in geometric asymmetry and thus may give rise to enantiomers, diastereomers, and other stereoisomers, defined in terms of absolute stereochemistry as (R)- or (S)-, or in the case of amino acids, (D)- or (L)-. Accordingly, the embodiments include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or may be resolved using conventional techniques, e.g., chromatography and fractional crystallization. Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of the racemates (or racemates of salts or derivatives), e.g., using chiral high-performance liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers, as well as all tautomers.

[0101] "Stereoisomers" refer to compounds consisting of the same atoms joined by the same bonds, but with different, incompatible three-dimensional structures. The present disclosure contemplates various stereoisomers and mixtures thereof, and includes "enantiomers," which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0102] "Diastereomers" are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.

[0103] "Tautomer" refers to the migration of a proton from one atom of a molecule to another atom of the same molecule. Thus, embodiments include tautomers of the compounds of the present disclosure.

[0104] The chemical naming protocol and structure diagrams used herein are modified IUPAC nomenclature using ACD / Name Version 9.07 software program and / or ChemDraw Ultra Version 11.0.1 software naming program (CambridgeSoft). In complex chemical names used herein, a substituent is typically named before the group to which it is attached. For example, cyclopropylethyl contains an ethyl skeleton with a cyclopropyl substituent. Except as noted below, in the chemical structure diagrams herein, all bonds are specified, except for all bonds on some carbon atoms, and it is assumed that the bonds on carbon atoms are attached to enough hydrogen atoms to satisfy the valence.

[0105] Although various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, although the invention may be described herein for clarity in the context of separate embodiments, the invention may also be implemented in a single embodiment.

[0106] compound In one aspect, provided herein is a compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof, L is a direct bond, -C(=O)-, -(CR a R b ) m -C(=O)-, -C(=O)-(CR a R b ) m - or -(CR a R b ) m - and Each R a and R b are independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, or C2 -C 6 is alkynyl, R 1a and R 1b are independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, halo, or C 6 -C 10 is arylalkyl, R 2 is H, oxo, or thioxo, R 3 is C 2 -C 6 Alkyl, C 3 -C 6 Alkenyl, C 3 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 6 Cycloalkylalkyl, C 6 -C 10 arylalkyl, 5- to 10-membered heteroarylalkyl, or 5- to 10-membered heterocyclylalkyl; wherein the 5- to 10-membered heteroarylalkyl or 5- to 10-membered heterocyclylalkyl contains 1 to 3 heteroatoms selected from nitrogen and oxygen; R 4 is C 6 -C 10 aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heterocyclyl; wherein the 5- to 10-membered heteroaryl or 5- to 10-membered heterocyclyl contains 1 to 3 heteroatoms selected from nitrogen and oxygen; Each R 5 is independently 1 -C 6 alkyl, oxo, or halo; R 6 , H, C 1 -C6 alkyl, or oxo; R 7 is H or oxo, m is 1 or 2; n is an integer from 0 to 3, Here, each C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 12 Cycloalkylalkyl, C 6 -C 10 Aryl, C 6 -C 10 Arylalkyl, 5-10 membered heteroaryl, 5-10 membered heteroarylalkyl, 5-10 membered heterocyclyl, and 5-10 membered heterocyclylalkyl are hydroxyl, halo, amino, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, cyano, -(C=O)NH 2 , Nitro, -SO 2 (C 1 -C 6 alkyl), and -CO 2 H.

[0107] In some embodiments, L is a direct bond. In some embodiments, L is -C(=O)- or -(CR a R b ) m In some embodiments, L is -C(=O)-. In some embodiments, L is -(CR a R b ) m In some embodiments, L is -(CR a R b ) m-C(=O)- or -C(=O)-(CR a R b ) m In some embodiments, L is -(CR a R b ) m In some embodiments, L is -C(=O)-(CR a R b ) m -It is.

[0108] In some embodiments, each R a and R b are independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, or C 2 -C 6 In some embodiments, each R a and R b are independently H, C 1 -C 3 Alkyl, C 2 -C 4 Alkenyl, or C 2 -C 4 In some embodiments, R a and R b are each H. In some embodiments, R a is H. In some embodiments, R a is C, such as methyl, ethyl, or propyl. 1 -C 6 In some embodiments, R a is vinyl or propenyl, etc. 2 -C 6 In some embodiments, R a is C, such as ethynyl or propynyl. 2 -C 6 In some embodiments, R b is H. In some embodiments, R bis C, such as methyl, ethyl, or propyl. 1 -C 6 In some embodiments, R b is vinyl or propenyl, etc. 2 -C 6 In some embodiments, R b is C, such as ethynyl or propynyl. 2 -C 6 It is alkynyl.

[0109] In some embodiments, R 1a and R 1b are independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, halo, or C 6 -C 10 In some embodiments, R 1a is H. In some embodiments, R 1a is C, such as methyl, ethyl, or propyl. 1 -C 6 In some embodiments, R 1a is vinyl or propenyl, etc. 2 -C 6 In some embodiments, R 1a is C, such as ethynyl or propynyl. 2 -C 6 In some embodiments, R 1a is C such as methoxy, ethoxy, or propoxy. 1 -C 6 In some embodiments, R 1a is halo, such as fluoro, chloro, or bromo. In some embodiments, R 1a C such as benzyl 6 -C 10In some embodiments, R 1b is H. In some embodiments, R 1b is C, such as methyl, ethyl, or propyl. 1 -C 6 In some embodiments, R 1b is vinyl or propenyl, etc. 2 -C 6 In some embodiments, R 1b is C, such as ethynyl or propynyl. 2 -C 6 In some embodiments, R 1b is C such as methoxy, ethoxy, or propoxy. 1 -C 6 In some embodiments, R 1b is halo, such as fluoro, chloro, or bromo. In some embodiments, R 1b C such as benzyl 6 -C 10 It is arylalkyl.

[0110] In some embodiments, R 1a and R 1b are each independently H; halo, -CO 2 H and -C(=O)NH 2 C optionally substituted with 1 to 3 substituents selected from 1 -C 6 Alkyl;C 1 -C 6 C optionally substituted with 1 to 3 substituents selected from alkoxy; halo; or halo and amino. 6 -C 10 In some embodiments, R 1a is C substituted with 1 to 3 halo such as fluoro or chloro. 1 -C 6 In some embodiments, R 1a 1 to 3 -CO 2 C substituted with H group1 -C 6 In some variations, R 1a 1-2 CO 2 C substituted with H group 1 -C 3 Alkyl, e.g., -CH 2 CO 2 H or -CH 2 CH 2 CO 2 H. In some embodiments, R 1a is 1 to 3 -C(=O)NH 2 C substituted with a group 1 -C 6 In some embodiments, R 1a is 1 or 2 -C(=O)NH 2 C substituted with a group 1 -C 3 Alkyl, e.g., -CH 2 C(=O)NH 2 or -CH 2 CH 2 C(=O)NH 2 In some embodiments, R 1a is C substituted with 1 to 3 substituents selected from halo and amino 6 -C 10 In some embodiments, R 1a is C substituted with 1-3 halo, such as fluoro, chloro, or bromo. 6 -C 10 In some embodiments, R 1a is C substituted with 1 to 3 amino groups 6 -C 10 In some embodiments, R 1b is C substituted with 1 to 3 halo such as fluoro or chloro. 1 -C 6 In some embodiments, R 1b 1 to 3 -CO 2 C substituted with H group 1 -C 6In some variations, R 1b 1-2 CO 2 C substituted with H group 1 -C 3 Alkyl, e.g., -CH 2 CO 2 H or -CH 2 CH 2 CO 2 H. In some embodiments, R 1b is 1 to 3 -C(=O)NH 2 C substituted with a group 1 -C 6 In some embodiments, R 1b is 1 or 2 -C(=O)NH 2 C substituted with a group 1 -C 3 Alkyl, e.g., -CH 2 C(=O)NH 2 or -CH 2 CH 2 C(=O)NH 2 In some embodiments, R 1b is C substituted with 1 to 3 substituents selected from halo and amino 6 -C 10 In some embodiments, R 1b is C substituted with 1-3 halo, such as fluoro, chloro, or bromo. 6 -C 10 In some embodiments, R 1b is C substituted with 1 to 3 amino groups 6 -C 10 In some embodiments, R 1a and R 1b are each independently H, methyl, fluoro, 2-methylbutyl, -CH 2 F, methoxy, -CH 2 CO 2 H, -CH 2 C(=O)NH 2 , benzyl, or 4-aminobenzyl. 1a and R1b are each independently H or C 1 -C 3 In some embodiments, R 1a is methyl, R 1b is H. In some embodiments, R 1a and R 1b are each H. In some embodiments, R 1a and R 1b One of the groups is H and the other is C, such as methyl. 1 -C 3 It is an alkyl.

[0111] In some embodiments, R 2 is H, oxo, or thioxo. In some embodiments, R 2 is H. In some embodiments, R 2 In some embodiments, R 2 is thioxo.

[0112] In some embodiments, R 3 is C 3 -C 6 Alkyl, C 3 -C 6 Alkenyl, C 3 -C 6 Alkynyl, C 3 -C 12 Cycloalkyl, C 3 -C 6 Cycloalkylalkyl, C 6 -C 10 arylalkyl, 5-10 membered heteroarylalkyl, or 5-10 membered heterocyclylalkyl, where the 5-10 membered heteroarylalkyl or 5-10 membered heterocyclylalkyl contains 1-3 heteroatoms selected from nitrogen and oxygen. 3 is propyl, butyl, pentyl, or hexyl, etc. 3 -C 6 In some embodiments, R 3 is C4 -C 6 In some embodiments, R 3 is C 3 -C 6 In some embodiments, R 3 is C 4 -C 6 In some embodiments, R 3 is C 3 -C 6 In some embodiments, R 3 is C 4 -C 6 In some embodiments, R 3 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, etc. 3 -C 12 In some embodiments, R 3 is C 3 -C 6 In some embodiments, R 3 is -(CH 2 ) 1-3 (C 3 -C 6 Cycloalkyl) etc. 3 -C 6 In some embodiments, R 3 C such as benzyl 6 -C 10 In some embodiments, R 3 is -(CH 2 ) 1-3 (5-10 membered heteroaryl) or -(CH 2 ) 1-3 (5-6 membered heteroaryl). In some embodiments, the 5-10 membered heteroarylalkyl contains 1-2 nitrogen atoms. In some embodiments, R 3 is -(CH 2 ) 1-3 (5-10 membered heterocyclyl) or -(CH 2 )1-2 (5- to 6-membered heterocyclyl). In some embodiments, the 5- to 10-membered heterocyclylalkyl contains 1 to 2 nitrogen atoms.

[0113] In some embodiments, R 3 is halo and -C(=O)NH 2 C optionally substituted with 1 to 3 substituents selected from 3 -C 6 Alkyl, C 2 -C 6 Alkenyl, or C 3 -C 6 In some embodiments, R 3 Halo, C 1 -C 3 Alkoxy, hydroxy, -NH 2 , -SO 2 (C 1 -C 3 alkyl), and -C(=O)NH 2 C optionally substituted with 1 to 3 substituents selected from 2 -C 6 Alkyl;C 2 -C 6 Alkenyl; C 3 -C 6 Cycloalkylalkyl; 5-6 membered heteroarylalkyl; 5-6 membered heterocyclylalkyl; or C 6 In some embodiments, R 3 is C 1 -C 3 Alkoxy, hydroxy, -NH 2 , and -SO 2 (C 1 -C 3 C substituted by 1 to 3 substituents selected from alkyl 2 In some embodiments, R 3 is: [ka] In some embodiments, R 3 is: [ka] In some embodiments, R 3 is 2-methylbutyl.

[0114] In some embodiments, R 4 is C 6 -C 10 aryl, 5-10 membered heteroaryl, or 5-10 membered heterocyclyl, where the 5-10 membered heteroaryl or 5-10 membered heterocyclyl contains 1-3 heteroatoms selected from nitrogen and oxygen. 4 C such as phenyl 6 -C 10 In some embodiments, R 4 is a 5-10 membered heteroaryl containing 1-2 nitrogen atoms. In some embodiments, R 4 is 5-10 membered heterocyclyl. In some embodiments, R 4 is a 5-9 membered heterocyclyl containing 1-2 nitrogen atoms. 4 is a 5-9 membered heterocyclyl containing 1-2 oxygen atoms. 4 is a 5-9 membered heterocyclyl containing one nitrogen atom and one oxygen atom.

[0115] In some embodiments, R 4 is halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 C optionally substituted with 1 to 3 substituents selected from haloalkoxy 6 -C 10 In some embodiments, R 4 -CF 3 , -OCHF 2, -OH, fluoro, and chloro. 4 is: [ka] In some embodiments, R 4 is: [ka]

[0116] In some embodiments, R 4 is halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 haloalkoxy. 4 is halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 In some embodiments, R is pyridyl or indolyl optionally substituted with 1 to 3 substituents selected from haloalkoxy. 4 teeth, [ka] In some embodiments, R 4 is halo, hydroxyl, C 1 -C 6 Haloalkyl and C 1 -C 6 In some embodiments, R is pyridyl substituted with 1 to 3 substituents selected from haloalkoxy. 4 teeth, [ka] In some embodiments, R 4 is halo, hydroxyl, C1 -C 6 Haloalkyl and C 1 -C 6 haloalkoxy. In some embodiments, R 4 is indolinyl. [ka]

[0117] In some embodiments, -LR 4 -CH 2 (phenyl) or -C(O)(phenyl), where phenyl is C 1 -C 3 Haloalkyl, C 1 -C 3 Substituted by 1 to 3 substituents selected from haloalkoxy, halo, and hydroxy. In some embodiments, -LR 4 -CH 2 (pyridyl) or -C(O)(pyridyl), where pyridyl is C 1 -C 3 Haloalkyl, C 1 -C 3 Substituted by 1 to 3 substituents selected from haloalkoxy, halo, and hydroxy. In some embodiments, -LR 4 is: [ka]

[0118] In some embodiments, each R 5 is independently 1 -C 6 In some embodiments, R is an alkyl, oxo, or halo. 5 is C, such as methyl, ethyl, or propyl. 1 -C 6 In some embodiments, R 5In some embodiments, R 5 is halo, such as fluoro, chloro, or bromo. In some embodiments, R 5 is oxo or halo. In some embodiments, R 5 is oxo or fluoro.

[0119] In some embodiments, R 6 , H, C 1 -C 6 In some embodiments, R 6 is H. In some embodiments, R 6 is C, such as methyl, ethyl, or propyl. 1 -C 6 In some embodiments, R 6 is oxo.

[0120] In some embodiments, R 7 is H or oxo. In some embodiments, R 7 is H. In some embodiments, R 7 is oxo.

[0121] In some embodiments, m is 1. In other embodiments, m is 2.

[0122] In some embodiments, n is 0. In other embodiments, n is an integer from 1 to 3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0123] In any embodiment of formula (I) or any variation thereof, each C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 3 -C12 Cycloalkyl, C 3 -C 12 Cycloalkylalkyl, C 6 -C 10 Aryl, C 6 -C 10 Arylalkyl, 5-10 membered heteroaryl, 5-10 membered heteroarylalkyl, 5-10 membered heterocyclyl, and 5-10 membered heterocyclylalkyl are hydroxyl, halo (such as fluoro, chloro, or bromo), amino, C 1 -C 6 Haloalkyl (-CF 3 or -CHF 2 etc.), C 1 -C 6 Alkoxy (such as methoxy or ethoxy), C 1 -C 6 Haloalkoxy (-OCHF 2 Or -OCF 3 etc.), and -(C=O)NH 2 is optionally substituted with 1 to 3 substituents selected from

[0124] In some embodiments, the compound of Formula (I) is a compound of Formula (II), (IIa), (IIb), (IIc), (IId), or (IIe): [ka] or a pharma- ceutically acceptable salt thereof, 1a , R 1b , R 3 , R 4 , R 5 , R 6 , R 7and n are as described for Formula (I). In some embodiments, the compound is represented by Formula (II) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by Formula (IIa) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by Formula (IIb) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by Formula (IIc) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by Formula (IId) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by Formula (IIe) or a pharma- ceutically acceptable salt thereof.

[0125] In some embodiments, the compound of Formula (I) is a compound of Formula (IIIa), (IIIb), (IIIc), or (IIId): [ka] or a pharma- ceutically acceptable salt thereof, wherein R 1a , R 1b , R 3 , R 5 , R 6 and n are as described for formula (I), and R is as described for formula (I), hydroxyl, halo, amino, C 1 -C 6 Haloalkyl, C 1 -C 6 In some embodiments, the compound is represented by formula (IIIa) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIIb) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIIc) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIId) or a pharma- ceutically acceptable salt thereof.

[0126] In some embodiments, the compound of Formula (I) is a compound of Formula (IVa), (IVb), (IVc), or (IVd): [ka] or a pharma- ceutically acceptable salt thereof, wherein R 5 and n are as described for formula (I), and R is as described for formula (I), including hydroxyl, halo, amino, C 1 -C 6 Haloalkyl, C 1 -C 6 In some embodiments, the compound is represented by formula (IVa) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IVb) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IVc) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IVd) or a pharma- ceutically acceptable salt thereof.

[0127] In some embodiments, the compound of formula (I) is a compound of formula (V): [ka] or a pharma- ceutically acceptable salt thereof, 1a , R 1b , R 3 , and R 4 is as described for formula (I). In some embodiments, L is -C(=O)- or -CH 2 - and R 1a and R 1b are independently H or -CO 2 C optionally substituted with H 1 -C 3 is alkyl, R 3 is C 4 -C 5 Alkyl, C 4 -C 5 Alkenyl, or C 3 -C 5 Cycloalkyl-substituted C 1 -C 3is alkyl, R 4 -CF 3 , -OCHF 2 , -OH, fluoro, and chloro. 1a and R 1b One of the groups is H and the other is C, such as methyl. 1 -C 3 It is an alkyl.

[0128] In the description herein, it is understood that any description, variation, embodiment, or aspect of a moiety can be combined with any description, variation, embodiment, or aspect of any other moiety, just as if each and every combination of descriptions were specifically and individually recited. For example, any description, variation, embodiment, or aspect provided herein with respect to L in formula (I) can be combined with R 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , R 7and n may be combined with any description, variation, embodiment, or aspect thereof as if each and every combination were specifically and individually listed. It is understood that all descriptions, variations, embodiments, or aspects of formula (I), where applicable, also apply and are similarly described to other formulas detailed herein as if each and every description, variation, embodiment, or aspect of all formulas were individually and independently listed. For example, any statement, variation, embodiment, or aspect with respect to formula (I) equally applies and is equally described, where applicable, to any of the formulas detailed herein, such as formulas (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIa), (IIIb), (IIIc), (IIId), (IVa), (IVb), (IVc), (IVd), and (V), as if each and every statement, variation, embodiment, or aspect with respect to all formulas was individually and independently listed.

[0129] In some embodiments, provided is a compound selected from the compounds of Table 1, or a pharma- ceutically acceptable salt thereof. Although some compounds described in this disclosure, including those in Table 1, are depicted as specific stereoisomeric and / or non-stereochemical forms, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomeric or other forms, of any of the compounds of this disclosure, including those in Table 1, are also described herein. TIFF2024546600000024.tif98165TIFF2024546600000025.tif233165TIFF2024546600000026.tif219165TIFF2024546600000027.tif227165TIFF2024546600000028.tif227165TIFF2024546600000029.tif233165TIFF2024546600000030.tif229165TIFF2024546600000031.tif110165, or a pharma- ceutically acceptable salt thereof.

[0130] In some embodiments, the compound of Formula (I) is not compound 3a, 3b, 9, 10, 13, 15, 16, 18, 21, 23-29, 31-41, 43-48, 50, 52, or 54.

[0131] In some embodiments, provided is a compound selected from the compounds of Table 1A, or a pharma- ceutically acceptable salt thereof. Although some compounds described in this disclosure, including those in Table 1A, are depicted as specific stereoisomeric and / or non-stereochemical forms, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomeric or other forms, of any of the compounds of this disclosure, including those in Table 1A, are also described herein. TIFF2024546600000032.tif60165TIFF2024546600000033.tif233165TIFF2024546600000034.tif227165 or a pharma- ceutically acceptable salt thereof.

[0132] In describing the present invention, it is understood that combinations of substituents and / or variables of the depicted formulae are permissible only if such combinations result in stable compounds.

[0133] Additionally, all compounds of formula (I) that exist in free base or free acid form can be converted to their pharma- ceutically acceptable salts by treatment with an appropriate inorganic or organic base or acid by methods known to those skilled in the art. Salts of compounds of formula (I) can be converted to their free base or free acid forms by standard techniques.

[0134] Synthesis method The compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer, can be prepared by using organic chemical synthesis methods known in the art. In general, the starting components can be obtained from commercial sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, or can be synthesized according to materials known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)), or can be prepared as described herein.

[0135] General reaction scheme 1. [ka] General Reaction Scheme 1 provides an exemplary method for preparing compounds of formula (I). 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , L, and n are as defined herein. X is a reactive moiety (e.g., halo) selected to facilitate the desired reaction. P 1 and P 2 is a suitable protecting group. L' is L'-R 4 The compound of structure A1 is either commercially available or prepared according to methods known in the art. A1 and A2 are coupled under suitable coupling conditions (e.g., T 3 P and base) to give the product A3 by the coupling reaction between A1 and A2. A3 and A4 are then reacted under suitable coupling conditions (e.g., T 3P and base) to give compound A5. Compound A5 is then cyclized (e.g., using formic acid) and deprotected (e.g., using piperidine) to give compound A6. Compound A6 is then reacted with compound A7 to give the final compound of formula (I) shown.

[0136] General reaction scheme 2. [ka] An alternative synthesis of compounds of formula (I) is shown in General Reaction Scheme 2. 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , L, and n are as defined herein. 2 is a suitable protecting group. Each X is a reactive moiety selected to facilitate the desired reaction (e.g., halo). L' is a group selected from the L'-R 4 Intermediate A5 is selected to yield the desired L moiety from the reaction between R 3 A removable protecting group P 3 (e.g., para-methoxybenzyl) to give intermediate A8. A8 is then cyclized (e.g., using formic acid) and deprotected (e.g., using piperidine) to give compound A9. Compound A9 is then reacted with A7 to give compound A10. Compound A10 is then deprotected (e.g., with ceric ammonium nitrate) to give compound A11. Compound A11 is then reacted with A12 to give the final compound of formula (I).

