PCSK9 inhibitor and method of use thereof

Abstract

To provide compounds for treating or preventing cardiovascular diseases.SOLUTION: The invention provides compounds of the formula (I) in the figure. In the formula: A is selected from H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR6, and -C(O)NR6R7; X is NR5 or O; and Y is selected from aryl, heteroaryl and heterocyclyl.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related applications This application claims priority to U.S. Patent Application No. 62 / 794,234, filed on 18 January 2019, which is incorporated herein by reference in whole. [Background technology]

[0002] PCSK9, also known as "proprotein convertase subtilisin / kexin 9," is a member of the secreted proprotein convertase family and plays a crucial role in cholesterol metabolism. PCSK9 increases circulating LDL cholesterol levels by enhancing the degradation of the LDL receptor independently of its catalytic activity. Secreted PCSK9 binds to the epidermal growth factor domain A (EGFA) of the LDL receptor (LDLR) on the cell surface, and the PCSK9 / LDL receptor complex is translocated into the endosomal / lysosomal compartment. The enhanced binding affinity of PCSK9 to the LDL receptor at the acidic pH of late endosomes / lysosomes reduces LDL receptor reuse and instead targets the LDL receptor for lysosomal degradation. Studies of genetic interactions have demonstrated that loss-of-function mutations in PCSK9 are associated with lower plasma LDL-C levels and a reduced incidence of adverse cardiovascular events.

[0003] Another biological pathway involving the action of PCSK9 on LDL receptors is the development of septic shock. Septic shock is often a fatal complication of severe microbial infection (sepsis) that induces an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis is caused by bacterial cell walls containing pathogenic lipid moieties such as lipopolysaccharide (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors [Tall-like receptors (TLRs)] and therefore plays a significant role in septic inflammatory responses (septic shock, or sepsis).

[0004] PCSK9 reduces LPS uptake by LDL receptors in the liver, resulting in free LPS excessively stimulating the body's immune response to pathogens and causing sepsis. Therefore, inhibiting PCSK9 is beneficial for preserving liver LDL receptors, which leads to systemic pathogen clearance and detoxification of the response to sepsis. However, currently there is no effective treatment for sepsis or septic shock other than antibiotic therapy.

[0005] Regarding cardiovascular disease, there are few options for inhibiting PCSK9. Statins do indeed upregulate PCSK9 in HepG2 cells and primary human hepatocytes by increasing the expression of SREBP-2, a transcription factor that upregulates both the LDLR and PCSK9 genes. Because elevated PCSK9 levels reduce the abundance of LDL receptors on the cell surface, increasing the dose of statins did not achieve a proportional reduction in LDL cholesterol.

[0006] Alirocumab and evolocumab, two monoclonal antibodies (mAbs) that selectively bind to extracellular PCSK9 and interfere with its interaction with the LDL receptor, have recently received FDA approval for lowering LDL-C levels. In clinical trials, alirocumab showed approximately a 50% reduction in LDL levels compared to placebo. (Non-patent document 1) Patients taking evolocumab showed approximately a 60–75% reduction in LDL levels. The efficacy of these drugs demonstrates the potential of PCSK9 inhibitors as an effective treatment for patients with hypercholesterolemia and other cardiovascular diseases. However, both antibody drugs require intravenous administration and may cause allergic reactions or other adverse immune responses in the body.

[0007] Cardiovascular diseases, unlike infections, are often temporary and require management throughout a person's life. Therefore, ease of administration and administration are crucial factors for patient compliance with maintenance medication. There is a need for highly effective and easily administered PCSK9 inhibitors, which can be achieved with small molecule PCSK9 inhibitors.

Prior Art Documents

Non-Patent Documents

[0008]

Non-Patent Document 1

Summary of the Invention

Means for Solving the Problems

[0009] In the present specification, a compound of formula (I)

Chemical Formula

[0010] In certain embodiments, the present invention provides a pharmaceutical composition suitable for use in a subject in the treatment or prevention of cardiovascular disease, comprising an effective amount of any of the compounds described herein (e.g., a compound of the present invention such as the compound of formula (I)) and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical may be for use in treating or preventing any of the conditions or diseases described herein.

