Bicyclic compounds and their use in the treatment of diseases

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] Cross-reference of related applications This application claims priority to U.S. Provisional Application No. 63 / 108,660, filed on 2 November 2020, the disclosure of which is incorporated herein by reference in its entirety.

[0002] This disclosure generally relates to compounds, compositions, and methods for preparing and using them for treating diseases such as neurodegenerative diseases. [Background technology]

[0003] Hepatocyte growth factor (HGF) is a multifaceted protein factor involved in numerous biological processes, including embryonic and organ development, regeneration, and inflammation. HGF plays a crucial role in the development and maturation of cortical, motor, sensory, sympathetic, and parasympathetic nerve cells. HGF is translated and secreted as inactive pro-HGF, but upon cleavage, the resulting α and β subunits are linked by disulfide bonds to form an active heterodimer. HGF expression primarily occurs in mesenchymal cells such as fibroblasts, chondrocytes, adipocytes, and endothelial cells. Expression has also been shown in the central nervous system (CNS), including neurons, astrocytes, and ependymal cells (Nakamura and Mizuno, 2010). All biological activity of HGF is mediated by MET, a transmembrane receptor tyrosine kinase that functions as the only known receptor for HGF. MET is known to be involved in various biological processes and has been shown to play a role in development, regeneration, and injury responses. When HGF binds to the extracellular domain of MET, the MET protein homodimerizes, leading to autophosphorylation of the intracellular domain. Phosphorylation of the intracellular domain of MET 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, particularly PI3K / Akt, Ras / Raf / MAPK, RAC1 / CDC42, and RAP / FAK, influencing various cellular components, including gene regulation, cytoskeletal rearrangement, cell cycle progression, cell adhesion, survival, and proliferation (Organ and Tsao, 2011).

[0004] Since HGF has been shown to play roles 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 system is an ideal target for therapeutic agents for various disease states. Treatments by modulating HGF activity have been proposed 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, compounds with highly effective and useful effects for modulating HGF / MET signaling activity have not yet been searched for or discovered.

[0005] Although progress has been made in the art, improved compounds and methods for the treatment of HGF-regulated diseases are still needed. Thus, in one aspect, compounds that modulate HGF for use in the treatment of neurodegenerative diseases are provided.

Summary of the Invention

[0006] In certain embodiments, described herein are compounds and compositions thereof for modulating hepatocyte growth factor (HGF) for the treatment of diseases. Non-limiting exemplary embodiments include the following.

[0007] Embodiment 1. A compound of formula (I):

Chemical formula

[0008] Embodiment 2.L is -C(=O)- or -(CR a R b ) m - The compound of Embodiment 1, or a pharmaceutically acceptable salt thereof.

[0009] Embodiment 3. The compound of Embodiment 1 or 2, or a pharmaceutically acceptable salt thereof, wherein L is -C(=O)-.

[0010] Embodiment 4.L is -(CR a R b ) m - The compound of Embodiment 1 or 2, or a pharmaceutically acceptable salt thereof.

[0011] Embodiment 5.R a and R b The compound of Embodiment 4, or a pharmaceutically acceptable salt thereof, wherein each of the elements is H and m is 1.

[0012] Embodiment 6.R 1a and R 1b However, each is independently a C1-C6 alkyl; C1-C6 alkoxy; halo; or a C6-C6 alkyl; C1-C6 alkoxy; halo; or C1 alkoxy; C1-C6 alkyl; C1-C6 alkoxy; halo; or C1-C 10 A compound from any one of Embodiments 1 to 5, which is an arylalkyl compound, or a pharmaceutically acceptable salt thereof.

[0013] Embodiment 7.R 1a and R1b The compounds of Embodiment 6, or pharmaceutically acceptable salts thereof, wherein each is independently H, methyl, fluoro, 2-methylbutyl, -CH2F, methoxy, -CH2CO2H, -CH2C(=O)NH2, benzyl, or 4-aminobenzyl.

[0014] Embodiment 8.R 1a and R 1b The compounds of Embodiment 6, or pharmaceutically acceptable salts thereof, wherein each is independently H or C1-C3 alkyl.

[0015] Embodiment 9.R 1a is methyl, and R 1b A compound of Embodiment 8, or a pharmaceutically acceptable salt thereof, wherein the compound is H.

[0016] Embodiment 10.R 1a and R 1b The compound of Embodiment 8, or a pharmaceutically acceptable salt thereof, wherein each of these is H.

[0017] Embodiment 11.R 2 A compound from any one of Embodiments 1 to 10, or a pharmaceutically acceptable salt thereof, wherein the compound is H.

[0018] Embodiment 12.R 2 A compound from any one of Embodiments 1 to 10, or a pharmaceutically acceptable salt thereof, wherein the compound is thioxo.

[0019] Embodiment 13.R 2 A compound from any one of Embodiments 1 to 10, or a pharmaceutically acceptable salt thereof, wherein the compound is an oxo.

[0020] Embodiment 14.R 3 However, C3-C6 alkyl, C3-C6 alkenyl, C3-C6 alkynyl, C3-C 12 Cycloalkyl, C3-C6 cycloalkylalkyl, C6-C 10An arylalkyl, a 5-10 membered heteroarylalkyl, or a 5-10 membered heterocyclylalkyl, where the alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, arylalkyl, heteroarylalkyl, or heterocyclylalkyl is optionally substituted with 1-5 substituents selected from hydroxyl, halo, amino, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, cyano, -(C=O)NH2, nitro, -SO2(C1-C6alkyl), and -CO2H, one of the compounds from Embodiments 1-13, or a pharmaceutically acceptable salt thereof.

[0021] Embodiment 15.R 3 The compound is one of the compounds from Embodiments 1 to 13, or a pharmaceutically acceptable salt thereof, which is a C2-C6 alkyl; C2-C6 alkenyl; C3-C6 cycloalkylalkyl; 5-6 member heteroarylalkyl; 5-6 member heterocyclylalkyl; or C6 arylalkyl, optionally substituted with 1 to 3 substituents selected from halo, C1-C3 alkoxy, hydroxy, -NH2, -SO2(C1-C3 alkyl), and -C(=O)NH2.

[0022] Embodiment 16.R 3 The compound of Embodiment 15, or a pharmaceutically acceptable salt thereof, is a C2 alkyl group substituted with 1 to 3 substituents selected from C1-C3 alkoxy, hydroxyl, -NH2, and -SO2(C1-C3 alkyl).

[0023] Embodiment 17.R 3 However, the following are any one compound from Embodiments 14 to 16, or a pharmaceutically acceptable salt thereof: [ka]

[0024] Embodiment 18.R 3 However, the following are the compounds of Embodiment 17, or pharmaceutically acceptable salts thereof: [ka]

[0025] Embodiment 19.R 4 However, C6-C is optionally substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy. 10 A compound from any one of Embodiments 1 to 18, or a pharmaceutically acceptable salt thereof, which is an aryl compound.

[0026] Embodiment 20.R 4 The compound of Embodiment 19, or a pharmaceutically acceptable salt thereof, is a phenyl compound substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro.

[0027] Embodiment 21.R 4 However, the following are the compounds of Embodiment 20, or pharmaceutically acceptable salts thereof: [ka]

[0028] Embodiment 22.R 4 However, the following are the compounds of Embodiment 21, or pharmaceutically acceptable salts thereof: [ka]

[0029] Embodiment 23.R 4 The compound is one of the compounds from Embodiments 1 to 18, or a pharmaceutically acceptable salt thereof, which is a 5- to 10-membered heteroaryl substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy.

[0030] Embodiment 24.R 4The compound of Embodiment 23, or a pharmaceutically acceptable salt thereof, is a pyridyl or indolyl compound optionally substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy.

[0031] Embodiment 25. R 4 but [ka] The compound of Embodiment 24, or a pharmaceutically acceptable salt thereof.

[0032] Embodiment 26. R 4 but [ka] The compound of Embodiment 25, or a pharmaceutically acceptable salt thereof.

[0033] Embodiment 27.R 4 The compound is one of the compounds from Embodiments 1 to 18, or a pharmaceutically acceptable salt thereof, which is a 5- to 10-membered heterocycline optionally substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy.

[0034] Embodiment 28.R 4 The compound of Embodiment 27, or a pharmaceutically acceptable salt thereof, wherein the compound is indolinyl.

[0035] Embodiment 29.R 4 but [ka] The compound of Embodiment 28, or a pharmaceutically acceptable salt thereof.

[0036] Embodiment 30.-LR 4 However, the following are any one compound from Embodiments 1 to 26, or a pharmaceutically acceptable salt thereof: [ka]

[0037] Embodiment 31. Any one of the compounds from Embodiments 1 to 30, or a pharmaceutically acceptable salt thereof, wherein n is 0.

[0038] Embodiment 32. A compound from any one of Embodiments 1 to 30, or a pharmaceutically acceptable salt thereof, wherein n is 1.

[0039] Embodiment 33.R 5 The compound of Embodiment 32, or a pharmaceutically acceptable salt thereof, is oxo or halo.

[0040] Embodiment 34.R 5 The compound of Embodiment 33, or a pharmaceutically acceptable salt thereof, is oxo or fluoro.

[0041] Embodiment 35.R 6 A compound from any one of Embodiments 1 to 34, or a pharmaceutically acceptable salt thereof, wherein the compound is H.

[0042] Embodiment 36.R 7 A compound from any one of Embodiments 1 to 35, or a pharmaceutically acceptable salt thereof, wherein the compound is an oxo.

[0043] Embodiment 37. The compound is one of the compounds of Embodiments 1-10, 13-31, 35, and 36, represented by formula (V), or a pharmaceutically acceptable salt thereof: [ka]

[0044] Embodiment 38. L is -C(=O)- or -CH2-, R 1a and R 1bHowever, independently, they are C1-C3 alkyl groups optionally substituted with H or -CO2H. R 3 However, it is a C1-C3 alkyl substituted with a C4-C5 alkyl, a C4-C5 alkenyl, or a C3-C5 cycloalkyl. R 4 The compound of Embodiment 37, or a pharmaceutically acceptable salt thereof, is a phenyl or pyridyl substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro.

[0045] Embodiment 39. A compound selected from the compounds in Table 1A and their pharmaceutically acceptable salts.

[0046] Embodiment 40. A pharmaceutical composition comprising one compound from any of Embodiments 1 to 39 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0047] Embodiment 41. A method for regulating hepatocyte growth factor in a subject requiring such regulation, comprising administering to the subject an effective amount of one compound from any one of Embodiments 1 to 39 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 40.

[0048] Embodiment 42. The method of Embodiment 41, wherein the adjustment includes treating a disease, condition, or injury.

[0049] Embodiment 43. The method of Embodiment 42, wherein the disease, condition, or injury is a neurodegenerative disease, spinal cord injury, traumatic brain injury, or sensorineural hearing loss.

[0050] Embodiment 44. The method of Embodiment 42 or 43, wherein the disease, condition, or injury is a neurodegenerative disease.

[0051] Embodiment 45. The method of Embodiment 44, wherein the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, Huntington's disease, or amyotrophic lateral sclerosis (ALS).

[0052] Embodiment 46. The method of Embodiment 45, wherein the neurodegenerative disease is Alzheimer's disease or Parkinson's disease.

[0053] Embodiment 47. A method for treating dementia or slowing its progression in a subject in need thereof, comprising administering to the subject an effective amount of one compound from any one of Embodiments 1 to 39 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 40.

[0054] Embodiment 48. The method of Embodiment 47, wherein the dementia is related to Alzheimer's disease or Parkinson's disease.

[0055] Embodiment 49. A method for preventing cognitive impairment in a subject in need thereof, comprising administering to the subject an effective amount of one compound from any one of Embodiments 1 to 39 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 40.

[0056] Embodiment 50. A method for treating, restoring, or preventing a disease, condition, or injury relating to nerve tissue in a subject in need thereof, comprising administering to the subject an effective amount of one compound from any one of Embodiments 1 to 39 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 40.

[0057] Embodiment 51. A method for treating or preventing a central nervous system disorder, a peripheral nervous system disorder, neuropathic pain, anxiety, or depression in a subject in need thereof, the method comprising administering to the subject an effective amount of one compound from any one of Embodiments 1 to 39 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Embodiment 40. [Modes for carrying out the invention]

[0058] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this disclosure belongs. Some specific details are provided in the following description to provide a complete understanding of the various embodiments of this disclosure. It should be understood that the above general description and the following detailed description are illustrative and descriptive only and do not limit any subject matter claimed. In the event of any conflict between any material incorporated herein by reference and the express content of this disclosure, the express content shall prevail. In this application, the use of the singular includes the plural unless otherwise specified. It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include multiple references unless otherwise clearly indicated by the context. In this application, the use of "or" means "and / or" unless otherwise specified. Furthermore, the use of the term "including," as well as other forms such as "include," "includes," and "included," is not limited to this.

[0059] Unless the context requires a different interpretation, the word “comprise” and its variations such as “comprises” and “comprising” throughout this specification and the claims shall be interpreted as having an open and comprehensive meaning, i.e., “including, but not limited to, ~”.

[0060] In the description of this invention, any concentration range, percentage range, ratio range, or integer range is understood to include any integer value within the enumerated range, and, where applicable, fractions thereof (such as 1 / 10 and 1 / 100 of an integer), unless otherwise specified. Similarly, any number range enumerated herein with respect to any physical characteristics of a polymer, such as subunits, size, or thickness, is understood to include any integer within the enumerated range, unless otherwise specified. Where used herein, the terms “about” and “approximately” mean ±20%, ±10%, ±5%, or ±1% of the indicated range, value, or structure, unless otherwise specified.

[0061] Any reference throughout this specification to "one embodiment" or "an embodiment" means that any particular feature, structure, or characteristic described in relation to an embodiment is included in at least one embodiment of this disclosure. Therefore, the phrases "one embodiment" or "an embodiment" appearing in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, any particular feature, structure, or characteristic can be combined in any preferred manner in one or more embodiments.

[0062] "Amino" refers to the -NH2 radical.

[0063] "Carboxy" or "carboxyl" refers to the -CO2H radical.

[0064] "Cyano" refers to the -CN radical.

[0065] "Hydroxy" or "hydroxyl" refers to the -OH radical.

[0066] "Nitro" refers to the -NO2 radical.

[0067] "Oxo" refers to an =O substituent.

[0068] "Thioxo" refers to the =S substituent.

[0069] "Thiol" refers to the -SH substituent.

[0070] "Alkyl" is a non-branched or branched saturated hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, with 1 to 12 carbon atoms (C1-C 12 Alkyl groups, preferably those having 1 to 8 carbon atoms (C1-C8 alkyl), 1 to 6 carbon atoms (C1-C6 alkyl), or 1 to 3 carbon atoms (C1-C3 alkyl), and bonded to the rest of the molecule by a single bond, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless otherwise specified herein, alkyl groups may be optionally substituted.

[0071] "Alkenyl" is a non-branched or branched unsaturated hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, containing one or more carbon-carbon double bonds and 2 to 12 carbon atoms (C2-C 12 Alkenyls) preferably have 2 to 8 carbon atoms (C2-C8 alkenyls) or 2 to 6 carbon atoms (C2-C6 alkenyls) and are bonded to the rest of the molecule by single bonds, such as ethenyl, propa-1-enyl, buta-1-enyl, pento-1-enyl, penta-1,4-dienyl, etc. Unless otherwise specified herein, the alkenyl group may be optionally substituted.

[0072] "Alkynyl" is a non-branched or branched unsaturated hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, containing one or more carbon-carbon triple bonds and 2 to 12 carbon atoms (C2-C 12Alkynnyl groups, preferably those having 2 to 8 carbon atoms (C2-C8 alkynyl) or 2 to 6 carbon atoms (C2-C6 alkynyl) and being bonded to the rest of the molecule by single bonds, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc. Unless otherwise specified herein, the alkynyl group may be optionally substituted.

[0073] "alkoxy" is the formula -OR a It refers to the radical of, in the formula, R a This is an alkyl radical as defined above, containing 1 to 12 carbon atoms. Preferred alkoxy groups have 1 to 6 carbon atoms (i.e., C1-C6 alkoxy) or 1 to 3 carbon atoms (i.e., C1-C3 alkoxy) in the alkyl radical. Unless otherwise specified herein, alkoxy groups are optionally substituted.

[0074] An "aromatic ring" refers to a cyclic planar portion (i.e., radical) of a molecule that has a ring of resonance bonds that exhibits improved stability compared to other bonding configurations with the same group of atoms. Generally, aromatic rings contain covalently bonded, coplanar atoms and an even number of π electrons that are not multiples of 4 (i.e., 4n + 2π electrons (where n = 0, 1, 2, 3, etc.)) (e.g., alternating double and single bonds). Aromatic rings include, but are not limited to, phenyl, naphthenyl, imidazolyl, pyrrolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridonyl, pyridadinyl, and pyrimidonyl. Unless otherwise specified herein, aromatic rings include all radicals that are optionally substituted.

[0075] "Aryl" consists of 6 to 18 carbon atoms and at least one aromatic ring (i.e., C6-C 18 It contains an aryl compound, preferably 6 to 10 carbon atoms (i.e., C6-C6). 10refers to a carbocyclic ring system radical having (aryl). For the purposes of embodiments of the present disclosure, an aryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include a fused or bridged ring system. 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, preiadene, pyrene, and triphenylene. Unless otherwise specified herein, an aryl group is optionally substituted.

[0076] “Arylalkyl” has the formula -R b -R c refers to a radical of the formula, where R b is an alkylene chain and R c is one or more aryl radicals as defined above, for example, benzyl, diphenylmethyl, etc. An arylalkyl group contains a C1-C 10 alkylene chain connected to the aryl radical (i.e., C6-C 10 arylalkyl). Unless otherwise specified herein, an arylalkyl group is optionally substituted.

[0077] “Cycloalkyl” is a stable non-aromatic monocyclic or polycyclic carbocyclic radical consisting of only carbon atoms and hydrogen atoms, which can include a fused or bridged ring system, having 3 to 15 carbon atoms (i.e., C3-C 15 cycloalkyl), preferably 3 to 10 carbon atoms (i.e., C3-C 10 ​​"(cycloalkyl) or having 3 to 6 carbon atoms (i.e., C3-C6 cycloalkyl), being saturated or unsaturated, and being bonded by a single bond to the rest of the molecule. 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 otherwise specified herein, cycloalkyl groups are optionally substituted."

[0078] "Cycloalkylalkyl" refers to the radical of the formula -R b -R c wherein, 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. The cycloalkylalkyl group may contain a C1-C 12 alkylene chain connected to a C3-C 10 cycloalkyl radical (i.e., C3-C 12 cycloalkylalkyl) or a C1-C 10 alkylene chain connected to a C3-C6 cycloalkyl radical (i.e., C3-C6 cycloalkylalkyl). Unless otherwise specified herein, cycloalkylalkyl groups are optionally substituted."

[0079] "Fused" refers to any ring structure described herein that is fused to an existing ring structure in the compounds 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 forms part of the fused heterocyclyl ring or fused heteroaryl ring is replaced by a nitrogen atom."

[0080] "Halo" or "halogen" refers to bromo, chloro, fluoro, or iodine.

[0081] "Haloalkyl" refers to an alkyl radical as defined above, which is substituted by one or more halo radicals as defined above, such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Preferred haloalkyls include alkyl groups having 1 to 6 carbon atoms and substituted by one or more halo radicals (i.e., C1-C6 haloalkyls). The halo radicals may all be the same or they may be different. Unless otherwise specified herein, haloalkyls are optionally substituted.

[0082] "Halalkoxy" is a formula -OR a It refers to the radical of, in the formula, R a This is a haloalkyl radical as defined herein, containing 1 to 12 carbon atoms. Preferred haloalkoxy groups include alkoxy groups having 1 to 6 carbon atoms (i.e., C1-C6 haloalkoxy) or 1 to 3 carbon atoms (C1-C3 haloalkoxy) and substituted with one or more halo radicals. The halo radicals may all be the same or may all be different. Unless otherwise specified herein, the haloalkoxy groups are optionally substituted.

[0083] "Heteroaryl" refers to an aromatic group having a monocyclic, polycyclic, or multiple fused rings containing one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur (e.g., a 5- to 14-membered ring system). As used herein, a heteroaryl contains 1 to 10 ring carbon atoms and 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur within the ring. Preferred heteroaryl groups have a 5- to 10-membered ring system containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur (i.e., a 5- to 10-membered heteroaryl) and a 5- to 6-membered ring system containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur (i.e., a 5- to 6-membered heteroaryl). For the purposes of the embodiments of this disclosure, heteroaryl radicals may be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which may include fused or bridged ring systems. Examples of heteroaryl groups include pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridadinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Heteroaryls may contain one or more N-oxide (NO-) moieties, such as pyridine-N-oxide. Unless otherwise specified herein, heteroaryl groups are optionally substituted.

[0084] "Heteroarylalkyl" is a compound of the formula -R b -R c It refers to the radical of, in the formula, R b It is an alkylene chain, and R c A heteroarylalkyl group is one or more heteroaryl radicals as defined above. A heteroarylalkyl group is a C1-C group connected to a 5-10 member heteroaryl group. 10 C1-C linked to an alkylene chain (i.e., a 5-10 member heteroarylalkyl) or a 5-6 member heteroaryl group 10 It may contain alkylene chains (i.e., 5-6 member heteroarylalkyl groups). Unless otherwise specified herein, heteroarylalkyl groups are optionally substituted.

[0085] A "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, condensed heterocyclyl groups, and spiroheterocyclyl groups. Heterocyclyls can be monocyclic or polycyclic, where the polycyclic may be condensed, bridged, or spiro and may 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 bonding (i.e., it can be bonded via carbon atoms or heteroatoms). Furthermore, the term heterocyclyl is intended to encompass any non-aromatic ring containing at least one heteroatom that can be condensed into an aryl ring or a heteroaryl ring, regardless of bonding to the rest of the molecule. As used herein, a heterocyclil 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 heterocyclils have a ring system containing 1 to 4 heteroatoms selected from nitrogen and oxygen with 5 to 10 members (i.e., a 5 to 10-membered heterocyclil) or a ring system containing 1 to 4 heteroatoms selected from nitrogen and oxygen with 5 to 8 members (i.e., a 5 to 8-membered heterocyclil).Examples of heterocyclyl groups include dioxolanil, thienyl[1,3]dithianil, decahydroisoquinolyl, imidazolinil, imidazolidinil, isothiazolidinil, isoxazolidinil, morpholinil, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinil, 2-oxopiperidinil, 2-oxopyrrolidinil, oxazolidinil, piperidinil, piperazinil, 4-piperidonil, pyrrolidinil, pyrazolidinil, quinuclidinil, thiazolidinil, tetrahydrofuryl, trithianil, tetrahydropyranil, thiomorpholinil, thiamorpholinil, 1-oxo-thiomorpholinil, and 1,1-dioxo-thiomorpholinil. Unless otherwise specified herein, heterocyclyl groups are substituted by choice.