[0137] General reaction scheme 3. [ka] A method related to that shown in General Reaction Scheme 2 is shown in General Reaction Scheme 3. In this method, the two amine nitrogen atoms of the bicyclic core are deprotected to give compound A10, which is then reacted with A7 to give compound A11, which is then reacted with A12 to give the final compound of formula (I).

[0138] It should be noted that various alternative strategies for the preparation of compounds of formula (I) are available to one skilled in the art. For example, other compounds of formula (I) can be prepared according to similar methods using appropriate starting materials.

[0139] It will be appreciated by those skilled in the art that in the process of preparing the compounds described herein, it may be necessary to protect the functional groups of intermediate compounds by suitable protecting groups. Such functional groups may include hydroxy, amino, and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino and amidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl, or arylalkyl esters. Protecting groups are optionally added or removed according to standard techniques known to those skilled in the art or described herein. The use of protecting groups is described in detail in Green, TW and PGM Hutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As will be appreciated by those skilled in the art, the protecting group may be a polymer resin such as a Wang resin, a Rink resin or a 2-chlorotrityl-chloride resin.

[0140] Pharmaceutical Compositions and Formulations In a further aspect, provided herein is a pharmaceutical composition. The pharmaceutical composition comprises any one (or more) of the aforementioned compounds and a pharma- ceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In still other embodiments, the pharmaceutical composition comprises a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and an additional therapeutic agent. Non-limiting examples of such therapeutic agents are described herein below.

[0141] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, otic, nasal, and topical administration. Additionally, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, and intramedullary injections, as well as intrathecal, intravenous, intraperitoneal, intralymphatic, and intranasal injections.

[0142] In certain embodiments, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered locally rather than systemically, for example, by injecting the compound directly into an organ, often in a depot preparation or sustained release formulation. In certain embodiments, a long-acting formulation is administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. In yet other embodiments, the drug is delivered in a targeted drug delivery system, for example, a liposome coated with an organ-specific antibody. In such embodiments, the liposome is targeted to the organ and selectively taken up by the organ. In still other embodiments, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is provided in the form of a fast release formulation, a sustained release formulation, or an intermediate release formulation. In still other embodiments, the compounds described herein are administered locally.

[0143] The compounds of formula (I), or pharma- ceutically acceptable salts, isotopic forms, or stereoisomers thereof, are effective over a wide dosage range. For example, in the treatment of adult humans, dosages of 0.01-1000 mg per day, 0.5-100 mg per day, 1-50 mg per day, and 5-40 mg per day are examples of dosages used in some embodiments. An exemplary dosage is 10-30 mg per day. The exact dosage may depend on the route of administration, the form in which the compound is administered, the subject to be treated, the weight of the subject to be treated, and the preference and experience of the attending physician.

[0144] In some embodiments, the compound of formula (I) or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is administered in a single dose.Typically, such administration is by injection, for example, intravenous injection, to rapidly introduce the agent.However, other routes are also used as appropriate.A single dose of the compound of the present disclosure can also be used to treat acute conditions (e.g., traumatic brain injury).

[0145] In some embodiments, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered in multiple doses. In some embodiments, dosing is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, dosing is about once a month, once every two weeks, once a week, or once every other day. In another embodiment, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and another therapeutic agent are administered together about once a day to about six times a day. In another embodiment, administration of the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and the therapeutic agent continues for less than about 7 days. In yet another embodiment, administration continues for more than about 6, 10, 14, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous administration is possible and is maintained as long as necessary.

[0146] Administration of the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer can be continued as long as necessary. In some embodiments, the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is administered continuously, for example, chronically, for the treatment of chronic effects (e.g., dementia).

[0147] In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt, isotopic form, or stereoisomer is administered in multiple doses.It is known in the art that due to the subject-to-subject variability in the pharmacokinetics of the compound, optimal treatment requires individualized administration regimen.The dosage of the compound can be found by routine experimentation in light of the present disclosure.

[0148] In some embodiments, the compound of formula (I) or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is formulated into a pharmaceutical composition.In a specific embodiment, the pharmaceutical composition is formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate the processing of active compounds into pharma- ceutically usable preparations.The appropriate formulation depends on the route of administration selected. Any pharma- ceutically acceptable technology, carrier, and excipient may be suitably used to formulate the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0149] Provided herein is a pharmaceutical composition comprising a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and a pharma- ceutically acceptable diluent(s), excipient(s), or carrier(s).Also provided herein is a method for administering a pharmaceutical composition comprising a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and a pharma- ceutically acceptable diluent(s), excipient(s), or carrier(s).

[0150] In certain embodiments, the compound is administered as a pharmaceutical composition in which the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer, is mixed with other therapeutic agents as a combination therapy.All combinations of active ingredients described in the method section below and throughout this disclosure are encompassed herein.In specific embodiments, the pharmaceutical composition comprises one or more compounds of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer.

[0151] A pharmaceutical composition, as used herein, refers to a mixture of a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments, to practice the methods of treatment or use provided herein, a therapeutically effective amount of a compound of formula (I) provided herein, or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered in a pharmaceutical composition to a mammal having a disease, disorder, or medical condition to be treated. In a specific embodiment, the mammal is a human. In certain embodiments, the therapeutically effective amount will vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds described herein are used alone or as components of a mixture in combination with one or more therapeutic agents.

[0152] In one embodiment, one or more compounds of formula (I), or pharma- ceutically acceptable salts, isotopic forms, or stereoisomers thereof, are formulated in an aqueous solution. In a specific embodiment, the aqueous solution is selected from physiologically compatible buffers, such as, by way of example only, Hank's solution, Ringer's solution, or saline buffer. In another embodiment, one or more compounds of formula (I), or pharma- ceutically acceptable salts, isotopic forms, or stereoisomers thereof, are formulated for transmucosal administration. In a specific embodiment, the transmucosal formulation comprises a penetrant appropriate for the barrier to be permeated (e.g., the blood-brain barrier). In yet another embodiment, where the compounds described herein are formulated for other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In a specific embodiment, such solutions comprise physiologically compatible buffers and / or excipients.

[0153] In another embodiment, the compound of formula (I) or its pharmaceutically acceptable salt, isotopic form, or stereoisomer is formulated for oral administration.The compound is formulated by combining the active compound with, for example, a pharmaceutically acceptable carrier or excipient.In various embodiments, the compound of formula (I) or its pharmaceutically acceptable salt, isotopic form, or stereoisomer is formulated into oral dosage forms, including, but not limited to, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, etc.

[0154] In certain embodiments, pharmaceutical preparations for oral use are obtained by mixing one or more solid excipients with the compound of formula (I) or one or more of its pharma- ceutically acceptable salts, isotopic forms, or stereoisomers, optionally grinding the mixture obtained, processing the mixture of granules, and optionally adding suitable auxiliaries to obtain tablets or dragee cores.Suitable excipients are in particular sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, etc.; or other fillers, such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate.In a specific embodiment, disintegrants are optionally added. Disintegrants include, by way of example only, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0155] In one embodiment, dosage forms such as dragee cores and tablets are provided with one or more suitable coatings. In a specific embodiment, concentrated sugar solutions are used to coat the dosage forms. The sugar solutions optionally contain additional components such as, by way of example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes and / or pigments are also optionally added to the coating for identification purposes. Furthermore, dyes and / or pigments are optionally used to characterize different combinations of active compound doses.

[0156] In certain embodiments, a therapeutically effective amount of a compound of formula (I), or at least one of its pharma- ceutically acceptable salts, isotopic forms, or stereoisomers, is formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin, and sealed soft capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. In specific embodiments, the push-fit capsules contain the active ingredient mixed with one or more fillers. The fillers include, by way of example only, binders, such as lactose, starch, and / or lubricants, such as talc or magnesium stearate, and optionally stabilizers. In other embodiments, the soft capsules contain one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

[0157] In other embodiments, a therapeutically effective amount of at least one of the compounds of formula (I) described herein, or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is formulated for buccal or sublingual administration. Formulations suitable for buccal or sublingual administration include, by way of example only, tablets, lozenges, or gels. In still other embodiments, the compounds of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof is formulated for parenteral injection, including formulations suitable for bolus injection or continuous infusion. In specific embodiments, the injectable formulation is provided in a unit dosage form (e.g., ampoules) or in multi-dose containers. Preservatives are optionally added to the injectable formulation. In still other embodiments, the pharmaceutical composition is formulated in a form suitable for parenteral injection, as a sterile suspension, solution, or emulsion in an oily or aqueous vehicle. Parenteral injection formulations optionally contain formulations such as suspending agents, stabilizing agents, and / or dispersing agents. In a specific embodiment, pharmaceutical preparations for parenteral administration include aqueous solutions of active compounds in water-soluble form. In additional embodiments, suspensions of one or more active compounds (e.g., compounds of formula (I), or pharma- ceutically acceptable salts, isotopic forms, or stereoisomers thereof) are prepared as appropriate oily injection suspensions. Lipophilic solvents or vehicles suitable for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. In certain specific embodiments, aqueous injection suspensions contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension contains suitable stabilizers or agents that increase the solubility of the compounds, allowing for the preparation of highly concentrated solutions. Alternatively, in other embodiments, the active ingredient is in powder form for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

[0158] In yet another embodiment, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered topically. The compound is formulated into various topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, or ointments. Such pharmaceutical compositions optionally contain solubilizers, stabilizers, tonicity enhancers, buffers, and preservatives.

[0159] In yet other embodiments, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is formulated for transdermal administration. In specific embodiments, transdermal formulations employ transdermal delivery devices and transdermal delivery patches, which may be lipophilic emulsions or buffered aqueous solutions dissolved and / or dispersed in a polymer or adhesive. In various embodiments, such patches may be configured for continuous pulsed or on-demand delivery of the drug. In further embodiments, transdermal delivery of the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is accomplished by means of iontophoretic patches or the like. In certain embodiments, the transdermal patch provides controlled delivery of the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof. In specific embodiments, the rate of absorption is slowed by using rate-controlling membranes or by trapping the compound within a polymer matrix or gel. In alternative embodiments, absorption enhancers are used to enhance absorption. Absorption enhancers or carriers include pharma- ceutically acceptable absorbent solvents that aid in passage through the skin. For example, in one embodiment, the transdermal device is in the form of a bandage comprising a support member, a reservoir containing the compound, optionally with a carrier, an optional rate-controlling barrier that delivers the compound to the host's skin at a controlled, pre-determined rate over an extended period of time, and a means for securing the device to the skin.

[0160] In other embodiments, the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer, is formulated for administration by inhalation.Various forms suitable for administration by inhalation include, but are not limited to, aerosol, mist, or powder.The pharmaceutical composition of the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer, is conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer by using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas).In a specific embodiment, the dosage unit of the pressurized aerosol is determined by providing a valve that delivers a metered amount.In certain embodiments, by way of example only, gelatin capsules and cartridges for use in inhalers or insufflators are formulated to contain a powder mixture of the compound and a suitable powder base, such as lactose or starch.

[0161] In yet other embodiments, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, is formulated into rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides, and synthetic polymers such as polyvinylpyrrolidone, PEG, and the like. In suppository forms of the composition, a low melting wax, such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with cocoa butter, is first melted.

[0162] In certain embodiments, pharmaceutical compositions are formulated in any conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate the processing of active compounds into medicament-usable preparations.The appropriate formulation depends on the route of administration selected.Any pharmaceutically acceptable technology, carrier, and excipient are optionally used suitably.The pharmaceutical composition containing the compound of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer, is prepared in a conventional manner, such as, for example, conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, encapsulating, or compressing processes.

[0163] A pharmaceutical composition comprises at least one pharma- ceutically acceptable carrier, diluent or excipient and at least one compound of formula (I) as described herein, or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, as an active ingredient. The active ingredient is in the form of a free acid or free base, or in the form of a pharma- ceutically acceptable salt. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), and active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein. Furthermore, the compounds of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, include unsolvated as well as solvated forms with pharma- ceutically acceptable solvents such as water, ethanol, and the like. Solvates of the compounds presented herein are also considered to be disclosed herein. In addition, pharmaceutical compositions optionally contain other medicinal or pharmaceutical agents, carriers, adjuvants, such as preservatives, stabilizing agents, wetting agents or emulsifying agents, solution promoters, salts for regulating osmotic pressure, buffers, and / or other therapeutically valuable substances.

[0164] Methods for preparing compositions containing a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, include formulating the compound with one or more pharma- ceutically acceptable inert excipients or carriers to form a solid, semi-solid, or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compound is dissolved, emulsions containing the compound, or solutions containing liposomes, micelles, or nanoparticles containing a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. The forms of the pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to use, or as emulsions. These compositions also optionally contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffers, and the like.

[0165] In some embodiments, the pharmaceutical composition comprising at least one compound of formula (I) or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is illustratively in the form of a liquid, in which the agent is in solution, in suspension, or both.Typically, when the composition is administered as a solution or suspension, a first portion of the agent is in solution, and a second portion of the agent is in particulate form that is in suspension in a liquid matrix.In some embodiments, the liquid composition comprises a gel formulation.In other embodiments, the liquid composition is aqueous.

[0166] In certain embodiments, useful aqueous suspensions contain one or more polymers, such as suspending agents. Useful polymers include water-soluble polymers, such as cellulosic polymers, e.g., hydroxypropylmethylcellulose, and water-insoluble polymers, such as water-crosslinkable carboxyl-containing polymers. Certain pharmaceutical compositions described herein include mucoadhesive polymers selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0167] Useful pharmaceutical compositions also optionally contain a solubilizer that aids in the dissolution of the compound of formula (I), or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer. "Solubilizer" generally includes agents that result in a micellar or true solution form of the agent. Certain acceptable non-ionic surfactants, such as polysorbate 80, are useful as solubilizers, as are ophthalmologically acceptable glycols, polyglycols, such as polyethylene glycol 400, and glycol ethers.

[0168] In addition, useful pharmaceutical compositions optionally include one or more pH adjusting or buffering agents, for example, acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane; and buffers such as citric acid / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition in an acceptable range.

[0169] Additionally, useful compositions also optionally contain one or more salts in an amount necessary to bring the osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions, with suitable salts including sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0170] Other useful pharmaceutical compositions optionally contain one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merphen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0171] Still other useful compositions contain one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkyl phenyl ethers, such as octoxynol 10, octoxynol 40.

[0172] Still other useful compositions optionally contain one or more antioxidants to enhance chemical stability. Suitable antioxidants include, by way of example only, ascorbic acid and sodium disulfite.

[0173] In certain embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multi-dose reclosable containers are used, in which case it is typical to include a preservative in the composition.

[0174] In alternative embodiments, other delivery systems for hydrophobic pharmaceutical compounds are employed. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also employed. In additional embodiments, the compound of formula (I), or its pharma-ceutically acceptable salt, isotopic form, or stereoisomer, is delivered using a sustained release system, such as a semipermeable matrix of hydrophobic solid polymers containing a therapeutic agent. A variety of sustained release materials are useful herein. In some embodiments, sustained release capsules release the compound for several weeks up to 100 days or more. Depending on the chemical nature and biological stability of the therapeutic agent, additional strategies for protein stability are employed.

[0175] In certain embodiments, the formulations described herein include one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizing agents. Examples of such stabilizers include, but are not limited to, (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% w / v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / v ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrins, (l) pentosan polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc, or (n) combinations thereof.

[0176] In some embodiments, the concentration of the compound of formula (I) provided in the pharmaceutical composition of the present disclosure is 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.08%, 0.09 ... less than 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v or v / v.

[0177] In some embodiments, the concentration of the compound of formula (I) provided in the pharmaceutical composition of the present disclosure is 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25%, 19%, 18.75%, 18.50%, 18.25%, 18%, 17.75%, 17.50%, 17.25%, 17%, 16.75%, 16.50%, 16.25%, 16%, 15.75%, 15.50%, %,15.25%,15%,14.75%,14.50%,14.25%,14%,13.75%,13.50%,13.25%,13%,12.75%,12.50%,12.25%,12%,11.75%,11.50%,11.25%,11%,10.75%,10.50%,10.25%,10%,9.75%,9.50%,9.25%,9%,8.75%,8.50%,8.25%,8%,7.75%,7 .50%,7.25%,7%,6.75%,6.50%,6.25%,6%,5.75%,5.50%,5.25%,5%,4.75%,4.50%,4.25%,4%,3.75%,3.50%,3.25%,3%,2.75%,2.50%,2.25%,2%,1.75%,1.50%,125%,1%,0.5%,0.4%,0.3%,0.2%,0.1%,0.09%,0.08%,0.07%,0.06% , 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or greater than 0.0001% w / w, w / v, or v / v.

[0178] In some embodiments, the concentration of the compound of Formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, provided in the pharmaceutical composition is from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02% to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, from about 0.07% to about 100%, from about 0.08% to about 120%, from about 0.09% to about 23%, from about 0.10% to about 25%, from about 0.11% to about 26%, from about 0.12% to about 27%, from about 0.13% to about 25%, from about 0.14% to about 25%, from about 0.15% to about 25%, from about 0.16% to about 25%, from about 0.17% to about 25%, from about 0.18% to about 25%, from about 0.19% to about 25%, from about 0.20% to about 25%, from about 0.25% to about 25%, from about 0.26% to about 25%, from about 0.27% to about 25%, from about 0.28% to about 25%, from about 0.29% to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, from about 0.29% to about 25%, from about 0.30% to about 25%, about 0.08% to about 23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, or about 1% to about 10% w / w, w / v or v / v.

[0179] In some embodiments, the concentration of the compound of Formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, provided in the pharmaceutical composition is in the range of about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, or about 0.1% to about 0.9% w / w, w / v, or v / v.

[0180] In some embodiments, the amount of the compound of Formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, provided in the pharmaceutical composition is 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35g, 0.3g, 0.25g, 0.2g, 0.15g, 0.1g, 0.09g, 0.08g, 0.07g, 0.06g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0.007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.0008g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g, or 0.0001g.

[0181] In some embodiments, the amount of the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, provided in the pharmaceutical composition of the present disclosure is 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, or 100 g. , 0.001g, 0.0015g, 0.002g, 0.0025g, 0.003g, 0.0035g, 0.004g, 0.0045g, 0.005g, 0.00 55g, 0.006g, 0.0065g, 0.007g, 0.0075g, 0.008g, 0.0085g, 0.009g, 0.0095g, 0.01g, 0.0 15g, 0.02g, 0.025g, 0.03g, 0.035g, 0.04g, 0.045g, 0.05g, 0.055g, 0.06g, 0.065g, 0.0 7g, 0.075g, 0.08g, 0.085g, 0.09g, 0.095g, 0.1g, 0.15g, 0.2g, 0.25g, 0.3g, 0.35g, 0.4g , 0.45g, 0.5g, 0.55g, 0.6g, 0.65g, 0.7g, 0.75g, 0.8g, 0.85g, 0.9g, 0.95g, 1g, 1.5g, 2g, 2.5g, 3g, 3.5g, 4g, 4.5g, 5g, 5.5g, 6g, 6.5g, 7g, 7.5g, 8g, 8.5g, 9g, 9.5g, or greater than 10g.

[0182] In some embodiments, the amount of the compound of Formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, provided in the pharmaceutical composition is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

[0183] Kits / Products Kits and articles of manufacture are also provided for use in the therapeutic applications described herein. In some embodiments, such kits include a case, package, or container that is partitioned to receive one or more containers, such as vials, tubes, etc., each of the container(s) containing one of the individual elements used in the methods described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers are formed from a variety of materials, such as glass or plastic.

[0184] The products provided herein contain packaging materials. Packaging materials used for packaging pharmaceutical products include, for example, those found in U.S. Patent Nos. 5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for the selected formulation and intended mode of administration and treatment. For example, the container(s) contain one or more compounds described herein, optionally in a composition or in combination with another agent disclosed herein. The container(s) optionally have a sterile access port (e.g., the container is an intravenous solution bag or vial with a stopper that can be pierced by a hypodermic needle). Such kits optionally contain the compound with identification information or labels or instructions for use in the methods described herein.

[0185] For example, the kit typically includes one or more additional containers each containing one or more of various materials (such as reagents (optionally concentrated forms) and / or devices) that are desirable from a commercial and user standpoint for use of the compound of formula (I), or a pharma-ceutically acceptable salt, isotopic form, or stereoisomer thereof. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, carriers, packages, containers, vials, and / or tube labels with contents and / or instructions for use, as well as package inserts with instructions for use. A set of instructions is also typically included. The label is optionally on or associated with the container. For example, the label is on the container when letters, numbers, or other characters forming the label are attached, molded, or etched into the container itself, and the label is associated with the container when the label is present in the receptacle or carrier that holds the container, e.g., as a package insert. In addition, the label is used to indicate that the contents are to be used for a particular therapeutic application. Additionally, the label indicates instructions for use of the contents, such as the methods described herein. In certain embodiments, the pharmaceutical composition is presented in a pack or dispenser device containing one or more unit dosage forms containing the compound provided herein. The pack contains, for example, metal or plastic foil, such as a blister pack. Alternatively, the pack or dispenser device is accompanied by instructions for administration. Alternatively, the pack or dispenser is accompanied by a label attached to the container in a format prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which reflects the agency's approval of the drug form for human or veterinary administration. Such a label is, for example, a label for prescription drugs approved by the U.S. Food and Drug Administration, or an approved product insert. In some embodiments, a composition containing a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, formulated in a compatible pharmaceutical carrier, is prepared, placed in a suitable container, and labeled for the treatment of a specified condition.

[0186] Method of Use / Treatment An embodiment of the present disclosure provides a method for modulating hepatocyte growth factor in a subject in need thereof, comprising administering to the subject an effective amount of a compound disclosed herein (e.g., a compound of Formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof). In some embodiments, the compounds described herein activate hepatocyte growth factor. Modulation (e.g., inhibition or activation) of hepatocyte growth factor can be assessed and demonstrated in a wide variety of ways known in the art. Kits and commercially available assays can be utilized to determine whether and to what extent hepatocyte growth factor is modulated (e.g., inhibited or activated).