[0011] This specification discloses methods for treating diseases and conditions that benefit from PCSK9 inhibition. These diseases include, but are not limited to, hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, abnormal lipoproteinemia, atherosclerosis, fatty liver, metabolic syndromes, and cardiovascular diseases such as coronary artery disease.

[0012] Other diseases and conditions that can be treated using the methods described herein include, but are not limited to, sepsis and septic shock.

[0013] This specification provides combination therapies of a compound of formula (I) and monoclonal antibodies, statins, and other cardiovascular agents that can enhance cardiovascular therapeutic effects beyond the capabilities of adjuvant therapy alone. Furthermore, this specification provides combination therapies of a compound of formula (I) and antibiotics that can reduce the incidence and severity of sepsis and septic shock beyond the capabilities of adjuvant therapy alone. [Modes for carrying out the invention]

[0014] In this specification, compounds of formula (I) [ka] (In the formula, A stands for H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 Selected from; B is selected from H, alkyl, and halo, or A and B, together with the carbon atoms to which they are bonded, form a 5- or 6-membered heteroaryl; X is NR 5 or O; R 1 and R 1’ Each is independently selected from H and alkyl; or If n is 0, R 1 and R 1’ These, together with the atoms to which they are bonded, form a 4- to 8-membered cycloalkyl or cycloalkenyl ring; R 2 is selected from H, halo, alkyl, alkoxy, amidealkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or R 1 and R 2 They, together with the atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; or R 1’ and R 2 These, together with the atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; R 2’ is selected from H, halo, alkyl, alkoxy, amidealkyl, aminoalkyl, and cyano; or R 2 and R 2’ These, together with the carbon atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; Each R 3 and R 4 is independently H or alkyl; or R 2 and R 3 They, together with the atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; and R 5 is H or alkyl; or R 1 and R 5They, together with the atoms to which they are bonded, form a 6-8 membered cycloalkyl or heterocyclyl ring; or R 2 and R 5 These, together with the atoms to which they are bonded, form a 5- to 8-membered cycloalkyl or heterocyclyl ring; Each R 6 and R 7 is independently H or alkyl; Y is selected from aryl, heteroaryl, and heterocyclyl; and n is either 0 or 1. It will be disclosed.

[0015] In this specification, compounds of formula (I') [ka] (In the formula, A stands for H, halo, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 Selected from; B is selected from H, alkyl, and halo; R 1 and R 1’ Each is independently selected from H and alkyl; or If n is 0, R 1 and R 1’ These, together with the atoms to which they are bonded, form a 4- to 8-membered cycloalkyl ring; R 2 is selected from H, halo, alkyl, alkoxy, amidealkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or R 1 and R 2 They, together with the atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; or R 1’ and R 2These, together with the atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; R 2’ is selected from H, halo, alkyl, alkoxy, amidealkyl, aminoalkyl, and cyano; or R 2 and R 2’ These, together with the carbon atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; Each R 3 and R 4 is independently H or alkyl; or R 2 and R 3 They, together with the atoms to which they are bonded, form a 3- to 8-membered cycloalkyl or heterocyclyl ring; and R 5 is H or alkyl; or R 1 and R 5 They, together with the atoms to which they are bonded, form a 6-8 membered cycloalkyl or heterocyclyl ring; or R 2 and R 5 These, together with the atoms to which they are bonded, form a 5- to 8-membered cycloalkyl or heterocyclyl ring; Each R 6 and R 7 is independently H or alkyl; Het is a heteroaryl or heterocyclyl; and n is either 0 or 1. It will be disclosed.

[0016] All embodiments described below and in this specification are understood to be embodiments of both formula (I) and formula (I').

[0017] In certain embodiments, A is H, hydroxy, thioalkyl, alkyl, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7The compound according to claim 1, selected from. In certain embodiments, A is H, but in other embodiments, A is an alkyl such as thioalkyl. In certain embodiments, A is selected from -SCH3, -SCHF2, and -OCHF2. In some embodiments, A is alkoxy. In other embodiments, A is cycloalkyl. In certain embodiments, B is H.

[0018] In certain embodiments, A and B together with the carbon atom to which they are attached form a pyrrolyl or thienyl ring, which is unsubstituted or substituted with one or more alkyls.

[0019] In certain embodiments, X is preferably NR5. In other embodiments, Y is preferably heteroaryl or heterocyclyl.