[0086] "Heterocyclylalkyl" is a compound of formulas -R b -R c It refers to the radical of, in the formula, R b It is an alkylene chain, and R c A is one or more heterocyclyl radicals as defined above. A heterocyclylalkyl group is a C1-C group connected to a 5- to 10-membered heterocyclyl radical. 10 C1-C connected to an alkylene chain (i.e., a 5-10 member heterocyclylalkyl) or a 5-8 member heterocyclyl radical 10 It may contain alkylene chains (i.e., 5- to 8-membered heterocyclylalkyl groups). Unless otherwise specified herein, the heterocyclylalkyl groups are optionally substituted.

[0087] In some embodiments, the term “substituted” as used herein refers to the above group or other substituents (e.g., C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 12 Cycloalkyl, C3-C 12In any of the cycloalkylalkyl, aryl, and heteroaryl groups, at least one hydrogen atom (e.g., 1, 2, 3, or all hydrogen atoms) is replaced by a bond to a halogen atom such as F, Cl, Br, and I (i.e., "halo"); an oxygen atom in a group such as a hydroxyl group or alkoxy group (e.g., alkoxy or haloalkoxy); a nitrogen atom in a group such as an amine (e.g., -NH2), an amide (e.g., -(C=O)NH2), and a nitro; an alkyl group containing one or more halogens such as F, Cl, Br, and I (e.g., haloalkyl); and a non-hydrogen atom such as a cyano.

[0088] L, R 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 Each selection regarding is understood to be substituted by arbitrary selection as described above, provided that all valencies are satisfied by substitution unless otherwise specified. Specifically, L, R 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 Each of the selections is made on the condition that such substitution results in a stable molecule (for example, groups such as H and halos are not substituted by choice), unless otherwise specified.

[0089] The “effective amount” or “therapeutically effective amount” of a compound or composition refers to the amount of the compound or composition that produces the desired intended result based on the disclosure herein. The effective amount is determined by standard pharmaceutical procedures in cell culture or experimental animals, for example, but is not limited to ED. 50 (Therapeutic dose effective in 50% of the population) and LD 50This can be determined by determining the lethal dose (for 50% of the population). In some embodiments, an effective dose of the compound results in symptom reduction or inhibition or prolonged survival in the subject (i.e., human patients). Multiple doses of the compound may be required to achieve the desired result.

[0090] "Treating" or "treating" a disease in question means 1) preventing the onset of the disease in patients who are predisposed to the disease or who have not yet exhibited symptoms of the disease, 2) inhibiting the disease or suppressing its onset, or 3) alleviating the disease or bringing it into remission. As used herein, "treatment" or "treating" refers to an approach to obtain beneficial or desirable outcomes, including clinical outcomes. For the purposes of this disclosure, beneficial or desirable outcomes include, but are not limited to, one or more of the following: reducing one or more symptoms of the disease or disorder in question; reducing the severity of the disease or disorder; stabilizing the disease or disorder (e.g., preventing or delaying the worsening of the disease or disorder); delaying the onset or recurrence of the disease or disorder; delaying or slowing the progression of the disease or disorder; improving the state of the disease or disorder; providing remission (whether partial or total) of the disease or disorder; reducing the dose of one or more other drugs required to treat the disease or disorder; enhancing the effect of another drug used to treat the disease or disorder; delaying the progression of the disease or disorder; improving quality of life; and / or extending survival. Mitigation of the pathological consequences of the disease or disorder is also included in “treatment.” The methods of the present invention aim to achieve one or more of these modes of treatment.

[0091] As used herein, the terms "individual(s)", "subject(s)", and "patient(s)" mean 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.

[0092] As used herein, the term "therapeutic effect" encompasses the therapeutic and / or prophylactic benefits described herein. A therapeutic effect includes delaying or eliminating the onset of a disease or condition; delaying or eliminating the development of symptoms of a disease or condition; slowing, halting, or reversing the progression of a disease or condition; effecting a partial or complete remission of a disease or condition; or any combination thereof.

[0093] The terms "co-administered", "administered in combination with", and their grammatical equivalents, as used herein, when referring to the administration of two or more agents to an animal, including a human, encompass the presence of both agents and / or their metabolites 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 a single composition in which both agents are present.

[0094] "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.

[0095] "Pharmaceutically acceptable salts" include both acid addition salts and base addition salts.

[0096] "Pharmacologically acceptable acid addition salts" refer to salts that retain the biological efficacy and properties of their free base and are not biologically or otherwise undesirable, and include, but are not limited to, those formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, and but are 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, carbonate, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, and glycerin. It is formed with organic acids such as thisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucinic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid.

[0097] A "pharmaceutically acceptable base addition salt" refers to a salt that retains the biological efficacy and properties of the free acid and is not biologically or otherwise undesirable. These salts are prepared by adding an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, salts of sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Preferred inorganic salts are salts of ammonium, sodium, potassium, calcium, and magnesium. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally 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, hydravamin, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0098] In some embodiments, pharmaceutically acceptable salts include quaternary ammonium compounds such as quaternary amine alkyl halide salts (e.g., methyl bromide).

[0099] As used herein, “therapeutic agent” refers to a biological, pharmaceutical, or chemical compound or other part. 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, such as small molecules and oligomers (e.g., oligopeptides and oligonucleotides), as well as synthetic organic compounds based on various core structures. In addition, compounds for screening can be obtained from various natural sources, such as plant or animal extracts.

[0100] The term "in vivo" refers to events that occur within the body of a subject.

[0101] Embodiments of the present disclosure are also intended to encompass all pharmaceutically acceptable compounds of formula (I) that are isotopically labeled by having one or more atoms replaced by atoms having different atomic masses or mass numbers (i.e., “isotope 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, for example, 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 is one example. These radiolabeled compounds may be useful in determining or measuring the efficacy of compounds by characterizing, for example, the site or mode of action, or the binding affinity to pharmacologically important sites of action. Compounds of formula (I) that are specific isotope-labeled, for example, those incorporating radioisotopes, are useful in tissue distribution studies of drugs and / or substrates. The radioisotope tritium, i.e., 3 H, and carbon-14, that is, 14 C is particularly useful for this purpose because it is easy to implement and has readily available detection methods.

[0102] Deuterium, that is, 2 Substitution with heavier isotopes, such as 1H, may be preferable in some situations because it can result in higher metabolic stability, for example, an extended in vivo half-life or a reduction in the required dose, thus offering certain therapeutic benefits.

[0103] 11 C, 18 F, 15 O and 13 Substitution with positron-emitting isotopes such as 1N may be useful in positron emission tomography (PET) studies to investigate substrate receptor occupancy. Isotope-labeled compounds of formula (I) can generally be prepared by prior art known to those skilled in the art, or by a process similar to that described in the examples below, using a suitable isotope-labeled reagent instead of the previously used unlabeled reagent.

[0104] Certain embodiments are also intended to include in vivo metabolites of the compounds of the Disclosure. Such products may arise from enzymatic processes, for example, oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound. Thus, embodiments include compounds resulting from processes that involve administering the compounds of the Disclosure to a mammal for a period of time sufficient for the metabolites to be produced. Such products are typically identified by administering a radiolabeled compound of the Disclosure to an animal such as a rat, mouse, guinea pig, monkey, or human in a detectable dose, allowing sufficient time for metabolism to occur, and then isolating the conversion product from urine, blood, or other biological samples.

[0105] "Stable compound" and "stable structure" refer to compounds that possess sufficient robustness to remain intact when isolated from a reaction mixture to a useful purity and formulated into an effective therapeutic agent.

[0106] In many cases, crystallization produces solvates of the compounds of this disclosure. As used herein, the term “solvate” refers to an aggregate comprising 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) may exist as a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate, and so on, as well as the corresponding solvates. In some embodiments, the compound of formula (I) is a true solvate, while in other embodiments, the compound of this disclosure simply holds incidental water or is a mixture of water and some incidental solvent.

[0107] "Optional" or "by optional choice" means that the event of the situation described thereafter may or may not occur, and that the description includes both cases in which such event or situation occurs and cases in which it does not occur. For example, "optionally substituted aryl" means that the aryl radical may or may not be substituted, and that the description includes both substituted and unsubstituted aryl radicals. No infinite polymer or similar structures achieved by defining substituents with an unlimited number of further substituents (e.g., a substituted aryl having a substituted alkyl, the substituted alkyl itself being substituted with a substituted aryl group, and the substituted aryl group being further substituted with a substituted heteroalkyl group) are intended to be included herein. Similarly, the above definitions are not intended to include unacceptable substitution patterns (e.g., a methyl group substituted with five fluorine atoms, or a heteroaryl group having two adjacent oxygen ring atoms). Such unacceptable substitution patterns are well known to those skilled in the art.

[0108] "Pharmaceutical composition" or "pharmaceutically acceptable composition" means a formulation of the compounds of this disclosure with a medium commonly accepted in the art for delivering the biologically active compounds to a mammal, such as a human. Such a medium may include any pharmaceutically acceptable carrier, diluent, or excipient for that purpose.

[0109] "Pharmacologically acceptable carriers, diluents or excipients" include, but are not limited to, any adjuvants, carriers, excipients, fluidizers, sweeteners, diluents, preservatives, colorants, flavor enhancers, surfactants, humectants, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers approved by the U.S. Food and Drug Administration as acceptable for use in humans or domesticated animals.

[0110] Compounds of formula (I), or their pharmaceutically acceptable salts or isotopic forms, may contain one or more centers resulting in geometric asymmetry, thus giving rise to enantiomers, diastereomers, and other stereoisomers defined from an absolute stereochemical standpoint as (R)- or (S)-, or in the case of amino acids, (D)- or (L)-. Therefore, 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 can be prepared using chiral synthons or chiral reagents, or they can be resolved using prior art, e.g., chromatography and fractional crystallization. Prior art for preparing / isolating individual enantiomers includes chiral synthesis from suitable optically pure precursors, or resolution of racemics (or racemics of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). Where a compound described herein contains an olefin double bond or other geometrically asymmetric center, unless otherwise specified, the compound is intended to include both E and Z geometric isomers. Similarly, all tautomers are also intended to be included.

[0111] A "stereoisomer" refers to a compound that consists of the same atoms bonded together by the same bonds but has different three-dimensional structures that are not interchangeable. This disclosure intends to cover a variety of stereoisomers and mixtures thereof, and includes "enantiomers," which refer to two stereoisomers that are mirror images of each other and whose molecules cannot be superimposed.

[0112] A "diastereomer" is a stereoisomer that has at least two chiral atoms but is not a mirror image of one another.

[0113] A "tautomer" refers to a molecule in which a proton moves from one atom to another atom in the same molecule. Therefore, embodiments include tautomers of the compounds of this disclosure.

[0114] The chemical nomenclature protocol and structural diagrams used in this specification are a modified version of the I.U.P.A.C. nomenclature using the ACD / Name Version 9.07 software program and / or the ChemDraw Ultra Version 11.0.1 software nomenclature program (CambridgeSoft). In the complex chemical names adopted in this specification, substituents are typically named before the group to which the substituent is attached. For example, cyclopropylethyl contains an ethyl backbone with a cyclopropyl substituent. Except as described below, in the chemical structural diagrams of this specification, all bonds except those on some carbon atoms are specified, and the bonds on carbon atoms are assumed to be bonded to hydrogen atoms sufficient to satisfy the valence.

[0115] Various features of the invention may be described in the context of a single embodiment, but the features may be provided separately or in any suitable combination. Conversely, the invention may be described herein in the context of separate embodiments for clarity, but the invention may be implemented in a single embodiment.

[0116] Compound In one aspect, provided herein is a compound of formula (I):

Chemical formula

[0117] 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 -C(=O)-. In some embodiments, L is -C(=O)-(CR a R b ) m - is

[0118] In some embodiments, each R a and R b R is independently H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. In some embodiments, each R a and R b R is independently H, C1-C3 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl. In some embodiments, R a and R b These are H, respectively. In some embodiments, Ra H is H. In some embodiments, R a is a C1-C6 alkyl such as methyl, ethyl, or propyl. In some embodiments, R a is a C2-C6 alkenyl such as vinyl or propenyl. In some embodiments, R a is a C2-C6 alkynyl such as ethinyl or propynyl. In some embodiments, R b H is H. In some embodiments, R b is a C1-C6 alkyl such as methyl, ethyl, or propyl. In some embodiments, R b is a C2-C6 alkenyl such as vinyl or propenyl. In some embodiments, R b These are C2-C6 alkynyl compounds such as ethinyl or propynyl.

[0119] In some embodiments, R 1a and R 1b These are independently H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo, or C6-C 10 It is an arylalkyl. In some embodiments, R 1a H is H. In some embodiments, R 1a is a C1-C6 alkyl such as methyl, ethyl, or propyl. In some embodiments, R 1a is a C2-C6 alkenyl such as vinyl or propenyl. In some embodiments, R 1a is a C2-C6 alkynyl such as ethinyl or propynyl. In some embodiments, R 1a R is a C1-C6 alkoxy such as methoxy, ethoxy, or propoxy. In some embodiments, R 1a is a halo such as fluoro, chloro, or bromo. In some embodiments, R 1a C6-C, such as benzyl 10 It is an arylalkyl. In some embodiments, R1b H is H. In some embodiments, R 1b is a C1-C6 alkyl such as methyl, ethyl, or propyl. In some embodiments, R 1b is a C2-C6 alkenyl such as vinyl or propenyl. In some embodiments, R 1b is a C2-C6 alkynyl such as ethinyl or propynyl. In some embodiments, R 1b R is a C1-C6 alkoxy such as methoxy, ethoxy, or propoxy. In some embodiments, R 1b is a halo such as fluoro, chloro, or bromo. In some embodiments, R 1b C6-C, such as benzyl 10 It is an arylalkyl.

[0120] In some embodiments, R 1a and R 1b Each of these is independently a C1-C6 alkyl; C1-C6 alkoxy; halo; or a C6-C alkyl; C1-C6 alkoxy; halo; or C6-C alkyl; C1-C6 alkoxy; halo; or C1-C6 alkyl; C1-C6 alkoxy; halo; or C1-C6 alkyl; C1-C6 alkoxy; halo; or C1-C6 alkyl; C1-C6 alkoxy; halo; or C1-C6 alkoxy; alkoxy; C1-C6 alkoxy; C1-C6 alkoxy; C1-C6 alk 10 It is an arylalkyl. In some embodiments, R 1a is a C1-C6 alkyl group substituted with 1-3 halos such as fluoro or chloro. In some embodiments, R 1a It is a C1-C6 alkyl group substituted with 1 to 3 -CO2H groups. In some variant forms, R 1a is a C1-C3 alkyl group substituted with 1-2 CO2H groups, for example, -CH2CO2H or -CH2CH2CO2H. In some embodiments, R 1a is a C1-C6 alkyl group substituted with 1 to 3 -C(=O)NH2 groups. In some embodiments, R 1ais a C1-C3 alkyl group substituted with 1-2 -C(=O)NH2 groups, for example, -CH2C(=O)NH2 or -CH2CH2C(=O)NH2. In some embodiments, R 1a C6-C is substituted with 1 to 3 substituents selected from halo and amino. 10 It is an arylalkyl. In some embodiments, R 1a C6-C is substituted with 1-3 halos such as fluoro, chloro, or bromo. 10 It is an arylalkyl. In some embodiments, R 1a This is a C6-C molecule substituted with 1-3 amino acids. 10 It is an arylalkyl. In some embodiments, R 1b is a C1-C6 alkyl group substituted with 1-3 halos such as fluoro or chloro. In some embodiments, R 1b It is a C1-C6 alkyl group substituted with 1 to 3 -CO2H groups. In some variant forms, R 1b is a C1-C3 alkyl group substituted with 1-2 CO2H groups, for example, -CH2CO2H or -CH2CH2CO2H. In some embodiments, R 1b is a C1-C6 alkyl group substituted with 1 to 3 -C(=O)NH2 groups. In some embodiments, R 1b is a C1-C3 alkyl group substituted with 1-2 -C(=O)NH2 groups, for example, -CH2C(=O)NH2 or -CH2CH2C(=O)NH2. In some embodiments, R 1b C6-C is substituted with 1 to 3 substituents selected from halo and amino. 10 It is an arylalkyl. In some embodiments, R 1b C6-C is substituted with 1-3 halos such as fluoro, chloro, or bromo. 10 It is an arylalkyl. In some embodiments, R 1b This is a C6-C molecule substituted with 1-3 amino acids. 10 It is an arylalkyl. In some embodiments, R 1a and R1b These are, independently, H, methyl, fluoro, 2-methylbutyl, -CH2F, methoxy, -CH2CO2H, -CH2C(=O)NH2, benzyl, or 4-aminobenzyl. In some embodiments, R 1a and R 1b Each is independently H or C1-C3 alkyl. In some embodiments, R 1a is methyl, and R 1b is H. In some embodiments, R 1a and R 1b These are H, respectively. In some embodiments, R 1a and R 1b One of the atoms is H, and the other is a C1-C3 alkyl group such as methyl.

[0121] In some embodiments, R 2 is H, oxo, or thioxo. In some embodiments, R 2 H is H. In some embodiments, R 2 is an oxo. In some embodiments, R 2 It is thioxo.

[0122] In some embodiments, R 3 C3-C6 alkyl, C3-C6 alkenyl, C3-C6 alkynyl, C3-C 12 Cycloalkyl, C3-C6 cycloalkylalkyl, C6-C 10 The 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. In some embodiments, R 3 is a C3-C6 alkyl such as propyl, butyl, pentyl, or hexyl. In some embodiments, R 3 is a C4-C6 alkyl group. In some embodiments, R 3 is a C3-C6 alkenyl. In some embodiments, R3 is a C4-C6 alkenyl. In some embodiments, R 3 is a C3-C6 alkynyl. In some embodiments, R 3 is a C4-C6 alkynyl. In some embodiments, R 3 These are C3-C compounds such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. 12 It is cycloalkyl. In some embodiments, R 3 is a C3-C6 cycloalkyl group. In some embodiments, R 3 is, -(CH2) 1-3 It is a C3-C6 cycloalkylalkyl such as (C3-C6 cycloalkyl). In some embodiments, R 3 C6-C, such as benzyl 10 It is an arylalkyl. In some embodiments, R 3 is, -(CH2) 1-3 (5-10 member heteroaryl) or -(CH2) 1-3 These are 5-10 member heteroarylalkyls such as (5-6 member heteroaryl). In some embodiments, the 5-10 member heteroarylalkyl contains 1-2 nitrogen atoms. In some embodiments, R 3 is, -(CH2) 1-3 (5-10 member heterocycline) or -(CH2) 1-2 These are 5-10 member heterocyclylalkyls, such as (5-6 member heterocyclyls). In some embodiments, the 5-10 member heterocyclylalkyl contains 1-2 nitrogen atoms.

[0123] In some embodiments, R 3 R is a C3-C6 alkyl, C2-C6 alkenyl, or C3-C6 cycloalkylalkyl molecule optionally substituted with 1 to 3 substituents selected from halo and -C(=O)NH2. In some embodiments, R 3is a C2-C6 alkyl; C2-C6 alkenyl; C3-C6 cycloalkylalkyl; 5-6 member heteroarylalkyl; 5-6 member heterocyclylalkyl; or C6 arylalkyl, optionally substituted with 1-3 substituents selected from halo, C1-C3 alkoxy, hydroxy, -NH2, -SO2(C1-C3 alkyl), and -C(=O)NH2. In some embodiments, R 3 R is a C2 alkyl group substituted with 1 to 3 substituents selected from C1-C3 alkoxy, hydroxyl, -NH2, and -SO2(C1-C3 alkyl). In some embodiments, R 3 The following is true: [ka] In some embodiments, R 3 The following is true: [ka] In some embodiments, R 3 It is 2-methylbutyl.

[0124] In some embodiments, R 4 C6-C 10 The aryl, 5-10 membered heteroaryl, or 5-10 membered heterocyclyl is a 5-10 membered heteroaryl or 5-10 membered heterocyclyl containing 1-3 heteroatoms selected from nitrogen and oxygen. In some embodiments, R 4 C6-C such as phenyl 10 It is an arrow. In some embodiments, R 4 R is a 5-10 membered heteroaryl compound containing 1-2 nitrogen atoms. In some embodiments, R 4 R is a 5-10 membered heterocycline. In some embodiments, R 4 R is a 5-9 membered heterocycline containing 1-2 nitrogen atoms. In some embodiments, R 4R is a 5-9 membered heterocycline containing 1-2 oxygen atoms. In some embodiments, R 4 It is a 5- to 9-membered heterocycline containing one nitrogen atom and one oxygen atom.

[0125] In some embodiments, R 4 C6-C is optionally substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy. 10 It is an arrow. In some embodiments, R 4 R is a phenyl compound substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro. In some embodiments, R 4 The following is true: [ka] In some embodiments, R 4 The following is true: [ka]

[0126] In some embodiments, R 4 R is a 5-10 member heteroaryl that is optionally substituted with 1-3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In some embodiments, R 4 R is a pyridyl or indolyl which is optionally substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In some embodiments, R 4 teeth, [ka] In some embodiments, R 4R is a pyridyl substituted with 1 to 3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In some embodiments, R 4 teeth, [ka] In some embodiments, R 4 R is a 5-10 membered heterocycline optionally substituted with 1-3 substituents selected from halo, hydroxyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In some embodiments, R 4 It is indlinil. [ka]

[0127] In some embodiments, -LR 4 is -CH2(phenyl) or -C(O)(phenyl), where phenyl is substituted with 1 to 3 substituents selected from C1-C3 haloalkyl, C1-C3 haloalkoxy, halo, and hydroxyl. In some embodiments, -LR 4 is -CH2(pyridyl) or -C(O)(pyridyl), where pyridyl is substituted with 1 to 3 substituents selected from C1-C3 haloalkyl, C1-C3 haloalkoxy, halo, and hydroxyl. In some embodiments, -LR 4 The following is true: [ka]

[0128] In some embodiments, each R 5 R is independently a C1-C6 alkyl, oxo, or halo. In some embodiments, R 5 is a C1-C6 alkyl such as methyl, ethyl, or propyl. In some embodiments, R 5is an oxo. In some embodiments, R 5 is a halo such as fluoro, chloro, or bromo. In some embodiments, R 5 is oxo or halo. In some embodiments, R 5 It is either oxo or fluoro.

[0129] In some embodiments, R 6 is H, C1-C6 alkyl, or oxo. In some embodiments, R 6 H is H. In some embodiments, R 6 is a C1-C6 alkyl such as methyl, ethyl, or propyl. In some embodiments, R 6 It is oxo.

[0130] In some embodiments, R 7 is H or oxo. In some embodiments, R 7 H is H. In some embodiments, R 7 It is oxo.

[0131] In some embodiments, m is 1. In other embodiments, m is 2.

[0132] 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.