[0187] In some embodiments, provided herein is a compound of formula (I), or a pharma- ceutically acceptable salt thereof, for use in modulating hepatocyte growth factor in a subject in need thereof. In some embodiments, provided herein is a compound of formula (I), or a pharma- ceutically acceptable salt thereof, for the manufacture of a medicament for modulating hepatocyte growth factor in a subject in need thereof.

[0188] Applicant has discovered that the compound of formula (I) shows promising activity in relation to certain diseases of interest. Thus, in one aspect, provided herein is a method for regulating hepatocyte growth factor in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma-ceutically acceptable salt, isotopic form, or stereoisomer thereof. In some embodiments, provided herein is a method for activating hepatocyte growth factor in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma-ceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0189] In more specific embodiments, modulating includes treating a disease, condition, or injury.

[0190] In some embodiments, the disease, condition, or injury is mild cognitive impairment, which may be a precursor to dementia due to neurodegenerative disease or may result from other factors, such as liver damage.

[0191] In some embodiments, there is provided a method of treating mild cognitive impairment in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0192] Also provided herein is a method for treating or slowing the progression of mild cognitive impairment in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof.Also provided herein is a method for treating or slowing the progression of dementia in a subject with mild cognitive impairment, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma-ceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0193] In a further aspect, provided herein is a method for preventing cognitive impairment in a subject with mild cognitive impairment, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof. In some embodiments, provided herein is a method for improving cognitive function and / or slowing the progression of cognitive impairment in a subject with mild cognitive impairment, comprising administering to the subject an effective amount of a compound of formula (I), or a pharma-ceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0194] The above method embodiments include administering to a mammal a therapeutically effective amount of a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof. The methods disclosed herein generally relate to the administration of a compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, to treat, protect, or reverse diseases and injuries associated with nerve cells or the nervous system. That is, embodiments of the present disclosure relate to the treatment, prevention, or reversal of neurodegenerative diseases, including the treatment of dementia; repair of traumatic damage; and / or prevention of cognitive dysfunction. Certain embodiments of the present disclosure are directed to the treatment, prevention, or reversal of mild cognitive impairment, including the treatment of dementia; repair of traumatic damage; improving cognitive function; and / or slowing the progression of cognitive dysfunction.

[0195] In some embodiments, the disclosure provides a method of modulating the activity of a protein (e.g., hepatocyte growth factor) in a subject, including but not limited to rodents and mammals (e.g., humans), by administering to the subject an effective amount of a compound of Formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof. In some embodiments, the modulation of hepatocyte growth factor is activation of hepatocyte growth factor. In some embodiments, the percentage of modulation is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some embodiments, the percentage of inhibition is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0196] In some embodiments, the disclosure provides a method of regulating hepatocyte growth factor activity in a cell by contacting the cell with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate the activity of hepatocyte growth factor. In some embodiments, the disclosure provides a method of regulating hepatocyte growth factor activity in a tissue by contacting the tissue with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate the activity of hepatocyte growth factor in the tissue. In some embodiments, the disclosure provides a method of regulating hepatocyte growth factor activity in an organism by contacting the organism with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate the activity of hepatocyte growth factor in the organism. In some embodiments, the disclosure provides a method of modulating hepatocyte growth factor activity in an animal by contacting the animal with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to modulate the activity of hepatocyte growth factor in the animal. In some embodiments, the disclosure provides a method of modulating hepatocyte growth factor activity in a mammal by contacting the mammal with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to modulate the activity of hepatocyte growth factor in the mammal. In some embodiments, the disclosure provides a method of modulating hepatocyte growth factor activity in a human by contacting the human with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to modulate the activity of hepatocyte growth factor in the human. In other embodiments, the disclosure provides a method of treating a disease mediated by hepatocyte growth factor activity in a subject in need of such treatment. In some variations, the modulation of hepatocyte growth factor by a compound of Formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof involves activation of hepatocyte growth factor.

[0197] Other embodiments provide methods for combination therapy in which therapeutic agents known to modulate other pathways or other elements of the same pathway or overlapping target enzymes are used in combination with a compound of formula (I) or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof. In one aspect, such therapy includes, but is not limited to, the combination of one or more compounds of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, with therapeutic agents, therapeutic antibodies, and other forms of therapy that provide a synergistic or additive therapeutic effect.

[0198] Currently, many therapeutic agents are known in the art and can be used in combination with the compounds of formula (I), or their pharma- ceutically acceptable salts, isotopic forms, or stereoisomers. In some embodiments, the therapeutic agent is selected from memantine, cholinesterase inhibitors, antidepressants, anxiolytics, and / or antipsychotics. Some embodiments include the use of therapeutic methods including reminiscence therapy, cognitive activation therapy, reality orientation training, physical activity, and the like.

[0199] Exemplary cholinesterase inhibitors include donepezil, galantamine, and rivastigmine, which help slow the breakdown of brain chemicals involved in memory and judgment. Memantine can help regulate another brain chemical necessary for learning and memory. In certain aspects, memantine can also be used with donepezil as a combination drug for moderate to severe dementia. Antidepressants can include, but are not limited to, selective serotonin reuptake inhibitors (SSRIs). Anti-anxiety medications can include, but are not limited to, lorazepam (Ativan) or oxazepam (Serax). Some embodiments of the methods described herein can include the use or administration of antipsychotics such as aripiprazole (Abilify), haloperidol (Haldol), olanzapine (Zyprexa), and risperidone (Risperdal).

[0200] In some embodiments, the compound of formula (I) or its pharma- ceutically acceptable salt, isotopic form, or stereoisomer is formulated or administered with a liquid or solid tissue barrier, also known as a lubricant. Examples of tissue barriers include, but are not limited to, polysaccharides, polyglycans, Seprafilm, Interceed, and hyaluronic acid.

[0201] In some embodiments, the therapeutic agent administered with a compound of Formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, includes any suitable therapeutic agent usefully delivered by inhalation, such as an analgesic, e.g., codeine, dihydromorphine, ergotamine, fentanyl, or morphine; an angina preparation, e.g., diltiazem; an antiallergic, e.g., cromoglycate, ketotifen, or nedocromil; an antiinfective, e.g., cephalosporin, penicillin, streptomycin, sulfonamide, tetracycline, or pentamidine; an antihistamine, e.g., methapyrilene; an anti-inflammatory, e.g., beclomethasone, flunisolide, budesonide, tipredane, triamcinolone acetonide, or fluticasone; an antitussive, e.g., noscapine; a bronchodilator, e.g., ephedrine, adrenaline, or the like. diuretics, such as amiloride; anticholinergics, such as ipratropium, atropine or oxitropium; hormones, such as cortisone, hydrocortisone or prednisolone; xanthines, such as aminophylline, choline theophyllinate, lysine theophyllinate or theophylline; and therapeutic proteins and peptides, such as insulin or glucagon. It will be apparent to those skilled in the art that therapeutic agents may be used, where appropriate, in the form of a salt (e.g., as an alkali metal or amine salt or an acid addition salt) or as an ester (e.g., a lower alkyl ester) or as a solvate (e.g., a hydrate) to optimize the activity and / or stability of the therapeutic agent.

[0202] Additional therapeutic agents which may be combined with the compounds of Formula (I), or pharma- ceutically acceptable salts, isotopic forms, or stereoisomers thereof, can be found in Goodman and Gilman's "The Pharmacological Basis of Therapeutics" Tenth Edition, edited by Hardman, Limbird, and Gilman, or in the Physician's Desk Reference, both of which are incorporated herein by reference in their entireties.

[0203] The compound of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer, can be used in combination with the therapeutic agents disclosed herein, depending on the condition to be treated. Thus, in some embodiments, one or more compounds of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer, are co-administered with other therapeutic agents as described above. When used in combination therapy, the compound of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer, is administered simultaneously or separately with the second therapeutic agent. This combined administration can include simultaneous administration in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compound of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer, and any of the therapeutic agents described above can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and any of the above therapeutic agents can both be present in separate formulations and administered simultaneously.In another alternative, any of the above therapeutic agents can be administered immediately after the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, or vice versa.In some embodiments of the separate administration protocol, the compound of formula (I), or a pharma- ceutically acceptable salt, isotopic form, or stereoisomer thereof, and any of the above therapeutic agents are administered minutes apart, or hours apart, or days apart.

[0204] The examples and preparations provided below further illustrate and demonstrate the compounds of formula (I), or pharma- ceutically acceptable salts, isotopic forms, or stereoisomers thereof, and methods of preparing such compounds. It is to be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples and preparations. In the following examples, and throughout the specification and claims, molecules with a single stereocenter exist as a racemic mixture unless otherwise noted. Molecules with two or more stereocenters exist as a racemic mixture of diastereomers unless otherwise noted. Single enantiomers / diastereomers can be obtained by methods known to those skilled in the art. EXAMPLES

[0205] The following examples are provided for illustrative purposes: Methods for the preparation of compounds of formula (I), or pharma-ceutically acceptable salts, isotopic forms, or stereoisomers thereof, are provided herein or can be derived by one skilled in the art.

[0206] The examples and preparations provided below further illustrate and demonstrate the compounds of the present disclosure and methods for testing such compounds. It is to be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples.

[0207] The chemical reactions of the examples described can be easily adapted to prepare some other compounds disclosed herein, and alternative methods for preparing the compounds of the present disclosure are considered to be within the scope of the present disclosure. For example, the synthesis of compounds according to the present disclosure that are not exemplified can be carried out by modifications obvious to those skilled in the art, such as by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art other than those described, or by routinely modifying reaction conditions, reagents, and starting materials. Alternatively, it will be recognized that other reactions disclosed herein or known in the art can be applied to the preparation of the compounds of the present disclosure.

[0208] Unless otherwise indicated in the examples below, the compounds are isolated as racemic mixtures.

[0209] In this application the following abbreviations apply: Abbreviation AcOH: acetic acid CAN: Cerium ammonium nitrate DAST: Diethylaminosulfur trifluoride DCM: dichloromethane DIPEA: N,N-diisopropylethylamine DMEM: Dulbecco's modified Eagle's medium DMF: Dimethylformamide DMSO: Dimethyl sulfoxide EMEM: Eagle's minimum essential medium EtOAc: ethyl acetate EtOH: Ethanol FBS: Fetal bovine serum Fmoc: fluorenylmethoxycarbonyl HATU: (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate LC / MS: Liquid chromatography mass spectrometry Me: Methyl MeOH: Methanol PBS: Phosphate-buffered saline Pic-BH 3 :Picoline borane PMB: para-methoxybenzyl ether Prep HPLC: Preparative High Performance Liquid Chromatography rt or RT: room temperature TFA: Trifluoroacetic acid TLC: Thin Layer Chromatography T 3 P: propanephosphonic anhydride.

[0210] Synthesis Example Example S1. Synthesis of (6S)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. The synthetic route for preparing this starting compound is shown in Scheme 1. Scheme 1. [ka] Step 1: Synthesis of (9H-fluoren-9-yl)methyl (2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylcarbamate. To a stirred solution of the compound (S)-2-(((9H-fluoren-9-yl)methoxy)carbonylamino)propanoic acid (5.0 g, 16.07) in dichloromethane (100 mL) was added T 3 P (15.2 mL, 24.1) and DIPEA (5.6 mL, 32.1 mmol) were added at room temperature. The reaction mixture was stirred at room temperature for 15 min, N-(2,2-dimethoxyethyl)-2-methylbutan-1-amine (2.81 g, 32.1 mmol.) was added and stirring was continued at room temperature for 8 h. The reaction was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (100 mL) and extracted with dichloromethane (2 x 100 mL). The combined organic layers were washed with anhydrous Na 2 SO 4 The crude compound was purified by flash column chromatography (100-200 mesh silica gel, eluted with 40% ethyl acetate / petroleum ether) to obtain the pure compound (9H-fluoren-9-yl)methyl (2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylcarbamate (5.2 g, 69.1%) as a gummy compound.

[0211] Step 2: Synthesis of (2S)-2-amino-N-(2,2-dimethoxyethyl)-N-(2-methylbutyl)propenamide. To a stirred solution of (9H-fluoren-9-yl)methyl (2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylcarbamate (34.0 g, 72.6 mmol) in DMF (230 mL) was added 20% piperidine in DMF (70 mL) at 0° C. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, excess DMF (100 mL) was added and then washed with excess n-hexane (3×200 mL). The DMF layer was collected, poured into ice-cold water (1000 mL) and extracted with 10% methanol-dichloromethane (3×500 mL). The combined organic layers were washed with anhydrous Na 2 SO 4 The mixture was dried at rt, filtered and concentrated under reduced pressure to give (2S)-2-amino-N-(2,2-dimethoxyethyl)-N-(2-methylbutyl)propanamide (20.4 g, 68.4%) as a gummy solid.

[0212] Step 3: Synthesis of (9H-fluoren-9-yl)methyl 3-((2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylamino)-3-oxopropylcarbamate. To a stirred solution of 3-(((9H-fluoren-9-yl)methoxy)carbonylamino)propanoic acid (20.2 g, 81.2 mmol) in dichloromethane (500 mL) at room temperature was added T 3 P (80 mL, 121.8 mmol) and DIPEA (28.6 mL, 160.4 mmol) were added and the mixture was stirred for 10 min. To this was added (2S)-2-amino-N-(2,2-dimethoxyethyl)-N-(2-methylbutyl)propanamide (25.5381.2 mmol) and stirring was continued at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with water (500 mL) and the mixture was extracted with dichloromethane (2×500 mL). The combined organic layers were washed with Na 2 SO 4The mixture was dried at 40° C., filtered and concentrated under reduced pressure to give the crude product. The crude compound was purified by flash column chromatography (100-200 mesh silica gel, eluted with 70% ethyl acetate / petroleum ether) to give the pure compound (9H-fluoren-9-yl)methyl 3-((2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylamino)-3-oxopropylcarbamate (21.2 g, 78.6%) as a gummy compound.

[0213] Step 4: Synthesis of (6S)-(9H-fluoren-9-yl)methyl 6-methyl-8-(2-methylbutyl)-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate. To a stirred solution of (9H-fluoren-9-yl)methyl 3-((2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylamino)-3-oxopropylcarbamate (21.0 g, 38.9 mmol) was added formic acid (105 mL). The reaction mixture was stirred at room temperature for 12 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude compound was purified by elution with saturated NaHCO 3 The combined organic layers were washed with brine solution (500 mL) and then extracted with anhydrous Na 2 SO 4 The crude compound was purified by flash column chromatography (100-200 mesh silica gel, eluted with 50% ethyl acetate / petroleum ether) to give the pure compound (6S)-(9H-fluoren-9-yl)methyl 6-methyl-8-(2-methylbutyl)-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate (25 g, 69.0%) as a gum.

[0214] Step 5: Synthesis of (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione. To a stirred solution of (6S)-(9H-fluoren-9-yl)methyl 6-methyl-8-(2-methylbutyl)-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate (14.0 g, 29.4 mmol) in DMF (70 mL) at 0° C. was added 20% piperidine in DMF (30 mL). The reaction mixture was allowed to warm to room temperature and stirred for 2 h. The reaction was monitored by TLC. Upon complete consumption of the starting material, additional DMF was added (50 mL) and the mixture was then washed with excess n-hexane (3×200 mL). The DMF layer was poured into ice-cold water (1000 mL) and extracted with 10% methanol-dichloromethane (3×500 mL). The combined organic layers were washed with anhydrous Na 2 SO 4 The mixture was dried at 40° C., filtered and concentrated under reduced pressure to give the crude desired compound (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (6.25 g, 83.8%) as a solid.

[0215] Example S2. Synthesis of Compound 1a The synthetic route for preparing compound 1a is shown in Scheme 2. Scheme 2. [ka] A solution of 4-(trifluoromethyl)benzoic acid (0.232 g, 0.91 mmol) in dichloromethane (20 mL) was stirred at room temperature. 3P (1.2 mL, 1.37 mmol) and DIPEA (0.42 mL, 1.82 mmol) were added and the mixture was stirred for 15 min. To this was added (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.310 g, 0.91 mmol) and stirring was continued for 8 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was quenched with water (50 mL) and extracted with dichloromethane (2×50 mL). The combined organic layers were washed with Na 2 SO 4 The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then lyophilized to give 1a (0.340 g, 65.3%) as a solid. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: X-Select Phenylhexyl (150×19 mm 5μ); Flow rate: 16 mL / min. MS (ESI) m / z [M+H] + :426.05.

[0216] Example S3. Synthesis of Compound 2a The synthetic route for preparing compound 2a is shown in Scheme 3. Scheme 3. [ka] A solution of 4-(difluoromethoxy)benzoic acid (0.37 g, 1.968 mmol) in dichloromethane (15 mL) at room temperature was added with DIPEA (0.8 ml, 5.904 mmol) and T 3P (2.0 mL, 3.936 mmol) was added. The mixture was stirred for 30 min, then (6S)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.4 g, 1.578 mmol) was added and stirring was continued for 16 h. The reaction progress was monitored by TLC and LC / MS. The reaction mixture was diluted with dichloromethane (100 mL), washed with water (50 mL) and saturated sodium chloride solution (50 mL), then dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by preparative HPLC. Pure fractions were collected and lyophilized to give 2a (380 mg 46%) as a solid. Preparative HPLC conditions: Mobile phase A: 10 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: Kromosil phenyl (150×25 mm 10μ); Flow rate: 25 mL / min. MS(ESI) m / z[M+H] + :424.11.

[0217] Example S4. Synthesis of Compound 3a The synthetic route for preparing compound 3a is shown in Scheme 4. Scheme 4. [ka] To a stirred solution of (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.500 g, 1.97 mmol) in methanol (20 mL) at room temperature was added 4-hydroxybenzaldehyde (0.289 g, 1.97 mmol) and acetic acid (0.23 mL, 3.95 mmol). The reaction mixture was stirred at room temperature for 5 minutes. To this was added picoline borane (0.253 g, 2.37 mmol) and stirring was continued for 48 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and the mixture was extracted with 10% methanol-dichloromethane (3×40 mL). The combined organic layers were washed with Na 2 SO 4The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then lyophilized to give 3a (0.180 g, 46.09%) as a solid. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: Kromosil phenyl (150×25 mm 10μ); Flow rate: 25 mL / min. MS (ESI) m / z [M+H] + :360.11.

[0218] Example S5. Synthesis of Compound 4a. The synthetic route for preparing compound 4a is shown in Scheme 5. Scheme 5. [ka] To a solution of 6-hydroxynicotinic acid (0.340 g 2.446 mmol) in DMF (15 mL) at room temperature was added DIPEA (1.30 mL, 7.338 mmol) and HATU (1.39 g, 3.669 mmol). The resulting reaction mixture was stirred for 30 min, then (6S)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.495 g, 1.956 mmol.) was added and the mixture was stirred for 16 h. The progress of the reaction was monitored by TLC and LC / MS (TLC system: 10% methanol / dichloromethane, Rf: 0.15, detection: UV). The reaction mixture was quenched with cold water (100 mL) and extracted with 10% methanol / dichloromethane (3 x 100 mL). The combined organic layers were washed with cold water (50 mL) and cold brine solution (50 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by preparative HPLC. Pure fractions were collected and lyophilized to give 4a (160 mg, 21.8%) as a solid. Preparative HPLC method: Mobile phase A: 0.01 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: X-Select Phenylhexyl (150×19 mm, 5μ); Flow rate: 15 mL / min. MS (ESI) m / z [M+H] + :375.05.

[0219] Example S6. Synthesis of Compound 5a. The synthetic route for preparing compound 5a is shown in Scheme 6. Scheme 6. [ka] A solution of (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.5 g, 1.97 mmol) and 1-(bromomethyl)-4-(trifluoromethyl)benzene (0.470 g, 1.97 mmol) in DMF (20 mL) was stirred at room temperature. 2 CO 3 (0.546 g, 3.95 mmol) was added and the mixture was stirred for 8 h. The progress of the reaction was monitored by TLC. After completion, the mixture was quenched with water (100 mL) and extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with Na 2 SO 4 The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then lyophilized to give 5a (0.270 g, 63.8%) as a gum. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: Kromosil C 18 (150×25mm 10μ);Flow rate: 25mL / min. MS(ESI)m / z[M+H] + :412.2.

[0220] Example S7. Synthesis of Compound 6a. The synthetic route for preparing compound 6a is shown in Scheme 7. Scheme 7. [ka] A solution of (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.500 g, 1.97 mmol) and 1-(bromomethyl)-4-(difluoromethoxy)benzene (0.466 g, 1.97 mmol) in DMF (20 mL) was stirred at room temperature. 2 CO 3 (0.546 g, 9.95 mmol) was added. The reaction mixture was stirred at room temperature for 18 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with Na 2 SO 4 The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then lyophilized to give 6a (0.178 g, 41.5%) as a semi-solid. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: X-Select C 18 (250×19mm, 5μ); flow rate: 18mL / min. MS(ESI)m / z[M+H] + :410.11.

[0221] Example S8. Synthesis of Compound 7a. The synthetic route for preparing compound 7a is shown in Scheme 8. Scheme 8. [ka] To a stirred solution of compound (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.500 g, 1.97 mmol) in methanol (20 mL) at room temperature, 6-hydroxynicotinaldehyde (0.243 g, 1.97 mmol) and acetic acid (0.25 mL, 3.95 mmol) were added and the mixture was stirred for 5 minutes. To this was added picoline borane (0.318 g, 2.96 mmol) and stirring was continued for 96 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and extracted with 10% methanol-dichloromethane (3×40 mL). The combined organic layers were washed with Na 2 SO 4 The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then lyophilized to give 7a (0.164 g, 42%) as a solid. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate in water; Mobile phase B: acetonitrile; Column: X-BRIDGE C 18 (250×19mm, 5μ); flow rate: 18mL / min. MS(ESI)m / z[M+H] + :361.11.