[0020] In certain embodiments, R 1 and R 1’ are each H. However, when n is 0, R 1 and R 1’ can together with the atom to which they are attached form a 4- to 8-membered cycloalkyl ring. In some embodiments, the cycloalkyl is monocyclic or bicyclic. In other embodiments, R 1 and R 1’ can together with the atom to which they are attached form a 4- to 8-membered cycloalkenyl ring. In some embodiments, the cycloalkyl ring is a cyclopentyl ring such as S,S-cyclopentyl. In some embodiments, the cycloalkyl ring is substituted with hydroxyl or hydroxyalkyl.

[0021] In certain embodiments, R 2 is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, alkylamino, cyano, and hydroxyl. In certain embodiments, R 2 is C 1~3 alkyl. R 2may be substituted with one or more substituents selected from amino, amide, cyano, hydroxy, and heterocyclyl. In some embodiments, R 2’ is C 1~3 alkyl, but in other embodiments, R 2’ is H.

[0022] In certain embodiments, R 1 and R 2 together with the atom to which they are attached form a 3- to 8-member cycloalkyl or heterocyclyl ring. In other embodiments, R 1’ and R 2 together with the atom to which they are attached form a 3- to 8-member cycloalkyl or heterocyclyl ring. In certain embodiments, R 2 and R 2’ together with the carbon atom to which they are attached form a 3- to 8-member cycloalkyl or heterocyclyl ring.

[0023] In certain embodiments, R 3 is C 1~3 alkyl, but in other embodiments, R 3 is H. In certain embodiments, R 4 is H.

[0024] In certain embodiments, R 2 and R 3 together with the atom to which they are attached form a 3- to 8-member cycloalkyl or heterocyclyl ring. In other embodiments, R 1 and R 5 together with the atom to which they are attached form a 6- to 8-member cycloalkyl or heterocyclyl ring. In other embodiments, R 2 and R 5 together with the atom to which they are attached form a 5- to 8-member cycloalkyl or heterocyclyl ring.

[0025] In certain embodiments, Y is a monocyclic heteroaryl, including but not limited to pyridinyl, pyrazinyl, pyrimidinyl, and thiazolyl. In other embodiments, Y is a monocyclic heteroaryl, including but not limited to pyridinyl, pyrazinyl, and pyrimidinyl. In some embodiments, Y is selected from triazenyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, and triazolyl. The monocyclic heteroaryl may not be substituted or may be substituted with one or more substituents selected from alkyl, thioalkylalkoxy, alkoxycarbonyl, amide, carboxy, cyano, halo, heteroaryl, nitro, sulfonamide, and thioalkyl. In another embodiment, Y is a monocyclic heteroaryl, including but not limited to alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxy, cyano, halo, aryl, heteroaryl, heterocyclyl, nitro, sulfonamide, and thioalkyl.

[0026] In certain embodiments, the monocyclic heteroaryl is substituted with an aryl, heteroaryl, or heterocyclil selected from phenyl, pyridinyl, 2-hydroxypyridinyl, piperidinonyl, 2-hydroxy-1-methylpyridinyl, triazolyl, imidazolidinonyl, pyrimidonyl, 2-hydroxyisoquinolinyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl, and morpholinonyl. In certain preferred embodiments, Y is a six-membered monocyclic heteroaryl. In some embodiments, the monocyclic heteroaryl is substituted with a heteroaryl or heterocyclil substituted with one or more substituents selected from halo, CN, alkyl, alkoxy, hydroxy, carboxy, -CO2 alkyl, and tetrazolyl. In certain preferred embodiments, the monocyclic heteroaryl is positioned para of A relative to X.

[0027] In other embodiments, Y is a bicyclic heteroaryl, including but not limited to benzothiazolyl, benzimidazolyl, benzoxazolyl, triazolopyridinyl, thiazolopyrindinyl, quinolinyl, and quinoxalinyl. The bicyclic heteroaryl may be unsubstituted or substituted with one or more substituents selected from alkyl, haloalkyl, hydroxyalkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxy, cyano, halo, heteroaryl, nitro, and sulfonamide. In certain embodiments, the bicyclic heteroaryl may be unsubstituted or substituted with one or more substituents selected from thioalkyl, alkoxycarbonyl, amide, carboxy, halo, and heteroaryl.