[0133] In any embodiment or variation of formula (I), each C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 12 Cycloalkyl, C3-C 12 Cycloalkylalkyl, C6-C 10 Ariel, C6-C 10Arylalkyls, 5-10 membered heteroaryls, 5-10 membered heteroarylalkyls, 5-10 membered heterocyclyls, and 5-10 membered heterocyclylalkyls are optionally substituted with 1-3 substituents selected from hydroxyl, halo (fluoro, chloro, or bromo, etc.), amino, C1-C6 haloalkyls (-CF3 or -CHF2, etc.), C1-C6 alkoxys (methoxy or ethoxy, etc.), C1-C6 haloalkoxys (-OCHF2 or -OCF3, etc.), and -(C=O)NH2.

[0134] In some embodiments, the compound of formula (I) is a compound of formula (II), (IIa), (IIb), (IIc), (IId), or (IIe): [ka] or a pharmaceutically acceptable salt thereof, where L, R 1a , R 1b , R 3 , R 4 , R 5 , R 6 , R 7 , and n are as described for formula (I). In some embodiments, the compound is represented by formula (II) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIa) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIb) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIc) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IId) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIe) or a pharmaceutically acceptable salt thereof.

[0135] In some embodiments, the compound of formula (I) is the compound of formula (IIIa), (IIIb), (IIIc), or (IIId): [ka] or a pharmaceutically acceptable salt thereof, where R 1a , R 1b , R 3 , R 5 , R 6 , and n are as described for formula (I), and R represents one or more arbitrary substituents such as hydroxyl, halo, amino, C1-C6 haloalkyl, C1-C6 haloalkoxy, etc., as described for formula (I). In some embodiments, the compound is represented by formula (IIIa) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIIb) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIIc) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IIId) or a pharmaceutically acceptable salt thereof.

[0136] In some embodiments, the compound of formula (I) is the compound of formula (IVa), (IVb), (IVc), or (IVd): [ka] or a pharmaceutically acceptable salt thereof, where R 5 n and n are as described for formula (I), and R represents one or more arbitrary substituents such as hydroxyl, halo, amino, C1-C6 haloalkyl, C1-C6 haloalkoxy, etc., as described for formula (I). In some embodiments, the compound is represented by formula (IVa) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IVb) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IVc) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is represented by formula (IVd) or a pharmaceutically acceptable salt thereof.

[0137] In some embodiments, the compound of formula (I) is the compound of formula (V): [ka] or a pharmaceutically acceptable salt thereof, where L, R 1a , R 1b , R 3 , and R 4 This is as described for formula (I). In some embodiments, L is -C(=O)- or -CH2-, and R 1a and R 1b R is independently a C1-C3 alkyl group optionally substituted with H or -CO2H, and 3 R is a C1-C3 alkyl substituted with a C4-C5 alkyl, a C4-C5 alkenyl, or a C3-C5 cycloalkyl, 4 R is a phenyl or pyridyl substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro. In some variant forms, R 1a and R 1b One of the atoms is H, and the other is a C1-C3 alkyl group such as methyl.

[0138] In this specification, it is understood that any description, variation, embodiment, or aspect relating to one part can be combined with any description, variation, embodiment, or aspect relating to another part, as if each and all combinations of descriptions were specifically and individually listed. For example, any description, variation, embodiment, or aspect provided herein relating to L of formula (I) can be combined with R 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , R 7Any description, variation, embodiment, or aspect relating to , and n can be combined as if each and every combination were specifically and individually listed. All descriptions, variations, embodiments, or aspects relating to formula (I) are understood to apply and be described in the same way to other formulas detailed herein, where applicable, as if each and every description, variation, embodiment, or aspect relating to all formulas were specifically and independently listed. For example, all descriptions, variations, embodiments, or aspects relating to formula (I) shall, where applicable, apply and be described in the same manner 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 description, variation, embodiment, or aspect relating to all formulas were listed individually and independently.

[0139] In some embodiments, compounds selected from the compounds in Table 1, or pharmaceutically acceptable salts thereof, are provided. While some of the compounds described herein, including those in Table 1, are shown as specific stereoisomers and / or non-stereochemical forms, any or all stereochemical forms, including any enantiomer or diastereomer of any of the compounds in this disclosure, including those in Table 1, and any tautomers or other forms, are understood to also be described herein. TIFF0007884275000024.tif218165TIFF0007884275000025.tif234165TIFF0007884275000026.tif232165TIFF00078842750 00027.tif235165TIFF0007884275000028.tif223165TIFF0007884275000029.tif225165TIFF0007884275000030.tif233165

[0140] 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.

[0141] In some embodiments, compounds selected from the compounds in Table 1A or pharmaceutically acceptable salts thereof are provided. While some of the compounds described herein, including those in Table 1A, are shown as specific stereoisomers and / or non-stereochemical forms, any or all stereochemical forms, including any enantiomer or diastereomer of any of the compounds in this disclosure, including those in Table 1A, and any tautomers or other forms, are understood to also be described herein. TIFF0007884275000031.tif223165TIFF0007884275000032.tif235165TIFF0007884275000033.tif91165

[0142] In the description of the present invention, it is understood that combinations of substituents and / or variables in the given formulas are permissible only if such involvement results in a stable compound.

[0143] Furthermore, all compounds of formula (I) existing in the form of a free base or free acid can be converted to their pharmaceutically acceptable salts by treatment with a suitable 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 form by standard techniques.

[0144] Synthesis method Compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, can be prepared by using organic chemical synthesis methods known in the art. Generally, starting components may be obtained from suppliers such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, 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 prepared as described herein.

[0145] General reaction scheme 1. [ka] General reaction scheme 1 provides an exemplary method for preparing the compound of formula (I). R in general reaction scheme 1 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 L, n, and n are as defined herein. X is a reactive moiety selected to promote the desired reaction (e.g., a halo). P1 and P2 are preferred protecting groups. L' is L'-R 4A compound is selected so that the desired L moiety is obtained from the reaction between the compound and a secondary amine. The compound of structure A1 is purchased or prepared according to methods known in the art. A1 and A2 are reacted under suitable coupling conditions (e.g., T3P and a base) to obtain product A3 from the coupling reaction between A1 and A2. Then A3 and A4 are reacted under suitable coupling conditions (e.g., T3P and a base) to obtain compound A5. Then compound A5 is cyclized (e.g., using formic acid) and deprotected (e.g., using piperidine) to obtain compound A6. Then compound A6 is reacted with compound A7 to obtain the final compound of formula (I) shown.

[0146] General reaction scheme 2. [ka] An alternative synthesis method for the compound of formula (I) is shown in general reaction scheme 2. R in general reaction scheme 2 1a , R 1b , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 L, n, and n are as defined herein. P2 is a preferred protecting group. Each X is a reactive moiety selected to facilitate the desired reaction (e.g., a halo). L' is L'-R 4 The reaction between the intermediate A5 and the secondary amine is selected to yield the desired L moiety. 3 Removable protective group P 3 The intermediate A8 is prepared using (e.g., para-methoxybenzyl). Then, A8 is cyclized (e.g., using formic acid) and deprotected (e.g., using piperidine) to obtain compound A9. Next, compound A9 is reacted with A7 to obtain compound A10. Next, compound A10 is deprotected (e.g., with cerium ammonium nitrate) to obtain compound A11. Finally, compound A11 is reacted with A12 to obtain the final compound of formula (I).

[0147] General reaction scheme 3. [ka] A method related to the one 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 obtain compound A10, which is then reacted with A7 to obtain compound A11. Subsequently, the reaction with A12 yields the final compound of formula (I).

[0148] It should be noted that various alternative strategies are available to those skilled in the art for the preparation of compounds of formula (I). For example, other compounds of formula (I) can be prepared according to similar methods using appropriate starting materials.

[0149] Those skilled in the art will understand that in the process of preparing the compounds described herein, it may be necessary to protect the functional groups of intermediate compounds with suitable protecting groups. Such functional groups may include hydroxy, amino, and carboxylic acids. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, and benzyl. Suitable protecting groups for amino and amidino include t-butoxycarbonyl and benzyloxycarbonyl. Suitable protecting groups for carboxylic acids include alkyl, aryl, or arylalkyl esters. Protecting groups are added or removed at will according to standard techniques known to those skilled in the art or described herein. The use of protecting groups is detailed in Green, TWand PGMWutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As those skilled in the art will understand, the protecting group may be a polymer resin such as Wang resin, Rink resin, or 2-chlorotrityl chloride resin.

[0150] Pharmaceutical compositions and preparations In further embodiments, pharmaceutical compositions are provided herein. A pharmaceutical composition comprises one (or more) of the aforementioned compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for infusion. In many more embodiments, the pharmaceutical composition comprises the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, and an additional therapeutic agent. Non-limiting examples of such therapeutic agents are described herein below.

[0151] Preferred routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, ophthalmic, nasal, and topical administration. In addition, parenteral delivery, though only examples, includes intramuscular, subcutaneous, intravenous, and intrathecal injections, as well as intrathecal, direct intravenous, intraperitoneal, intralymphatic, and intranasal injections.

[0152] In certain embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered topically rather than systemically, for example, often in depot preparations or sustained-release formulations, by direct injection of the compound into an organ. In certain embodiments, long-acting formulations are administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. Furthermore, in other embodiments, the drug is delivered by a targeted drug delivery system, for example, by liposomes coated with organ-specific antibodies. In such embodiments, the liposomes are targeted to the organ and selectively taken up by the organ. In yet another embodiment, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is provided in the form of an immediate-release formulation, a sustained-release formulation, or an intermediate-release formulation. In yet another embodiment, the compounds described herein are administered topically.

[0153] The compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is effective over a wide range of doses. For example, in the treatment of adults, doses of 0.01–1000 mg, 0.5–100 mg, 1–50 mg, and 5–40 mg per day are examples of doses used in some embodiments. An exemplary dose is 10–30 mg per day. The exact dose may depend on the route of administration, the form in which the compound is administered, the patient being treated, the patient's body weight, and the preference and experience of the attending physician.

[0154] In some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered in a single dose. Typically, such administration is by injection, e.g., intravenous injection, to rapidly introduce the agent. However, other routes may also be used as appropriate. Single doses of the compounds of this disclosure may also be used in the treatment of acute conditions (e.g., traumatic brain injury).

[0155] In some embodiments, the compound of formula (I), or its pharmaceutically acceptable salts, isotopes, or stereoisomers, is administered in multiple doses. In some embodiments, administration is approximately once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, administration is approximately once a month, once every two weeks, once a week, or once every other day. In yet another embodiment, the compound of formula (I), or its pharmaceutically acceptable salts, isotopes, or stereoisomers, and another therapeutic agent are administered together approximately once to six times per day. In yet another embodiment, administration of the compound of formula (I), or its pharmaceutically acceptable salts, isotopes, or stereoisomers, and the therapeutic agent continues for less than approximately seven days. In yet another embodiment, administration continues for approximately six, ten, fourteen, or twenty-eight days, two months, six months, or more than one year. In some cases, continuous administration is possible and maintained for as long as necessary.

[0156] The administration of the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be continued for as long as necessary. In some embodiments, the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be 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 pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be 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 pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be administered continuously, for example, for the treatment of chronic conditions (e.g., dementia).

[0157] In some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered in multiple doses. Due to inter-subject variability in the pharmacokinetics of the compound, it is known in the art that individualization of the administration regimen is necessary for optimal treatment. The dosage of the compound can be determined by routine experiments in light of this disclosure.

[0158] In some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is formulated into a pharmaceutical composition. In specific embodiments, the pharmaceutical composition is formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and adjuvants that facilitate the processing of the active compound into a pharmaceutically usable preparation. The appropriate formulation depends on the selected route of administration. Any pharmaceutically acceptable techniques, elements, and excipients are suitably used to formulate the pharmaceutical compositions described in this specification: 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).

[0159] Provided herein are pharmaceutical compositions comprising a compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, and a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s). Also provided herein are methods for administering a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, and a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s).

[0160] In certain embodiments, the compound is administered as a combination therapy, in which the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is mixed with other therapeutic agents in a pharmaceutical composition. All combinations of active ingredients described in the following Methods section and throughout this disclosure are incorporated herein. In specific embodiments, the pharmaceutical composition comprises one or more compounds of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0161] When used herein, a pharmaceutical composition refers to a mixture of a compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates the administration of the compound to a living organism. In some embodiments, to carry out the treatment or method of use provided herein, a therapeutically effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof provided herein, is administered in the pharmaceutical composition to a mammal having the disease, disorder, or medical condition to be treated. In specific embodiments, the mammal is a human. In certain embodiments, the therapeutically effective amount varies 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.

[0162] In one embodiment, one or more compounds of formula (I), or pharmaceutically 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 Hank's solution, Ringer's solution, or physiological saline buffer, but these are just examples. In another embodiment, one or more compounds of formula (I), or pharmaceutically acceptable salts, isotopic forms, or stereoisomers thereof, are formulated for transmucosal administration. In a specific embodiment, the transmucosal formulation includes a permeabilizing agent suitable for the barrier to be permeated (e.g., the blood-brain barrier). In yet another embodiment in which the compounds described herein are formulated for other parenteral injection, suitable formulations include aqueous or non-aqueous solutions. In a specific embodiment, such solutions include physiologically compatible buffers and / or excipients.

[0163] In another embodiment, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, 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 a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is formulated in oral dosage forms, including, but not limited to, tablets, powders, pills, sugar-coated tablets, capsules, liquids, gels, syrups, elixirs, slurries, and suspensions.

[0164] In certain embodiments, pharmaceutical preparations for oral use are obtained by mixing one or more solid excipients with one or more compounds of formula (I), or pharmaceutically acceptable salts, isotopic forms, or stereoisomers thereof; optionally, grinding the resulting mixture, processing the granular mixture, adding optionally suitable excipients, and then obtaining tablets or sugar-coated tablet cores. Suitable excipients include, in particular, sugars including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, e.g., corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.; or other fillers, e.g., polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. In specific embodiments, disintegrants are optionally added. Disintegrants include, but are not limited to, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or salts thereof such as alginic acid or sodium alginate.

[0165] In one embodiment, the dosage form, such as a sugar-coated tablet core and a tablet, comprises one or more suitable coatings. In a specific embodiment, a concentrated sugar solution is used for coating the dosage form. The sugar solution optionally contains additional components, such as gum arabic, talc, polyvinylpyrrolidone, Carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solution, and suitable organic solvents or solvent mixtures, but these are just examples. Dyes and / or pigments are also optionally added to the coating for identification. Furthermore, dyes and / or pigments are optionally used to characterize different combinations of active compounds in different doses.

[0166] In certain embodiments, a therapeutically effective amount of the compound of formula (I), or at least one of its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is formulated into other oral dosage forms. These oral dosage forms include push-fit capsules made of gelatin, and sealable 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, but are not limited to, binders such as lactose or 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, but are not limited to, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, optionally, stabilizers are added.

[0167] In other embodiments, at least one of the compounds of formula (I) described herein, or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, in a therapeutically effective amount, is formulated for buccal or sublingual administration. Formulations suitable for buccal or sublingual administration include, but are not limited to, tablets, lozenges, or gels. In yet another embodiment, the compounds of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, are 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., an ampoule) or a multi-dose container. Preservatives are optionally added to the injectable formulation. In yet another embodiment, 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. The parenteral injectable formulation optionally contains formulations such as suspending agents, stabilizers, and / or dispersants. In specific embodiments, the parenteral pharmaceutical formulation comprises an aqueous solution of the active compound in a water-soluble form. In additional embodiments, a suspension of one or more active compounds (e.g., the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof) is prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles for use in the pharmaceutical compositions described herein include, but are not limited to, fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. In certain specific embodiments, the aqueous injection suspension contains a substance that increases the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension contains a suitable stabilizer or an agent that increases the solubility of the compound, enabling the preparation of a highly concentrated solution. Alternatively, in other embodiments, the active ingredient is in powder form for use with a suitable vehicle, e.g., sterile pyrogen-free water.

[0168] In further embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is administered topically. The compound is formulated into a variety of topically administerable compositions, such as solutions, suspensions, lotions, gels, pastes, medicinal sticks, ointments, creams, or ointments. Such pharmaceutical compositions optionally contain solvents, stabilizers, tonicity enhancers, buffers, and preservatives.

[0169] In further embodiments, the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is formulated for transdermal administration. In specific embodiments, the transdermal formulation may employ a transdermal delivery device and / or a transdermal delivery patch, and may be a lipophilic emulsion or buffered aqueous solution dissolved and / or dispersed in a polymer or adhesive. In various embodiments, such patches may be configured for continuous pulsed delivery or on-demand delivery of the drug. In further embodiments, transdermal delivery of the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is achieved by means such as iontophoresis patches. In certain embodiments, the transdermal patch provides controlled delivery of the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers. In specific embodiments, the absorption rate is slowed by using a rate-controlled membrane or by encapsulating the compound within a polymer matrix or gel. In alternative embodiments, absorption enhancers are used to increase absorption. The absorption enhancer or carrier comprises a pharmaceutically acceptable absorbent solvent that facilitates 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 optionally containing the compound together with the carrier, an optional rate-controlled barrier for delivering the compound to the host's skin at a controlled rate over a long period of time, and means for securing the device to the skin.

[0170] In other embodiments, the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is formulated for administration by inhalation. Various forms suitable for administration by inhalation include, but are not limited to, aerosols, mists, or powders. Any pharmaceutical composition of the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is conveniently delivered in the form of presentation by aerosol spray from a pressurized pack or nebulizer, using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas). In specific embodiments, the dosage unit of the pressurized aerosol is determined by providing a valve that delivers a measured amount. In certain embodiments, for example, gelatin capsules and cartridges for use in inhalers or blowers are formulated to contain a powder mixture of the compound and a suitable powder base material such as lactose or starch.

[0171] In further embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is formulated into transrectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retained enemas, containing conventional suppository bases such as cocoa butter or other glycerides, and synthetic polymers such as polyvinylpyrrolidone or PEG. In the suppository form of the composition, a low-melting-point wax, such as a mixture of fatty acid glycerides combined with cocoa butter (optional), melts first.

[0172] In certain embodiments, the pharmaceutical composition is formulated in any conventional manner using one or more physiologically acceptable carriers, including excipients and adjuvants that facilitate the processing of the active compound into a pharmaceutically usable preparation. The appropriate formulation depends on the selected route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are preferably used at will. Pharmaceutical compositions comprising the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, are manufactured in conventional manner, such as conventional mixing, dissolution, granulation, sugar-coated tablet manufacturing, polishing, emulsification, encapsulation, encapsulation, or compression processes, but are not limited to these examples.

[0173] A pharmaceutical composition comprises at least one pharmaceutically acceptable carrier, diluent, or excipient and at least one compound of formula (I) described herein, or a pharmaceutically 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 a pharmaceutically 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 fall within the scope of compounds presented herein. Furthermore, the compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, include not only solvates with pharmaceutically acceptable solvents such as water and ethanol, but also non-solvates. Solvates of the compounds presented herein are also considered to be disclosed herein. In addition, the pharmaceutical composition may optionally include other medical or pharmaceutical agents, carriers, and adjuvants, such as preservatives, stabilizers, humectants or emulsifiers, solution enhancers, salts for adjusting osmotic pressure, buffers, and / or other substances of therapeutic value.

[0174] A method for preparing a composition comprising the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, comprises formulating the compound with one or more pharmaceutically 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 of the compound, emulsions containing the compound, or solutions containing liposomes, micelles, or nanoparticles comprising the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. The forms of pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for being converted into solutions or suspensions in liquid before use, or emulsions. These compositions also optionally contain trace amounts of non-toxic auxiliary substances such as wetting agents or emulsifiers or pH buffers.

[0175] In some embodiments, a pharmaceutical composition comprising at least one compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof takes the form of a liquid, exemplarially, in which the drug exists in a solution, a suspension, or both. Typically, when the composition is administered as a solution or suspension, the first part of the drug exists in a particulate form, with the second part of the drug being a suspension in a liquid matrix. In some embodiments, the liquid composition comprises a gel formulation. In other embodiments, the liquid composition is aqueous.

[0176] In certain embodiments, a useful aqueous suspension contains one or more polymers, such as suspending agents. Useful polymers include cellulosic polymers, such as water-soluble polymers like hydroxypropyl methylcellulose, and non-water-soluble polymers such as water-crosslinked carboxyl-containing polymers. Certain pharmaceutical compositions described herein include, for example, mucosal adhesive polymers selected from carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0177] Useful pharmaceutical compositions also optionally include a solubilizer that assists in the dissolution of the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers. “Solubilizer” generally includes agents that result in a micelle solution or true solution of the agent. Certain acceptable nonionic surfactants, such as polysorbate 80, are useful as solubilizers, and glycols and polyglycols acceptable in the ophthalmic field, such as polyethylene glycol 400, and glycol ethers are also possible.

[0178] Furthermore, useful pharmaceutical compositions may optionally include one or more pH adjusters or buffers, such as acids like acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases like sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane; and buffers like citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition within an acceptable range.

[0179] Furthermore, useful compositions may also optionally include one or more salts in amounts necessary to bring the osmolality of the composition into an acceptable range. Such salts may include those having sodium, potassium, or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions, and preferred salts may include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0180] Other useful pharmaceutical compositions may optionally include one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0181] Further useful compositions include one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; as well as polyoxyethylene alkyl ethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40.

[0182] Further useful compositions may include, if necessary, one or more antioxidants to enhance chemical stability. Suitable antioxidants include, but are not limited to, ascorbic acid and sodium disulfite.

[0183] In certain embodiments, the aqueous suspension composition is packaged in a single-dose, non-resealable container. Alternatively, a resealable container for multiple doses is used, in which case the composition typically contains a preservative.

[0184] 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 pharmaceutically acceptable salts, isotopic forms, or stereoisomers, is delivered using a sustained-release system such as a semipermeable matrix of a hydrophobic solid polymer containing the therapeutic agent. Various sustained-release materials are useful herein. In some embodiments, sustained-release capsules release the compound over several weeks to up to 100 days or more. Depending on the chemical properties and biological stability of the therapeutic agent, additional strategies for protein stability are employed.

[0185] In certain embodiments, the formulations described herein include one or more antioxidants, metal chelating agents, thiol-containing compounds, and / or other common stabilizers. Examples of such stabilizers include, but are not limited to, (a) glycerol at approximately 0.5% to 2% w / v, (b) methionine at approximately 0.1% to 1% w / v, (c) monothioglycerol at approximately 0.1% to 2% w / v, (d) EDTA at approximately 1 mM to 10 mM, (e) ascorbic acid at approximately 0.01% to 2% w / v, (f) polysorbate 80 at approximately 0.003% to 0.02% w / v, (g) polysorbate 20 at approximately 0.001% to 0.05% w / v, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparin analogs, (m) divalent cations such as magnesium and zinc, or (n) combinations thereof.

[0186] In some embodiments, the concentrations of the compound of formula (I) provided in the pharmaceutical composition of the present disclosure are 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. It is 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 less than 0.0001% w / w, w / v, or v / v.

[0187] In some embodiments, the concentrations of the compound of formula (I) provided in the pharmaceutical composition of the present disclosure are 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%, and 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 0.50%, 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.