[0222] Example S9. Synthesis of Compound 8a. The synthetic route for preparing compound 8a is shown in Scheme 9. Scheme 9. [ka] Step 1: Synthesis of (6S)-1-(4-(benzyloxy)benzoyl)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. To a stirred solution of 4-(benzyloxy)benzoic acid (0.360 g, 1.42 mmol) in dichloromethane (20 mL) at room temperature was added T 3P (1.2 mL, 1.7 mmol) and DIPEA (0.55 mL, 2.84 mmol) were added and the mixture was stirred for 15 min. To this was added (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.400 g, 1.42 mmol) and stirring was continued at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (2×50 mL). The combined organic layers were washed with Na 2 SO 4 The crude material was dried at 40° C., filtered and concentrated under reduced pressure to give 0.9 g of crude material. The crude material was analyzed by LC / MS to show 54.59% of the desired product. The crude material was used in the next step without purification.

[0223] Step 2: Synthesis of compound 8a. A solution of (6S)-1-(4-(benzyloxy)benzoyl)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.900 g) in methanol (20 mL) was stirred at room temperature and 10% Pd-C (0.200 g) was added to the solution. 2 The reaction mixture was added under H 2 The mixture was stirred under a balloon for 8 hours at room temperature. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered through Celite and evaporated under reduced pressure to give the crude compound. The crude compound was dissolved in dichloromethane (50 mL) and added with NaHCO 3 The filtrate was washed with aqueous solution (20 mL) and brine solution (20 mL). 2 SO 4 The crude compound was triturated with diethyl ether to give 8a (0.330 g, 82%) as a solid. MS (ESI) m / z [M+H] + :374.11.

[0224] Example S10. Synthesis of Compound 9. The synthetic route for preparing compound 9 is shown in Scheme 10. Scheme 10. [ka] Step 1: Synthesis of (9H-fluoren-9-yl)methyl 2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylcarbamate. To a stirred solution of 2-(((9H-fluoren-9-yl)methoxy)carbonylamino)acetic acid (10 g, 33.6 mmol) in dichloromethane (100 mL) cooled to 0 °C was added DIPEA (11.88 mL, 67.3 mmol), N-(2,2-dimethoxyethyl)butan-2-amine (10.84 g, 67.3 mmol) and T 3 P (53.0 mL, 84.1 mmol) was added and the reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, ice-cold water (100 mL) was added and extracted with ethyl acetate (2×150 mL). The combined organic layer was washed with anhydrous Na 2 SO 4 The crude compound was purified by flash column chromatography (100-200 mesh silica gel) eluting with 20-25% ethyl acetate / petroleum ether to give (9H-fluoren-9-yl)methyl 2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylcarbamate (10.8 g, 72.9%) as a solid.

[0225] Step 2: Synthesis of 2-amino-N-sec-butyl-N-(2,2-dimethoxyethyl)acetamide. To a solution of (9H-fluoren-9-yl)methyl 2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylcarbamate (10.8 g, 24.5 mmol) in DMF (20 mL) cooled to 0° C., piperidine (2.4 mL) was added and the reaction mixture was stirred at room temperature for 2 h. The progress of the reaction was monitored by TLC. When TLC showed complete consumption of the starting material, the reaction mixture was diluted with petroleum ether (2×100 mL), then water was added and the mixture was separated. The aqueous layer was extracted with dichloromethane (2×150 mL). The combined organic layers were washed with anhydrous Na 2 SO 4The mixture was dried at 40° C., filtered and concentrated under reduced pressure to give the desired pure product 2-amino-N-sec-butyl-N-(2,2-dimethoxyethyl)acetamide (3.6 g, 67.2%) as a solid.

[0226] Step 3: Synthesis of (9H-fluoren-9-yl)methyl-3-(2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylamino)-3-oxopropylcarbamate. To a stirred solution of 2-amino-N-sec-butyl-N-(2,2-dimethoxyethyl)acetamide (3.6 g, 16.5 mmol) in dichloromethane (40 mL) was added DIPEA (31.91 mL, 49.5 mmol), 3-(((9H-fluoren-9-yl)methoxy)carbonylamino)propanoic acid (5.14 g, 16.5 mmol) and T 3 P (39.13 g, 33 mmol) was added at 0° C. The reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, reaction water (100 mL) was added and the organic phase was separated. The aqueous phase was extracted with dichloromethane (2×150 mL). The combined organic layers were washed with Na 2 SO 4 The mixture was dried at 40° C., filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography using silica (230-400 mesh; 23-25% ethyl acetate / petroleum ether as eluent). The collected pure fractions were concentrated under reduced pressure to give (9H-fluoren-9-yl)methyl-3-(2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylamino)-3-oxopropylcarbamate (4.1 g, 48.6%) as a gum.

[0227] Step 4: Synthesis of (9H-fluoren-9-yl)methyl 8-sec-butyl-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate. A solution of (9H-fluoren-9-yl)methyl-3-(2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylamino)-3-oxopropylcarbamate (4.1 g, 8.01 mmol) in acetic acid (2 mL) was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. When TLC showed complete consumption of starting material, the reaction mixture was concentrated and the resulting mass was diluted with water and extracted with dichloromethane (2×100 mL). The combined organic layers were washed with Na 2 SO 4 Drying at rt, filtration and concentration under reduced pressure gave the product (9H-fluoren-9-yl)methyl 8-sec-butyl-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate (3.2 g, 89.3%) as a gum.

[0228] Step 5: Synthesis of 8-sec-butyltetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione. To a solution of (9H-fluoren-9-yl)methyl 8-sec-butyl-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate (3.2 g, 7.1 mmol) in DMF (20 mL) cooled to 0° C., piperidine (0.7 mL, 1.0 equiv.) was added and the reaction mixture was stirred at room temperature for 2 h. The progress of the reaction was monitored by TLC. When TLC showed complete consumption of starting material, the reaction mixture was washed with petroleum ether (2×50 mL) to remove non-polar impurities. Cold water was added and extracted with dichloromethane (2×100 mL). The combined organic layers were washed with Na 2 SO 4 The mixture was dried at rt, filtered and concentrated under reduced pressure to give the pure product 8-sec-butyltetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (900 mg, 55.9%) as a solid.

[0229] Step 6: Synthesis of compound 9. To a stirred solution of (8-(sec-butyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.500 g, 2.2 mmol) and 4-hydroxybenzaldehyde (0.271 g, 2.2 mmol) in methanol (10 mL) was added acetic acid (0.27 mL, 2.0 equiv.) and picoline borane (0.285 g, 2.6 mmol) at room temperature. The reaction mixture was stirred at room temperature for 48 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (10 mL) and extracted with ethyl acetate (2×20 mL). The combined organic layers were washed with Na 2 SO 4 The crude compound was analyzed by LC / MS. The crude LC / MS data showed 8.28% of the desired mass. The crude compound was purified by column chromatography on silica gel (100-200) to elute the desired compound with 50-70% ethyl acetate / petroleum ether. LC / MS of the eluted fraction showed 72.16% of the desired mass, which was further purified by preparative HPLC. After preparative HPLC purification, the fractions were collected, concentrated under reduced pressure, and then lyophilized to give 9 (0.168 g, 22.8%) as a solid. Preparative HPLC Method: Mobile Phase A: 10 mM ammonium bicarbonate in water; Mobile Phase B: acetonitrile; Column: X-BRIDGE C 18 (150×19mm 5μ);Flow rate: 18mL / min. MS(ESI)m / z[M+H] + :332.2.

[0230] Example S11. Synthesis of Compound 10. The synthetic route for preparing compound 10 is shown in Scheme 11. Scheme 11. [ka] To a solution of 6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (250 mg, 0.98 mmol) and 4-chlorobenzoic acid (170 mg, 1.09 mmol) in DMF (4 mL) at 0° C., HATU (413 mg, 1.08 mmol) was added followed by DIPEA (0.35 mL, 1.97 mmol). The reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and the aqueous layer was extracted with EtOAc (50 mL×2). The organic layer was washed with cold H 2 2H2O (30 mL), followed by saturated brine (30 mL), and 2 SO 4 The resulting crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / hexanes) to give 1-(4-chlorobenzoyl)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione 10 (150 mg, 0.383 mmol, 39.2% yield) as a solid. MS (ESI) m / z [M+H] + :392.05. 1 H NMR (400MHz, DMSO-d 6 )δ0.66-0.89(m,6H)0.91-1.42(m,4H)1.57-1.78(m,1H)2.16-2.35(m,2H)2.55-2.65(m,2H)3.08-3. 23(m,2H)3.28-3.40(m,1H)3.51-3.64(m,2H)4.76-4.89(m,1H)5.88-6.02(m,1H)7.46-7.56(m,4H).

[0231] Example S12. Synthesis of Compound 11. The synthetic route for preparing compound 11 is shown in Scheme 12. Scheme 12. [ka] To a solution of 6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (250 mg, 0.98 mmol) and 4-fluorobenzoic acid (153 mg, 1.09 mmol) in DMF (4 mL) at 0° C., HATU (413 mg, 1.08 mmol) was added followed by DIPEA (0.35 mL, 1.97 mmol). The reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and the aqueous layer was extracted with EtOAc (50 mL×2). The organic layer was washed with cold H 2 2H2O (30 mL), followed by saturated brine (30 mL), and 2 SO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The resulting crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / hexanes) to give 1-(4-fluorobenzoyl)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione 11 (140 mg, 0.37 mmol, 38.0% yield) as a solid. MS (ESI) m / z [M+H] + :376.05. 1 H NMR(400MHz,DMSO-d6)δ0.69-0.81(m,3H)0.86(t,J=7.23Hz,3H)0.95-1.14(m,2H)1.20-1.43(m,4H)1.59-1.80(m,2H)2.26(d,J=16.95Hz,1H) 2.55-2.72(m,1H)3.20-3.31(m,2H)3.35-3.39(m,1H)3.52-3.70(m,2H) 4.73-4.89(m,1H)7.33(t,J=8.73Hz,2H)7.61(dd,J=8.23,5.73Hz,2H).

[0232] Example S13. Synthesis of Compound 12. The synthetic route for preparing compound 12 is shown in Scheme 13. Scheme 13. [ka] To a solution of 6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (250 mg, 0.98 mmol) and 3-chloro-4-(trifluoromethyl)benzoic acid (242 mg, 1.09 mmol) in DMF (4 mL) at 0° C., HATU (413 mg, 1.08 mmol) was added followed by DIPEA (0.35 mL, 1.97 mmol). The reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and the aqueous layer was extracted with EtOAc (50 mL×2). The organic layer was washed with cold H 2 2H2O (30 mL), followed by saturated brine (30 mL), and 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / hexanes) to give 1-(3-chloro-4-(trifluoromethyl)benzoyl)-6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione 12 (250 mg, 0.55 mmol, 55.2% yield) as a solid. MS (ESI) m / z [M+H] + :460.0. 1 H NMR(400MHz,DMSO-d6)δ0.74-0.93(m,6H)0.98-1.19(m,2H)1.28-1.46(m, 3H)1.64-1.81(m,1H)2.22(d,J=17.45Hz,1H)2.57-2.70(m,1H)3.14(dd,J= 13.21,6.23Hz,1H)3.25-3.31(m,2H)3.44-3.57(m,1H)3.61-3.87(m,2H)4. 78-4.90(m,1H)5.89-6.05(m,1H)7.72(d,J=7.98Hz,1H)7.90-8.02(m,2H).

[0233] Example S14. Synthesis of intermediate compound 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. The synthetic route for preparing this intermediate compound is shown in Scheme 14. Scheme 14. [ka] Step 1: Synthesis of 2,2-diethoxy-N-(4-methoxybenzyl)ethan-1-amine. A 500 mL round bottom flask was charged with anisaldehyde (12 mL, 90.22 mmol) and 2,2-diethoxyethanamine (10 g, 75.18 mmol). The reaction mixture was heated at 100° C. for 1 h. The reaction mixture was cooled to room temperature and added EtOH (100 mL) followed by NaBH 4 (4.28 g, 112.7 mmol) was added. The resulting reaction mixture was stirred at room temperature for 16 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The resulting crude was dissolved in EtOAc (300 mL). The organic layer was washed with brine (100 mL) and diluted with Na 2 SO 4 The mixture was dried at rt and concentrated in vacuo to give the crude product, which was purified by column chromatography (silica 100-200 mesh; 70% EtOAc / hexanes) to give 2,2-diethoxy-N-(4-methoxybenzyl)ethan-1-amine (15 g, 59.28 mmol, 78% yield) as a liquid. MS (ESI) m / z [M+H] + :254.3.

[0234] Step 2: (9H-Fluoren-9-yl)methyl = (1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)carbamate. To a stirred solution of (((9H-fluoren-9-yl)methoxy)carbonyl)alanine (32 g, 102.76 mmol) in anhydrous DMF (140 mL) maintained at 0 °C was added HATU (42 g, 110.67 mmol), DIPEA (21.06 mL, 118.57 mmol) followed by 2,2-diethoxy-N-(4-methoxybenzyl)ethan-1-amine (20 g, 79.05 mmol). The reaction mixture was stirred at room temperature for 16 h. Upon complete consumption of the starting material, the reaction mixture was quenched with ice-cold water (300 mL) and the aqueous layer was extracted with EtOAc (200 mL x 2). The organic layer was cooled 2O (200 mL), followed by brine (100 mL), and 2 SO 4 The mixture was dried at 40° C. and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (silica 100-200 mesh; 50% EtOAc / hexanes) to give (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)carbamate (28 g, 51.22 mmol, 64.8% yield) as a gummy liquid. MS (ESI) m / z [M+H-EtOH] + :501.2.

[0235] Step 3: Synthesis of 2-amino-N-(2,2-diethoxyethyl)-N-(4-methoxybenzyl)propanamide. CH 2 Cl 2 To a solution of (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)carbamate (28 g, 51.22 mmol) in (30 mL) was added diethylamine (200 mL). The reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated and the resulting crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 2-amino-N-(2,2-diethoxyethyl)-N-(4-methoxybenzyl)propanamide (14.5 g, 44.75 mmol, 87% yield) as a viscous liquid. MS(ESI) m / z[M+H-EtOH] + :279.05.

[0236] Step 4: Synthesis of (9H-fluoren-9-yl)methyl (3-((1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)amino)-3-oxopropyl)carbamate. To a stirred solution of 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanoic acid (14.78 g, 47.53 mmol) in anhydrous DMF (120 mL) maintained at 0° C. was added HATU (18.06 g, 47.53 mmol), DIPEA (9.21 mL, 51.85 mmol) followed by 2-amino-N-(2,2-diethoxyethyl)-N-(4-methoxybenzyl)propanamide (14 g, 43.20 mmol). The reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was quenched with ice-cold water (200 mL) and the aqueous layer was extracted with EtOAc (200 mL×2). The organic layer was washed with cold H 2 20 (500 mL), followed by saturated brine (200 mL), and 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / hexanes) to give (9H-fluoren-9-yl)methyl (3-((1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)amino)-3-oxopropyl)carbamate (18 g, 29.14 mmol, 67.44% yield) as a viscous liquid. MS (ESI) m / z [M+H-EtOH] + :572.

[0237] Step 5: Synthesis of (9H-fluoren-9-yl)methyl 8-(4-methoxybenzyl)-6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. A solution of (9H-fluoren-9-yl)methyl (3-((1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)amino)-3-oxopropyl)carbamate (18 g, 29.14 mmol) in formic acid (120 mL) was stirred at room temperature for 12 h. Upon completion, the reaction mixture was concentrated and the resulting crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / Hexanes) to give (9H-fluoren-9-yl)methyl 8-(4-methoxybenzyl)-6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (14.5 g, 27.58 mmol, 94% yield) as a solid. MS (ESI) m / z [M+H] + :526.

[0238] Step 6: Synthesis of 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. CH 2 Cl 2 To a solution of (9H-fluoren-9-yl)methyl 8-(4-methoxybenzyl)-6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (14 g, 26.63 mmol) in 150 mL of ethyl acetate was added diethylamine (100 mL) and the reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated and the resulting crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (7 g, 23.07 mmol, 87% yield) as a sticky solid. MS (ESI) m / z [M+H] + :304.

[0239] Example S15. Synthesis of intermediate compound 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. The synthetic route for preparing this intermediate compound is shown in Scheme 15. Scheme 15. [ka] Step 1: Synthesis of 8-(4-methoxybenzyl)-6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. To a solution of 4-(trifluoromethyl)benzoic acid (5.26 g, 27.69 mmol) in DMF (100 mL) maintained at 0° C. was added HATU (10.52 g, 27.69 mmol), DIPEA (12.30 mL, 69.23 mmol) followed by 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (7 g, 23.07 mmol) and the reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was quenched with ice-cold water (200 mL) and the aqueous layer was extracted with EtOAc (200 mL×2). The organic layer was washed with cold H 2 200 mL, followed by saturated brine (150 mL) and 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / hexanes) to give 8-(4-methoxybenzyl)-6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (9 g, 18.92 mmol, 82.04% yield) as a solid. MS (ESI) m / z [M+H] + : 476.15 and MS (ESI) m / z [M + Na] + :498.05.

[0240] Step 2: Synthesis of 6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. CH 3 CN:H 2 To a solution of 8-(4-methoxybenzyl)-6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (9 g, 18.92 mmol) in 2H2O (2:1, 150 mL) was added CAN (31.15 g, 56.82 mmol) and the reaction mixture was stirred at room temperature for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with NaHCO 3 (200 mL) of saturated aqueous solution of 1,2-dichloromethane and extracted with EtOAc (200 mL×2). The combined organic layers were washed with H 2 200 mL, followed by saturated brine solution (150 mL) and Na 2 SO 4 The resulting crude was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to give 6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.5 g, 9.85 mmol, 52.8% yield) as a solid. MS (ESI) m / z [M+H+CH 3 CN] + :397.0. 1 H NMR(400MHz,DMSO-d6)δ1.25-1.46(m,3H)2.15-2.30(m,1H)2.56-2.69(m,1H)3.16(d,J=4.99Hz,1H)3.22-3.30(m,1 H)3.42-3.72(m,2H)4.70-4.87(m,1H)5.85-5.95(m,1H)7.75(d,J=7.98Hz,2H)7.86(d,J=7.98Hz,2H)8.11(brs,1H).

[0241] Example S16. General Procedure A for the synthesis of final compounds. To a solution of 6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (200 mg, 0.56 mmol) in DMF (2 mL), KO t Bu (1M in THF, 1.69 mmol, 1.69 mL) was added followed by alkyl halide (1.12 mmol) and the reaction mixture was exposed to microwave irradiation at 120° C. for 1 h. The reaction mixture was cooled to room temperature and diluted with H 2 Quenched with O (25 mL). The aqueous layer was extracted with EtOAc (10 mL×3). The combined organic layers were washed with brine and concentrated. The crude product was purified by CombiFlash.

[0242] Example S17. Synthesis of compound 15. Compound 15 was synthesized by general procedure A using (bromomethyl)cyclopentane as the alkyl halide. MS (ESI) m / z [M+H] + :438.65. 1 H NMR (400MHz, DMSO-d 6 )δ1.02-1.26(m,3H)1.28-1.42(m,2H)1.44-1.76(m,6H)1.80-2.08-2.33( m,2H)2.55-2.71(m,1H)3.22(dd,J=12.96,7.48Hz,1H)3.26-3.32(m,1H)3 .39(d,J=6.98Hz,1H)3.49-3.57(m,1H)3.59-3.74(m,1H)3.76-3.91(m,1H )4.80-4.90(m,1H)5.95-6.05(m,1H)7.72-7.79(m,2H)7.84-7.91(m,2H).

[0243] Example S18. Synthesis of compound 16. Compound 16 was synthesized by general procedure A using bromomethylcyclobutane as the alkyl halide. MS (ESI) m / z [M+H] + :424.15. 1 H NMR (400MHz, DMSO-d 6)δ1.29-1.44(m,2H)1.58-1.89(m,4H)1.90-2.08(m,2H)2.16-2.31(m,1H)2.55-2.70(m,2H)3.18-3.31(m,1H)3.25-3.26(m,1H)3.34-3.42 (m,1H)3.36-3.57(m,2H)3.60-3.69(n,1H)3.71-3.83(m,1H)4.75-4.89(m,1H)5.90-6.05(m,1H)7.70-7.79(m,2H)7.87(d,J=8.31Hz,2H).

[0244] Example S19. Synthesis of compound 19. Compound 19 was synthesized by general procedure A using (2-bromoethyl)cyclopentane as the alkyl halide. MS (ESI) m / z [M+H] + :452.35. 1 H NMR (400MHz, DMSO-d 6 )δ0.94-1.18(m,3H)1.26-1.61(m,9H)1.66-1.83(m,2H)2.16-2.31(m,1H)2.56-2.70(m,1H)3.16-3.28(m,1H)3.35-3.56(m ,3H)3.60-3.73(m,1H)3.77-3.90(m,1H)4.72-4.92(m,1H)5.94-6.06(m,1H)7.77(d,J=7.98Hz,2H)7.87(d,J=7.98Hz,2H).

[0245] Example S20. Synthesis of compound 20. Compound 20 was synthesized by general procedure A using (2-bromoethyl)cyclobutane as the alkyl halide. MS (ESI) m / z [M+H] + :438.25. 1 H NMR (400MHz, DMSO-d 6)δ1.27-1.44(m,3H)1.50-1.71(m,4H)1.71-1.88(m,2H)1.93-2.09(m,2H)2.13-2.34(m,2H)2.56-2.70(m,2H)3.25-3.32(m,1H)3.35-3.42 (m,1H)3.45-3.55(m,1H)3.59-3.72(m,1H)3.74-3.90(m,1H)4.75-4.89(m,1H)5.94-6.05(m,1H)7.71-7.79(m,2H)7.87(d,J=8.31Hz,2H).

[0246] Example S21. Synthesis of compound 21. Compound 21 was synthesized by general procedure A using 1-bromobutane as the alkyl halide. MS (ESI) m / z [M+H] + :412.20. 1 H NMR (400MHz, DMSO-d 6 )δ0.81-0.97(m,3H)1.15-1.57(m,7H)2.15-2.31(m,1H)2.57-2.69(m,1H)3.14-3.28(m,1H)3.35-3.60(m,3H)3.6 2-3.73(m,1H)3.74-3.92(m,1H)4.75-4.91(m,1H)5.94-6.06(m,1H)7.76(d,J=7.34Hz,2H)7.87(d,J=7.83Hz,2H).