[0028] In certain embodiments, Y is given by formula -C(O)NR 8 R 9 or -NR 9 C(O)R 10 Substituted with an amide substituent, R 8 and R 9 Each of these is independently selected from H, alkyl, heterocyclyl, and heteroaryl; or R 8 and R 9 Together with the nitrogen atoms to which they are bonded, Forms a 4, 5, 6, or 7-membered heterocyclic ring or heteroaryl ring; and R 10 It is an alkyl group.

[0029] In certain embodiments, Y is given by formula -S(O)2NR 8 R 9 or -NR 9 S(O)2R 10 Substituted with a sulfonamide substituent; R 8 and R 9 Each of these is independently selected from H, alkyl, and heteroaryl; or R 8 and R 9Together with the nitrogen atoms to which they are bonded, Forms a 4, 5, 6, or 7-membered heterocyclic ring; and R 10 It is an alkyl group.

[0030] It is understood that all the aforementioned embodiments of the variable element Y in equation (I) are also embodiments of the variable element Het in equation (I').

[0031] In a particular embodiment, R 8 and R 9 Each of these is independently selected from H, methyl, ethyl, triazolyl, and pyrazolyl. 8 and R 9 In embodiments where one or both of are alkyl, each alkyl is independently either unsubstituted or substituted with one or more substituents selected from methyl, methoxy, carboxy, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, oxadiazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl, and 2,4-dihydro-3H-1,2,4-triazole-3-onyl. In certain embodiments, R 8 and R 9 These, together with the nitrogen atom to which they are bonded, form a heterocycle selected from azirazine, isothiazolidine-1,1-dioxide, azetidine, thiazole-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, and 2-oxa-6-azaspiro[3.3]heptane. In some embodiments, R 8 and R 9These, along with the nitrogen atom to which they are bound, are 2,8-diazaspiro[5,5]undecene, tetrahydroimidazo[1,2-a]pyrazine, octahydropyrazino[2,1-c][1,4]oxazine, tetrahydropyrido[3,4-d]pyrimidine, 2-oxa-8-azaspiro[4.5]decane, tetrahydropyrrolo[3,4-c]pyrazole, thiomorpholine, 2-oxa-7-azaspiro[3.5]nonane, 2,8-diazaspiro[4.5]decane-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9-diaza Spiro[5.5]undecane-3-one, tetrahydropyrrolo[3,4-d]imidazole, pyrimidine, 8-oxa-2-azaspiro[4.5]decane, hexahydro-3H-oxazolo[3,4-a]pyrazine-3-one, 1-oxa-7-azaspiro[3.5]nonane, octahydrocyclopenta[c]pyrrole, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine, 2,7-diazaspiro[4.4]nonane, 2,6-diazaspiro[3.4]octane, 7-oxa-2-azaspiro[3.5]nonane, 1-oxa-8λ 2 -Forms a heterocycle selected from azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, oxadiazole, triazole, pyridinone, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3(2H)-one, piperidinone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5]nonane, pyrazole, and pyridazine-3(2H)-one.

[0032] In some embodiments, the heterocycle is either unsubstituted or substituted with one or more substituents selected from alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxy, and heterocyclyl. In certain embodiments, the heterocycle is either unsubstituted or substituted with one or more substituents selected from methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxy, and oxetanyl.

[0033] In certain embodiments, the present invention provides a pharmaceutical product suitable for use in human patients, comprising any of the compounds shown above (e.g., compounds of the present invention such as the compound of formula (I)) and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical product may be used to treat or prevent any of the conditions or diseases described herein.

[0034] Any of the disclosed compounds may be used in the manufacture of a pharmacopoeia for the treatment of any of the diseases or conditions disclosed herein.

[0035] definition Unless otherwise defined, all technical and scientific terms used herein have the meanings generally understood by those skilled in the art. The following references provide general definitions of many of the terms used herein: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, unless otherwise specified, the following terms have the meanings attributed to them below.

[0036] In the present disclosure, terms such as "comprises", "comprising", "contains", and "has" can have the meanings ascribed to them in United States patent law and can mean "includes", "including", etc.; "consisting essentially of" or "consisting of" likewise have the meanings ascribed to them in United States patent law, and the term is non-limiting and allows for entities beyond those recited provided that the basic or novel feature being described is not changed by the entities beyond those recited, except for prior art embodiments.