[0188] In some embodiments, the concentrations of the compound of formula (I), or its pharmaceutically acceptable salts, isotopes, or stereoisomers provided in the pharmaceutical composition are approximately 0.0001% to approximately 50%, approximately 0.001% to approximately 40%, approximately 0.01% to approximately 30%, approximately 0.02% to approximately 29%, approximately 0.03% to approximately 28%, approximately 0.04% to approximately 27%, approximately 0.05% to approximately 26%, approximately 0.06% to approximately 25%, and approximately 0.07%. The ranges are approximately 24%, 0.08% to 23%, 0.09% to 22%, 0.1% to 21%, 0.2% to 20%, 0.3% to 19%, 0.4% to 18%, 0.5% to 17%, 0.6% to 16%, 0.7% to 15%, 0.8% to 14%, 0.9% to 12%, or approximately 1% to 10% in the range of w / w, w / v, or v / v.

[0189] In some embodiments, the concentration of the compound of formula (I), or its pharmaceutically acceptable salt, isotope, or stereoisomer provided in the pharmaceutical composition is in the range of approximately 0.001% to approximately 10%, approximately 0.01% to approximately 5%, approximately 0.02% to approximately 4.5%, approximately 0.03% to approximately 4%, approximately 0.04% to approximately 3.5%, approximately 0.05% to approximately 3%, approximately 0.06% to approximately 2.5%, approximately 0.07% to approximately 2%, approximately 0.08% to approximately 1.5%, approximately 0.09% to approximately 1%, or approximately 0.1% to approximately 0.9% w / w, w / v, or v / v.

[0190] In some embodiments, the amounts of the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers provided in the pharmaceutical composition are 10g, 9.5g, 9.0g, 8.5g, 8.0g, 7.5g, 7.0g, 6.5g, 6.0g, 5.5g, 5.0g, 4.5g, 4.0g, 3.5g, 3.0g, 2.5g, 2.0g, 1.5g, 1.0g, 0.95g, 0.9g, 0.85g, 0.8g, 0.75g, 0.7g, 0.65g, 0.6g, 0.55g, 0.5g, 0.45g, 0.4g, 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 or less.

[0191] In some embodiments, the amounts of the compound of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer provided in the pharmaceutical composition of the present disclosure are 0.0001g, 0.0002g, 0.0003g, 0.0004g, 0.0005g, 0.0006g, 0.0007g, 0.0008g, and 0.0009g. , 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 more than 10g.

[0192] In some embodiments, the amount of the compound of formula (I), or its pharmaceutically acceptable salt, isotopic form, or stereoisomer provided in the pharmaceutical composition is in the range of 0.0001 to 10 g, 0.0005 to 9 g, 0.001 to 8 g, 0.005 to 7 g, 0.01 to 6 g, 0.05 to 5 g, 0.1 to 4 g, 0.5 to 4 g, or 1 to 3 g.

[0193] Kit / Product Kits and products are also provided for use in the therapeutic applications described herein. In some embodiments, such kits comprise a case, package, or container partitioned to receive one or more containers, such as vials, tubes, etc., each of which contains one of the individual elements used in the manner described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. Containers are formed from a variety of materials, such as glass or plastic.

[0194] The products provided herein include packaging materials. Packaging materials used for packaging pharmaceuticals include, for example, those found in U.S. Patents No. 5,323,907, No. 5,052,558 and No. 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 materials suitable for the selected formulation and intended mode of administration and treatment. For example, a container(s) may optionally contain one or more compounds described herein, either in the composition or in combination with another agent disclosed herein. A container(s) may optionally have a sterile access port (for example, a container may be an intravenous solution bag or vial with a stopper that can be pierced by a subcutaneous needle). Such a kit may optionally contain the compounds together with identifying information or a label or instructions for use in the manner described herein.

[0195] For example, a kit typically includes one or more additional containers, each containing one or more materials (such as reagents (optionally concentrated forms) and / or devices) that are commercially and user-desirable for the use of the compound of formula (I) or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, carriers, packaging, containers, vials, and / or tube labels describing the contents and / or instructions for use, as well as accompanying documents describing the contents. A set of instructions is also typically included. Labels may optionally be on or attached to the container. For example, if the letters, numbers, or other characters forming the label are attached to, molded into, or etched onto the container itself, the label is on the container; if the label is in the container or carrier holding the container, for example, as part of accompanying documents, the label is attached to the container. In addition, labels are used to indicate that the contents are intended for a specific therapeutic use. In addition, the label provides instructions for the 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 compounds 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 labeling in the form prescribed by the government agency that regulates the manufacture, use, or sale of pharmaceuticals, which reflects the agency's approval for the form of the drug for human or veterinary administration. Such labeling is, for example, a label for a prescription drug approved by the U.S. Food and Drug Administration, or a package insert for an approved product. In some embodiments, a composition containing the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, formulated on a suitable pharmaceutical carrier, is prepared, placed in a suitable container, and labeled for the treatment of a specified condition.

[0196] Instructions for use / treatment Embodiments of this disclosure provide a method for modulating hepatocyte growth factor in a subject requiring such modification, comprising administering to the subject an effective amount of a compound disclosed herein (e.g., the compound of formula (I), or a pharmaceutically 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 evaluated and demonstrated by a wide variety of methods known in the art. Kits and commercially available assays can be used to determine whether and to what extent hepatocyte growth factor has been modified (e.g., inhibited or activated).

[0197] In some embodiments, provided herein are compounds of formula (I) or pharmaceutically acceptable salts thereof for use in modifying hepatocyte growth factor in subjects requiring such modification. In some embodiments, provided herein are compounds of formula (I) or pharmaceutically acceptable salts thereof for manufacturing pharmaceuticals for modifying hepatocyte growth factor in subjects requiring such modification.

[0198] The applicant has discovered that the compound of formula (I) exhibits promising activity in relation to specific diseases of interest. Accordingly, in one embodiment, the herein provides a method for modulating hepatocyte growth factor in a subject requiring such modification, comprising administering to the subject an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof. In some embodiments, the herein provides a method for activating hepatocyte growth factor in a subject requiring such modification, comprising administering to the subject an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0199] In more specific embodiments, modification includes treating a disease, condition, or injury (e.g., traumatic brain injury). Non-limiting examples include neurodegenerative diseases, traumatic brain injury, memory loss or functional impairment, spinal cord injury, sensorineural hearing loss, and nerve damage. In some embodiments, the disease, condition, or injury is a neurodegenerative disease, spinal cord injury, traumatic brain injury, or sensorineural hearing loss.

[0200] In a more specific embodiment, the disease, condition, or injury is a neurodegenerative disease. For example, in some embodiments, the neurodegenerative disease is Alzheimer's disease, dementia, Parkinson's disease, Huntington's disease, or amyotrophic lateral sclerosis (ALS). In a more specific embodiment, the neurodegenerative disease is Alzheimer's disease or Parkinson's disease.

[0201] Furthermore, provided herein are methods for treating or slowing the progression of dementia in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof. In one specific embodiment, dementia is related to Alzheimer's disease or Parkinson's disease.

[0202] In a further embodiment, the foregoing provides a method for preventing cognitive impairment in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0203] Further related embodiments provide a method for treating, restoring, or preventing a disease, condition, or injury relating to nerve tissue in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof.

[0204] In other embodiments, the foregoing provides a method for treating a neuropsychiatric disorder or condition, comprising administering an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, to a subject in need thereof. Non-limiting examples of neuropsychiatric disorders or conditions include, but are not limited to, depression and anxiety.

[0205] In further embodiments, the foregoing provides a method for treating a disease or disorder of the central nervous system, comprising administering an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, to a subject in need. In some embodiments, the foregoing provides a method for preventing a disease or disorder of the central nervous system, comprising administering an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, to a subject in need. A non-limiting example of a disease or disorder of the central nervous system is traumatic brain injury.

[0206] In further other embodiments, provided herein are methods for treating a disease or disorder of the central nervous system, comprising administering an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, to a subject in need. In some embodiments, provided herein are methods for preventing a disease or disorder of the peripheral nervous system, comprising administering an effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, to a subject in need. A non-limiting example of a disease or disorder of the peripheral nervous system is neuropathic pain.

[0207] Embodiments of the above methods involve administering to a mammal a therapeutically effective amount of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof. The methods disclosed herein generally relate to the administration of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, for the treatment, protection, or reversal of diseases and injuries related to nerve cells or the nervous system. Specifically, embodiments of the disclosure relate to the treatment, prevention, or reversal of neurodegenerative diseases, including the treatment of dementia; the repair of traumatic injuries; and / or the prevention of cognitive impairment.

[0208] In some embodiments, the Disclosure provides methods for modulating the activity of a protein (e.g., hepatocyte growth factor) in subjects including, but not limited to, rodents and mammals (e.g., humans) by administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer of the compound or mammal in the subject. 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%.

[0209] In some embodiments, the Disclosure provides a method for regulating hepatocyte growth factor (HGR) activity in cells by contacting the cells with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate HGR activity. In some embodiments, the Disclosure provides a method for regulating HGR activity in tissues by contacting the tissue with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate HGR activity in the tissue. In some embodiments, the Disclosure provides a method for regulating HGR activity in organisms by contacting the organism with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate HGR activity in the organism. In some embodiments, the Disclosure provides a method for regulating hepatocyte growth factor (HGR) activity in animals by contacting the animals with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate HGR activity in animals. In some embodiments, the Disclosure provides a method for regulating HGR activity in mammals by contacting the mammals with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate HGR activity in mammals. In some embodiments, the Disclosure provides a method for regulating HGR activity in humans by contacting the humans with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof in an amount sufficient to regulate HGR activity in humans. In other embodiments, the Disclosure provides a method for treating diseases mediated by HGR activity in subjects requiring such treatment. In several variant forms, the regulation of hepatocyte growth factor by the compound of formula (I) or its pharmaceutically acceptable salts, isotopes, or stereoisomers is involved in the activation of hepatocyte growth factor.

[0210] Other embodiments provide methods for combination therapy in which therapeutic agents known to modulate other pathways or other elements or overlapping target enzyme groups of the same pathway are used in combination with a compound of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof. In one embodiment, such a therapy includes, but is not limited to, a combination of one or more compounds of formula (I) or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof with therapeutic agents, therapeutic antibodies, and other therapeutic forms that produce a synergistic or additive therapeutic effect.

[0211] Currently, many therapeutic agents are known in the art and can be used in combination with compounds of formula (I), or their pharmaceutically 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 involve the use of therapies including reminiscence therapy, cognitive activation therapy, reality orientation training, and physical activity.

[0212] Examples of cholinesterase inhibitors include donepezyl, galantamine, and rivastigmine, which help slow the breakdown of brain chemicals involved in memory and judgment. Memantine may help control other brain chemicals necessary for learning and memory. In certain embodiments, memantine may also be used with donepezyl as an adjunct agent for moderate to severe dementia. Antidepressants may include, but are not limited to, selective serotonin reuptake inhibitors (SSRIs). Anxiolytics may include, but are not limited to, lorazepam (Ativan) or oxazepam (Serax). Some embodiments of the methods described herein may involve the use or administration of antipsychotics such as aripiprazole (Abilify), haloperidol (Haldol), olanzapine (Zyprexa), and risperidone (Risperdal).

[0213] In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers are formulated or administered together with liquid or solid tissue barriers also known as lubricants. Examples of tissue barriers include, but are not limited to, polysaccharides, polyglycans, Seprafilm, Interseed, and hyaluronic acid.

[0214] In some embodiments, the therapeutic agent administered with the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, is any suitable therapeutic agent that is usefully delivered by inhalation, e.g., analgesics, e.g., codeine, dihydromorphine, ergotamine, fentanyl, or morphine; preparations for angina, e.g., diltiazem; antiallergic agents, e.g., cromoglycate, ketotifen, or nedocromil; antiinfective agents, e.g., cephalosporins, penicillin, streptomycin, sulfonamides, tetracycline, or pentamidine; antihistamines, e.g., metapyrylene; anti-inflammatory agents, e.g., beclomethasone, flunisolide, budesonide, typredan, triamcinolone acetonide, or fluticasone; antitussives, e.g., noscapine; bronchodilators, e.g., ephedrine, adrenaline These include phenoterol, formoterol, isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pyrbuterol, reproterol, limiterol, salbutamol, salmeterol, terbutaline, isoethaline, tulobuterol, orciprenaline or (-)-4-amino-3,5-dichloro-α-[[[6-[2-(2-pyridinyl)ethoxy]hexyl]-amino]methyl]benzenemethanol; diuretics, e.g., amiloride; anticholinergics, e.g., ipratropium, atropine or oxytropium; hormones, e.g., cortisone, hydrocortisone or prednisolone; xanthines, e.g., aminophylline, choline theophylline, lysine theophylline or theophylline; and therapeutic proteins and peptides, e.g., insulin or glucagon. It will be apparent to those skilled in the art that therapeutic agents may, where appropriate, be used in the form of salts (e.g., alkali metal salts, amine salts, or acid addition salts), esters (e.g., lower alkyl esters), or solvates (e.g., hydrates) to optimize the activity and / or stability of the therapeutic agent.

[0215] Further therapeutic agents that can be combined with the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, 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 entirety.

[0216] Compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be used in combination with the therapeutic agents disclosed herein, depending on the condition being treated. Therefore, in some embodiments, one or more compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be co-administered with the other therapeutic agents described above. When used in combination therapy, compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, may be administered simultaneously with or separately from the second therapeutic agent. This co-administration may include simultaneous administration in the same dosage form, simultaneous administration in different dosage forms, and separate administration. That is, compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, and any of the therapeutic agents described above may be formulated together in the same dosage form and administered simultaneously. Alternatively, compounds of formula (I), or their pharmaceutically acceptable salts, isotopic forms, or stereoisomers, and any of the therapeutic agents described above may both exist in separate formulations and be administered simultaneously. Alternatively, one of the above therapeutic agents may be administered immediately after administration of the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, and vice versa. In some embodiments of separate administration protocols, the compound of formula (I), or a pharmaceutically acceptable salt, isotopic form, or stereoisomer thereof, and one of the above therapeutic agents are administered at intervals of several minutes, several hours, or several days.

[0217] The examples and preparations provided below further illustrate and demonstrate the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers, and methods for preparing such compounds. It should be understood that the scope of this disclosure is not limited in any way by the scope of the following examples and preparations. In the following examples, and throughout this specification and the claims, molecules having a single stereocenter exist as racemic mixtures unless otherwise specified. Molecules having two or more stereocenters exist as racemic mixtures of diastereomers unless otherwise specified. Single enantiomers / diastereomers can be obtained by methods known to those skilled in the art. [Examples]

[0218] The following examples are provided for illustrative purposes only. Methods for preparing the compound of formula (I), or its pharmaceutically acceptable salts, isotopic forms, or stereoisomers are provided herein or can be derived by those skilled in the art.

[0219] The examples and preparations provided below further illustrate and demonstrate the compounds of the Disclosure and the methods for testing such compounds. It should be understood that the scope of the Disclosure is not limited in any way by the scope of the following examples.

[0220] The chemical reactions of the examples described herein can be readily adapted to prepare several other compounds disclosed herein, and alternative methods for preparing the compounds of this disclosure are considered to be within the scope of this disclosure. For example, the synthesis of compounds according to this disclosure that are not illustrated may be carried out by modifications obvious to those skilled in the art, for example, by appropriately protecting interfering groups, by using other suitable reagents known in the art other than those described, or by modifying the reaction conditions, reagents, and starting materials to those of a more conventional method. Alternatively, other reactions disclosed herein or known in the art will be recognized as applicable to the preparation of the compounds of this disclosure.

[0221] Unless otherwise specified in the following examples, the compounds are isolated as a racemic mixture.

[0222] In this application, the following abbreviations apply. Abbreviation Acetic acid (TOH) CAN: Cerium ammonium nitrate DAST: Diethylaminosulfur trifluoride DCM: Dichloromethane DIPEA: N,N-diisopropylethylamine DMEM: Dulbecco's modified Eagle medium DMF: Dimethylformamide DMSO: Dimethyl sulfoxide EMEM: Eagle's Minimum Essential Medium æ:ethyl acetate EtOH: Ethanol FBS: Fetal Bovine Serum Fmoc: Fluorenyl Methoxycarbonyl 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-BH3: Picolinboran PMB: Para-methoxybenzyl ether Preparative HPLC: Preparative High-Performance Liquid Chromatography rt or RT: Room temperature TFA: Trifluoroacetic acid TLC: Thin-layer chromatography T3P: Propanephosphonic anhydride

[0223] Example of synthesis 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-oxopropane-2-ylcarbamate. To a stirred solution of compound (S)-2-(((9H-fluoren-9-yl)methoxy)carbonylamino)propanoic acid (5.0 g, 16.0 g) in dichloromethane (100 mL), T3P (15.2 mL, 24.1 g) and DIPEA (5.6 mL, 32.1 mmol) were added at room temperature. The reaction mixture was stirred at room temperature for 15 minutes, and 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 hours. The reaction was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (100 mL) and extracted with dichloromethane (2 × 100 mL). The combined organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by flash column chromatography (eluted with 100-200 mesh silica gel, 40% ethyl acetate / petroleum ether) to obtain the pure compound (9H-fluoren-9-yl)methyl(2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropane-2-ylcarbamate (5.2 g, 69.1%) as a rubbery compound.

[0224] Step 2: Synthesis of (2S)-2-amino-N-(2,2-dimethoxyethyl)-N-(2-methylbutyl)propenamide. 20% piperidine (70 mL) in DMF was added at 0°C to a stirred solution of (9H-fluoren-9-yl)methyl(2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropane-2-ylcarbamate (34.0 g, 72.6 mmol) in DMF (230 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC. After the reaction was complete, excess DMF (100 mL) was added, followed by washing 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 dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain (2S)-2-amino-N-(2,2-dimethoxyethyl)-N-(2-methylbutyl)propanamide (20.4 g, 68.4%) as a rubbery solid.

[0225] Step 3: Synthesis of (9H-fluoren-9-yl)methyl 3-((2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropan-2-ylamino)-3-oxopropylcarbamate. 3-(((9H-fluoren-9-yl)methoxy)carbonylamino)propanoic acid (20.2 g, 81.2 mmol) was stirred in dichloromethane at room temperature (500 mL). T3P (80 mL, 121.8 mmol) and DIPEA (28.6 mL, 160.4 mmol) were added, and the mixture was stirred for 10 minutes. (2S)-2-amino-N-(2,2-dimethoxyethyl)-N-(2-methylbutyl)propanamide (25.5381.2 mmol) was added, and stirring was continued at room temperature for 16 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with water (500 mL) and extracted with dichloromethane (2 × 500 mL). The combined organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product. The crude compound was purified by flash column chromatography (eluted with 100-200 mesh silica gel, 70% ethyl acetate / petroleum ether) to obtain 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 the rubbery compound.

[0226] Step 4: Synthesis of (6S)-(9H-fluoren-9-yl)methyl6-methyl-8-(2-methylbutyl)-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate. Formic acid (105 mL) was added to a stirred solution of (9H-fluoren-9-yl)methyl3-((2S)-1-((2,2-dimethoxyethyl)(2-methylbutyl)amino)-1-oxopropane-2-ylamino)-3-oxopropylcarbamate (21.0 g, 38.9 mmol). 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 obtain the crude compound. The crude compound was placed in a saturated aqueous solution of NaHCO3 (200 mL) and then extracted with ethyl acetate (3 × 500 mL). The combined organic layers were washed with brine solution (500 mL), then dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude compound was purified by flash column chromatography (eluted with 100-200 mesh silica gel, 50% ethyl acetate / petroleum ether) to obtain the pure compound (6S)-(9H-fluoren-9-yl)methyl6-methyl-8-(2-methylbutyl)-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate (25 g, 69.0%) as a rubbery substance.

[0227] Step 5: Synthesis of (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione. 20% piperidine (30 mL) in DMF was added to a stirred solution of (6S)-(9H-fluoren-9-yl)methyl6-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. The reaction mixture was heated to room temperature and stirred for 2 hours. The reaction was monitored by TLC. Once the starting materials were completely consumed, additional DMF (50 mL) was added, 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 dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the desired crude 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.

[0228] Example S2. Synthesis of Compound 1a. The synthesis route for preparing Compound 1a is shown in Scheme 2. Scheme 2. [ka] To a solution of 4-(trifluoromethyl)benzoic acid (0.232 g, 0.91 mmol) in dichloromethane (20 mL) stirred at room temperature, T3P (1.2 mL, 1.37 mmol) and DIPEA (0.42 mL, 1.82 mmol) were added, and the mixture was stirred for 15 minutes. (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.310 g, 0.91 mmol) was added, and stirring was continued for 8 hours. The progress of the reaction was monitored by TLC. After the reaction was complete, the mixture was quenched with water (50 mL) and extracted with dichloromethane (2 × 50 mL). The combined organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then freeze-dried to obtain 1a (0.340 g, 65.3%) as a solid. Preparative HPLC: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: Acetonitrile; Column: X-Select phenylhexyl (150 × 19 mm 5 μm); Flow rate: 16 mL / min. MS (ESI) m / z [M+H] + : 426.05.

[0229] Example S3. Synthesis of compound 2a. The synthesis route for preparing compound 2a is shown in Scheme 3. Scheme 3. [ka] To a solution of 4-(difluoromethoxy)benzoic acid (0.37 g, 1.968 mmol) in dichloromethane (15 mL) at room temperature, DIPEA (0.8 mL, 5.904 mmol) and T3P (2.0 mL, 3.936 mmol) were added. The mixture was stirred for 30 minutes, 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 hours. The progress of the reaction 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. The pure fraction was recovered and freeze-dried to obtain 2a (380 mg 46%) as a solid. Preparative HPLC conditions: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: acetonitrile; Column: Kromosil phenyl (150 × 25 mm 10 μm); Flow rate: 25 mL / min. MS (ESI) m / z [M+H] + : 424.11.

[0230] Example S4. Synthesis of compound 3a. The synthesis route for preparing compound 3a is shown in Scheme 4. Scheme 4. [ka] (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.500 g, 1.97 mmol) was mixed with methanol (20 mL) at room temperature and stirred. 4-hydroxybenzaldehyde (0.289 g, 1.97 mmol) and acetic acid (0.23 mL, 3.95 mmol) were added to the mixture. The reaction mixture was stirred at room temperature for 5 minutes. Picoline borane (0.253 g, 2.37 mmol) was added, 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 layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then freeze-dried to obtain 3a (0.180 g, 46.09%) as a solid. Preparative HPLC: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: acetonitrile; Column: Kromosil phenyl (150 × 25 mm, 10 μm); Flow rate: 25 mL / min. MS (ESI) m / z [M+H] + : 360.11.