[0247] Example S22. Synthesis of compound 22. Compound 22 was synthesized by general procedure A using 4-bromobut-1-ene as the alkyl halide. MS (ESI) m / z [M+H] + :410.20. 1 H NMR (400MHz, DMSO-d 6)δ1.28-1.45(m,3H)2.14-2.38(m,3H)2.55-2.69(m,1H)3.36-3.57(m,4H)3.58-3.72(m,1H)3.75-3.89(m,1H)4.7 5-4.90(m,1H)4.98-5.19(m,2H)5.69-5.84(m,1H)5.93-6.05(m,1H)7.76(d,J=7.98Hz,2H)7.88(d,J=7.98Hz,2H).

[0248] Example S23. Synthesis of compound 23. Compound 23 was synthesized by general procedure A using 1-bromo-2-methylpropane as the alkyl halide. MS (ESI) m / z [M+H] + :412.25. 1 H NMR (400MHz, DMSO-d 6 )δ0.80-0.96(m,6H)1.30-1.48(m,3H)1.85-2.03(m,1H)2.15-2.31(m, 1H)2.57-2.70(m,1H)3.06-3.16(m,1H)3.18-3.28(m,1H)3.36-3.45(m ,1H)3.44-3.57(m,1H)3.60-3.74(m,1H)3.73-3.87(m,1H)4.77-4.92( m,1H)5.93-6.07(m,1H)7.76(d,J=7.48Hz,2H)7.87(d,J=7.48Hz,2H).

[0249] Example S24. Synthesis of compound 24. Compound 24 was synthesized by general procedure A using 2-bromopropane as the alkyl halide. MS (ESI) m / z [M+H] + :398.55. 1 H NMR (400MHz, DMSO-d 6 )δ1.10(d,J=5.49Hz,6H)1.28-1.45(m,3H)2.16-2.24(m,1H)2.56-2.71(m,1H)3.34-3.40(m,1H)3.44-3. 79(m,3H)4.59-4.72(m,1H)4.75-4.90(m,1H)5.86-6.00(m,1H)7.79(d,J=7.98Hz,2H)7.83-7.92(m,2H).

[0250] Example S25. Synthesis of intermediate compound 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. The synthetic route for preparing this intermediate compound is shown in Scheme 16. Scheme 16. [ka] Step 1: Synthesis of 1-(4-(difluoromethoxy)benzoyl)-8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. To a solution of 4-(difluoromethoxy)benzoic acid (1.71 g, 9.08 mmol) in DMF (25 mL) maintained at 0° C. was added HATU (3.45 g, 9.08 mmol), DIPEA (4.34 mL, 24.8 mmol) followed by 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (2.5 g, 8.25 mmol) and the reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and the aqueous layer was extracted with EtOAc (100 mL×2). The organic layer was washed with cold H 2 2H2O (100 mL), followed by saturated brine (100 mL), and 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 30% EtOAc / hexanes) to give 1-(4-(difluoromethoxy)benzoyl)-8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.5 g, 7.38 mmol, 89.5% yield) as a solid. MS (ESI) m / z [M+H] + :474.12.

[0251] Step 2: Synthesis of 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. 3 CN:H 2 To a solution of 1-(4-(difluoromethoxy)benzoyl)-8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.0 g, 6.34 mmol) in 2H2O (2:1, 45 mL) was added CAN (12.0 g, 21.90 mmol) and the reaction mixture was stirred at room temperature for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with NaHCO 3 (100 mL) of saturated aqueous solution of ethyl acetate and extracted with EtOAc (200 mL x 2). The combined organic layers were 2 250 mL, followed by saturated brine solution (250 mL) and Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to give 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (2.0 g, 5.66 mmol, 89.6% yield) as a solid. MS (ESI) m / z [M+H] + :353.95. 1 H NMR (400MHz, DMSO-d 6 )δ1.10-1.39(m,3H)2.17-2.18(m,1H)2.52-2.68(m,1H)3.18-3.27(m,2H)3.44-3.71(m,2H)4.69-4.83(m ,1H)5.75-5.92(m,1H)7.24(d,J=7.83Hz,2H)7.32(t,J=72.0Hz,1H)7.57(d,J=8.31Hz,2H)8.04(brs,1H).

[0252] Example S26. General procedure B for the synthesis of final compounds. To a solution of 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (200 mg, 0.56 mmol) in DMF (4 mL) maintained at 0° C., NaH (122 mg, 2.8 mmol, 55% mineral oil dispersion) was added and the reaction mixture was stirred at the same temperature for 15 min. To this reaction mixture, alkyl halide (1.6 mmol) was added and the reaction mixture was allowed to warm to room temperature and stirred for 3 h. After completion, the reaction mixture was immersed in ice-cold H 2 It was quenched with O (15 mL) and the aqueous layer was extracted with EtOAc (15 mL×3). The combined organic layers were washed with brine and concentrated. The crude product was purified by CombiFlash.

[0253] Example S27. Synthesis of compound 13. Compound 13 was synthesized by general procedure B using (bromomethyl)cyclopentane as the alkyl halide. MS (ESI) m / z [M+H] + :436.05. 1 H NMR (400MHz, DMSO-d 6 )δ1.07-1.16(m,3H)1.32(d,J=6.48Hz,3H)1.41-1.73(m,7H)2.06-2.21( m,1H)2.21-2.34(m,1H)2.54-2.70(m,1H)3.14-3.29(m,1H)3.35-3.45(m ,1H)3.52-3.69(m,1H)3.75-3.93(m,1H)4.75-4.91(m,1H)5.88-5.99(m, 1H)7.27(d,J=8.48Hz,2H)7.35(t,J=72.0Hz,1H)7.61(d,J=8.98Hz,2H).

[0254] Example S28. Synthesis of compound 14. Compound 14 was synthesized by general procedure B using (bromomethyl)cyclobutane as the alkyl halide. MS (ESI) m / z [M+H] + :421.14. 1 H NMR (400MHz, DMSO-d 6)δ1.16-1.25(m,1H)1.27-1.43(m,3H)1.54-1.73(m,2H)1.73-1.86(m,2H)1.8 9-2.03(m,2H)2.24(d,J=17.12Hz,1H)2.53-2.69(m,2H)3.20-3.28(m,1H)3.2 9-3.40(m,1H)3.40-3.66(m,2H)3.69-3.87(m,1H)4.75-4.86(m,1H)5.74-6.0 2(m,1H)7.26(d,J=8.31Hz,2H))7.33(t,J=72.0Hz,1H)7.59(d,J=8.31Hz,2H).

[0255] Example S29. Synthesis of compound 17. Compound 20 was synthesized by general procedure B using 1-bromobutane as the alkyl halide. MS(ESI) m / z[M+H] + :410.0. 1 H NMR (400MHz, DMSO-d 6 )δ0.81-0.96(m,3H)1.15-1.39(m,4H)1.40-1.55(m,2H)2.26(d,J=16.95Hz,1H)2.53-2.70(m,2H)3.12-3.30(m,2H)3.38-3.46(m,1H)3.56- 3.74(m,2H)3.75-3.92(m,1H)4.84(q,J=6.81Hz,1H)5.86-6.06(m,1H)7.28(d,J=7.98Hz,2H)7.36(t,J=72.0Hz,1H)7.62(d,J=8.48Hz,2H).

[0256] Example S30. Synthesis of compound 18. Compound 18 was synthesized by general procedure B using 4-bromobut-1-ene as the alkyl halide. MS (ESI) m / z [M+H] + :408.06. 1 H NMR (400MHz, DMSO-d 6)δ1.16-1.45(m,3H)2.18-2.33(m,3H)2.53-2.70(m,1H)3.36-3.46(m,3H)3.51-3.72(m,2H)3.74-3.90(m,1H)4.84(q,J=6.65Hz ,1H)4.91-5.15(m,2H)5.67-5.84(m,1H)5.86-6.03(m,1H)7.29(d,J=8.48Hz,2H)7.36(t,J=72.0Hz,1H)7.61(d,J=8.48Hz,2H).

[0257] Example S31. Synthesis of compound 27. Compound 27 was synthesized by general procedure B using 2-(bromomethyl)tetrahydrofuran as the alkyl halide. MS (ESI) m / z [M+H] + :438.1. 1 H NMR (400 MHz, CDCl 3 )δ7.48-7.55(m,2H),7.20-7.30(m,2H),6.40-6.76(m,1H),5.90-6.20(m,1H),5.16-5.26(m,1H),4.06-4.17(m, 2H),3.82-3.92(m,4H),3.61-3.77(m,2H),2.83-2.99(m,1H),2.47-2.59(m,2H),2.01-2.12(m,4H),1.49(s,3H).

[0258] Example S32. Synthesis of compound 28. Compound 28 was synthesized by general procedure B using (2-bromoethyl)benzene as the alkyl halide. MS (ESI) m / z [M+H] + :458.10. 1 H NMR (400 MHz, CDCl 3)δ7.40-7.50(m,2H),7.20-7.28(m,2H),7.33-7.43(m,5H),6.40-6.76(m,1H),5.90-6.20(m,1H),5.16-5.26(m,1H),3.72-3.96(m, 2H),3.44-3.52(m,1H),3.25-3.35(m,2H),2.83-2.99(m,2H),2.47-2.59(m,1H),2.42-2.60(m,1H),2.30-2.57(m,1H),1.49(s,3H).

[0259] Example S33. Synthesis of compound 29. Compound 29 was synthesized by general procedure B using 4-(2-bromoethyl)pyridine as the alkyl halide. MS (ESI) m / z [M+H] + :459.10. 1 H NMR (400 MHz, CDCl 3 )δ8.50-8.58(m,2H),7.24-7.46(m,4H),7.18(d,J=7.99Hz,2H),6.40-6.76(m,1H),5.90-6.20(m,1H),5.16-5.26(m,1H),3.72-3.96( m,2H),3.44-3.52(m,1H),3.25-3.35(m,2H),2.83-2.99(m,2H),2.47-2.59(m,1H),2.42-2.60(m,1H),2.30-2.57(m,1H),1.49(s,3H).

[0260] Example S34. Synthesis of compound 30. Compound 30 was synthesized by general procedure B using (3-bromopropyl)cyclopropane as the alkyl halide. MS (ESI) m / z [M+H] + :459.10. 1 H NMR (400 MHz, CDCl 3)δ8.50-8.58(m,2H),7.24-7.46(m,4H),7.18(d,J=7.99Hz,2H),6.40-6.76(m,1H),5.90-6.20(m,1H),5.16-5.26(m,1H),3.72-3.96( m,2H),3.44-3.52(m,1H),3.25-3.35(m,2H),2.83-2.99(m,2H),2.47-2.59(m,1H),2.42-2.60(m,1H),2.30-2.57(m,1H),1.49(s,3H).

[0261] Example S35. Synthesis of compound 31. Compound 31 was synthesized by general procedure B using (2-bromoethyl)cyclopropane as the alkyl halide. MS (ESI) m / z [M+H] + :422.2. 1 H NMR (400 MHz, CDCl 3 )δ7.48(d,J=8.01Hz,2H),7.20-7.28(m,2H),6.40-6.76(m,1H),5.90-6.20(m,1H),5.16-5.26(m,1H),3.72- 3.96(m,1H),3.46-3.64(m,5H),2.46-2.64(m,2H),1.43-1.56(m,5H),0.43-0.65(m,2H),0.75-0.85(m,2H).

[0262] Example S36. Synthesis of compound 32. Compound 32 was synthesized by general procedure B using 1-bromo-2-methoxyethane as the alkyl halide. MS (ESI) m / z [M+H] + :412.1. 1 H NMR (400MHz, DMSO-d 6 )δ7.52-7.62(m,2H),7.16-7.34(m,3H),5.85-5.95(m,1H),4.80-4.90(m,1H),3.85-3.95(m,1H),3. 70-3.80(m,2H),3.25-3.46(m,5H),3.22(s,3H),2.62-2.72(m,1H),2.20-2.30(m,1H),1.49(s,3H).

[0263] Example S37. Synthesis of compound 33. Compound 33 was synthesized by general procedure B using 1-bromo-3-methoxypropane as the alkyl halide. MS (ESI) m / z [M+H] + :426.20. 1 H NMR (400MHz, DMSO-d 6 )δ7.52-7.62(m,2H),7.16-7.34(m,3H),5.85-5.95(m,1H),4.80-4.90(m,1H),3.85-3.95(m,1H),3.70-3.80( m,2H),3.58-3.68(m,2H),3.45-3.55(m,4H),3.22(s,3H),2.62-2.72(m,1H),2.20-2.30(m,2H),1.49(s,3H).

[0264] Example S38. Synthesis of compound 36. Compound 36 was synthesized by general procedure B using (2-bromoethyl)methylsulfone as the alkyl halide. MS (ESI) m / z [M+H] + :459.95. 1 H NMR (400MHz, chloroform) δ7.49(d,J=8.01Hz,2H),7.15-7.26(m,2H),6.40-6.76(m,1H),5.90-6.20(m,1H),5.15-5. 25(m,1H),3.86-3.97(m,3H),3.66-3.77(m,2H),3.38-3.49(m,3H),2.97(s,3H),2.59-2.69(m,2H),1.49(s,3H).

[0265] Example S39. Synthesis of compound 34. Step 1. To a solution of 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.300 g, 0.849 mmol) in DMF (6 mL), add Cs 2 CO 3(0.827 g, 2.547 mmol) followed by (2-bromoethoxy)(tert-butyl)dimethylsilane (0.243 g, 1.018 mmol) were added at 0° C. and the reaction mixture was heated in a sealed tube at 120° C. for 1 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched slowly with ice-cold water (30 mL) and extracted with EtOAc (50 mL). The combined organic layers were washed with ice-cold brine solution (3×30 mL) and diluted with Na 2 SO 4 After drying at 40° C. and concentrating under reduced pressure, 8-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.250 g, crude) was obtained. The crude compound was used directly in the next reaction without further purification. MS (ESI) m / z [M+H] + :512.10.

[0266] Step 2. To a solution of 8-(2-((tert-butyldimethylsilyl)oxy)ethyl)-1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.250 g, 0.4886 mmol) in THF (5 mL) was added TBAF (3 mL) at a temperature of 0° C. The reaction mixture was allowed to warm to room temperature and stirred for 6 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched slowly with ice-cold water (5 mL) and extracted with EtOAc (2×10 mL). The combined organic layers were washed with ice-cold brine solution (10 mL) and diluted with Na 2 SO 4 The crude compound was dried at 40° C. and concentrated under reduced pressure to give the crude compound. The crude compound was purified by column chromatography (silica gel 60-120 mesh; 10% MeOH / DCM) to give 1-(4-(difluoromethoxy)benzoyl)-8-(2-hydroxyethyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.102 g, 52% yield) as a white solid. MS(ESI) m / z[M+H] + :398.2.1 H NMR (400MHz, DMSO-d 6 )δ7.52-7.62(m,2H),7.16-7.34(m,3H),5.92-6.02(m,1H),6.78-6.88(m,2H),3.86-3.92(m, 1H), 3.47-3.62 (m, 6H), 3.21-3.31 (m, 1H), 2.57-2.67 (m, 1H), 2.25-3.35 (m, 1H), 1.49 (s, 3H).

[0267] Example S40. Synthesis of compound 35. Step 1. To a solution of 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.300 g, 0.849 mmol) in DMF (6 mL) was added NaH (0.050 g, 1.274 mmol) followed by 2-bromoacetonitrile (0.112 g, 0.933 mmol) at 0° C. and the reaction mixture was allowed to stand at room temperature for 1 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched slowly with ice-cold water (70 mL) and extracted with EtOAc (100 mL). The combined organic layers were washed with ice-cold brine solution (100 mL) and the Na 2 SO 4 The crude compound was dried at 40° C. and concentrated under reduced pressure to give the crude compound. The crude compound was purified by column chromatography (silica gel 60-120 mesh; 10% MeOH / DCM) to give 2-(1-(4-(difluoromethoxy)benzoyl)-6-methyl-4,7-dioxooctahydro-8H-pyrazino[1,2-a]pyrimidin-8-yl)acetonitrile (0.120 g, 36% yield) as a white solid. MS(ESI) m / z[M+H] + :393.05.

[0268] Step 2. To a solution of 2-(1-(4-(difluoromethoxy)benzoyl)-6-methyl-4,7-dioxooctahydro-8H-pyrazino[1,2-a]pyrimidin-8-yl)acetonitrile (0.120 g, 0.305 mmol) in ethanol (5 mL), concentrated HCl (0.100 mL) was added followed by platinum oxide (0.012 g, 0.030 mmol) at room temperature and the reaction mixture was heated under hydrogen gas atmosphere for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered through a Celite pad. The Celite pad was washed with ethanol (20 mL) and the filtrate was concentrated under reduced pressure to give the crude compound. The crude compound was triturated with n-pentane to give 8-(2-aminoethyl)-1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.110 g, 90% yield) as a white solid. MS(ESI) m / z[M+H] + :397.05. 1 H NMR (400MHz, DMSO d 6 )δ7.96(s,2H),7.55-7.65(m,2H),7.20-7.35(m,3H),5.90-6.20(m,1H),4.85-4.95(m,1H),3.82-3.92(m,1H) ,3.55.-3.85(m,2H),3.35-3.45(m,3H),2.95-3.05(m,2H),2.60-2.70(m,1H),2.20-2.30(m,1H),1.35(s,3H).

[0269] Example S41. General procedure C for the synthesis of final compounds. To a solution of 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.200 g, 0.566 mmol) in DMF (5 mL) was added Cs 2 CO 3(0.735 g, 2.264 mmol, 4 equiv.) followed by alkyl halide (0.679 mmol, 1.2 equiv.) were added at 0° C. and the reaction mixture was heated at 50° C. under microwave irradiation for 1 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was slowly quenched with ice-cold water (6 mL) and extracted with EtOAc (20 mL×3). The combined organic layers were washed with saturated brine solution (10 mL) and diluted with Na 2 SO 4 The crude compound was purified by column chromatography to give the final compound.

[0270] Example S42. Synthesis of compound 25. Compound 25 was synthesized by general procedure C using 2-(2-iodoethyl)furan as the alkyl halide. MS (ESI) m / z [M+H] + :448.10. 1 H NMR:δ7.40-7.50(m,2H),7.28-7.38(m,1H),7.15-7.25(m,2H),6.39-6.78(m,1H),6.25-6.35(m,1H),5.90-6.12(m,2H),5.25-5.35(m ,1H),5.10-5.20(m,1H),3.70-3.80(m,1H),3.50-3.60(m,1H),3.20-3.40(m,2H),2.95-3.05(m,3H),2.45-2.60(m,2H),1.59(s,3H).

[0271] Example S43. Synthesis of compound 26. Compound 26 was synthesized by general procedure C using 2-(2-bromoethyl)thiophene as the alkyl halide. MS (ESI) m / z [M+H] + :464.1. 1 H NMR (400 MHz, CDCl 3)δ7.40-7.48(m,2H),7.15-7.26(m,3H),6.85-6.95(m,2H),6.39-6.95(m,2H),5.90-6.20(m,1H),5.15-5.25(m, 1H),3.72-3.96(m,2H),3.47-3.54(m,1H),3.32-3.42(m,3H),3.10-3.20(m,2H),2.42-2.56(m,2H),1.49(s,3H).

[0272] Example S44. Synthesis of intermediate compound 1-(4-(difluoromethoxy)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. [ka] Step 1: Synthesis of (9H-fluoren-9-yl)methyl 6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. A solution of (9H-fluoren-9-yl)methyl 8-(4-methoxybenzyl)-6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (1.0 g, 26.63 mmol) in TFA (10 mL) was stirred in a microwave at 130° C. for 2 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under vacuum and the crude product was extracted with ethyl acetate (100 ml) and a saturated solution of sodium bicarbonate. The organic layer was washed with anhydrous Na 2 SO 4 The mixture was dried at rt, concentrated in vacuo and purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give (9H-fluoren-9-yl)methyl 6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (300 mg, 42% yield) as a sticky solid. MS (ESI) m / z [M+H] + :406.

[0273] Step 2: Synthesis of 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. CH 2 Cl 2 To a solution of (9H-fluoren-9-yl)methyl 6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (300 mg, 0.74 mmol) in 1H-tetrahydrofuran (5 mL) was added diethylamine (6 mL). The reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated and the crude product was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to give 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (120 mg, 92% yield) as a white solid. MS (ESI) m / z [M+H] + :184.

[0274] Step 3: Synthesis of 1-(4-(difluoromethoxy)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. To a solution of 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.700 g, 3.820 mmol) in DMF (8.0 mL) was added K 2 CO 3 (1.58 g, 11.46 mmol) was added at room temperature and stirred for 10 min. To the resulting reaction mixture was added 1-(bromomethyl)-4-(difluoromethoxy)benzene (1.086 g, 4.584 mmol) and the reaction mixture was heated at 80° C. for 6 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, quenched with water (50 mL) and extracted with EtOAc (50 mL×2). The combined organic layer was washed with saturated brine solution (20 mL) and diluted with Na 2 SO 4The mixture was dried at 40° C. and concentrated under reduced pressure. The crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 1-(4-(difluoromethoxy)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.550 g, 43.0% yield) as an off-white solid. MS (ESI) m / z [M+H] + :340.34.

[0275] Example S45. General procedure D for the synthesis of final compounds. To a solution of 1-(4-(difluoromethoxy)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.100 g, 0.2949 mmol) in DMF (2 mL) was added NaH (0.021 g, 0.8847 mmol) at 0° C., followed by the appropriate alkyl halide (2 eq.) and the reaction mixture was allowed to warm to room temperature and stirred for 5 h. The progress of the reaction was monitored by TLC. Upon completion, the reaction mixture was diluted with NaHCO 3 The mixture was slowly quenched with a saturated aqueous solution of (2 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were 2 2H2O (5 mL), followed by a saturated brine solution (5 mL) and washed with Na 2 SO 4 The crude material was purified by Combiflash column chromatography (5% MeOH / DCM) to give the final product.