[0037] Unless otherwise specified or clear from the context, as used herein, the term "or" is understood to be inclusive. Unless otherwise specified or clear from the context, as used herein, the terms "a", "an", and "the" are understood to be singular or plural.

[0038] The term "acyl" is recognized in the art and refers to a group represented by the general formula hydrocarbyl C(O)-, preferably alkyl C(O)-.

[0039] The term "acylamino" is recognized in the art and refers to an amino group substituted with an acyl group and can be represented, for example, by the formula hydrocarbyl C(O)NH-.

[0040] The term "acyloxy" is recognized in the art and refers to a group represented by the general formula hydrocarbyl C(O)O-, preferably alkyl C(O)O-.

[0041] The term "alkoxy" refers to an alkyl group, preferably a lower alkyl group, to which oxygen is attached. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0042] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula alkyl-O-alkyl.

[0043] As used herein, the term "alkenyl" refers to an aliphatic group containing at least one double bond and is intended to include both "unsubstituted alkenyl" and "substituted alkenyl", the latter referring to an alkenyl moiety having a substituent that replaces a hydrogen on one or more carbons of the alkenyl group. Such substituents can be present on one or more carbons that are or are not included in one or more double bonds. Further, such substituents include all those contemplated for an alkyl group as described below, provided that stability is not impaired. For example, substitution of an alkenyl group by one or more alkyl, carbocyclic, aryl, heterocyclic, or heteroaryl groups is contemplated.

[0044] An "alkyl" group or "alkane" is a fully saturated straight-chain or branched-chain non-aromatic hydrocarbon. Typically, a straight-chain or branched-chain alkyl group has 1 to about 20 carbon atoms, preferably 1 to about 10 carbon atoms, unless otherwise defined. Examples of straight-chain and branched-chain alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. A C1-C6 straight-chain or branched-chain alkyl group is also referred to as a "lower alkyl" group.

[0045] Furthermore, as used throughout this specification, the examples, and the claims, the term “alkyl” (or “lower alkyl”) is intended to include both “unsubstituted alkyl” and “substituted alkyl,” the latter referring to an alkyl moiety having substituents that substitute hydrogens on one or more carbons of a hydrocarbon skeleton. Such substituents, unless otherwise specified, may include, for example, halogens, hydroxyls, carbonyls (such as carboxyls, alkoxycarbonyls, formyls, or acyls), thiocarbonyls (such as thioesters, thioacetates, or thioformates), alkoxys, phosphoryls, phosphates, phosphonates, phosphinates, aminos, amides, amidines, imines, cyanos, nitros, azides, sulfhydryls, alkylthios, sulfates, sulfonates, sulfamoyls, sulfonamides, sulfonyls, heterocyclyls, aralkyls, or aromatic or heteroaromatic ring moieties. It will be understood by those skilled in the art that any moiety substituted on a hydrocarbon chain may be substituted itself, if necessary. For example, substituents on substituted alkyls may include amino, azide, imino, amide, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl and sulfonate), and silyl groups, as well as substituted and unsubstituted forms such as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, and -CN. Exemplary substituted alkyls are described later. Cycloalkyls can be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyls, -CF3, and -CN.

[0046] "C x~y The term "C" means a group containing x to y carbon atoms in the chain when used with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy. For example, "C x~yThe term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including linear and branched alkyl groups containing x to y carbon atoms in the chain, such as haloalkyl groups like trifluoromethyl and 2,2,2-trifluoroethyl. C0 alkyl indicates hydrogen when the group is at the terminal and a bond when it is in the middle. 2~y "Alkenil" and "C 2~y The term "alkynyl" refers to a substituted or unsubstituted unsaturated aliphatic group that is similar to the alkyl group described above in terms of length and possible substitutions, but each contains at least one double or triple bond.

[0047] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0048] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group, and may be represented by the general formula alkylS-.

[0049] As used herein, the term “alkynyl” refers to an aliphatic group containing at least one triple bond and is intended to include both “unsubstituted alkynyl” and “substituted alkynyl,” the latter referring to an alkynyl moiety having substituents that substitute hydrogens on one or more carbons of the alkynyl group. Such substituents may be located on one or more carbons that are not included in the one or more triple bonds. Furthermore, such substituents include all those conceivable for alkyl groups as described above, provided that stability is not compromised. For example, substitution of an alkynyl group with one or more alkyl, carbocykryl, aryl, heterocyclyl, or heteroaryl groups is conceivable.