[0231] Example S5. Synthesis of compound 4a. The synthesis 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, DIPEA (1.30 mL, 7.338 mmol) and HATU (1.39 g, 3.669 mmol) were added. The resulting reaction mixture was stirred for 30 minutes, 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 hours. 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 × 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. The pure fraction was recovered and lyophilized to obtain 4a (160 mg, 21.8%) as a solid. Preparative HPLC method: Mobile phase A: 0.01 mM ammonium bicarbonate aqueous solution; Mobile phase B: Acetonitrile; Column: X-Select phenylhexyl (150 × 19 mm, 5 μm); Flow rate: 15 mL / min. MS (ESI) m / z [M+H] + : 375.05.

[0232] Example S6. Synthesis of compound 5a. The synthesis route for preparing compound 5a is shown in Scheme 6. Scheme 6. [ka] (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) were stirred in DMF (20 mL) at room temperature. K2CO3 (0.546 g, 3.95 mmol) was added to the solution, and the mixture was stirred for 8 hours. 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 layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then freeze-dried to obtain 5a (0.270 g, 63.8%) as a rubbery substance. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: Acetonitrile; Column: Kromosil C 18 (150×25mm 10μ);Flow rate: 25mL / min. MS (ESI) m / z [M+H] + : 412.2.

[0233] Example S7. Synthesis of compound 6a. The synthesis route for preparing compound 6a is shown in Scheme 7. Scheme 7. [ka] (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) were stirred at room temperature in DMF (20 mL) and K2CO3 (0.546 g, 9.95 mmol) was added. The reaction mixture was stirred at room temperature for 18 hours. 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 layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then freeze-dried to obtain 6a (0.178 g, 41.5%) as a semi-solid. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: Acetonitrile; Column: X-Select C 18 (250×19mm, 5μ); flow rate: 18mL / min. MS (ESI) m / z [M+H] + : 410.11.

[0234] Example S8. Synthesis of compound 7a. The synthetic route for preparing compound 7a is shown in Scheme 8. Scheme 8. [ka] To 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) in methanol (20 mL) stirred 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. Picoline borane (0.318 g, 2.96 mmol) was added, 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 layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude compound was purified by preparative HPLC. The pure fractions were combined, concentrated under reduced pressure, and then freeze-dried to obtain 7a (0.164 g, 42%) as a solid. Preparative HPLC: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: Acetonitrile; Column: X-BRIDGE C 18 (250×19mm, 5μ); flow rate: 18mL / min. MS (ESI) m / z [M+H] + : 361.11.

[0235] Example S9. Synthesis of compound 8a. The synthesis 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. 4-(benzyloxy)benzoic acid (0.360 g, 1.42 mmol) was mixed in dichloromethane (20 mL) at room temperature. T3P (1.2 mL, 1.7 mmol) and DIPEA (0.55 mL, 2.84 mmol) were added, and the mixture was stirred for 15 minutes. (6S)-6-methyl-8-(2-methylbutyl)tetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (0.400 g, 1.42 mmol) was added, and stirring was continued at room temperature for 16 hours. 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 layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain 0.9 g of crude material. Analysis of the crude material by LC / MS showed 54.59% of the desired product. The crude material was used in the next step without purification.

[0236] Step 2: Synthesis of compound 8a. (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) was mixed with methanol (20 mL) at room temperature and stirred. 10% Pd-C (0.200 g) was added under an N2 atmosphere. The reaction mixture was stirred under an H2 balloon at room temperature for 8 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered through Celite and evaporated under reduced pressure to obtain the crude compound. The crude compound was dissolved in dichloromethane (50 mL) and washed with aqueous NaHCO3 solution (20 mL) and brine solution (20 mL). The filtrate was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude compound was triturated with diethyl ether to obtain 8a (0.330 g, 82%) as a solid. MS (ESI) m / z [M+H] + : 374.11.

[0237] Example S10. Synthesis of Compound 9. The synthesis 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-oxoethyl carbamate. 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, DIPEA (11.88 mL, 67.3 mmol), N-(2,2-dimethoxyethyl)butan-2-amine (10.84 g, 67.3 mmol) and T3P (53.0 mL, 84.1 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC. After the completion of the reaction, ice-cold water (100 mL) was added, and the mixture was extracted with ethyl acetate (2 × 150 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the desired crude product. The crude compound was purified by flash column chromatography (100-200 mesh silica gel) and eluted with 20-25% ethyl acetate / petroleum ether to obtain (9H-fluoren-9-yl)methyl 2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethyl carbamate (10.8 g, 72.9%) as a solid.

[0238] Step 2: Synthesis of 2-amino-N-sec-butyl-N-(2,2-dimethoxyethyl)acetamide. Piperidine (2.4 mL) was added to a solution of (9H-fluoren-9-yl)methyl 2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethyl carbamate (10.8 g, 24.5 mmol) in DMF (20 mL) cooled to 0°C, and the reaction mixture was stirred at room temperature for 2 hours. The progress of the reaction was monitored by TLC. Once TLC indicated the complete consumption of the starting materials, 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 dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the desired pure product 2-amino-N-sec-butyl-N-(2,2-dimethoxyethyl)acetamide (3.6 g, 67.2%) as a solid.

[0239] Step 3: Synthesis of (9H-fluoren-9-yl)methyl-3-(2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylamino)-3-oxopropylcarbamate. DIPEA (31.91 mL, 49.5 mmol), 3-(((9H-fluoren-9-yl)methoxy)carbonylamino)propanoic acid (5.14 g, 16.5 mmol), and T3P (39.13 g, 33 mmol) were added at 0°C 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). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC. After the 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 dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography using silica (230-400 mesh; eluent: 23-25% ethyl acetate / petroleum ether). The recovered pure fraction was concentrated under reduced pressure to obtain (9H-fluoren-9-yl)methyl-3-(2-(sec-butyl(2,2-dimethoxyethyl)amino)-2-oxoethylamino)-3-oxopropylcarbamate (4.1 g, 48.6%) as a rubbery substance.

[0240] Step 4: Synthesis of (9H-fluoren-9-yl)methyl 8-sec-butyl-4,7-dioxooctahydro-1H-pyrazino[1,2-a]pyrimidine-1-carboxylate. The mixture was stirred at room temperature for 16 hours in 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). The reaction was monitored by TLC. Once TLC indicated complete consumption of the starting materials, the reaction mixture was concentrated, the resulting mass was diluted with water, and extracted with dichloromethane (2 × 100 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 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 rubbery substance.

[0241] 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 equivalent) was added, and the reaction mixture was stirred at room temperature for 2 hours. The progress of the reaction was monitored by TLC. Once TLC indicated complete consumption of the starting materials, the reaction mixture was washed with petroleum ether (2 × 50 mL) to remove nonpolar impurities. Cold water was added, and the mixture was extracted with dichloromethane (2 × 100 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the pure product 8-sec-butyltetrahydro-1H-pyrazino[1,2-a]pyrimidine-4,7(6H,8H)-dione (900 mg, 55.9%) as a solid.

[0242] 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), acetic acid (0.27 mL, 2.0 equivalents) and picoline borane (0.285 g, 2.6 mmol) were added at room temperature. The reaction mixture was stirred at room temperature for 48 hours. 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 layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product. The prepared compound was analyzed by LC / MS. Crude LC / MS data showed a desired mass of 8.28%. The crude compound was purified by column chromatography on silica gel (100-200), and the desired compound was eluted with 50-70% ethyl acetate / petroleum ether. LC / MS of the eluted fraction showed a desired mass of 72.16%, which was further purified by preparative HPLC. After preparative HPLC purification, the fraction was collected, concentrated under reduced pressure, and then lyophilized to obtain 9 (0.168 g, 22.8%) as a solid. Preparative HPLC method: Mobile phase A: 10 mM ammonium bicarbonate aqueous solution; Mobile phase B: acetonitrile; Column: X-BRIDGE C 18 (150×19mm 5μ); Flow rate: 18 mL / min. MS (ESI) m / z [M+H] + : 332.2.

[0243] Example S11. Synthesis of Compound 10. The synthesis 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 hours. After completion, the reaction mixture was quenched with ice-cold water (50 mL), and the aqueous layer was extracted with SiO2 (50 mL x 2). The organic layer was washed with cold H2O (30 mL), followed by saturated brine (30 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% siRNA / hexane) to obtain 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, yield 39.2%) as a solid. MS (ESI) m / z [M+H] + : 392.05. 1 H NMR (400 MHz, DMSO-d6) δ 0.66 - 0.89 (m, 6 H) 0.91 - 1.42 (m, 4 H) 1.57 - 1.78 (m, 1 H) 2.16 - 2.35 (m, 2 H) 2.55-2.65 (m, 2 H) 3.08-3.23 (m, 2 H) 3.28-3.40 (m, 1 H) 3.51-3.64 (m, 2 H) 4.76-4.89 (m, 1 H) 5.88-6.02 (m, 1 H) 7.46-7.56 (m, 4 H).

[0244] 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 hours. After completion, the reaction mixture was quenched with ice-cold water (50 mL), and the aqueous layer was extracted with SiO2 (50 mL x 2). The organic layer was washed with cold H2O (30 mL), followed by saturated brine (30 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% HCl / hexane) to obtain 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, yield 38.0%) as a solid. MS (ESI) m / z [M+H] + : 376.05. 1 H NMR (400 MHz, DMSO-d6) δ 0.69 - 0.81 (m, 3 H) 0.86 (t, J=7.23 Hz, 3 H) 0.95 - 1.14 (m, 2 H) 1.20 - 1.43 (m, 4 H) 1.59 - 1.80 (m, 2 H) 2.26 (d, J=16.95 Hz, 1 H) 2.55 - 2.72 (m, 1 H) 3.20 - 3.31 (m, 2 H) 3.35 - 3.39 (m, 1 H) 3.52 - 3.70 (m, 2 H) 4.73 - 4.89 (m, 1 H) 7.33 (t, J=8.73Hz, 2H) 7.61 (dd, J=8.23, 5.73 Hz, 2H).

[0245] 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 hours. After completion, the reaction mixture was quenched with ice-cold water (50 mL), and the aqueous layer was extracted with SiO2 (50 mL x 2). The organic layer was washed with cold H2O (30 mL), followed by saturated brine (30 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% ethyl acetate / hexane) to obtain 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, yield 55.2%) as a solid. MS (ESI) m / z [M+H] + : 460.0. 1 H NMR (400 MHz, DMSO-d6) δ 0.74 - 0.93 (m, 6 H) 0.98 - 1.19 (m, 2 H) 1.28 - 1.46 (m, 3 H) 1.64 - 1.81 (m, 1 H) 2.22 (d, J=17.45 Hz, 1 H) 2.57 - 2.70 (m, 1 H) 3.14 (dd, J=13.21, 6.23 Hz, 1 H) 3.25 - 3.31 (m, 2 H) 3.44 - 3.57 (m, 1 H) 3.61 - 3.87 (m, 2 H) 4.78 - 4.90 (m, 1 H) 5.89 - 6.05 (m, 1 H) 7.72 (d, J=7.98 Hz, 1 H) 7.90 - 8.02 (m, 2 H).

[0246] Example S14. Synthesis of the 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)ethane-1-amine. Anisaldehyde (12 mL, 90.22 mmol) and 2,2-diethoxyethaneamine (10 g, 75.18 mmol) were charged into a 500 mL round-bottom flask. The reaction mixture was heated at 100 °C for 1 hour. The reaction mixture was cooled to room temperature, and EtOH (100 mL) was added, followed by NaBH4 (4.28 g, 112.7 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The resulting crude product was dissolved in Depositphotos (300 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4, and concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (silica 100-200 mesh; 70% siRNA / hexane) to obtain 2,2-diethoxy-N-(4-methoxybenzyl)ethane-1-amine (15 g, 59.28 mmol, 78% yield) in liquid form. MS (ESI) m / z [M+H] + : 254.3.

[0247] 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, HATU (42 g, 110.67 mmol), DIPEA (21.06 mL, 118.57 mmol), followed by 2,2-diethoxy-N-(4-methoxybenzyl)ethane-1-amine (20 g, 79.05 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. Once the starting materials were completely consumed, the reaction mixture was quenched with ice-cold water (300 mL), and the aqueous layer was extracted with ELISA (200 mL x 2). The organic layer was washed with cold H2O (200 mL), followed by brine (100 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. The obtained crude product was purified by column chromatography (silica 100-200 mesh; 50% siRNA / hexane) to obtain (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(4-methoxybenzyl)amino)-1-oxopropan-2-yl)carbamate (28 g, 51.22 mmol, yield 64.8%) as a rubbery liquid. MS (ESI) m / z [M+H-EtOH] + : 501.2.

[0248] Step 3: Synthesis of 2-amino-N-(2,2-diethoxyethyl)-N-(4-methoxybenzyl)propanamide. Diethylamine (200 mL) was added 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 CH2Cl2 (30 mL). The reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was concentrated, and the resulting crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 2-amino-N-(2,2-diethoxyethyl)-N-(4-methoxybenzyl)propanamide (14.5 g, 44.75 mmol, yield 87%) as a viscous liquid. MS (ESI) m / z [M+H-EtOH] + : 279.05.

[0249] 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, HATU (18.06 g, 47.53 mmol), DIPEA (9.21 mL, 51.85 mmol), and then 2-amino-N-(2,2-diethoxyethyl)-N-(4-methoxybenzyl)propanamide (14 g, 43.20 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. After completion, the reaction mixture was quenched with ice-cold water (200 mL), and the aqueous layer was extracted with ELISA (200 mL x 2). The organic layer was washed with cold H₂O (500 mL), followed by saturated brine (200 mL), dried over Na₂SO₄, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% ELISA / hexane) to obtain (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, yield 67.44%) as a viscous liquid. MS (ESI) m / z [M+H-EtOH] + : 572.

[0250] 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 hours. After completion, the reaction mixture was concentrated, and the resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% siRNA / hexane) to obtain (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, yield 94%) as a solid. MS (ESI) m / z [M+H] + : 526.

[0251] Step 6: Synthesis of 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. Diethylamine (100 mL) was added 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 CH2Cl2 (150 mL), and the reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was concentrated, and the resulting crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (7g, 23.07 mmol, yield 87%) as a sticky solid. MS (ESI) m / z [M+H] + : 304.

[0252] Example S15. Synthesis of the 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, HATU (10.52 g, 27.69 mmol), DIPEA (12.30 mL, 69.23 mmol), and then 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (7 g, 23.07 mmol) were added, and the reaction mixture was stirred at room temperature for 12 hours. After completion, the reaction mixture was quenched with ice-cold water (200 mL), and the aqueous layer was extracted with SiO2 (200 mL x 2). The organic layer was washed with cold H2O (200 mL), followed by saturated brine (150 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% SiO2 / hexane) to obtain 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, yield 82.04%) as a solid. MS (ESI) m / z [M+H] + : 476.15 and MS (ESI) m / z [M+Na] + : 498.05.

[0253] Step 2: Synthesis of 6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. 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 CH3CN:H2O (2:1, 150 mL) maintained at 0°C, CAN (31.15 g, 56.82 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with a saturated aqueous solution of NaHCO3 (200 mL) and extracted with ELISA (200 mL x 2). The combined organic layers were washed with H2O (200 mL), followed by saturated brine solution (150 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to obtain 6-methyl-1-(4-(trifluoromethyl)benzoyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.5 g, 9.85 mmol, yield 52.8%) as a solid. MS (ESI) m / z [M+H+CH3CN] + : 397.0. 1 H NMR (400 MHz, DMSO-d6) δ 1.25 - 1.46 (m, 3 H) 2.15-2.30 (m, 1 H) 2.56 - 2.69 (m, 1 H) 3.16 (d, J=4.99 Hz, 1 H) 3.22-3.30 (m, 1 H) 3.42 - 3.72 (m, 2 H) 4.70 - 4.87 (m, 1 H) 5.85-5.95 (m, 1 H) 7.75 (d, J=7.98 Hz, 2 H) 7.86 (d, J=7.98 Hz, 2 H) 8.11 (brs, 1 H).

[0254] Example S16. General procedure A for the synthesis of the final compound. In 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 (1.69 mmol, 1.69 mL, 1 M in THF), followed by alkyl halide (1.12 mmol), was added, and the reaction mixture was exposed to microwave irradiation at 120°C for 1 hour. The reaction mixture was cooled to room temperature and quenched with H2O (25 mL). The aqueous layer was extracted with ELISA (10 mL x 3). The combined organic layers were washed with brine and concentrated. The crude product was purified by CombiFlash.

[0255] Example S17. Synthesis of compound 15. Compound 15 was synthesized using (bromomethyl)cyclopentane as the alkyl halide according to general procedure A. MS (ESI) m / z [M+H] + : 438.65. 1 H NMR (400 MHz, DMSO-d6) δ 1.02 - 1.26 (m, 3 H) 1.28 - 1.42 (m, 2 H) 1.44 - 1.76 (m, 6 H) 1.80 - 2.08 - 2.33 (m, 2 H) 2.55 - 2.71 (m, 1 H) 3.22 (dd, J=12.96, 7.48 Hz, 1 H) 3.26 - 3.32 (m, 1 H) 3.39 (d, J=6.98 Hz, 1 H) 3.49-3.57 (m, 1 H) 3.59-3.74 (m, 1 H) 3.76-3.91 (m, 1 H) 4.80-4.90 (m, 1 H) 5.95-6.05 (m, 1 H) 7.72 - 7.79 (m, 2 H) 7.84 - 7.91 (m, 2 H).

[0256] Example S18. Synthesis of compound 16. Compound 16 was synthesized using bromomethylcyclobutane as the alkyl halide according to general procedure A. MS (ESI) m / z [M+H] + : 424.15. 1H NMR (400 MHz, DMSO-d6) δ 1.29 - 1.44 (m, 2 H) 1.58 - 1.89 (m, 4 H) 1.90 - 2.08 (m, 2 H) 2.16-2.31 (m, 1 H) 2.55 - 2.70 (m, 2 H) 3.18 - 3.31 (m, 1 H) 3.25 - 3.26 (m, 1 H) 3.34 - 3.42 (m, 1 H) 3.36 - 3.57 (m, 2 H) 3.60-3.69 (n, 1 H) 3.71-3.83 (m, 1 H) 4.75-4.89 (m, 1 H) 5.90-6.05 (m, 1 H) 7.70 - 7.79 (m, 2 H) 7.87 (d, J=8.31 ​​Hz, 2 H).

[0257] Example S19. Synthesis of compound 19. Compound 19 was synthesized using (2-bromoethyl)cyclopentane as the alkyl halide according to general procedure A. MS (ESI) m / z [M+H] + : 452.35. 1 H NMR (400 MHz, DMSO-d6) δ 0.94 - 1.18 (m, 3 H) 1.26 - 1.61 (m, 9 H) 1.66-1.83 (m, 2 H) 2.16-2.31 (m, 1 H) 2.56 - 2.70 (m, 1 H) 3.16 - 3.28 (m, 1 H) 3.35 - 3.56 (m, 3 H) 3.60-3.73 (m, 1 H) 3.77-3.90 (m, 1 H) 4.72 - 4.92 (m, 1 H) 5.94-6.06 (m, 1 H) 7.77 (d, J=7.98 Hz, 2 H) 7.87 (d, J = 7.98 Hz, 2 H).

[0258] Example S20. Synthesis of compound 20. Compound 20 was synthesized using (2-bromoethyl)cyclobutane as the alkyl halide according to general procedure A. MS (ESI) m / z [M+H] + : 438.25. 1H NMR (400 MHz, DMSO-d6) δ 1.27 - 1.44 (m, 3 H) 1.50-1.71 (m, 4 H) 1.71-1.88 (m, 2 H) 1.93 - 2.09 (m, 2 H) 2.13 - 2.34 (m, 2 H) 2.56 - 2.70 (m, 2 H) 3.25 - 3.32 (m, 1 H) 3.35 - 3.42 (m, 1 H) 3.45-3.55 (m, 1 H) 3.59 - 3.72 (m, 1 H) 3.74-3.90 (m, 1 H) 4.75-4.89 (m, 1 H) 5.94-6.05 (m, 1 H) 7.71 - 7.79 (m, 2 H) 7.87 (d, J=8.31 ​​Hz, 2 H).

[0259] Example S21. Synthesis of compound 21. Compound 21 was synthesized using 1-bromobutane as an alkyl halide according to general procedure A. MS (ESI) m / z [M+H] + : 412.20. 1 H NMR (400 MHz, DMSO-d6) δ 0.81-0.97 (m, 3 H) 1.15 - 1.57 (m, 7 H) 2.15-2.31 (m, 1 H) 2.57 - 2.69 (m, 1 H) 3.14 - 3.28 (m, 1 H) 3.35 - 3.60 (m, 3 H) 3.62-3.73 (m, 1 H) 3.74 - 3.92 (m, 1 H) 4.75 - 4.91 (m, 1 H) 5.94-6.06 (m, 1 H) 7.76 (d, J=7.34 Hz, 2 H) 7.87 (d, J=7.83 Hz, 2H).

[0260] Example S22. Synthesis of compound 22. Compound 22 was synthesized using general procedure A with 4-bromobuta-1-ene as the alkyl halide. MS (ESI) m / z [M+H] + : 410.20. 1H NMR (400 MHz, DMSO-d6) δ 1.28-1.45 (m, 3 H) 2.14-2.38 (m, 3 H) 2.55 - 2.69 (m, 1 H) 3.36 - 3.57 (m, 4 H) 3.58-3.72 (m, 1 H) 3.75-3.89 (m, 1 H) 4.75 - 4.90 (m, 1 H) 4.98 - 5.19 (m, 2 H) 5.69-5.84 (m, 1 H) 5.93-6.05 (m, 1 H) 7.76 (d, J=7.98 Hz, 2 H) 7.88 (d, J=7.98 Hz, 2 H).

[0261] 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 (400 MHz, DMSO-d6) δ 0.80-0.96 (m, 6 H) 1.30 - 1.48 (m, 3 H) 1.85-2.03 (m, 1 H) 2.15-2.31 (m, 1 H) 2.57 - 2.70 (m, 1 H) 3.06-3.16 (m, 1 H) 3.18-3.28 (m, 1 H) 3.36-3.45 (m, 1 H) 3.44 - 3.57 (m, 1 H) 3.60-3.74 (m, 1 H) 3.73 - 3.87 (m, 1 H) 4.77-4.92 (m, 1 H) 5.93-6.07 (m, 1H) 7.76 (d, J=7.48 Hz, 2 H) 7.87 (d, J=7.48 Hz, 2 H).

[0262] Example S24. Synthesis of compound 24. Compound 24 was synthesized using general procedure A with 2-bromopropane as the alkyl halide. MS (ESI) m / z [M+H] + : 398.55. 1H NMR (400 MHz, DMSO-d6) δ 1.10 (d, J=5.49 Hz, 6 H) 1.28-1.45 (m, 3 H) 2.16-2.24 (m, 1 H) 2.56 - 2.71 (m, 1 H) 3.34-3.40 (m, 1 H) 3.44 - 3.79 (m, 3 H) 4.59-4.72 (m, 1 H) 4.75-4.90 (m, 1 H) 5.86-6.00 (m, 1 H) 7.79 (d, J=7.98 Hz, 2 H) 7.83 - 7.92 (m, 2 H).