[0276] Example S46. Synthesis of compound 37. Compound 37 was synthesized by general procedure D using 4-bromo-1,1,1-trifluorobutane as the alkyl halide. MS (ESI) m / z [M+H] + :354.2. 1 H NMR (400 MHz, CDCl 3):δ1.41(d,J=7.13Hz,3H),1.71-1.86(m,2H),2.01-2.15(m,2H),2.26-2. 35(m,1H),2.60-2.67(m,1H),2.89-3.01(m,1H),3.07-3.15(m,1H),3.21-3 .34(m,2H),3.46-3.65(m,2H),3.81-3.95(m,2H),4.35-4.41(m,1H),5.20- 5.29(m,1H),6.53(t,J=72.0Hz,1H),7.08-7.16(m,2H),7.30-7.36(m,2H).

[0277] Example S47. Synthesis of compound 38. Compound 38 was synthesized by general procedure D using (2-bromoethyl)cyclopentane as the alkyl halide. MS (ESI) m / z [M+H] + :436.2. 1 H NMR (400 MHz, CDCl 3 )δ1.01-1.15(m,2H),1.41(d,J=7.13Hz,3H),1.45-1.62(m,8 H),1.66-1.80(m,2H),2.23-2.34(m,1H),2.58-2.72(m,1H),2.89-2.9 8(m,1H),3.04-3.18(m,2H),3.23-3.33(m,1H),3.43-3.54(m,1H),3.55 -3.65(m,1H),3.78-3.93(m,1H),4.31-4.39(m,1H),5.15-5.26(m,1H), 6.53(t,J=72.0Hz,1H),7.13(d,J=8.50Hz,2H),7.34(d,J=8.50Hz,2H).

[0278] Example S48. Synthesis of compound 39. Compound 39 was synthesized by general procedure D using 4-bromobut-1-ene as the alkyl halide. MS (ESI) m / z [M+H] + :394.2. 1 H NMR (400 MHz, CDCl 3)δ1.41(d,J=7.13Hz,3H),2.23-2.35(m,3H),2.60-2.71(m,1H),2.92-3.0 1(m,1H),3.06-3.14(m,1H),3.22-3.46(m,3H),3.53-3.64(m,1H),3.79-3. 93(m,2H),4.28-4.38(m,1H),4.91-5.00(m,2H),5.16-5.26(m,1H),5.64- 5.76(m,1H),6.52(t,J=72.0Hz,1H),7.10-7.16(m,2H),7.30-7.36(m,2H).

[0279] Example S49. Synthesis of compound 40. Compound 40 was synthesized by general procedure D using (2-bromoethyl)cyclobutene as the alkyl halide. MS (ESI) m / z [M+H] + :422.25. 1 H NMR (400 MHz, CDCl 3 )δ1.41(d,J=7.13Hz,3H),1.56-1.65(m,4H),1.73-1.92(m,2H),1.95-2.07(m,2H),2. 14-2.25(m,1H),2.26-2.35(m,1H),2.59-2.72(m,1H),2.91-2.99(m,1H),3.04-3.14(m ,2H),3.23-3.43(m,2H),3.53-3.63(m,1H),3.87(q,J=13.38Hz,2H),4.29-4.39(m,1H) ,5.17-5.24(m,1H),6.53(t,J=72.0Hz,1H),7.13(d,J=8.63Hz,2H),7.30-7.37(m,2H).

[0280] Example S50. Synthesis of compound 41. Compound 41 was synthesized by general procedure D using 1-bromobutane as the alkyl halide. MS(ESI) m / z[M+H] + :396.05. 1 H NMR (400MHz, DMSO-d 6)δ0.86(t,J=7.34Hz,3H),1.14-1.24(m,2H),1.24-1.30(m,2H),1.38-1.50(m,2H) ),1.98-2.10(m,1H),2.53-2.61(m,2H),2.64-2.77(m,2H),3.07-3.25(m,3H),3.3 2-3.41(m,1H),3.62-3.73(m,1H),3.87-3.93(m,2H),4.49-4.58(m,1H),4.84-4. 94(m,1H),7.15(d,J=8.56Hz,2H),7.22(t,J=72.0Hz,1H),7.43(d,J=8.56Hz,1H).

[0281] Example S51. Synthesis of compound 52. Compound 52 was synthesized by general procedure D using 2-trifluoromethyl-1-bromoethane as the alkyl halide. MS (ESI) m / z [M+H] + :420.16. 1 H NMR (400 MHz, CDCl 3 )δppm 7.31-7.38(m,2H),7.11-7.16(m,2H),6.31-6.73(m,1H),5.26(q,J=7.21Hz,1H),4.23 -4.44(m,2H),3.98-4.13(m,1H),3.80-3.93(m,3H),3.59(t,J=11.07Hz,1H),3.10(dd ,J=11.51,3.75Hz,1H),2.90-2.99(m,1H),2.62-2.72(m,1H),2.32(dd,J=4.38,2.38H z,1H),2.28(dd,J=4.31,2.31Hz,1H),1.48(d,J=7.25Hz,1H),1.41(d,J=7.13Hz,3H).

[0282] Example S52. General procedure E for the synthesis of final compounds. To a stirred solution of 6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.300 g, 1.184 mmol) in DMF (6 mL) in a flask immersed in an ice / water bath, cesium carbonate (0.771 g, 2.368 mmol, 2 equiv.) was added followed by the appropriate alkyl halide (1.1 equiv.). The flask was removed from the bath and stirred until TLC showed complete consumption of starting material. The reaction mixture was poured into ice-cold water (70 mL) and the aqueous layer was extracted with EtOAc (100 mL). The organic layer was washed with ice-cold brine (50 mL x 3) and anhydrous Na 2 SO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give the final compound.

[0283] Example S53. Synthesis of compound 42. Compound 42 was synthesized by general procedure E using 4-(bromomethyl)-2-chloro-1-(trifluoromethyl)benzene as the alkyl halide. MS (ESI) m / z [M+H] + :362.2. 1 H NMR (400MHz, DMSO-d 6 )δ0.75-0.89(m,3H),0.82-0.87(m,3H),0.96-1.13(m,1H),1.23-1.31(m,4H),1. 64-1.75(m,1H),2.06-2.09(m,1H),2.55-2.62(m,1H),2.65-2.76(m,1H),3.05-3 .15(m,1H),3.15-3.26(m,3H),3.64-3.74(m,1H),3.84-3.95(m,2H),4.52-4.60( m,1H),4.86-4.94(m,1H),7.17(t,J=8.76Hz,2H),7.41(dd,J=8.19,5.82Hz,2H).

[0284] Example S54. Synthesis of compound 43. Compound 43 was synthesized by general procedure E using 4-(bromomethyl)-2-chloro-1-(trifluoromethyl)benzene as the alkyl halide. MS (ESI) m / z [M+H]+ :446.2. 1 H NMR (400MHz, DMSO-d 6 )0.72-0.80(m,3H),0.80-0.87(m,3H),0.96-1.10(m,1H),1.21-1.27(m,1H),1.28-1 .34(m,3H),1.62-1.79(m,1H),2.00-2.13(m,1H),2.53-2.65(m,1H),2.66-2.76(m,1 H),3.00-3.10(m,1H),3.17-3.29(m,3H),3.62-3.72(m,1H),4.00-4.08(m,2H),4.55 -4.65(m,1H),4.85-4.95(m,1H),7.52-7.60(m,1H),7.73(s,1H),7.80-7.88(m,1H).

[0285] Example S55. Synthesis of compound 44. To a solution of 6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.420 g, 1.657 mmol) and 1H-indole-3-carbaldehyde (0.264 g, 1.823 mmol) in DCE (15 mL) was added acetic acid (1 mL, 1.657 mmol) and the reaction mixture was heated at 80° C. for 1 h. The resulting reaction mixture was treated with NaBH 4 (0.188 g, 4.973 mmol) was added in portions and the reaction mixture was heated at 80° C. and stirred for 4 h. When TLC analysis (5% MeOH / DCM) showed complete consumption of starting material, the reaction mixture was diluted with water (40 mL) and the aqueous layer was extracted with DCM (100 mL). The organic layer was washed with brine (50 mL) and diluted with anhydrous Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM), followed by washing with water (30 mL) and drying under reduced pressure to give compound 44 (0.250 g, 39% yield) as an off-white solid. MS (ESI) m / z [M+H] + :383.4. 1 H NMR (400MHz, DMSO-d 6)δ0.70(t,J=7.09Hz,3H),0.75-0.82(m,3H),0.91-1.11(m,1H),1.22-1.31(m,3H),1.57-1.72(m,1H ),1.97-2.07(m,1H),2.55-2.70(m,2H),2.83(dt,J=10.91,2.74Hz,1H),2.95-3.07(m,1H),3.10-3. 26(m,3H),3.54-3.69(m,1H),3.96-4.04(m,1H),4.06-4.15(m,1H),4.54-4.64(m,1H),4.84-4.95(m ,1H),6.94-7.02(m,1H),7.04-7.13(m,1H),7.29-7.40(m,2H),7.65(d,J=7.95Hz,1H),10.95(s,1H).

[0286] Example S56. Synthesis of intermediate compound 1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)dione. To a solution of 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (250 mg, 1.40 mmol) in DMF (3 mL), potassium carbonate (580 mg, 4.20 mmol) was added followed by 4-fluorobenzyl bromide (0.320 g, 1.70 mmol) and stirred at a temperature of 80° C. for 3 h. After completion, the reaction mixture was monitored by TLC (5% MeOH / DCM). The reaction mixture was poured into ice-cold water (50 mL) and the aqueous layer was extracted with EtOAc (50 mL). The organic layer was washed with anhydrous Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)dione (160 mg, 70% yield) as a white solid. MS (ESI) m / z [M+H] + :292.

[0287] Example S56. Synthesis of compound 45. To a solution of 1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)dione (80 mg, 0.2739 mmol) in DMF (3 mL) at 0° C. under ice-cooling, NaH (20 mg, 0.2739 mmol) was added and stirred for 20 min, then after 3 h, (2-bromoethyl)cyclobutane (67 mg, 0.41 mmol) was added. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was quenched with ice-cold water and extracted with ethyl acetate. The organic layer was washed with anhydrous Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 8-(2-cyclobutylethyl)-1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (13 mg, 16% yield) as a gummy liquid. MS(ESI) m / z[M+H] + :374. 1 H NMR (400MHz, CD 3 Cl 3 ):δ7.30-7.40(m,2H),7.00-7.10(m,2H),5.15-5.25(m,1H),4.25-4.35(m,1H),3.80-3.95(m,2H),3.55-3.65(m,1H),3.25-3.45(m,2H) ),3.05-3.20(m,2H),2.90-3.0(m,1H),2.60-2.70(m,1H),2.15-2.40(m,2H),1.75-2.10(m,4H),1.55-1.65(m,4H),1.20-1.30(m,3H).

[0288] Example S57. Synthesis of compound 46. To a solution of 1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)dione (80 mg, 0.2739 mmol) in DMF (3 mL) at 0° C. under ice-cooling, NaH (20 mg, 0.2739 mmol) was added and stirred for 20 min, then after 3 h, (2-bromoethyl)cyclopentane (72 mg, 0.41 mmol) was added and the completion of starting material was monitored by TLC, the reaction mixture was quenched with ice-cold water and extracted with ethyl acetate. The organic layer was washed with anhydrous Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 8-(2-cyclopentylethyl)-1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione as a gummy liquid.

[0289] Example S58. Synthesis of intermediate compound 6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione hydrochloride. Step 1: Synthesis of N-(2,2-diethoxyethyl)-2-methylbutan-1-amine. To stirred untreated 2,2-diethoxyethan-1-amine (20.0 g, 0.137 mmol) was added 2-methylbutanal (11.60 g, 0.137 mmol) at room temperature, and the reaction mixture was heated to 100° C. for 3 h. The resulting reaction mixture was added with ethanol (200 mL), followed by NaBH 4 (15.40 g, 0.413 mmol) was slowly added at room temperature and the reaction mixture was stirred for 16 h. Once the starting material was completely consumed (monitored by TLC), the reaction mixture was cooled to room temperature and NH 4 The mixture was slowly quenched with a saturated solution of NaCl (100 mL). The aqueous layer was extracted with EtOAc (200 mL×2). The combined organic layers were washed with brine (400 mL) and diluted with Na 2 SO 4The crude compound was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to give N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (25.8 g, 88% yield) as a colorless liquid. MS(ESI) m / z[M+H] + :204.3. 1 H NMR(400MHz,DMSO-d6)δ0.80-0.89(m,6H)1.11(t,J=6.98Hz,6H)1.35-1.48(m,2H)2.28-2.32(m,1H) 2.41-2.45(m,1H)2.55(d,J=5.49Hz,2H)3.42-3.52(m,2H)3.57-3.65(m,2H)4.49(t,J=5.49Hz,1H).

[0290] Step 2: (9H-Fluoren-9-yl)methyl (1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)carbamate. To a stirred solution of (((9H-fluoren-9-yl)methoxy)carbonyl)serine (15.0 g, 45.81 mmol) in anhydrous DMF (150 mL) maintained at 0° C. was added HATU (26.0 g, 68.80 mmol), DIPEA (23.92 mL, 137.61 mmol) followed by N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (12.10 g, 59.63 mmol). The reaction mixture was stirred at room temperature for 4 h. After complete consumption of the starting material, the reaction mixture was quenched with ice-cold water (500 mL) and the aqueous layer was extracted with EtOAc (250 mL×2). The combined organic layers were washed with cold H 2 O (200 mL), followed by brine (200 mL), and 2 SO 4The crude material was purified by column chromatography (silica 100-200 mesh; 80% EtOAc / hexanes) to give (9H-fluoren-9-yl)methyl (1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)carbamate (21.0 g, 89.43% yield) as a yellow sticky solid. MS (ESI) m / z [M+Na] + :535.35.

[0291] Step 3: Synthesis of 2-amino-N-(2,2-diethoxyethyl)-3-hydroxy-N-(2-methylbutyl)propenamide. To a stirred solution of (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)carbamate (21.0 g, 41.01 mmol) in anhydrous DCM (110 mL) maintained at 0° C. was added diethylamine (58 mL, 2.80 vol.) and the reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under reduced pressure to give the crude product. The resulting crude material was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 2-amino-N-(2,2-diethoxyethyl)-3-hydroxy-N-(2-methylbutyl)propenamide (9.50 g, 80% yield) as a yellow sticky solid. MS (ESI) m / z [M+H] + :291.4.

[0292] Step 4: Synthesis of (9H-fluoren-9-yl)methyl (3-((1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-3-oxopropyl)carbamate. To a stirred solution of 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanoic acid (9.50 g, 30.54 mmol) in anhydrous DMF (95 mL) maintained at 0° C. was added HATU (17.40 g, 45.81 mmol), DIPEA (16.0 mL, 91.62 mmol) followed by 2-amino-N-(2,2-diethoxyethyl)-3-hydroxy-N-(2-methylbutyl)propanamide (13.20 g, 45.81 mmol) at room temperature and the reaction mixture was stirred for 16 h. After completion, the reaction mixture was quenched with ice-cold water (200 mL) and the aqueous layer was extracted with EtOAc (200 mL×2). The organic layer was washed with cold H 2 20 (500 mL), followed by saturated brine (200 mL), and 2 SO 4 The crude compound was purified by column chromatography (silica 100-200 mesh; 80% EtOAc / hexanes) to give (9H-fluoren-9-yl)methyl (3-((1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-3-oxopropyl)carbamate (8.0 g, 31.0% yield) as a yellow viscous oil. MS(ESI) m / z[MH] - :582.2.

[0293] Step 5: Synthesis of (9H-fluoren-9-yl)methyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. A stirred solution of (9H-fluoren-9-yl)methyl (3-((1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-3-oxopropyl)carbamate (8.0 g, 13.77 mmol) in formic acid (48.0 mL, 6.0 vol) at room temperature and the reaction mixture was stirred for 16 h. After completion, the reaction mixture was concentrated under reduced pressure to give (9H-fluoren-9-yl)methyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (6.0 g, crude) as a brown semi-solid. The crude compound was used directly in the next reaction without further purification. MS (ESI) m / z [M+H] + :492.2.

[0294] Step 6: Synthesis of 6-(hydroxymethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. CH 2 Cl 2 To a solution of (9H-fluoren-9-yl)methyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (6.0 g, 12.20 mmol) in hexanes (36.0 mL) was added diethylamine (18.0 mL) at 0° C. and the reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude material was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 6-(hydroxymethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.0 g, 93.75% yield) as a colorless viscous oil. MS(ESI)m / z[M+H] + :270.20.

[0295] Step 7: Synthesis of tert-butyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. CH 2 Cl 2 To a solution of 6-(hydroxymethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.0 g, 11.15 mmol) in 1,2-dimethylphenylsulfonyl ether (60 mL) was added triethylamine (4.5 mL, 33.45 mmol) followed by Boc anhydride (3.78 mL, 16.72 mmol) at 0° C. and the reaction mixture was stirred at room temperature for 16 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was slowly quenched with ice-cold water (30 mL) and extracted with DCM (40 mL). The organic layer was washed with brine (30 mL) and concentrated to dryness with NaCl. 2 SO 4 The crude compound was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to give tert-butyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (8.0 g, 31.0% yield) as a yellow viscous oil. MS (ESI) m / z [M+H] + :370.25.

[0296] Step 8: Synthesis of tert-butyl 6-(fluoromethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. To a solution of 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (1.50 g, 4.065 mmol) in DCM (30 mL) was added DAST (1.97 g, 12.19 mmol) at -78°C and stirred for 15 min. The reaction mixture was allowed to warm to room temperature and stirred for 3 h. After completion of the reaction (monitored by TLC), the reaction mixture was washed with saturated NaHCO 3 The mixture was quenched with ethyl acetate (15 mL) and the aqueous layer was extracted with EtOAc (100 mL×2). The combined organic layers were washed with saturated brine (50 mL) and diluted with Na 2 SO 4 The crude compound was dried at 40° C. and concentrated under reduced pressure to give the crude compound. The crude material was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give tert-butyl 6-(fluoromethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (0.800 g, 72.0% yield) as a colorless viscous oil. MS (ESI) m / z [M+H] + :372.2.

[0297] Step 9: Synthesis of 6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione hydrochloride. To a stirred solution of tert-butyl 6-(fluoromethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (1.0 g, 2.695 mmol) in 1,4-dioxane (5 mL) was added 4M HCl in dioxane (5 mL) at 0° C. and the reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was quenched with a saturated solution of sodium bicarbonate (10 mL) and extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with saturated brine (10 mL) and concentrated to 100° C. with 100 mL of Na HCl. 2 SO 4 Drying at 40° C. and concentration under reduced pressure gave 6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione hydrochloride (0.630 g, crude) as a brown sticky oil. MS (ESI) m / z [M+H] + Free base: 271.00.

[0298] Example S59. Synthesis of compound 47. To a solution of 6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione hydrochloride (0.150 g, 0.550 mmol) in DMF (1.5 mL), 2 CO 3 (0.381 g, 2.760 mmol) followed by 1-(bromomethyl)-4-(difluoromethoxy)benzene (0.261 g, 1.100 mmol) were added and the reaction mixture was stirred at room temperature for 16 h. After completion (monitored by TLC), the reaction mixture was slowly quenched with ice-cold water (6 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with saturated brine solution (10 mL) and diluted with Na 2 SO 4The crude compound was dried at 40° C. and concentrated under reduced pressure to give the crude compound. The crude compound was purified by preparative HPLC to give 1-(4-(difluoromethoxy)benzyl)-6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.040 g, 17.0% yield) as a white solid. MS(ESI)m / z[M+H] + :428.10. 1 H NMR (400 MHz, CDCl 3 )δ7.34(d,J=8.01,2H),7.11(d,J=8.01,2H),6.32-6.69(m,1H),5.14-5. 25(m,2H),4.60-4.76(m,2H),3.84-3.97(m,2H),3.35-3.45(m,2H),3.12- 3.40(m,4H),2.85-3.05(m,1H),2.65-2.75(m,1H),2.29-2.34(m,1H),1. 65-1.75(m,1H),1.30-1.40(m,1H),1.05-1.18(m,1H),0.80-0.90(m,6H).

[0299] Example S60. Synthesis of compound 48. To a solution of 6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione hydrochloride (0.340 g, 1.253 mmol) in DMF (3.4 mL) was added Cs 2 CO 3 (0.814 g, 2.506 mmol) followed by 1-(bromomethyl)-4-(trifluoromethyl)benzene (0.598 g, 2.506 mmol) were added and the reaction mixture was stirred at room temperature for 16 h. After completion (monitored by TLC), the reaction mixture was slowly quenched with ice-cold water (6 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with saturated brine solution (10 mL) and diluted with Na 2 SO 4The crude compound was dried at 40° C. and concentrated under reduced pressure to give the crude compound. The crude compound was purified by preparative HPLC to give 6-(fluoromethyl)-8-(2-methylbutyl)-1-(4-(trifluoromethyl)benzyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.045 g, 8.0% yield) as a white solid. MS (ESI) m / z [M+H] + :430.10. 1 H NMR (400 MHz, CDCl 3 )δ7.34(d,J=8.01,2H),7.11(d,J=8.01,2H),5.14-5.25(m,2H),4.60-4.76(m,2H),3.84-3.97(m,2H),3.35-3.45(m,2H),3.12-3.40(m,4 H),2.85-3.05(m,1H),2.65-2.75(m,1H),2.29-2.34(m,1H),1.65-1.75(m,1H),1.30-1.40(m,1H),1.05-1.18(m,1H),0.80-0.90(m,6H).

[0300] Example S61. Synthesis of intermediate compound methyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate. Step 1: Synthesis of methyl 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate. To a stirred solution of 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-methoxy-4-oxobutanoic acid (1.90 g, 9.475 mmol) in anhydrous DMF (30 mL) at 0° C., HATU (3.60 g, 1.137 mmol) was added followed by DIPEA (2.70 mL, 1.895 mmol) and the reaction mixture was stirred at the same temperature for 10 min. To the resulting reaction mixture was added N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (3.50 g, 9.475 mmol), and then the mixture was allowed to warm to room temperature and stirred for 6 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was quenched with ice-cold water (100 mL) and the aqueous layer was extracted with EtOAc (50 mL×2). The combined organic layers were washed with cold H 2 2H2O (50 mL), followed by brine (50 mL), and then washed with Na 2 SO 4 The mixture was dried at 40° C. and concentrated under reduced pressure to give the crude product. The crude material was purified by CombiFlash column chromatography using 50% EtOAc / n-hexane to give methyl 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (4.30 g, 83.0% yield) as a white solid. MS (ESI) m / z [M+H-EtOH] + :509.2.

[0301] Step 2: Synthesis of methyl 3-amino-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate. CH 2 Cl 2To a solution of methyl 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (1.36 g, 2.451 mmol) in 1H-hexanediamine (27.0 mL) was added diethylamine (1.53 mL, 14.71 mmol) at room temperature and the reaction mixture was stirred for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude compound was purified by CombiFlash column chromatography using 5% MeOH / DCM to give methyl 3-amino-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.700 g, 86% yield) as a yellow viscous liquid. MS(ESI)m / z[M+H-EtOH] + :287.68.

[0302] Step 3: Synthesis of methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate. To a stirred solution of 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanoic acid (0.490 g, 1.594 mmol) in anhydrous DMF (10 mL) maintained at 0° C. was added HATU (0.720 g, 1.913 mmol), DIPEA (0.555 mL, 3.188 mmol), followed by methyl 3-amino-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.530 g, 1.594 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 6 h. After completion, the reaction mixture was quenched with ice-cold water (20 mL) and the aqueous layer was extracted with EtOAc (20 mL×2). The organic layer was washed with cold H 2 20 mL, followed by washing with saturated brine (20 mL) and 2 SO 4The crude compound was purified by Combiflash column chromatography using 5% MeOH / DCM to give methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.630 g, 70% yield) as an off-white solid. MS (ESI) m / z [M+H-EtOH] + :580.20.

[0303] Step 4: Synthesis of (9H-fluoren-9-yl)methyl 6-(2-methoxy-2-oxoethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. To a stirred solution of methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.300 g, 0.4794 mmol) was added formic acid (1.5 mL) at room temperature and the reaction mixture was stirred for 16 h. Upon completion, the reaction mixture was concentrated and the resulting crude was purified by column chromatography (silica 100-200 mesh; 0-5% MeOH / DCM) to give (9H-fluoren-9-yl)methyl 6-(2-methoxy-2-oxoethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (0.200 g, 80% yield) as a yellow solid. MS (ESI) m / z [M+H] + :534.67.

[0304] Step 5: Synthesis of methyl 2-(8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate. CH 2 Cl 2To a solution of (9H-fluoren-9-yl)methyl 6-(2-methoxy-2-oxoethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (0.240 g, 0.4499 mmol) in 1H-fluoren-9-yl (0.5 mL) was added diethylamine (0.280 mL) and the reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated and the crude material was purified by Combiflash column chromatography using 0-5% MeOH / DCM to give methyl 2-(8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate (0.130 g, 93% yield) as a white solid. MS (ESI) m / z [MH] + :310.4.

[0305] Step 6: Synthesis of methyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate. To a solution of methyl 2-(8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate (3.08 g, 9.890 mmol) in DMF (30 mL) was added K 2 CO 3 (4.10 g, 29.66 mmol) was added at room temperature and the reaction mixture was stirred at 80° C. for 15 min. To the resulting reaction mixture was added 1-(bromomethyl)-4-(difluoromethoxy)benzene (3.48 g, 14.36 mmol) and the stirred mixture was heated to 80° C. for 2 h. After completion, the reaction mixture was quenched with ice-cold water (200 mL) and the aqueous layer was extracted with EtOAc (200 mL×2). The organic layer was washed with cold H 2 200 mL, followed by saturated brine (150 mL) and 2 SO 4The crude compound was purified by Combiflash column chromatography (5% MeOH / DCM) to give methyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate (2.20 g, 48% yield) as a yellow solid. MS (ESI) m / z [M-CH 3 ] + :454.10.

[0306] Example S61. Synthesis of compound 49. To a solution of methyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetate (2.20 g, 4.705 mmol) in THF (22.0 mL) was added NaOH (0.560 g, 14.11 mmol) followed by water (4 mL) and the reaction mixture was stirred at room temperature for 3 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude residue was dissolved in water (10 mL) and slowly acidified with 6N HCl (10 mL) and stirred for 5 min. The resulting solid precipitate was filtered through a Buchner funnel and dried under reduced pressure to give 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetic acid (0.85 g, 40% yield) as a white solid. MS (ESI) m / z [M+H] + :454.10. 1 H NMR (400 MHz, CDCl 3)δ7.28-7.38(m,2H),7.11(d,J=7.99Hz,2H),6.33-6.71(m,1H),5.36-5.40(m ,1H),4.70-4.80(m,1H),4.65-4.75(m,1H),3.80-4.00(m,2H),3.55-3.65(m, 1H),3.35-3.45(m,1H),2.85-3.30(m,6H),2.70-2.80(m,1H),2.25-2.35(m,1 H),1.65-1.76(m,1H),1.25-1.35(m,1H),1.10-1.20(m,1H),0.8-0.9(m,6H).

[0307] Example S62. Synthesis of compound 50. To a solution of 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetic acid (0.470 g, 1.036 mmol) in THF (5 mL) was added 1,1'-carbonyldiimidazole (0.500 g, 3.109 mmol) at room temperature and the reaction mixture was stirred for 15 min. The resulting reaction mixture was treated with NH 3 (10 mL) was added and the reaction mixture was stirred at room temperature for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was slowly quenched with ice-cold water (6 mL) and extracted with EtOAc (20 mL×3). The combined organic layers were washed with saturated brine solution (10 mL) and diluted with Na 2 SO 4 The crude compound was dried at 40° C. and concentrated under reduced pressure to give the crude compound. The crude compound was purified by Combiflash column chromatography using 5% MeOH / DCM followed by preparative HPLC to give 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidin-6-yl)acetamide (0.070 g, 15% yield) as a white solid. MS (ESI) m / z [M+H] + :453.20. 1 H NMR (400 MHz, CDCl 3)δ7.30-7.40(m,2H),7.05-7.15(m,2H),6.39-6.70(m,1H),5.20-5.40(m,2H),4. 75-4.85(m,1H),3.95-4.05(m,1H),3.75-3.85(m,1H),3.50-3.60(m,1H),3.30-3. 40(m,1H),3.05-3.25(m,2H),2.85-2.95(m,2H),2.55-2.70(m,1H),2.25-2.35(m ,1H),1.70-1.80(m,2H),1.30-1.40(m,2H),1.05-1.20(m,2H),0.75-0.90(m,6H).

[0308] Example S63. Synthesis of intermediate compound 1-(3-chloro-4-(trifluoromethyl)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. To a solution of 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (500 mg, 2.732 mmol) in DMF (7 mL), potassium carbonate (1.13 g, 8.196 mmol) was added followed by 4-(bromomethyl)-2-chloro-1-(trifluoromethyl)benzene (0.894 g, 3.278 mmol) and stirred at a temperature of 80° C. for 12 hours. After completion of the reaction, it was monitored by TLC (5% MeOH / DCM). The reaction mixture was poured into ice-cold water (50 mL) and the aqueous layer was extracted with EtOAc (50 mL). The organic layer was washed with anhydrous Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give 1-(3-chloro-4-(trifluoromethyl)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (320 mg, 42% yield) as a white solid. MS (ESI) m / z [M+H] + :376.34.

[0309] Example S64. General procedure F for the synthesis of final compounds. To a solution of 1-(3-chloro-4-(trifluoromethyl)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (150 mg, 0.400 mmol) in DMF (2 mL) at 0 °C, 2 CO 3 (4 eq.) was added and stirred for 20 min, then the appropriate alkyl halide (1.2 eq.) was added at room temperature and the reaction mixture was heated at 80° C. and stirred for 12 h. Once the starting material was consumed (monitored by TLC), the reaction mixture was quenched with ice-cold water and extracted with ethyl acetate. The organic layer was washed with anhydrous Na 2 SO 4 The crude was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to give the final compound.

[0310] Example S65. Synthesis of compound 51. Compound 51 was synthesized by general procedure F using (2-bromoethyl)cyclobutane as the alkyl halide. MS (ESI) m / z [M+H] + :458.2. 1 H NMR (400 MHz, CDCl 3 )δppm7.70(d,J=8.07Hz,1H),7.54(s,1H),7.33(d,J=8.68Hz,1H),4.34(dd,J=10 .64,3.55Hz,1H),3.87-3.99(m,2H),3.61(t,J=11.13Hz,1H),3.25-3.42(m,2H), 3.08-3.19(m,2H),2.89-2.98(m,1H),2.64-2.74(m,1H)2.29-2.38(m,1H)2.17-2 .27(m,1H)1.97-2.09(m,2H)1.72-1.92(m,3H)1.58-1.66(m,4H)1.55(br.s,3H).

[0311] Example S66. Synthesis of compound 54. Compound 54 was synthesized by general procedure F using (2-bromoethyl)cyclopentane as the alkyl halide. MS (ESI) m / z [M+H] +:472.15. 1 H NMR (400 MHz, CDCl 3 )δppm7.69(d,J=8.11Hz,1H)7.52-7.56(m,1H)7.33(d,J=7.89Hz,1H),5.23(q,J=7.23Hz,1H),4.36(dd,J= 10.52,3.29Hz,1H),3.87-3.98(m,2H),3.63(t,J=11.07Hz,1H),3.46-3.56(m,1H),3.25-3.35(m,1H),3.1 2-3.23(m,2H),2.87-2.97(m,1H),2.60-2.70(m,1H),2.29-2.37(m,1H),1.66-1.82(m,2H),1.54-1.63(m, 1H),1.51(d,J=,2.63Hz,2H),1.42(d,J=7.23Hz,3H),1.26(br.s,2H)1.04-1.16(m,2H)0.80-0.92(m,2H).

[0312] Example S67. Synthesis of compound 53. Step 1: Synthesis of (9H-fluoren-9-yl)methyl (1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)carbamate. To a stirred solution of (((9H-fluoren-9-yl)methoxy)carbonyl)leucine (20.0 g, 56.58 mmol) in anhydrous DMF (200 mL), HATU (21.50 g, 56.58 mmol) was added followed by DIPEA (10.62 mL, 61.10 mmol) at 0° C. and the reaction mixture was stirred at the same temperature for 10 min. To the resulting reaction mixture, N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (11.48 g, 56.58 mmol) was added at room temperature and the reaction mixture was stirred for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was quenched with ice-cold water (100 mL) and the aqueous layer was extracted with EtOAc (50 mL×2). The combined organic layers were washed with cold H 2 2×50 mL, followed by brine (50 mL) and Na 2 SO 4The mixture was dried at 40° C. and concentrated under reduced pressure to give the crude product. The crude product was purified by CombiFlash column chromatography using 5% MeOH / DCM to give (9H-fluoren-9-yl)methyl = (1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)carbamate (14.5 g, 47.57% yield) as a white solid. MS (ESI) m / z [M+H] + :539.04.

[0313] Step 2: Synthesis of 2-amino-N-(2,2-diethoxyethyl)-4-methyl-N-(2-methylbutyl)pentanamide. CH 2 Cl 2 To a solution of (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)carbamate (8.50 g, 15.77 mmol) in (50 mL) was added diethylamine (16 mL, 157.7 mmol) and the reaction mixture was stirred at room temperature for 3 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude compound was purified by CombiFlash column chromatography using 5% MeOH / DCM to give 2-amino-N-(2,2-diethoxyethyl)-4-methyl-N-(2-methylbutyl)pentanamide (3.60 g, 72% yield) as a yellow viscous liquid. MS(ESI) m / z[M+H-EtOH] + :272.10.

[0314] Step 3: Synthesis of (9H-fluoren-9-yl)methyl = (3-((1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)amino)-3-oxopropyl)carbamate. To a stirred solution of 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanoic acid (3.80 g, 12.28 mmol) in anhydrous DMF (35 mL) maintained at 0 °C was added HATU (6.48 g, 17.05 mmol) and DIPEA (4.90 mL, 28.42 mmol), followed by 2-amino-N-(2,2-diethoxyethyl)-4-methyl-N-(2-methylbutyl)pentanamide (3.60 g, 11.37 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 3 h. After completion, the reaction mixture was quenched with ice-cold water (20 mL) and the aqueous layer was extracted with EtOAc (30 mL×2). The organic layer was washed with cold H 2 20 mL, followed by washing with saturated brine (20 mL) and 2 SO 4 The crude compound was purified by Combiflash column chromatography using 5% MeOH / DCM to give (9H-fluoren-9-yl)methyl(3-((1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)amino)-3-oxopropyl)carbamate (3.8 g, 55% yield) as an off-white solid. MS(ESI) m / z[M+H-EtOH] + :565.30.

[0315] Step 4: Synthesis of (9H-fluoren-9-yl)methyl 6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. To a stirred solution of (9H-fluoren-9-yl)methyl 6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (3.80 g, 6.231 mmol) was added formic acid (20 mL) at room temperature and the reaction mixture was stirred for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure. The crude compound was purified by column chromatography (silica 100-200 mesh; 0-5% MeOH / DCM) to give (9H-fluoren-9-yl)methyl 6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (3.60 g, 94% yield) as a yellow solid. MS (ESI) m / z [M+H] + :518.23.

[0316] Step 5: Synthesis of 6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. CH 2 Cl 2 To a solution of (9H-fluoren-9-yl)methyl 6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (3.60 g, 6.954 mmol) in 1H-fluoren-9-yl (36 mL) was added diethylamine (6.8 mL, 69.54 mmol) and the reaction mixture was stirred at room temperature for 16 h. Upon complete consumption of the starting material (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the crude product was purified by Combiflash column chromatography using 10-50% ethyl acetate / n-hexane to give 6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (1.20 g, 60% yield) as a white solid. MS (ESI) m / z [M+H] + :296.10.

[0317] Step 6: Synthesis of 1-(4-(difluoromethoxy)benzyl)-6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. To a solution of 6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.170 g, 0.576 mmol) in DMF (5 mL) was added K 2 CO 3 (0.159 g, 1.152 mmol) was added at 0° C. and the reaction mixture was stirred for 10 min. To the resulting reaction mixture was added 1-(bromomethyl)-4-(difluoromethoxy)benzene (0.150 g, 0.632 mmol) at room temperature and stirred for 3 h. After completion, the reaction mixture was quenched with ice-cold water (200 mL) and the aqueous layer was extracted with EtOAc (20 mL×2). The organic layer was washed with cold H 2 20 mL, followed by saturated brine (15 mL) and Na 2 SO 4 The resulting crude compound was purified by preparative HPLC to give 1-(4-(difluoromethoxy)benzyl)-6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.103 g, 40% yield) as a white solid. MS(ESI) m / z[M+H] + :452.3. 1 H NMR (400MHz, DMSO d 6 )δ7.42(d,J=8.8Hz,2H),7.14-7.24(m,3H),5.0-5.10(m,1H),4.50-4.60(m,1H),3.90-4.00(m,2H),3.60-3.70(m,1H),3.02- 3.40(m,4H),2.70-2.85(m,2H),2.0-2.10(m,1H),1.50-1.70(m,4H),1.20-1.35(m,1H),1.0-1.10(m,1H),0.70-0.98(m,12H).

[0318] Biological Examples Example B1. Phospho-MET ELISA. Compounds were screened for efficacy against the HGF / MET system using a phospho-MET (pMET) ELISA kit (Cell Signaling). pMET levels were detected in samples with low (1 ng / mL) and high (10 ng / mL) concentrations of HGF.

[0319] HEK293 cells were subcultured in 6-well multiplates and incubated in DMEM + 10% FBS at 37°C and 5% CO 2 Cells were grown in 5% CO at 4°C for 12 h until approximately 90% confluent. Cells were then starved in serum-free growth medium for at least 8 h.

[0320] Exemplary compounds were prepared and diluted in DMEM + 0.1% FBS and added to treatment media containing 1 ng / mL recombinant HGF protein (R&D Systems). Cells were incubated at 37°C and 5% CO 2 The samples were incubated in triplicate at 4°C for 15 min. Samples were then treated with 180 μL of ice-cold RIPA (radioimmunoprecipitation assay) buffer and cells were lysed for 15 min on ice. The lysates were clarified by centrifugation at 16,000 g for 15 min and the supernatant was retained. Samples were normalized using a BCA assay of the lysates to determine protein concentration in the whole sample.

[0321] 50-100 μg of total protein lysate was loaded into the ELISA wells of the pMET Sandwich ELISA kit (Cell Signaling catalog no. 7227C) to ensure equal loading of protein in each well. The ELISA was processed according to the manufacturer's instructions. After color development, the absorbance was read at 450 nm on an optical plate reader.

[0322] Potency measures were determined using peak efficacy by scaling treatments with test compound between HGF doses of 1 ng / mL and 10 ng / mL along a scale of 1 to 10 according to the following formula: y = 1 + (xA) * (10-1) / (BA) where y is the normalized data point, x is the raw data point, A is the mean HGF at 1 ng / mL, and B is the mean HGF at 10 ng / mL. The calculated potency results are shown in Table 2. TIFF2024546600000055.tif206165

[0323] Example B2. Cell spreading behavior assay MDCK cells were cultured under normal conditions and observed to spontaneously form robust colonies as they proliferated. MDCK cells responded to HGF treatment by moving away from each other (diffusing), which can be quantified to assess the amount of HGF / MET activation in a cell population. In this experiment, MDCK cells were plated in a 96-well format, treated with HGF and an exemplary compound, fluorescently stained, imaged in a wide field, and the spreading behavior quantified. Quantitation was determined by analyzing the number of contiguous cell clusters (normalized particle count) compared to the total stained area imaged.

[0324] MDCK cells were plated at low density in black-walled imaging plates and cultured in DMEM + 10% FBS at 37°C and 5% CO. 2 The cells were allowed to adhere overnight at 4° C. The cells were then starved for 2 hours in DMEM without FBS ("starvation medium"). Samples containing exemplary compounds were prepared in DMEM without FBS with 5 ng / mL HGF protein ("treatment medium"). Control curves for each plate were also generated using HGF concentrations of 0, 5, 10, and 20 ng / mL. The starvation medium was replaced with treatment medium, and the cells were incubated at 37° C. and 5% CO. 2 The mixture was incubated at RT for 24 hours.

[0325] After incubation, the cells were fixed by replacing the treatment medium with cold ethanol and incubating at 4° C. for 20 minutes. The cells were then rehydrated by washing with PBS and then with staining solution (fluorescent wheat germ agglutinin; WGA488, 20 μg / mL in PBS). The cells were incubated with the staining solution for 30 minutes at room temperature, after which the staining solution was replaced with fresh PBS.

[0326] Cellular areas were imaged using an iCyte High Content Imager at green wavelength. Images were binary converted and analyzed for particle size and particle count. For analysis, individual cells or isolated cell colonies not in contact with other cells were identified as particles, and particle counts were normalized by total signal area to account for differences in cell numbers. An increase in particle count indicates that individual cells moved away from each other in response to diffusion behavior. Compound potency was assessed by a statistical increase in normalized particle count compared to HGF treatment alone. Results are shown in Table 3. TIFF2024546600000056.tif212165

[0327] Example B3. Solubility Assay. Water solubility is an important drug property that helps predict bioavailability. Generally, compounds with water solubility below 100 μg / ml are poor drugs. To assess the solubility of compounds, turbidimetric solubility assays were performed using exemplary compounds in the concentration range of 3-300 μM.

[0328] To assess the solubility of compounds by turbidimetry, test compounds were first dissolved in an organic solvent (DMSO) at a concentration of 10 mM. The compound solutions were then diluted in an aqueous solvent (PBS) in a dilution series ranging from 3 to 300 μM in a 96-well assay plate. The solutions were incubated at 37°C for 2 hours.

[0329] Wells containing test compound in excess of their solubility limit would experience an increase in UV absorbance signal at 620 nm wavelength as the compound precipitated, effectively blocking the passage of light. Compounds were considered insoluble at the test concentrations if turbidity caused the absorbance to increase by more than 10% above the control reading. Results are shown in Table 4. TIFF2024546600000057.tif207165

[0330] Example B4. Permeability assay. To be bioavailable, a drug must be able to permeate the cell membranes lining the gastrointestinal tract. To estimate the permeability of exemplary compounds, a parallel artificial membrane permeability assay (PAMPA) was performed in vitro.

[0331] Test compounds should have a standard curve on the final read plate to determine the concentration of each drug fractionated. For each compound, a six-point standard curve was generated from 0 to 200 μM in phosphate buffered saline (PBS).

[0332] Test compound solution (300 μL in PBS) was added to the donor (bottom) wells of the PAMPA plate in replicates of five, and PBS vehicle (200 μL) was added to the acceptor (top) wells of the appropriate wells to match the loading volume of the donor plate. The bottom and top of the PAMPA plate were then clamped together. The PAMPA plate was then incubated for 5 hours at room temperature. After incubation, 150 μL of the donor solution was added to a UV-compatible plate with the corresponding standard curve. 150 μL of the acceptor well solution was added adjacent to the corresponding standard curve and donor well samples for the compound. The plate was then read using a UV plate reader.

[0333] The permeability and membrane retention were then calculated based on the following formula: Transparency (cm / s):(Pe)(cm / s)={-ln[1-CA(t) / Ceq]} / [A*(1 / VD+1 / VA)*t] (Equation 1) During the ceremony: A = filter area (0.3 cm 2 ); VD = donor well volume (0.3 mL); VA = acceptor well volume (0.2 mL); t = incubation time (sec); CA(t) = compound concentration in acceptor wells at time t; CD(t) = compound concentration in the donor well at time t; and Ceq = [CD(t)*VD+CA(t)*VA] / (VD+VA) Membrane retention rate (R)=1-[CD(t)*VD+CA(t)*VA] / (C0*VD) (Equation 2) During the ceremony: CD(t), VD, CA(t), and VA are as defined for Equation 1; and C0 = initial concentration in donor well (200uM).

[0334] The results are shown in Table 5. TIFF2024546600000058.tif211165

[0335] Example B5. Cytotoxicity Assay. This experiment was designed to obtain a preliminary assessment of cytotoxicity: compounds were tested at high concentrations to determine whether any cytotoxic effects were observed in hepatocyte (HepG2) cultures by measuring the release of lactate dehydrogenase (LDH) into the medium as a measure of lysed / dead cells.