[0263] Example S25. Synthesis of the 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, HATU (3.45 g, 9.08 mmol), DIPEA (4.34 mL, 24.8 mmol), and then 8-(4-methoxybenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (2.5 g, 8.25 mmol) were added, and the reaction mixture was stirred at room temperature for 12 hours. After completion, the reaction mixture was quenched with ice-cold water (50 mL), and the aqueous layer was extracted with SiO2 (100 mL x 2). The organic layer was washed with cold H2O (100 mL), followed by saturated brine (100 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 30% SiO2 / hexane) to obtain 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, yield 89.5%) as a solid. MS (ESI) m / z [M+H] + : 474.12.

[0264] Step 2: Synthesis of 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. 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 CH3CN:H2O (2:1, 45 mL) maintained at 0°C, CAN (12.0 g, 21.90 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was quenched with a saturated aqueous solution of NaHCO3 (100 mL) and extracted with ELISA (200 mL x 2). The combined organic layers were washed with H2O (250 mL), followed by saturated brine solution (250 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to obtain 1-(4-(difluoromethoxy)benzoyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (2.0 g, 5.66 mmol, yield 89.6%) as a solid. MS (ESI) m / z [M+H] + : 353.95. 1 H NMR (400 MHz, DMSO-d6) δ 1.10 - 1.39 (m, 3 H) 2.17-2.18 (m, 1 H) 2.52 - 2.68 (m, 1 H) 3.18 - 3.27 (m, 2 H) 3.44 - 3.71 (m, 2 H) 4.69 - 4.83 (m, 1 H) 5.75 - 5.92 (m, 1 H) 7.24 (d, J=7.83 Hz, 2 H) 7.32 (t, J=72.0 Hz, 1 H) 7.57 (d, J=8.31 ​​Hz, 2 H) 8.04 (brs, 1 H).

[0265] Example S26. General procedure B for the synthesis of the final compound. 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 minutes. Alkyl halide (1.6 mmol) was added to this reaction mixture, and the reaction mixture was heated to room temperature and stirred for 3 hours. After completion, the reaction mixture was quenched with ice-cold H2O (15 mL), and the aqueous layer was extracted with ELISA (15 mL x 3). The combined organic layers were washed with brine and concentrated. The crude product was purified by CombiFlash.

[0266] Example S27. Synthesis of compound 13. Compound 13 was synthesized using (bromomethyl)cyclopentane as the alkyl halide according to general procedure B. MS (ESI) m / z [M+H] + : 436.05. 1 H NMR (400 MHz, DMSO-d6) δ 1.07-1.16 (m, 3 H) 1.32 (d, J=6.48 Hz, 3 H) 1.41 - 1.73 (m, 7 H) 2.06 - 2.21 (m, 1 H) 2.21 - 2.34 (m, 1 H) 2.54 - 2.70 (m, 1 H) 3.14 - 3.29 (m, 1 H) 3.35 - 3.45 (m, 1 H) 3.52 - 3.69 (m, 1 H) 3.75 - 3.93 (m, 1 H) 4.75 - 4.91 (m, 1 H) 5.88-5.99 (m, 1 H) 7.27 (d, J=8.48 Hz, 2 H) 7.35 (t, J=72.0 Hz, 1 H) 7.61 (d, J=8.98 Hz, 2 H).

[0267] Example S28. Synthesis of compound 14. Compound 14 was synthesized using (bromomethyl)cyclobutane as the alkyl halide according to general procedure B. MS (ESI) m / z [M+H] + : 421.14.1 H NMR (400 MHz, DMSO-d6) δ 1.16-1.25 (m, 1 H) 1.27-1.43 (m, 3 H) 1.54 - 1.73 (m, 2 H) 1.73 - 1.86 (m, 2 H) 1.89-2.03 (m, 2 H) 2.24 (d, J=17.12 Hz, 1 H) 2.53 - 2.69 (m, 2 H) 3.20-3.28 (m, 1 H) 3.29-3.40 (m, 1 H) 3.40 - 3.66 (m, 2 H) 3.69 - 3.87 (m, 1 H) 4.75-4.86 (m, 1 H) 5.74 - 6.02 (m, 1 H) 7.26 (d, J=8.31 ​​Hz, 2 H) ) 7.33 (t, J=72.0 Hz, 1 H) 7.59 (d, J=8.31 ​​Hz, 2 H).

[0268] Example S29. Synthesis of compound 17. Compound 20 was synthesized using 1-bromobutane as an alkyl halide according to general procedure B. MS (ESI) m / z [M+H] + : 410.0. 1 H NMR (400 MHz, DMSO-d6) δ 0.81 - 0.96 (m, 3 H) 1.15 - 1.39 (m, 4 H) 1.40-1.55 (m, 2 H) 2.26 (d, J=16.95 Hz, 1 H) 2.53 - 2.70 (m, 2 H) 3.12 - 3.30 (m, 2 H) 3.38 - 3.46 (m, 1 H) 3.56-3.74 (m, 2 H) 3.75-3.92 (m, 1 H) 4.84 (q, J=6.81 Hz, 1 H) 5.86-6.06 (m, 1 H) 7.28 (d, J=7.98 Hz, 2H) 7.36 (t, J=72.0 Hz, 1 H) 7.62 (d, J=8.48 Hz, 2 H).

[0269] Example S30. Synthesis of compound 18. Compound 18 was synthesized by general procedure B using 4-bromobuta-1-ene as the alkyl halide. MS (ESI) m / z [M+H] + : 408.06. 1 H NMR (400 MHz, DMSO-d6) δ 1.16 - 1.45 (m, 3 H) 2.18 - 2.33 (m, 3 H) 2.53 - 2.70 (m, 1 H) 3.36 - 3.46 (m, 3 H) 3.51 - 3.72 (m, 2 H) 3.74-3.90 (m, 1 H) 4.84 (q, J=6.65 Hz, 1 H) 4.91-5.15 (m, 2 H) 5.67-5.84 (m, 1 H) 5.86 - 6.03 (m, 1 H) 7.29 (d, J=8.48 Hz, 2 H) 7.36 (t, J=72.0Hz, 1H) 7.61 (d, J=8.48 Hz, 2 H).

[0270] Example S31. Synthesis of compound 27. Compound 27 was synthesized using 2-(bromomethyl)tetrahydrofuran as the alkyl halide according to general procedure B. MS (ESI) m / z [M+H] + : 438.1. 1 H NMR (400 MHz, CDCl3) δ 7.48 - 7.55 (m, 2 H), 7.20 - 7.30 (m, 2 H), 6.40 - 6.76 (m, 1 H), 5.90 - 6.20 (m, 1 H), 5.16 - 5.26 (m, 1 H), 4.06 - 4.17 (m, 2H), 3.82 - 3.92 (m, 4 H), 3.61 - 3.77 (m, 2 H), 2.83 - 2.99 (m, 1 H), 2.47 - 2.59 (m, 2 H), 2.01 - 2.12 (m, 4 H), 1.49 (s, 3 H).

[0271] Example S32. Synthesis of compound 28. Compound 28 was synthesized using (2-bromoethyl)benzene as the alkyl halide according to general procedure B. MS (ESI) m / z [M+H] + : 458.10. 1 H NMR (400 MHz, CDCl3) δ 7.40-7.50 (m, 2 H), 7.20 - 7.28 (m, 2 H), 7.33 - 7.43 (m, 5 H), 6.40 - 6.76 (m, 1 H), 5.90 - 6.20 (m, 1 H), 5.16 - 5.26 (m, 1 H), 3.72 - 3.96 (m, 2H), 3.44 - 3.52 (m, 1 H), 3.25 - 3.35 (m, 2 H), 2.83 - 2.99 (m, 2 H), 2.47 - 2.59 (m, 1 H), 2.42 - 2.60 (m, 1 H), 2.30 - 2.57 (m, 1H), 1.49 (s, 3H).

[0272] Example S33. Synthesis of compound 29. Compound 29 was synthesized using general procedure B with 4-(2-bromoethyl)pyridine as the alkyl halide. MS (ESI) m / z [M+H] + : 459.10. 1 H NMR (400 MHz, CDCl3) δ 8.50 - 8.58 (m, 2 H), 7.24 - 7.46 (m, 4 H), 7.18 (d, J = 7.99Hz, 2 H), 6.40 - 6.76 (m, 1 H), 5.90 - 6.20 (m, 1 H), 5.16 - 5.26 (m, 1 H), 3.72 - 3.96 (m, 2H), 3.44 - 3.52 (m, 1 H), 3.25 - 3.35 (m, 2 H), 2.83 - 2.99 (m, 2 H), 2.47 - 2.59 (m, 1 H), 2.42 - 2.60 (m, 1H), 2.30 - 2.57 (m, 1H), 1.49 (s, 3H).

[0273] 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, CDCl3) δ 8.50 - 8.58 (m, 2 H), 7.24 - 7.46 (m, 4 H), 7.18 (d, J = 7.99Hz, 2 H), 6.40 - 6.76 (m, 1 H), 5.90 - 6.20 (m, 1 H), 5.16 - 5.26 (m, 1 H), 3.72 - 3.96 (m, 2H), 3.44 - 3.52 (m, 1 H), 3.25 - 3.35 (m, 2 H), 2.83 - 2.99 (m, 2 H), 2.47 - 2.59 (m, 1 H), 2.42 - 2.60 (m, 1H), 2.30 - 2.57 (m, 1H), 1.49 (s, 3H).

[0274] 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, CDCl3) δ 7.48 (d, J = 8.01 Hz, 2H), 7.20 - 7.28 (m, 2 H), 6.40 - 6.76 (m, 1 H), 5.90 - 6.20 (m, 1 H), 5.16 - 5.26 (m, 1 H), 3.72 - 3.96 (m, 1H), 3.46 - 3.64 (m, 5 H), 2.46 - 2.64 (m, 2 H), 1.43 - 1.56 (m, 5 H), 0.43-0.65 (m, 2H), 0.75-0.85 (m, 2 H).

[0275] Example S36. Synthesis of compound 32. Compound 32 was synthesized using 1-bromo-2-methoxyethane as the alkyl halide according to general procedure B. MS (ESI) m / z [M+H] + : 412.1. 1 H NMR (400 MHz, DMSO-d6) δ 7.52-7.62 (m, 2 H), 7.16 - 7.34 (m, 3 H), 5.85 - 5.95 (m, 1 H), 4.80 - 4.90 (m, 1 H), 3.85 - 3.95 (m, 1 H), 3.70 - 3.80 (m, 2 H), 3.25 - 3.46 (m, 5H), 3.22 (s, 3 H), 2.62 - 2.72 (m, 1 H), 2.20 - 2.30 (m, 1 H), 1.49 (s, 3 H).

[0276] 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 (400 MHz, DMSO-d6) δ 7.52-7.62 (m, 2 H), 7.16 - 7.34 (m, 3 H), 5.85 - 5.95 (m, 1 H), 4.80 - 4.90 (m, 1 H), 3.85 - 3.95 (m, 1 H), 3.70 - 3.80 (m, 2 H), 3.58 - 3.68 (m, 2H), 3.45 - 3.55 (m, 4H), 3.22 (s, 3 H), 2.62 - 2.72 (m, 1 H), 2.20 - 2.30 (m, 2 H), 1.49 (s, 3 H).

[0277] 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. 1H NMR (400 MHz, chloroform) δ 7.49 (d, J = 8.01 Hz, 2 H), 7.15 - 7.26 (m, 2 H), 6.40 - 6.76 (m, 1 H), 5.90 - 6.20 (m, 1 H), 5.15 - 5.25 (m, 1 H), 3.86 - 3.97 (m, 3 H), 3.66 - 3.77 (m, 2 H), 3.38 - 3.49 (m, 3 H), 2.97 (s, 3 H), 2.59 - 2.69 (m, 2 H), 1.49 (s, 3 H).

[0278] 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), Cs2CO3 (0.827 g, 2.547 mmol) was added, followed by (2-bromoethoxy)(tert-butyl)dimethylsilane (0.243 g, 1.018 mmol) at 0°C. The reaction mixture was heated in a sealed tube at 120°C for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was slowly quenched with ice-cold water (30 mL) and extracted with SiO2 (50 mL). The combined organic layers were washed with ice-cold brine solution (3 × 30 mL), dried over Na₂SO₄, and concentrated under reduced pressure to obtain 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). The crude compound was used directly in the next reaction without further purification. MS (ESI) m / z [M+H] + : 512.10.

[0279] 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), TBAF (3 mL) was added at 0°C. The reaction mixture was allowed to return to room temperature and stirred for 6 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was slowly quenched with ice-cold water (5 mL) and extracted with ELISA (2 × 10 mL). The combined organic layer was washed with ice-cold brine solution (10 mL), dried over Na₂SO₄, and concentrated under reduced pressure to obtain the crude compound. The obtained crude compound was purified by column chromatography (silica gel 60-120 mesh; 10% MeOH / DCM) to obtain 1-(4-(difluoromethoxy)benzoyl)-8-(2-hydroxyethyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.102 g, yield 52%) as a white solid. MS (ESI) m / z [M+H] + : 398.2. 1 H NMR (400 MHz, DMSO-d6) δ 7.52-7.62 (m, 2 H), 7.16 - 7.34 (m, 3 H), 5.92 - 6.02 (m, 1 H), 6.78 - 6.88 (m, 2 H), 3.86 - 3.92 (m, 1 H), 3.47 - 3.62 (m, 6 H), 3.21 - 3.31 (m, 1H), 2.57 - 2.67 (m, 1 H), 2.25 - 3.35 (m, 1 H), 1.49 (s, 3 H).

[0280] 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), NaH (0.050 g, 1.274 mmol), followed by 2-bromoacetonitrile (0.112 g, 0.933 mmol) was added at 0°C, and the reaction mixture was allowed to stand at room temperature for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was slowly quenched with ice-cold water (70 mL) and extracted with SiO2 (100 mL). The combined organic layer was washed with ice-cold brine solution (100 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The obtained crude compound was purified by column chromatography (silica gel 60-120 mesh; 10% MeOH / DCM) to obtain 2-(1-(4-(difluoromethoxy)benzoyl)-6-methyl-4,7-dioxooctahydro-8H-pyrazino[1,2-a]pyrimidine-8-yl)acetonitrile (0.120 g, yield 36%) as a white solid. MS (ESI) m / z [M+H] + : 393.05.

[0281] Step 2. To a solution of 2-(1-(4-(difluoromethoxy)benzoyl)-6-methyl-4,7-dioxooctahydro-8H-pyrazino[1,2-a]pyrimidine-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. The reaction mixture was heated under a hydrogen gas atmosphere for 3 hours. 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 obtain the crude compound. The crude compound was triturated with n-pentane to obtain 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 (400 MHz, DMSO d6) δ 7.96 (s, 2 H), 7.55 - 7.65 (m, 2 H), 7.20 - 7.35 (m, 3 H), 5.90 - 6.20 (m, 1 H), 4.85 - 4.95 (m, 1 H), 3.82 - 3.92 (m, 1H), 3.55. - 3.85 (m, 2 H), 3.35 - 3.45 (m, 3 H), 2.95 - 3.05 (m, 2 H), 2.60 - 2.70 (m, 1H), 2.20 - 2.30 (m, 1 H), 1.35 (s, 3 H).

[0282] Example S41. General procedure C for the synthesis of the final compound. 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), Cs2CO3 (0.735 g, 2.264 mmol, 4 equivalents) and then alkyl halide (0.679 mmol, 1.2 equivalents) were added at 0°C. The reaction mixture was heated at 50°C for 1 hour under microwave irradiation. 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 RINKAN (20 mL x 3). The combined organic layers were washed with saturated brine solution (10 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The obtained crude compound was purified by column chromatography to obtain the final compound.

[0283] Example S42. Synthesis of compound 25. Compound 25 was synthesized using 2-(2-iodoethyl)furan as the alkyl halide according to general procedure C. MS (ESI) m / z [M+H] + : 448.10. 1H NMR: δ 7.40 - 7.50 (m, 2 H), 7.28 - 7.38 (m, 1 H), 7.15-7.25 (m, 2 H), 6.39 - 6.78 (m, 1 H), 6.25-6.35 (m, 1 H), 5.90 - 6.12 (m, 2 H), 5.25 - 5.35 (m, 1 H), 5.10 - 5.20 (m, 1 H), 3.70 - 3.80 (m, 1 H), 3.50 - 3.60 (m, 1 H), 3.20 - 3.40 (m, 2 H), 2.95 - 3.05 (m, 3 H), 2.45 - 2.60 (m, 2 H), 1.59 (s, 3 H).

[0284] 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, CDCl3) δ 7.40 - 7.48 (m, 2 H), 7.15-7.26 (m, 3 H), 6.85 - 6.95 (m, 2 H), 6.39 - 6.95 (m, 2 H), 5.90 - 6.20 (m, 1 H), 5.15 - 5.25 (m, 1 H), 3.72 - 3.96 (m, 2 H), 3.47 - 3.54 (m, 1 H), 3.32 - 3.42 (m, 3 H), 3.10 - 3.20 (m, 2 H), 2.42 - 2.56 (m, 2 H), 1.49 (s, 3 H).

[0285] Example S44. Synthesis of the 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 with microwaves at 130°C for 2 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was concentrated under vacuum, and the crude product was extracted with saturated solutions of ethyl acetate (100 mL) and sodium bicarbonate. The organic layer was dried over anhydrous Na2SO4, concentrated under vacuum, and purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain (9H-fluoren-9-yl)methyl6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (300 mg, yield 42%) as a sticky solid. MS (ESI) m / z [M+H] + : 406.

[0286] Step 2: Synthesis of 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. Diethylamine (6 mL) was added to a solution of (9H-fluoren-9-yl)methyl6-methyl-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (300 mg, 0.74 mmol) in CH2Cl2 (5 mL). The reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (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 obtain 6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (120 mg, yield 92%) as a white solid. MS (ESI) m / z [M+H] + : 184.

[0287] 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), K2CO3 (1.58 g, 11.46 mmol) was added at room temperature and the mixture was stirred for 10 minutes. 1-(bromomethyl)-4-(difluoromethoxy)benzene (1.086 g, 4.584 mmol) was added to the resulting reaction mixture, and the reaction mixture was heated at 80 °C for 6 hours. 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 ELISA (50 mL x 2). The combined organic layers were washed with saturated brine (20 mL), dried over Na₂SO₄, and concentrated under reduced pressure. The crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 1-(4-(difluoromethoxy)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.550 g, yield 43.0%) as an off-white solid. MS (ESI) m / z [M+H] + : 340.34.

[0288] Example S45. General procedure D for the synthesis of the final compound. 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), NaH (0.021 g, 0.8847 mmol) was added at 0°C, followed by the addition of a suitable alkyl halide (2 equivalents). The reaction mixture was heated to room temperature and stirred for 5 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was slowly quenched with a saturated aqueous solution of NaHCO3 (2 mL) and extracted with ELISA (10 mL x 2). The combined organic layer was washed with H2O (5 mL), followed by a saturated brine solution (5 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude material was purified by Combiflash column chromatography (5% MeOH / DCM) to obtain the final product.

[0289] Example S46. Synthesis of compound 37. Compound 37 was synthesized using 4-bromo-1,1,1-trifluorobutane as the alkyl halide according to general procedure D. MS (ESI) m / z [M+H] + : 354.2. 1 H NMR (400 MHz, CDCl3): δ 1.41 (d, J = 7.13 Hz, 3 H), 1.71 - 1.86 (m, 2 H), 2.01 - 2.15 (m, 2 H), 2.26 - 2.35 (m, 1 H), 2.60 - 2.67 (m, 1 H), 2.89 - 3.01 (m, 1 H), 3.07 - 3.15 (m, 1 H), 3.21 - 3.34 (m, 2 H), 3.46 - 3.65 (m, 2 H), 3.81 - 3.95 (m, 2 H), 4.35 - 4.41 (m, 1 H), 5.20 - 5.29 (m, 1 H), 6.53 (t, J = 72.0 Hz, 1 H), 7.08 - 7.16 (m, 2 H), 7.30 - 7.36 (m, 2 H).

[0290] 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, CDCl3) δ 1.01 - 1.15 (m, 2 H), 1.41 (d, J = 7.13 Hz, 3 H), 1.45 - 1.62 (m, 8 H), 1.66 - 1.80 (m, 2 H), 2.23 - 2.34 (m, 1 H), 2.58 - 2.72 (m, 1 H), 2.89 - 2.98 (m, 1 H), 3.04 - 3.18 (m, 2 H), 3.23 - 3.33 (m, 1 H), 3.43 - 3.54 (m, 1 H), 3.55 - 3.65 (m, 1 H), 3.78 - 3.93 (m, 1 H), 4.31 - 4.39 (m, 1 H), 5.15 - 5.26 (m, 1 H), 6.53 (t, J = 72.0 Hz, 1 H), 7.13 (d, J = 8.50 Hz, 2 H), 7.34 (d, J = 8.50 Hz, 2 H).

[0291] Example S48. Synthesis of compound 39. Compound 39 was synthesized by general procedure D using 4-bromobuta-1-ene as the alkyl halide. MS (ESI) m / z [M+H] + : 394.2. 1H NMR (400 MHz, CDCl3) δ 1.41 (d, J = 7.13 Hz, 3 H), 2.23 - 2.35 (m, 3 H), 2.60 - 2.71 (m, 1 H), 2.92 - 3.01 (m, 1 H), 3.06 - 3.14 (m, 1 H), 3.22 - 3.46 (m, 3 H), 3.53 - 3.64 (m, 1 H), 3.79 - 3.93 (m, 2 H), 4.28 - 4.38 (m, 1 H), 4.91 - 5.00 (m, 2 H), 5.16 - 5.26 (m, 1 H), 5.64 - 5.76 (m, 1 H), 6.52 (t, J = 72.0 Hz, 1 H), 7.10 - 7.16 (m, 2 H), 7.30 - 7.36 (m, 2 H).

[0292] 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, CDCl3) δ 1.41 (d, J = 7.13 Hz, 3 H), 1.56 - 1.65 (m, 4 H), 1.73 - 1.92 (m, 2 H), 1.95 - 2.07 (m, 2 H), 2.14 - 2.25 (m, 1 H), 2.26 - 2.35 (m, 1 H), 2.59 - 2.72 (m, 1 H), 2.91 - 2.99 (m, 1 H), 3.04 - 3.14 (m, 2 H), 3.23 - 3.43 (m, 2 H), 3.53 - 3.63 (m, 1 H), 3.87 (q, J = 13.38 Hz, 2H), 4.29 - 4.39 (m, 1 H), 5.17 - 5.24 (m, 1 H), 6.53 (t, J = 72.0 Hz, 1 H), 7.13 (d, J = 8.63 Hz, 2 H), 7.30 - 7.37 (m, 2 H).