[0336] HepG2 cells were plated in 96-well cell culture plates and cultured in EMEM + 10% FBS at 37 °C and 5% CO. 2 Cells were allowed to adhere overnight at 4°C. Treatments were performed in complete medium (EMEM + 10% FBS) and included a dilution series of test compounds ranging from 0.1 to 100 μM. Cerivastatin, a known cytotoxin, was used as a positive assay control and was prepared at a final concentration of 0.5 μM.

[0337] The growth medium was replaced with treatment medium (EMEM+10% FBS containing test compound dissolved in DMSO) and the cells were incubated with the test compound for 48 hours. After the incubation period, the supernatant medium from each well was transferred to a new plate and LDH assay working solution was added. The LDH assay solution produces a colorimetric reaction proportional to the amount of lactate dehydrogenase (an intracellular protein found in the medium only in the presence of lysed cells) in the medium. The color reaction was quantified by measuring the absorbance at a wavelength of 490 nm.

[0338] The signal range of the assay was determined by no manipulation in the negative control treatment and by complete lysis of all cells in the lysis control samples. Compounds that increased the level of cytotoxicity by more than 20% over the negative control samples were considered cytotoxic in this assay. The results are shown in Table 6. TIFF2024546600000059.tif204165

[0339] Example B6. In vitro stability assay. Compound stability when exposed to body conditions allows for the estimation of bioavailability. As an initial assessment of stability characteristics under various conditions present in animals, the stability of exemplary compounds was tested in an artificial body fluid compartment battery. Compounds were tested for stability in the following solutions: artificial gastric fluid (SGF: 34.2 mM NaCl, pH 1.2), artificial gastric fluid with the digestive enzyme pepsin (SGF+enzymes: SGF with 3.2 mg / ml pepsin), artificial intestinal fluid with porcine pancreatin enzyme mixture (SIF+enzymes: 28.7 mM NaH 2 PO 4 , 105.7 mM NaCl, pH 6.8, 10 mg / ml pancreatin), rat plasma, and human plasma.

[0340] Test compounds were incubated in the above solutions at 37°C at a final concentration of 5 μM, and samples were withdrawn at the following time points: 0, 1, 2, and 4 hours. The reaction was stopped and prepared for quantification by adding an excess of quench solution containing an internal standard (acetonitrile, 200 ng / mL bucetin). Test compounds and internal standard in each sample were quantified by LC-MS / MS, and the concentration of test compounds was expressed as a percentage of the concentration at 0 hours after internal normalization to bucetin. The stability of the corresponding test solution was then determined by the remaining percentage at 4 hours. The results are shown in Table 7. TIFF2024546600000060.tif183165

[0341] Example B7. In vivo pharmacokinetics. Exemplary compounds were administered by selected routes followed by blood sampling and quantification of the compound in plasma to determine the pharmacokinetic (PK) profile of the compound. Test compounds were administered to mixed-sex Sprague-Dawley rats of at least 250 grams by dissolving the compounds in DMSO and then diluting the compounds in the appropriate vehicle, either saline or saline and polyethylene glycol. Dosing was performed either by tail vein puncture (IV) or oral gavage (PO), with animals receiving the compound at 1 mL / kg according to body weight. Blood was collected by tail vein sampling at selected intervals (10, 20, 40, 60, 120, and 360 min) after dosing. Whole blood was then processed by centrifugation to obtain plasma. Compound content in plasma samples was quantified by LC-MS / MS and concentrations were accurately determined by comparison with internal standards and standard curves.

[0342] The plasma concentrations were then averaged at each time point and plotted as a function of time. The area under the curve was calculated by integrating the curve, Cmax was the maximum concentration reached in plasma, and Tmax was determined by the time point of Cmax. The results are shown in Table 8. TIFF2024546600000061.tif228165

[0343] Example B8. Oral availability calculation. Oral bioavailability is important in the development of small molecule therapeutics for oral administration. Oral bioavailability (%F) is calculated by using IV administration as the maximum possible exposure, determining the exposure rate after PO administration, and comparing with in vivo pharmacokinetic data (Example B7). In these studies, the dose-corrected AUC from PO administration was divided by the dose-corrected AUC from IV administration and multiplied by 100 to obtain %F. The results are shown in Table 9. TIFF2024546600000062.tif183165

[0344] Example B9. Non-specific protein binding. Plasma and tissue exposure of exemplary compounds was scaled by the nonspecific affinity for protein binding in the target tissue or fluid to determine the fraction of compound available for target interaction. Nonspecific binding was measured in plasma and brain homogenates from mixed sex Sprague-Dawley rats.

[0345] Known concentrations of test compounds were mixed with plasma or brain homogenate and incubated in the donor chamber of a rapid equilibrium dialysis (RED) device with empty PBS buffer in the receiving chamber. After 4 hours of incubation at 37 °C in an orbital shaker incubator, compounds in each chamber were quantified by LC-MS / MS. The unbound fraction (f u,組織 ) was calculated as:

number

[0346] The results are shown in Table 10. TIFF2024546600000064.tif188165

[0347] Example B10. In vivo tissue distribution. The distribution rate to target tissue is an important characteristic of therapeutic molecules. Tissue distribution of exemplary compounds was performed in mixed-sex Sprague-Dawley rats. Test compounds were delivered via tail vein injection (IV) and tissues were harvested at Tmax (10 min after administration). Animals were deeply anesthetized with isoflurane, whole blood was collected from the right atrium and processed by centrifugation to obtain plasma. Animals were then fully perfused with PBS administered into the left ventricle to prevent blood contamination of tissues.

[0348] Tissues were harvested and homogenized, and the compound content in the target tissues was quantified by LC-MS / MS. The tissue distribution percentage was determined by dividing the tissue concentration of the compound by the plasma concentration and multiplying by 100. The results are shown in Table 11. TIFF2024546600000065.tif195165

[0349] Example B11. In vivo efficacy: Scopolamine-induced spatial memory impairment in the Morris water maze. Exemplary compounds 2a and 6a were evaluated in the Morris water maze for their ability to reverse chemically induced spatial memory impairment in rats. The water maze consisted of a large circular tank (2.1 m diameter) filled with 26-28°C water to a depth of approximately 30 cm, and the water was made cloudy with white paint. A circular platform (13 cm diameter) was fixed to be 2-3 cm below the water surface. High contrast visual cues were placed around the tank to aid in spatial orientation of the test animals. Testing consisted of placing the animal in the water facing the tank wall at one of three randomly assigned starting points and allowing the animal to swim for up to 120 seconds to search for the hidden platform. The time it took the animal to locate the platform was recorded as escape latency. Animals were tested five times per day with a 30 second break between trials. Testing was performed for a total of eight consecutive days.

[0350] Animals were divided into groups (N=8 / group) according to treatment. Control animals received only empty vehicle. Scopolamine group received 3 mg / kg scopolamine dissolved in sterile saline by intraperitoneal (IP) injection 30 min before testing. Test compound group received various concentrations of test compound dissolved in 48% sterile saline, 50% polyethylene glycol (PEG-400), and 2% DMSO by oral gavage (PO) 40 min before testing. For each animal, escape latency was recorded for 8 consecutive days with 5 trials per day. Curve changes in escape latency were statistically analyzed by two-way ANOVA and Bonferroni post-hoc test. The results are shown in Table 12.

[0351] Exemplary compound 1a was evaluated in the Morris water maze for its ability to reverse chemically induced spatial memory impairment in rats. The water maze consisted of a large circular tank (1.5 m diameter) filled with 23-26°C water to a depth of approximately 30 cm, and the water was made cloudy with white paint. A circular platform was fixed 2-3 cm below the water surface. High-contrast visual cues were placed around the tank to aid in the spatial orientation of the test animals. Testing consisted of placing the animal in the water facing the tank wall at one of three randomly assigned starting points and allowing the animal to swim for up to 90 seconds to search for the hidden platform. The time it took the animal to locate the platform was recorded as escape latency. Animals were tested five times per day with a 30-second break between trials. Testing was performed for a total of five consecutive days.

[0352] Animals were divided into groups (N=12 / group) according to treatment. Control animals received only empty vehicle. Scopolamine group received 2 mg / kg scopolamine dissolved in sterile saline by intraperitoneal (IP) injection 30 min before testing. Test compound group received various concentrations of test compound dissolved in 78% sterile saline, 20% polyethylene glycol (PEG-400), and 2% DMSO by oral gavage (PO) 40 min before testing. For each animal, escape latency was recorded for 5 consecutive days with 5 trials per day. Curve changes in escape latency were statistically analyzed by two-way ANOVA and Bonferroni post-hoc test. The results are shown in Table 12. TIFF2024546600000066.tif92165

[0353] Example B12. Broad protection from neurotoxicity by compound 1a in vitro. Activation of HGF / MET signaling is expected to protect cells, including neurons, from cell death. To demonstrate the HGF / MET enhancing properties of the example compounds, we tested the ability of compound 1a to improve the viability of cultured primary neurons exposed to chemical insults that induce neuronal death by various mechanisms. 2 O 2) treatment produces toxic oxidative stress. Bacteria-derived lipopolysaccharide (LPS) treatment induces inflammatory cell death. Bacteria-derived lipopolysaccharide (LPS) treatment induces inflammatory cell death. Excessive glutamate (Glut) treatment causes excitotoxicity in neuronal cultures. 1-Methyl-4-phenylpyridinium (MPP + ) treatment causes cell death by inhibiting mitochondrial function.

[0354] Primary rat cortical neuron cultures were grown to maturity in complete neurobasal medium supplemented with B27 / GDNF / BDNF containing 10% FBS at 37°C and 5% CO2 and seeded at 5,000 cells / well in 384-well plates. Cultured cells were then treated with 1uM, 100nM, 10nM, 1nM, or 0.1nM of compound 1a for 15 minutes. Cells were then subjected to the following individual conditional insults for 24 hours: H 2 O 2 The mice were subjected to 1 uM LPS, 1 uM glutamate, and 500 uM MPP+. All treatments included recombinant human HGF (R&D Systems) at a concentration of 5 ng / ml.

[0355] To quantify neuroprotection, cell viability was measured using the Cell Titer-Glo Luminescent Cell Viability Assay (Promega, Cat #G7571). Data were normalized to DMSO+HGF control, which was defined as 100% cell viability. Statistical analysis for each group (n=4) was performed using one-way ANOVA with Tukey's multiple comparison test.

[0356] The results are shown in Table 13. Data are reported as statistically significant difference versus injured control, NS=not significant, +=p<0.05, ++=p<0.01, +++=p<0.001. TIFF2024546600000067.tif75165

[0357] In this study, administration of compound 1a acted as an effective neuroprotective treatment across a wide range of concentrations against all insults tested.The results indicate that enhancement of HGF / MET signaling by compound 1a exerts a potent neuroprotective effect on cultured rat neurons.

[0358] Example B13: Suppression of scopolamine-induced cognitive impairment. Mild cognitive impairment can be assessed in rodent models by administering various compounds that inhibit normal neurotransmitter signaling. Scopolamine is a well-known compound that antagonizes not only glutamatergic receptors such as NMDA receptors, but also muscarinic acetylcholine receptors and nicotinic receptors. Thus, scopolamine administration leads to cognitive impairment, which can be assessed with various cognitive tasks. Positive modulation of the HGF / MET system promotes NMDA receptor activation and has pro-cognitive effects. Two test compounds, compound 1a and compound 5a, were evaluated for their protective ability against scopolamine-induced cognitive impairment.

[0359] In this study, mild cognitive impairment was modeled by acute administration of 0.89 mg / kg scopolamine (dissolved in saline) to otherwise healthy male CD-1 mice (4–5 weeks old at the start of the study). Test compounds were administered orally 0.5 h before scopolamine exposure, and cognitive performance was assessed with a T-maze spontaneous alternation task 0.5 h after scopolamine exposure (1 h after test compound exposure). The experimental protocol consisted of one session, starting with one "forced choice" trial followed by 14 "free choice" trials. In the first "forced choice" trial, the animal was restrained in the start arm for 5 s and then released while either the left or right goal arm was blocked by a horizontal door. After the mouse entered the open goal arm and returned to the start position, the left and right gold doors were opened, and the animal was allowed to freely choose the left or right goal arm ("free choice trial"). An arm was considered entered when the animal placed all four forepaws into the arm. Normal animals choose to explore a previously unexplored arm, which is recorded as a spontaneous alternation. Exploring a previously entered arm indicates that the animal no longer remembers the previously explored arm and is considered a cognitive deficit behavior. The session was terminated and the animal removed from the maze after 14 free-choice trials or 10 min, whichever came first.

[0360] Analytical variables included percentage of spontaneous alternations (equation), the results of which are presented in Table 14. This percentage was defined as entering different goal arms of the T-maze on consecutive trials (e.g., left-right-left-right) and was calculated using the formula: (percentage of alternations = number of alternations / 14*100). Both compounds attenuated the degree of scopolamine-induced cognitive impairment as indicated by an increased percentage of spontaneous alternations compared to the scopolamine-only control group. Statistical analysis was performed by one-way ANOVA with Dunnett's multiple comparison post-hoc test. TIFF2024546600000068.tif73165

[0361] Although the present invention has been described in some detail by illustration and example for purposes of clarity of understanding, these descriptions and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are hereby incorporated by reference in their entireties.

Claims

1. A method of treating mild cognitive impairment in a subject in need thereof, comprising administering to a subject an effective amount of a compound of formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, L is a direct bond, -C(=O)-, -(CR ａ R ｂ ) ｍ -C(=O)-, -C(=O)-(CR ａ R ｂ ) ｍ - or - (CR ａ R ｂ ) ｍ - and Each R ａ and R ｂ are independently H, C １ -C ６ Alkyl, C ２ -C ６ Alkenyl, or C ２ -C ６ is alkynyl, R １ａ and R １ｂ are independently H, C １ -C ６ Alkyl, C ２ -C ６ Alkenyl, C ２ -C ６ Alkynyl, C １ -C ６ Alkoxy, halo, or C ６ -C １０ is arylalkyl, R ２ is H, oxo, or thioxo; R ３ is C ２ -C ６ Alkyl, C ３ -C ６ Alkenyl, C ３ -C ６ Alkynyl, C ３ -C １２ Cycloalkyl, C ３ -C ６ Cycloalkylalkyl, C ６ -C １０ arylalkyl, 5- to 10-membered heteroarylalkyl, or 5- to 10-membered heterocyclylalkyl; wherein the 5- to 10-membered heteroarylalkyl or 5- to 10-membered heterocyclylalkyl contains 1 to 3 heteroatoms selected from nitrogen and oxygen; R ４ is C ６ -C １０ aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heterocyclyl; wherein the 5- to 10-membered heteroaryl or 5- to 10-membered heterocyclyl contains 1 to 3 heteroatoms selected from nitrogen and oxygen; Each R ５ is independently １ -C ６ alkyl, oxo, or halo; R ６ is H, C １ -C ６ alkyl, or oxo; R ７ is H or oxo, m is 1 or 2; n is an integer from 0 to 3, Here, each C １ -C ６ Alkyl, C ２ -C ６ Alkenyl, C ２ -C ６ Alkynyl, C ３ -C １２ Cycloalkyl, C ３ -C １２ Cycloalkylalkyl, C ６ -C １０ Aryl, C ６ -C １０ Arylalkyl, 5- to 10-membered heteroaryl, 5- to 10-membered heteroarylalkyl, 5- to 10-membered heterocyclyl, and 5- to 10-membered heterocyclylalkyl are hydroxyl, halo, amino, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, cyano, -(C=O)NH ２ , nitro, -SO ２ (C １ -C ６ alkyl), and -CO ２ H).

2. L is -C(=O)- or -(CR ａ R ｂ ) ｍ The method of claim 1, wherein

3. The method of claim 1 or 2, wherein L is -C(=O)-.

4. L is - (CR ａ R ｂ ) ｍ The method according to claim 1 or 2, wherein

5. R ａ and R ｂ The method of claim 4 , wherein each is H and m is 1.

6. R １ａ and R １ｂ each independently represents H; halo, —CO ２ H, and -C(=O)NH ２ C optionally substituted with 1 to 3 substituents selected from １ -C ６ Alkyl; C １ -C ６ C optionally substituted with 1 to 3 substituents selected from alkoxy; halo; or halo and amino ６ -C １０ The method according to any one of claims 1 to 5, wherein the alkyl group is arylalkyl.

7. R １ａ and R １ｂ are each independently H, methyl, fluoro, 2-methylbutyl, -CH ２ F, methoxy, -CH ２ CO ２ H, -CH ２ C(=O)NH ２ 7. The method of claim 6, wherein the aryl group is 4-aminobenzyl, benzyl, or 4-aminobenzyl.

8. R １ａ and R １ｂ are each independently H or C １ -C ３ The method of claim 6, wherein the alkyl group is an alkyl group.

9. R １ａ is methyl, R １ｂ The method of claim 8 , wherein

10. R １ａ and R １ｂ The method of claim 8 , wherein each is H.

11. R ２ The method of any one of claims 1 to 10, wherein is H.

12. R ２ The method of any one of claims 1 to 10, wherein is thioxo.

13. R ２ The method of any one of claims 1 to 10, wherein is oxo.

14. R ３ But, C ３ -C ６ Alkyl, C ３ -C ６ Alkenyl, C ３ -C ６ Alkynyl, C ３ -C １２ Cycloalkyl, C ３ -C ６ Cycloalkylalkyl, C ６ -C １０ arylalkyl, 5- to 10-membered heteroarylalkyl, or 5- to 10-membered heterocyclylalkyl, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, arylalkyl, heteroarylalkyl, or heterocyclylalkyl is selected from the group consisting of hydroxyl, halo, amino, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, cyano, -(C=O)NH ２ , nitro, -SO ２ (C １ -C ６ alkyl), and -CO ２ The method of any one of claims 1 to 13, optionally substituted with 1 to 5 substituents selected from H.

15. R ３ But, Halo, C １ -C ３ Alkoxy, hydroxy, -NH ２ , -SO ２ (C １ -C ３ alkyl), and -C(=O)NH ２ C optionally substituted with 1 to 3 substituents selected from ２ -C ６ Alkyl; C ２ -C ６ Alkenyl; C ３ -C ６ cycloalkylalkyl; 5- to 6-membered heteroarylalkyl; 5- to 6-membered heterocyclylalkyl; or C ６ The method according to any one of claims 1 to 13, wherein the alkyl group is arylalkyl.

16. R ３ But, C １ -C ３ Alkoxy, hydroxy, -NH ２ , and -SO ２ (C １ -C ３ C substituted by 1 to 3 substituents selected from ２ The method of claim 15, wherein the alkyl group is an alkyl group.

17. R ３ The method according to any one of claims 14 to 16, wherein: 【Chemistry 2】

18. R ３ 18. The method of claim 17, wherein: 【Chemistry 3】

19. R ４ But halo, hydroxyl, C １ -C ６ Haloalkyl, and C １ -C ６ C optionally substituted with 1 to 3 substituents selected from haloalkoxy ６ -C １０ The method according to any one of claims 1 to 18, wherein the aryl is aryl.

20. R ４ But -CF ３ , -OCHF ２ 20. The method of claim 19, wherein the aryl group is phenyl substituted with 1 to 3 substituents selected from -OH, fluoro, and chloro.

21. R ４ 21. The method of claim 20, wherein: 【Chemistry 4】

22. R ４ 22. The method of claim 21 , wherein: 【Chemistry 5】

23. R ４ But halo, hydroxyl, C １ -C ６ Haloalkyl and C １ -C ６ The method of any one of claims 1 to 18, wherein the heteroaryl is 5 to 10 membered optionally substituted with 1 to 3 substituents selected from haloalkoxy.

24. R ４ But halo, hydroxyl, C １ -C ６ Haloalkyl, and C １ -C ６ 24. The method of claim 23, which is pyridyl or indolyl optionally substituted with 1 to 3 substituents selected from haloalkoxy.

25. R ４ but, 【Chemistry 6】 25. The method of claim 24, wherein:

26. R ４ but, 【Chemistry 7】 26. The method of claim 25, wherein:

27. R ４ But halo, hydroxyl, C １ -C ６ Haloalkyl and C １ -C ６ The method of any one of claims 1 to 18, wherein the heterocyclyl is a 5-10 membered heterocyclyl optionally substituted with 1 to 3 substituents selected from haloalkoxy.

28. R ４ 28. The method of claim 27, wherein is indolinyl.

29. R ４ but, 【Chemistry 8】 29. The method of claim 28, wherein:

30. -L-R ４ The method according to any one of claims 1 to 26, wherein: 【Chemistry 9】

31. The method of any one of claims 1 to 30, wherein n is 0.

32. The method of any one of claims 1 to 30, wherein n is 1.

33. R ５ The method of claim 32, wherein is oxo or halo.

34. R ５ The method of claim 33, wherein is oxo or fluoro.

35. R ６ The method of any one of claims 1 to 34, wherein is H.

36. R ７ The method of any one of claims 1 to 35, wherein is oxo.

37. 37. The method of any one of claims 1-10, 13-31, 35, and 36, wherein the compound is of formula (V) or a pharma- ceutically acceptable salt thereof: 【Chemistry 10】

38. L is -C(=O)- or -CH ２ - and R １ａ and R １ｂ are independently H or -CO ２ C optionally substituted with H １ -C ３ is alkyl, R ３ But, C ４ -C ５ Alkyl, C ４ -C ５ Alkenyl, or C ３ -C ５ Cycloalkyl-substituted C １ -C ３ is alkyl, R ４ But -CF ３ , -OCHF ２ 38. The method of claim 37, wherein the aryl group is phenyl or pyridyl substituted with 1 to 3 substituents selected from -OH, fluoro, and chloro.

39. A method of treating mild cognitive impairment, comprising administering to a subject in need thereof an effective amount of a compound selected from the compounds of Table 1A and pharma- ceutically acceptable salts thereof.

40. 2. The method of any one of the preceding claims, wherein the progression of dementia in the subject is slowed.

41. 2. The method of any one of the preceding claims, wherein the method improves cognitive function or slows the progression of cognitive impairment in the subject.

42. 2. The method of any one of the preceding claims, wherein the compound is administered orally or intravenously.

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