[0293] 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 (400 MHz, DMSO-d6) δ 0.86 (t, J = 7.34 Hz, 3 H), 1.14 - 1.24 (m, 2 H), 1.24 - 1.30 (m, 2 H), 1.38 - 1.50 (m, 2 H), 1.98 - 2.10 (m, 1 H), 2.53 - 2.61 (m, 2 H), 2.64 - 2.77 (m, 2 H), 3.07 - 3.25 (m, 3 H), 3.32 - 3.41 (m, 1 H), 3.62 - 3.73 (m, 1 H), 3.87 - 3.93 (m, 2 H), 4.49 - 4.58 (m, 1 H), 4.84 - 4.94 (m, 1 H), 7.15 (d, J = 8.56 Hz, 2 H), 7.22 (t, J = 72.0 Hz, 1 H), 7.43 (d, J = 8.56 Hz, 1 H).

[0294] Example S51. Synthesis of compound 52. Compound 52 was synthesized by general procedure D using 2-trifluromethyl-1-bromoethane as the alkyl halide. MS (ESI) m / z [M+H] + : 420.16. 1H NMR (400 MHz, CDCl3) δ ppm 7.31 - 7.38 (m, 2 H), 7.11 - 7.16 (m, 2 H), 6.31 - 6.73 (m, 1 H), 5.26 (q, J = 7.21 Hz, 1 H), 4.23 - 4.44 (m, 2 H), 3.98 - 4.13 (m, 1 H), 3.80 - 3.93 (m, 3 H), 3.59 (t, J = 11.07 Hz, 1 H), 3.10 (dd, J = 11.51, 3.75 Hz, 1 H), 2.90 - 2.99 (m, 1 H), 2.62 - 2.72 (m, 1 H), 2.32 (dd, J = 4.38, 2.38 Hz, 1 H), 2.28 (dd, J = 4.31, 2.31 Hz, 1 H), 1.48 (d, J = 7.25 Hz, 1 H), 1.41 (d, J = 7.13 Hz, 3 H).

[0295] Example S52. General procedure E for the synthesis of the final compound. In a flask immersed in an ice / water bath, 6-methyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.300 g, 1.184 mmol) was stirred in DMF (6 mL) to a solution to which cesium carbonate (0.771 g, 2.368 mmol, 2 equivalents) was added, followed by a suitable alkyl halide (1.1 equivalents). The flask was removed from the bath and stirred until complete consumption of the starting materials was indicated by TLC. The reaction mixture was poured into ice-cold water (70 mL) and the aqueous layer was extracted with SiO2 (100 mL). The organic layer was washed with ice-cold brine (50 mL x 3), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude product was purified by preparative HPLC to obtain the final compound.

[0296] 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 (400 MHz, DMSO-d6) δ 0.75 - 0.89 (m, 3 H), 0.82 - 0.87 (m, 3 H), 0.96 - 1.13 (m, 1 H), 1.23 - 1.31 (m, 4 H), 1.64 - 1.75 (m, 1 H), 2.06 - 2.09 (m, 1 H), 2.55 - 2.62 (m, 1 H), 2.65 - 2.76 (m, 1 H), 3.05 - 3.15 (m, 1 H), 3.15 - 3.26 (m, 3 H), 3.64 - 3.74 (m, 1 H), 3.84 - 3.95 (m, 2H), 4.52-4.60 (m, 1 H), 4.86 - 4.94 (m, 1 H), 7.17 (t, J = 8.76 Hz, 2 H), 7.41 (dd, J = 8.19, 5.82 Hz, 2 H).

[0297] 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. 1H NMR (400 MHz, DMSO-d6) 0.72-0.80 (m, 3 H), 0.80 - 0.87 (m, 3 H), 0.96 - 1.10 (m, 1 H),1.21 - 1.27 (m, 1 H), 1.28 - 1.34 (m, 3 H), 1.62 - 1.79 (m, 1 H), 2.00 - 2.13 (m, 1 H), 2.53 - 2.65 (m, 1 H), 2.66 - 2.76 (m, 1 H), 3.00 - 3.10 (m, 1 H), 3.17 - 3.29 (m, 3 H), 3.62 - 3.72 (m, 1 H), 4.00 - 4.08 (m, 2 H), 4.55 - 4.65 (m, 1 H), 4.85 - 4.95 (m, 1 H), 7.52 - 7.60 (m, 1 H), 7.73 (s, 1 H), 7.80 - 7.88 (m, 1 H).

[0298] 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), acetic acid (1 mL, 1.657 mmol) was added, and the reaction mixture was heated at 80°C for 1 hour. To the resulting reaction mixture, NaBH4 (0.188 g, 4.973 mmol) was added in portions, and the reaction mixture was heated at 80°C and stirred for 4 hours. Once complete consumption of the starting materials was shown by TLC analysis (5% MeOH / DCM), 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), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM), subsequently washed with water (30 mL), and dried under reduced pressure to obtain compound 44 (0.250 g, yield 39%) as an off-white solid. MS (ESI) m / z [M+H] + : 383.4. 1H NMR (400 MHz, DMSO-d6) δ 0.70 (t, J = 7.09 Hz, 3 H), 0.75 - 0.82 (m, 3 H), 0.91 - 1.11 (m, 1 H), 1.22 - 1.31 (m, 3 H), 1.57 - 1.72 (m, 1 H), 1.97 - 2.07 (m, 1 H), 2.55 - 2.70 (m, 2 H), 2.83 (dt, J = 10.91, 2.74 Hz, 1 H), 2.95 - 3.07 (m, 1 H), 3.10 - 3.26 (m, 3 H), 3.54 - 3.69 (m, 1H), 3.96 - 4.04 (m, 1 H), 4.06 - 4.15 (m, 1 H), 4.54 - 4.64 (m, 1 H), 4.84 - 4.95 (m, 1 H), 6.94 - 7.02 (m, 1 H), 7.04 - 7.13 (m, 1 H), 7.29 - 7.40 (m, 2 H), 7.65 (d, J = 7.95 Hz, 1 H), 10.95 (s, 1 H).

[0299] Example S55. Synthesis of the 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 the mixture was stirred at 80°C for 3 hours. 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 ELISA (50 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The obtained crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)dione (160 mg, yield 70%) as a white solid. MS (ESI) m / z [M+H] + : 292.

[0300] Example S56. Synthesis of compound 45. 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 an ice bath was mixed with NaH (20 mg, 0.2739 mmol), stirred for 20 minutes, and then, after 3 hours, (2-bromoethyl)cyclobutane (67 mg, 0.41 mmol) was added. Once the starting material was completely consumed (monitored by TLC), the reaction mixture was quenched with ice-cold water and extracted with ethyl acetate. The organic layer was dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 8-(2-cyclobutylethyl)-1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (13 mg, yield 16%) as a rubbery liquid. MS (ESI) m / z [M+H] + : 374. 1H NMR (400 MHz, CD3Cl3): δ 7.30 - 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).

[0301] 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 an ice bath, NaH (20 mg, 0.2739 mmol) was added and the mixture was stirred for 20 minutes. After 3 hours, (2-bromoethyl)cyclopentane (72 mg, 0.41 mmol) was added, and the completion of the starting material was monitored by TLC. The reaction mixture was quenched with ice-cold water and extracted with ethyl acetate. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The obtained crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 8-(2-cyclopentylethyl)-1-(4-fluorobenzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione as a rubbery liquid.

[0302] Example S58. Synthesis of the 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. 2-methylbutanal (11.60 g, 0.137 mmol) was added to stirred untreated 2,2-diethoxyethane-1-amine (20.0 g, 0.137 mmol) at room temperature, and the reaction mixture was heated to 100°C for 3 hours. Ethanol (200 mL), followed by NaBH4 (15.40 g, 0.413 mmol) was slowly added to the resulting reaction mixture at room temperature, and the reaction mixture was stirred for 16 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was cooled to room temperature and slowly quenched with a saturated solution of NH4Cl (100 mL). The aqueous layer was extracted with ELISA (200 mL x 2). The combined organic layers were washed with brine (400 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude compound. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to obtain N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (25.8 g, yield 88%) as a colorless liquid. MS (ESI) m / z [M+H] + : 204.3. 1 H NMR (400 MHz, DMSO-d6) δ 0.80 - 0.89 (m, 6 H) 1.11 (t, J=6.98 Hz, 6H) 1.35 - 1.48 (m, 2 H) 2.28-2.32 (m, 1 H) 2.41-2.45 (m, 1 H) 2.55 (d, J=5.49 Hz, 2 H) 3.42 - 3.52 (m, 2 H) 3.57 - 3.65 (m, 2 H) 4.49 (t, J=5.49 Hz, 1 H).

[0303] 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, HATU (26.0 g, 68.80 mmol), DIPEA (23.92 mL, 137.61 mmol), and then N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (12.10 g, 59.63 mmol) were added. The reaction mixture was stirred at room temperature for 4 hours. Once the starting materials were completely consumed, the reaction mixture was quenched with ice-cold water (500 mL), and the aqueous layer was extracted with siRNA (250 mL x 2). The combined organic layer was washed with cold H₂O (200 mL), followed by brine (200 mL), dried over Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude material was purified by column chromatography (silica 100-200 mesh; 80% siRNA / hexane) to obtain (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)carbamate (21.0 g, yield 89.43%) as a yellow viscous solid. MS (ESI) m / z [M+Na] + : 535.35.

[0304] Step 3: Synthesis of 2-amino-N-(2,2-diethoxyethyl)-3-hydroxy-N-(2-methylbutyl)propenamide. Diethylamine (58 mL, 2.80 times the volume) was added to a stirred solution of (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-3-hydroxy-1-oxopropan-2-yl)carbamate in anhydrous DCM (110 mL) maintained at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitoring by TLC), the reaction mixture was concentrated under reduced pressure to obtain the crude product. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 2-amino-N-(2,2-diethoxyethyl)-3-hydroxy-N-(2-methylbutyl)propenamide (9.50 g, yield 80%) as a yellow, sticky solid. MS(ESI)m / z[M+H] + :291.4.

[0305] 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, 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) were added at room temperature, and the reaction mixture was stirred for 16 hours. After completion, the reaction mixture was quenched with ice-cold water (200 mL), and the aqueous layer was extracted with ethyl acetate (200 mL x 2). The organic layer was washed with cold H2O (500 mL), followed by saturated brine (200 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by column chromatography (silica 100-200 mesh; 80% ethyl acetate / hexane) to obtain (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, yield 31.0%) as a yellow, viscous oil. MS (ESI) m / z [MH] - : 582.2.

[0306] Step 5: Synthesis of (9H-fluoren-9-yl)methyl6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. The 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 times volume) at room temperature and the reaction mixture were stirred for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure to obtain (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.

[0307] Step 6: Synthesis of 6-(hydroxymethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. Diethylamine (18.0 mL) was added at 0°C to a solution of (9H-fluoren-9-yl)methyl6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (6.0 g, 12.20 mmol) in CH2Cl2 (36.0 mL), and the reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitoring by TLC), the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude material was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 6-(hydroxymethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (3.0 g, yield 93.75%) as a colorless, viscous oil. MS (ESI) m / z [M+H] + : 270.20.

[0308] Step 7: Synthesis of tert-butyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. 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 CH2Cl2 (60 mL), triethylamine (4.5 mL, 33.45 mmol), followed by Boc anhydride (3.78 mL, 16.72 mmol) was added at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. Once the starting materials were completely consumed (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), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by column chromatography (silica 100-200 mesh; 10% MeOH / DCM) to obtain tert-butyl 6-(hydroxymethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (8.0 g, yield 31.0%) as a yellow, viscous oil. MS (ESI) m / z [M+H] + : 370.25.

[0309] Step 8: Synthesis of tert-butyl 6-(fluoromethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. DAST (1.97 g, 12.19 mmol) was added at -78°C 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) and the mixture was stirred for 15 minutes. The reaction mixture was raised to room temperature and stirred for 3 hours. After the reaction was complete (monitored by TLC), the reaction mixture was quenched with saturated NaHCO3 solution (15 mL), and the aqueous layer was extracted with SiO2 (100 mL x 2). The combined organic layers were washed with saturated brine (50 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The crude material was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain tert-butyl 6-(fluoromethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (0.800 g, yield 72.0%) as a colorless, viscous oil. MS (ESI) m / z [M+H] + : 372.2.

[0310] 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), 4 M HCl (5 mL) in dioxane was added at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was quenched with a saturated solution of sodium bicarbonate (10 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (10 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain 6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione hydrochloride (0.630 g, crude) as a brown, viscous oil. MS (ESI) m / z [M+H] + Free base: 271.00.

[0311] 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), K2CO3 (0.381 g, 2.760 mmol) was added, followed by 1-(bromomethyl)-4-(difluoromethoxy)benzene (0.261 g, 1.100 mmol). The reaction mixture was stirred at room temperature for 16 hours. After completion (monitoring by TLC), the reaction mixture was slowly quenched with ice-cold water (6 mL) and extracted with SiO2 (20 mL x 3). The combined organic layer was washed with saturated brine solution (10 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by preparative HPLC to obtain 1-(4-(difluoromethoxy)benzyl)-6-(fluoromethyl)-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.040 g, yield 17.0%) as a white solid. MS (ESI) m / z [M+H] + : 428.10. 1 H NMR (400 MHz, CDCl3) δ 7.34 (d, J =8.01, 2 H), 7.11 (d, J =8.01, 2 H), 6.32 - 6.69 (m, 1 H), 5.14-5.25 (m, 2 H), 4.60 - 4.76 (m, 2 H), 3.84 - 3.97 (m, 2 H), 3.35 - 3.45 (m, 2 H), 3.12 - 3.40 (m, 4 H), 2.85 - 3.05 (m, 1 H), 2.65 - 2.75 (m, 1 H), 2.29 - 2.34 (m, 1 H), 1.65 - 1.75 (m, 1H), 1.30 - 1.40 (m, 1 H), 1.05 -1.18 (m, 1 H), 0.80 - 0.90 (m, 6 H).

[0312] 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), Cs2CO3 (0.814 g, 2.506 mmol) was added, followed by 1-(bromomethyl)-4-(trifluoromethyl)benzene (0.598 g, 2.506 mmol). The reaction mixture was stirred at room temperature for 16 hours. After completion (monitoring by TLC), the reaction mixture was slowly quenched with ice-cold water (6 mL) and extracted with SiO2 (20 mL x 3). The combined organic layers were washed with saturated brine solution (10 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by preparative HPLC to obtain 6-(fluoromethyl)-8-(2-methylbutyl)-1-(4-(trifluoromethyl)benzyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.045 g, yield 8.0%) as a white solid. MS (ESI) m / z [M+H] + : 430.10. 1 H NMR (400 MHz, CDCl3) δ 7.34 (d, J =8.01, 2 H), 7.11 (d, J =8.01, 2 H), 5.14-5.25 (m, 2 H), 4.60 - 4.76 (m, 2 H), 3.84 - 3.97 (m, 2 H), 3.35 - 3.45 (m, 2 H), 3.12 - 3.40 (m, 4 H), 2.85 - 3.05 (m, 1 H), 2.65 - 2.75 (m, 1 H), 2.29 - 2.34 (m, 1 H), 1.65 - 1.75 (m, 1H), 1.30 - 1.40 (m, 1 H), 1.05 -1.18 (m, 1 H), 0.80 - 0.90 (m, 6 H).

[0313] Example S61. Synthesis of the intermediate compound methyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-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. 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-methoxy-4-oxobutanoic acid (1.90 g, 9.475 mmol) was stirred 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 minutes. N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (3.50 g, 9.475 mmol) was added to the resulting reaction mixture, and the mixture was then heated to room temperature and stirred for 6 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was quenched with ice-cold water (100 mL), and the aqueous layer was extracted with RINKAN (50 mL x 2). The combined organic layer was washed with cold H2O (50 mL), followed by brine (50 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. The crude material was purified by CombiFlash column chromatography using 50% RINKAN / n-hexane to obtain methyl 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (4.30 g, yield 83.0%) as a white solid. MS (ESI) m / z [M+H-EtOH] + : 509.2.

[0314] Step 2: Synthesis of methyl 3-amino-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate. To 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 CH2Cl2 (27.0 mL), diethylamine (1.53 mL, 14.71 mmol) was added at room temperature, and the reaction mixture was stirred for 3 hours. Once the starting materials were completely consumed (monitoring by TLC), the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by CombiFlash column chromatography using 5% MeOH / DCM to obtain methyl 3-amino-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.700 g, yield 86%) as a yellow viscous liquid. MS (ESI) m / z [M+H-EtOH] + : 287.68.

[0315] Step 3: Synthesis of methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamide)-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, HATU (0.720 g, 1.913 mmol) and DIPEA (0.555 mL, 3.188 mmol) were added, followed by methyl 3-amino-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.530 g, 1.594 mmol). The reaction mixture was heated to room temperature and stirred for 6 hours. After completion, the reaction mixture was quenched with ice-cold water (20 mL), and the aqueous layer was extracted with ELISA (20 mL x 2). The organic layer was washed with cold H2O (10 mL), followed by saturated brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by Combiflash column chromatography using 5% MeOH / DCM to obtain methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamide)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.630 g, yield 70%) as an off-white solid. MS (ESI) m / z [M+H-EtOH] + : 580.20.

[0316] Step 4: Synthesis of (9H-fluoren-9-yl)methyl6-(2-methoxy-2-oxoethyl)-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. Formic acid (1.5 mL) was added to a stirred solution of methyl3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamide)-4-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-oxobutanoate (0.300 g, 0.4794 mmol) at room temperature, and the reaction mixture was stirred for 16 hours. After completion, the reaction mixture was concentrated, and the resulting crude product was purified by column chromatography (silica 100-200 mesh; 0-5% MeOH / DCM) to obtain (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, yield 80%) as a yellow solid. MS (ESI) m / z [M+H] + : 534.67.

[0317] Step 5: Synthesis of methyl 2-(8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-6-yl) acetate. Diethylamine (0.280 mL) was added to 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 CH2Cl2 (0.5 mL), and the reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was concentrated, and the crude material was purified by Combiflash column chromatography using 0-5% MeOH / DCM to obtain methyl 2-(8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-6-yl) acetate (0.130 g, yield 93%) as a white solid. MS (ESI) m / z [MH] +: 310.4.

[0318] Step 6: Synthesis of methylmethyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-6-yl) acetate. To a solution of methyl 2-(8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-6-yl) acetate (3.08 g, 9.890 mmol) in DMF (30 mL), K2CO3 (4.10 g, 29.66 mmol) was added at room temperature, and the reaction mixture was stirred at 80°C for 15 minutes. 1-(bromomethyl)-4-(difluoromethoxy)benzene (3.48 g, 14.36 mmol) was added to the resulting reaction mixture, and the stirred mixture was heated to 80°C for 2 hours. After completion, the reaction mixture was quenched with ice-cold water (200 mL), and the aqueous layer was extracted with ELISA (200 mL x 2). The organic layer was washed with cold H2O (200 mL), followed by saturated brine (150 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by Combiflash column chromatography (5% MeOH / DCM) to obtain methyl 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-6-yl) acetate (2.20 g, yield 48%) as a yellow solid. MS (ESI) m / z [M-CH3] + : 454.10.

[0319] 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]pyrimidine-6-yl) acetate (2.20 g, 4.705 mmol) in THF (22.0 mL), NaOH (0.560 g, 14.11 mmol), followed by water (4 mL), was added, and the reaction mixture was stirred at room temperature for 3 hours. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude residue was dissolved in water (10 mL), slowly acidified with 6N HCl (10 mL), and stirred for 5 minutes. The resulting solid precipitate was filtered through a Buchner funnel and dried under reduced pressure to obtain 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-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, CDCl3) δ 7.28 - 7.38 (m, 2 H), 7.11 (d, J = 7.99 Hz, 2 H), 6.33 - 6.71 (m, 1 H), 5.36 - 5.40 (m, 1 H), 4.70 - 4.80 (m, 1 H), 4.65 - 4.75 (m, 1 H), 3.80 - 4.00 (m, 2 H), 3.55 - 3.65 (m, 1 H), 3.35 - 3.45 (m, 1 H), 2.85 - 3.30 (m, 6 H), 2.70 - 2.80 (m, 1 H), 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).

[0320] 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]pyrimidine-6-yl)acetic acid (0.470 g, 1.036 mmol) in THF (5 mL), 1,1'-carbonyldiimidazole (0.500 g, 3.109 mmol) was added at room temperature, and the reaction mixture was stirred for 15 minutes. NH3 (10 mL) was added to the resulting reaction mixture, and the reaction mixture was stirred at room temperature for 3 hours. 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 ELISA (20 mL x 3). The combined organic layers were washed with saturated brine solution (10 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The resulting crude compound was purified by Combiflash column chromatography using 5% MeOH / DCM, followed by preparative HPLC, to obtain 2-(1-(4-(difluoromethoxy)benzyl)-8-(2-methylbutyl)-4,7-dioxooctahydro-2H-pyrazino[1,2-a]pyrimidine-6-yl)acetamide (0.070 g, yield 15%) as a white solid. MS (ESI) m / z [M+H] + : 453.20. 1 H NMR (400 MHz, CDCl3) δ 7.30 - 7.40 (m, 2 H), 7.05-7.15 (m, 2 H), 6.39 - 6.70 (m, 1 H), 5.20 - 5.40 (m, 2 H), 4.75 - 4.85 (m, 1 H), 3.95 - 4.05 (m, 1 H), 3.75 - 3.85 (m, 1 H), 3.50 - 3.60 (m, 1 H), 3.30 - 3.40 (m, 1 H), 3.05 - 3.25 (m, 2 H), 2.85 - 2.95 (m, 2 H), 2.55 - 2.70 (m, 1 H), 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).

[0321] Example S63. Synthesis of the 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 the mixture was stirred at 80°C for 12 hours. After the reaction was complete, the 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 SiO2 (50 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The obtained crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain 1-(3-chloro-4-(trifluoromethyl)benzyl)-6-methylhexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (320 mg, yield 42%) as a white solid. MS (ESI) m / z [M+H] + : 376.34.

[0322] Example S64. General procedure F for the synthesis of the final compound. 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 2 mL of DMF at 0°C, Cs2CO3 (4 equivalents) was added and the mixture was stirred for 20 minutes. Then, a suitable alkyl halide (1.2 equivalents) was added at room temperature, and the reaction mixture was heated to 80°C and stirred for 12 hours. 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 dried over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica 100-200 mesh; 5% MeOH / DCM) to obtain the final compound.

[0323] 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, CDCl3) δ ppm 7.70 (d, J = 8.07 Hz, 1 H), 7.54 (s, 1 H), 7.33 (d, J = 8.68 Hz, 1 H), 4.34 (dd, J = 10.64, 3.55 Hz, 1 H), 3.87 - 3.99 (m, 2 H), 3.61 (t, J = 11.13 Hz, 1 H), 3.25 - 3.42 (m, 2 H), 3.08 - 3.19 (m, 2 H), 2.89 - 2.98 (m, 1 H), 2.64 - 2.74 (m, 1 H) 2.29 - 2.38 (m, 1H) 2.17 - 2.27 (m, 1 H) 1.97 - 2.09 (m, 2 H) 1.72 - 1.92 (m, 3 H) 1.58 - 1.66 (m, 4 H) 1.55 (br. s, 3 H).

[0324] Example S66. Synthesis of compound 54. Compound 54 was synthesized using (2-bromoethyl)cyclopentane as the alkyl halide according to general procedure F. MS (ESI) m / z [M+H] + : 472.15. 1H NMR (400 MHz, CDCl3) δ ppm 7.69 (d, J = 8.11 Hz, 1 H) 7.52 - 7.56 (m, 1 H) 7.33 (d, J = 7.89 Hz, 1 H), 5.23 (q, J = 7.23 Hz, 1 H), 4.36 (dd, J = 10.52, 3.29 Hz, 1 H), 3.87 - 3.98 (m, 2 H), 3.63 (t, J = 11.07 Hz, 1 H), 3.46 - 3.56 (m, 1 H), 3.25 - 3.35 (m, 1 H), 3.12 - 3.23 (m, 2 H), 2.87 - 2.97 (m, 1 H), 2.60 - 2.70 (m, 1 H), 2.29 - 2.37 (m, 1 H), 1.66 - 1.82 (m, 2 H), 1.54 - 1.63 (m, 1 H), 1.51 (d, J =,2.63 Hz, 2 H), 1.42 (d, J = 7.23 Hz, 3 H), 1.26 (br. s, 2 H) 1.04 - 1.16 (m, 2 H) 0.80 - 0.92 (m, 2 H).

[0325] 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), followed by DIPEA (10.62 mL, 61.10 mmol) was added at 0°C, and the reaction mixture was stirred at the same temperature for 10 minutes. N-(2,2-diethoxyethyl)-2-methylbutan-1-amine (11.48 g, 56.58 mmol) was added to the resulting reaction mixture at room temperature, and the reaction mixture was stirred for 3 hours. Once the starting materials were completely consumed (monitored by TLC), the reaction mixture was quenched with ice-cold water (100 mL), and the aqueous layer was extracted with RINKAN (50 mL x 4). The combined organic layer was washed with cold H2O (50 mL x 2), followed by brine (50 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by CombiFlash column chromatography using 5% MeOH / DCM to obtain (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)carbamate (14.5 g, yield 47.57%) as a white solid. MS (ESI) m / z [M+H] + : 539.04.

[0326] Step 2: Synthesis of 2-amino-N-(2,2-diethoxyethyl)-4-methyl-N-(2-methylbutyl)pentanamide. Diethylamine (16 mL, 157.7 mmol) was added to a solution of (9H-fluoren-9-yl)methyl(1-((2,2-diethoxyethyl)(2-methylbutyl)amino)-4-methyl-1-oxopentan-2-yl)carbamate in CH2Cl2 (50 mL), and the reaction mixture was stirred at room temperature for 3 hours. Once the starting materials were completely consumed (monitoring by TLC), the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by CombiFlash column chromatography using 5% MeOH / DCM to obtain 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.

[0327] 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. 3.80 g, 12.28 mmol of 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanoic acid (3.80 g, 12.28 mmol) was stirred in anhydrous DMF (35 mL) maintained at 0°C. HATU (6.48 g, 17.05 mmol) and DIPEA (4.90 mL, 28.42 mmol) were added, followed by the addition of 2-amino-N-(2,2-diethoxyethyl)-4-methyl-N-(2-methylbutyl)pentanamide (3.60 g, 11.37 mmol). The reaction mixture was allowed to return to room temperature and stirred for 3 hours. After completion, the reaction mixture was quenched with ice-cold water (20 mL), and the aqueous layer was extracted with ELISA (30 mL x 2). The organic layer was washed with cold H2O (10 mL), followed by saturated brine (20 mL), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by Combiflash column chromatography using 5% MeOH / DCM to obtain (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, yield 55%) as an off-white solid. MS (ESI) m / z [M+H-EtOH] + : 565.30.

[0328] Step 4: Synthesis of (9H-fluoren-9-yl)methyl6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate. Formic acid (20 mL) was added to a stirred solution of (9H-fluoren-9-yl)methyl6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (3.80 g, 6.231 mmol) 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 obtain (9H-fluoren-9-yl)methyl6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (3.60 g, yield 94%) as a yellow solid. MS (ESI) m / z [M+H] + : 518.23.

[0329] Step 5: Synthesis of 6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione. Diethylamine (6.8 mL, 69.54 mmol) was added to a solution of (9H-fluoren-9-yl)methyl6-isobutyl-8-(2-methylbutyl)-4,7-dioxohexahydro-2H-pyrazino[1,2-a]pyrimidine-1(6H)-carboxylate (3.60 g, 6.954 mmol) in CH2Cl2 (36 mL), and the reaction mixture was stirred at room temperature for 16 hours. Once the starting materials were completely consumed (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 obtain 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.

[0330] 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), K2CO3 (0.159 g, 1.152 mmol) was added at 0°C, and the reaction mixture was stirred for 10 minutes. To the resulting reaction mixture, 1-(bromomethyl)-4-(difluoromethoxy)benzene (0.150 g, 0.632 mmol) was added at room temperature, and the mixture was stirred for 3 hours. After completion, the reaction mixture was quenched with ice-cold water (200 mL), and the aqueous layer was extracted with SiO2 (20 mL x 2). The organic layer was washed with cold H2O (20 mL), followed by saturated brine (15 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting crude compound was purified by preparative HPLC to obtain 1-(4-(difluoromethoxy)benzyl)-6-isobutyl-8-(2-methylbutyl)hexahydro-4H-pyrazino[1,2-a]pyrimidine-4,7(6H)-dione (0.103 g, yield 40%) as a white solid. MS (ESI) m / z [M+H] + : 452.3. 1 H NMR (400 MHz, DMSO d6) δ 7.42 (d, J = 8.8 Hz, 2 H), 7.14 - 7.24 (m, 3 H), 5.0 - 5.10 (m, 1 H), 4.50 - 4.60 (m, 1H), 3.90 - 4.00 (m, 2 H), 3.60 - 3.70 (m, 1 H), 3.02 - 3.40 (m, 4 H), 2.70 - 2.85 (m, 2 H), 2.0 - 2.10 (m, 1 H), 1.50 - 1.70 (m, 4H), 1.20 - 1.35 (m, 1H), 1.0 - 1.10 (m, 1H), 0.70 - 0.98 (m, 12H).

[0331] Biological examples Example B1. Phospho-MET ELISA. The efficacy of compounds against the HGF / MET system was screened 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.

[0332] HEK293 cells were subcultured into 6-well multiplates and grown in DMEM + 10% FBS at 37°C and 5% CO2 until approximately 90% confluence. The cells were then starved in serum-free growth medium for at least 8 hours.

[0333] Exemplary compounds were prepared in DMEM + 0.1% FBS, diluted, and added to a treatment medium containing 1 ng / mL of recombinant HGF protein (R&D Systems). Cells were incubated in triplicates at 37°C and 5% CO2 for 15 minutes. The samples were then treated with 180 μL of ice-cold RIPA (radioimmunoprecipitation assay) buffer, and the cells were lysed on ice for 15 minutes. The lysates were clarified by centrifugation at 16,000 g for 15 minutes, and the supernatant was retained. The samples were normalized using a BCA assay of the lysates to determine the overall protein concentration of the samples.

[0334] 50–100 μg of total protein lysate was loaded into each well of the pMET Sandwich ELISA kit (Cell Signaling catalog number 7227C), ensuring that the protein was evenly distributed in each well. The ELISA was processed according to the manufacturer's instructions. After color development, the absorbance at 450 nm was read using an optical plate reader.

[0335] Efficacy measurements were determined using the peak effect, by scaling the administration treatment with the 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) In the formula, y is the normalized data point, x is the raw data point, A is the average HGF at 1 ng / mL, and B is the average HGF at 10 ng / mL. The calculated efficacy results are shown in Table 2. TIFF0007884275000054.tif202165

[0336] Example B2. Cell diffusion behavior assay. MDCK cells were cultured under normal conditions, and the spontaneous formation of robust colonies during proliferation was observed. Since MDCK cells respond to HGF treatment by moving away from each other (diffusing), the amount of HGF / MET activation in the cell population can be evaluated by quantifying this diffusion. In this experiment, MDCK cells were plated in a 96-well format, treated with HGF and exemplary compounds, fluorescently stained, and imaged in a wide field to quantify the diffusion behavior. Quantification was determined by analyzing the number of consecutive cell clusters (normalized particle count) compared to the total stained area imaged.

[0337] MDCK cells were plated at low density onto black-walled imaging plates and incubated overnight in DMEM + 10% FBS at 37°C and 5% CO2. The cells were then starved for 2 hours in FBS-free DMEM ("starvation medium"). Samples containing exemplary compounds were prepared in FBS-free DMEM ("treated medium") containing 5 ng / mL of HGF protein. Control curves were also created for each plate using HGF concentrations of 0, 5, 10, and 20 ng / mL. The starvation medium was replaced with treated medium, and the cells were incubated at 37°C and 5% CO2 for 24 hours.

[0338] 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 washed with PBS and rehydrated by washing with staining solution (fluorescent wheat germ agglutinin; WGA488, 20 μg / mL in PBS). After incubating the cells with the staining solution at room temperature for 30 minutes, the staining solution was replaced with fresh PBS.

[0339] Cellular regions were imaged using an iCyte high-content imager at a green wavelength. Images were converted to binary, and particle size and particle count were analyzed. For analysis, individual cells not in contact with other cells or isolated cell colonies were identified as particles, and particle counts were normalized by the total signal area to account for differences in cell number. An increase in particle count indicates that individual cells moved away from each other in the diffusion behavior response. The potency of the compound was evaluated by the statistical increase in normalized particle count compared to HGF treatment alone. The results are shown in Table 3. TIFF0007884275000055.tif208165

[0340] Example B3. Solubility assay. Water solubility is an important drug property that helps predict bioavailability. Generally, compounds with water solubility of less than 100 μg / ml are unsuitable as drugs. To evaluate the solubility of compounds, turbidimetric solubility assays were performed using representative compounds in the concentration range of 3–300 μM.

[0341] To evaluate the solubility of the compound by turbidimetric analysis, the test compound was first dissolved in an organic solvent (DMSO) at a concentration of 10 mM. Next, this solution was diluted in aqueous solvent (PBS) in a 96-well assay plate in a dilution series of 3–300 μM. The solutions were incubated at 37°C for 2 hours.

[0342] In wells containing test compounds exceeding their solubility limit, the compound precipitates, effectively blocking the passage of light, thus increasing the absorbance signal for ultraviolet light at a wavelength of 620 nm. If the absorbance increased by more than 10% compared to the control reading due to turbidity, the compound was considered insoluble at the test concentration. The results are shown in Table 4. TIFF0007884275000056.tif203165

[0343] Example B4. Permeability assay. Bioavailable drugs need to permeate the cell membranes of the inner wall of the gastrointestinal tract. Parallel membrane permeability assays (PAMPA) were performed in vitro to estimate the permeability of exemplary compounds.

[0344] The test compounds need to have standard curves on the final reading plate to determine the concentration of each fractionated drug. For each compound, a 6-point standard curve was created in phosphate-buffered saline (PBS) at concentrations from 0 to 200 μM.

[0345] 300 μL of the test compound solution (in PBS) was added to the donor (bottom) well of a PAMPA plate in 5 replicates, and 200 μL of PBS vehicle was added to the acceptor (top) well in an appropriate well to match the loading volume of the donor plate. The bottom and top of the PAMPA plate were then sandwiched together. The PAMPA plate was then incubated at room temperature for 5 hours. After incubation, 150 μL of the donor solution was added to a UV-readable plate with the corresponding standard curve. 150 μL of the acceptor well solution was added adjacent to the corresponding standard curve and the donor well sample of the compound. The plate was then read using a UV plate reader.

[0346] Next, the permeability and film retention rate were 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 cm²) 2 ), VD = Donnerwell volume (0.3 mL) VA = acceptor well volume (0.2 mL) t = incubation time (seconds) CA(t) = compound concentration in the acceptor well at time t. CD(t) = Donnerwell compound concentration 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 defined as follows for Equation 1, and C0 = Donnerwell's initial concentration (200 μM).

[0347] The results are shown in Table 5. TIFF0007884275000057.tif206165

[0348] Example B5. Cytotoxicity assay. This experiment was designed to obtain a preliminary assessment of cytotoxicity. The compound was tested at high concentrations, and the release of lactate dehydrogenase (LDH) into the culture medium was measured as a measure of lysed / dead cells to determine whether any cytotoxic effects were observed in hepatocyte (HepG2) cultures.

[0349] HepG2 cells were plated into 96-well cell culture plates and adhered overnight in EMEM + 10% FBS at 37°C and 5% CO2. Treatment was carried out in complete medium (EMEM + 10% FBS) and included a series of test compounds diluted from 0.1 to 100 μM. Cerivastatin, a known cytotoxic, was used as a positive assay control and prepared at a final concentration of 0.5 μM.

[0350] The growth medium was replaced with a treatment medium (EMEM + 10% FBS containing the 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 the LDH assay 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.

[0351] The signal range of the assay was determined by performing no manipulation in the negative control treatment and by completely lysing all cells in the lysis control sample. In this assay, compounds were considered cytotoxic if their cytotoxicity level increased by more than 20% compared to the negative control sample. The results are shown in Table 6. TIFF0007884275000058.tif208165

[0352] Example B6. In vitro stability assay. The bioavailability of a compound can be estimated by its stability when exposed to internal biological conditions. As an initial assessment of stability properties under various conditions present in animals, the stability of exemplary compounds was tested in an artificial body fluid compartment battery. The stability of compounds was tested in the following solutions: artificial gastric juice (SGF: 34.2 mM NaCl, pH 1.2), artificial gastric juice containing the digestive enzyme pepsin (SGF + enzyme: SGF with 3.2 mg / ml pepsin), artificial intestinal juice containing a porcine pancreatin enzyme mixture (SIF + enzyme: 28.7 mM NaH2PO4, 105.7 mM NaCl, pH 6.8, 10 mg / ml pancreatin), rat plasma, and human plasma.

[0353] The test compound was incubated in the above solution at a final concentration of 5 μM at 37°C, and samples were withdrawn at the following time points: 0, 1, 2, and 4 hours. The reaction was stopped, and for quantification, an excess quench solution containing an internal standard (acetonitrile, 200 ng / mL of busetin) was added. The test compound and internal standard in each sample were quantified by LC-MS / MS, and after internal normalization with respect to busetin, the concentration of the test compound was expressed as a percentage of the concentration at time 0. The stability of the corresponding test solution was then determined by the residual percentage at time 4. The results are shown in Table 7. TIFF0007884275000059.tif180165

[0354] Example B7. In vivo pharmacokinetics. The pharmacokinetic (PK) profiles of exemplary compounds were determined by administering them via a selected route, collecting blood, and quantifying the compounds in plasma. At least 250 grams of mixed male and female Sprague Dolly rats were administered the compounds by dissolving them in DMSO and then diluting them in a suitable vehicle of either physiological saline or physiological saline and polyethylene glycol. Administration was performed either by tail vein puncture (IV) or forced oral administration (PO), with animals receiving 1 mL / kg of the compound according to body weight. Following administration, blood was collected by tail vein sampling at selected intervals (10, 20, 40, 60, 120, and 360 minutes). 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.

[0355] Next, the plasma concentrations were averaged at each time point and plotted as a function of time. The area under the curve was calculated by integrating the curve, with Cmax being the highest concentration reached in plasma and Tmax being determined by the time point at which Cmax occurred. The results are shown in Table 8. TIFF0007884275000060.tif222165

[0356] Example B8. Calculation of oral availability. Oral bioavailability is crucial for the development of small molecule therapeutics for oral administration. Oral bioavailability (%F) is calculated by using IV administration as the maximum possible exposure dose, determining the exposure rate after PO administration, and comparing it with in vivo pharmacokinetic data (Example B7). In these studies, %F was obtained by dividing the dose-adjusted AUC from PO administration by the dose-adjusted AUC from IV administration and multiplying by 100. The results are shown in Table 9. TIFF0007884275000061.tif180165

[0357] Example B9. Non-specific protein binding. The plasma and tissue exposures of exemplary compounds were scaled by nonspecific affinity for protein binding in target tissues or body fluids to determine the fractions of compounds that can be utilized for interaction with targets. Nonspecific binding was measured in plasma and brain homogenates collected from mixed-sex Sprague Dolly rats.

[0358] Test compounds of known concentrations were mixed with plasma or brain homogenate and incubated in the donor chamber of a rapid equilibrium dialysis (RED) device with an empty PBS buffer in the receiving chamber. After incubation at 37°C for 4 hours in an orbital shaker incubator, the compounds in each chamber were quantified by LC-MS / MS. The unbound fraction (f) was determined using the following formula. u,組織 ) was calculated:

number

[0359] The results are shown in Table 10. TIFF0007884275000063.tif205165

[0360] Example B10. In vivo tissue distribution. Distribution rate to target tissue is an important characteristic of therapeutic molecules. Tissue distribution of exemplary compounds was performed in mixed-sex Sprague Dolly rats. Test compounds were delivered via tail vein injection (IV), and tissue was collected at Tmax (10 minutes post-administration). Animals were deeply anesthetized with isoflurane, whole blood was collected from the right atrium, and processed by centrifugation to obtain plasma. Then, to prevent tissue hematopoietic contamination, the animals were completely perfused by administering PBS to the left ventricle.

[0361] Tissue samples were collected, homogenized, and the compound content in the target tissue was quantified by LC-MS / MS. The tissue distribution rate 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. TIFF0007884275000064.tif186165

[0362] Example B11. In vivo efficacy: Scopolamine-induced spatial memory impairment in Morris water maze. Exemplary compounds 2a and 6a were evaluated in a Morris water maze for their ability to restore spatial memory impairment in chemically induced rats. The water maze consisted of a large circular tank (2.1 m in diameter) filled with water at 26–28°C to a depth of approximately 30 cm, and the water was clouded with white paint. A circular platform (13 cm in diameter) was fixed 2–3 cm below the water surface. High-contrast visual cues were placed around the tank to assist the test animals' spatial orientation. The test involved placing the animals in the water facing the tank wall at one of three randomly assigned starting points and allowing them to swim for up to 120 seconds to find the hidden platform. The time it took the animals to find the platform was recorded as escape latency. The animals were tested five times a day, with a 30-second rest between tests. The test was conducted for a total of eight consecutive days.

[0363] The animals were divided into groups (N=8 / group) according to the treatment. Control animals were given only empty vehicles. The scopolamine group received 3 mg / kg of scopolamine dissolved in sterile saline by intraperitoneal (IP) injection 30 minutes before the test. The test compound group received various concentrations of the test compound dissolved in 48% sterile saline, 50% polyethylene glycol (PEG-400), and 2% DMSO by forced oral (PO) administration 40 minutes before the test. For each animal, escape latency was recorded daily for 5 tests over 8 consecutive days. Changes in the escape latency curve were statistically analyzed using two-way ANOVA and Bonferroni post-hoc tests. The results are shown in Table 12.

[0364] Exemplary compound 1a was evaluated in a Morris water maze for its ability to restore spatial memory impairment in chemically induced rats. The water maze consisted of a large circular tank (1.5 m in diameter) filled with water at 23–26°C to a depth of approximately 30 cm, and the water was clouded 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 assist the test animals' spatial orientation. The test involved placing the animals in the water facing the tank wall at one of three randomly assigned starting points and allowing them to swim for up to 90 seconds to find the hidden platform. The time it took the animals to find the platform was recorded as escape latency. The animals were tested five times a day, with a 30-second rest between tests. The test was conducted for a total of five consecutive days.

[0365] The animals were divided into groups (N=12 / group) according to the treatment. Control animals were given only empty vehicles. The scopolamine group received 2 mg / kg of scopolamine dissolved in sterile saline by intraperitoneal (IP) injection 30 minutes before the test. The test compound group received various concentrations of the test compound dissolved in 78% sterile saline, 20% polyethylene glycol (PEG-400), and 2% DMSO by forced oral (PO) administration 40 minutes before the test. For each animal, escape latency was recorded daily for 5 consecutive days across 5 tests. Changes in the escape latency curve were statistically analyzed using two-way ANOVA and Bonferroni post-hoc tests. The results are shown in Table 12. TIFF0007884275000065.tif91165

[0366] For the purpose of clarifying the present invention, it has been described in some detail with reference to illustrations and examples, but these descriptions and examples should not be construed as limiting the scope of the invention. All patent and scientific document disclosures cited herein are incorporated herein by reference in their entirety.

Claims

1. Compound of formula (V): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof (In the formula, L is -C(=O)- or -(CH2)-, R １ａ and R １ｂ C may be independently substituted with H, or -CO2H. １ - It is a C3 alkyl group, R ３ This is a C1-C3 alkyl substituted with a C4-C5 alkyl, a C4-C5 alkenyl, or a C3-C5 cycloalkyl. R ４ (This refers to phenyl substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro, or pyridyl which may be substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro.)

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R1a and R1b are each independently H or C1-C3 alkyl.

3. R １ａ is methyl, R １ｂ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein is H.

4. R ３ However, the compound according to claim 1, or a pharmaceutically acceptable salt thereof, is as follows: 【Chemistry 2】

5. R ４ is -CF ３ , -OCHF ２ Phenyl substituted with one to three substituents selected from -OH, fluoro, and chloro, or Pyridyls substituted with 1 to 3 substituents selected from -CF3, -OCHF2, -OH, fluoro, and chloro The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

6. R ４ However, the compound according to claim 5, or a pharmaceutically acceptable salt thereof, is as follows: 【Transformation 3】

7. -L-R ４ However, the compound according to claim 1, or a pharmaceutically acceptable salt thereof, is as follows: 【Chemistry 4】

8. the below described: 【Transformation 5】 【change】 【change】 A compound selected from, or a pharmaceutically acceptable salt thereof.

9. The following formula: 【Transformation 6】 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, represented by [formula].

10. The following formula: 【Transformation 7】 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, represented by [formula].

11. The following formula: 【Transformation 8】 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, represented by [formula].

12. The following formula: 【Chemistry 9】 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, represented by [formula].

13. The following formula: 【Chemistry 10】 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, represented by [formula].

14. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition for regulating hepatocyte growth factor, comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition for treating a disease, condition, or injury that is a neurodegenerative disease, spinal cord injury, traumatic brain injury, or sensorineural hearing loss, comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

17. The pharmaceutical composition according to claim 16, wherein the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, Huntington's disease, or amyotrophic lateral sclerosis (ALS).

18. A pharmaceutical composition for treating dementia, slowing its progression, or preventing cognitive impairment, comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

19. A pharmaceutical composition for treating, restoring, or preventing a disease, condition, or injury relating to nerve tissue, comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

20. A pharmaceutical composition for treating or preventing a disease or disorder of the central nervous system, a disease or disorder of the peripheral nervous system, neuropathic pain, anxiety, or depression, comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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