[0050] As used herein, the term "amide" means base [ka] This refers to, and in the formula, each R 11 R independently represents a hydrogen or a hydrocarbyl group, or two R 11These, along with the N atoms to which they are bonded, complete a heterocycle with 4 to 8 atoms in the ring structure.

[0051] The terms "amine" and "amino" are recognized in the art and refer to both unsubstituted and substituted amines and their salts, for example, [ka] This refers to the part that can be represented by, and in the formula, each R 11 R independently represents a hydrogen or a hydrocarbyl group, or two R 11 These, along with the N atoms to which they are bonded, complete a heterocycle having 4 to 8 atoms in the ring structure. As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0052] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0053] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of which is aromatic, and for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Examples of aryl groups include benzene, naphthalene, phenanthrene, phenol, and aniline.

[0054] The term "carbamate" is recognized in the art and is based on [ka] It refers to, and in the formula, R 11 and R 12 This independently represents a hydrogen or a hydrocarbyl group such as an alkyl group, or R 11and R 12 Together with the intervening atoms, it completes a heterocycle having 4 to 8 atoms in its ring structure.

[0055] As used herein, the terms “carbocyclic ring” and “carbocyclic formula” refer to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocyclic ring includes both aromatic and non-aromatic carbocyclic rings. Non-aromatic carbocyclic rings include both cycloalkane rings in which all carbon atoms are saturated and cycloalkene rings containing at least one double bond.

[0056] The term “carbocyclic” includes monocyclic rings with 5 to 7 members and bicyclic rings with 8 to 12 members. Each ring in a bicyclic carbocyclic may be selected from saturated, unsaturated, and aromatic rings. A carbocyclic includes a bicyclic molecule in which one, two, or three or more atoms are shared between two rings. The term “condensed carbocyclic” refers to a bicyclic carbocyclic in which each ring shares two adjacent atoms with the other ring. Each ring in a condensed carbocyclic may be selected from saturated, unsaturated, and aromatic rings. In exemplary embodiments, an aromatic ring, e.g., phenyl, may be condensed to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of a carbocyclic formula, provided the valence allows it. Examples of “carbocyclic rings” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octa-3-ene, naphthalene, and adamantane. Examples of condensed carbocyclic rings include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hepta-3-ene. The “carbocyclic rings” can be substituted at any one or more positions capable of holding a hydrogen atom.

[0057] A "cycloalkyl" group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms, unless otherwise defined. The second ring of a bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Cycloalkyl includes bicyclic molecules in which one, two, or more atoms are shared between the two rings. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each ring shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. A "cycloalkenyl" group is a cyclic hydrocarbon containing one or more double bonds.

[0058] As used herein, the term "carbocyclic alkyl" refers to an alkyl group substituted with a carbocyclic group.

[0059] The term "carbonate" is recognized in the art and refers to the group -OCO2-R 10 (wherein R 10 represents a hydrocarbyl group).

[0060] As used herein, the term "carboxy" refers to the group represented by the formula -CO2H.

[0061] As used herein, the term "ester" refers to the group -C(O)OR 10 (wherein R 10 represents a hydrocarbyl group).

[0062] As used herein, the term “ether” refers to a hydrocarbyl group linked to another hydrocarbyl group via oxygen. Therefore, the ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. Ethers can be symmetric or asymmetric. Examples of ethers, but not limited to, include heterocyclic-O-heterocyclic and aryl-O-heterocyclic groups. Ethers contain an “alkoxyalkyl” group, which can be represented by the general formula alkyl-O-alkyl.

[0063] As used herein, the terms "halo" and "halogen" mean halogens, including chloro, fluoro, bromo, and iodine.

[0064] As used herein, the terms "hetallalkyl" and "hetallalkyl" refer to alkyl groups substituted with hetalil groups.

[0065] As used herein, the term "heteroalkyl" means a saturated or unsaturated chain of carbon atoms and at least one heteroatom, wherein no two heteroatoms are adjacent.

[0066] The terms "hetaliarl" and "hetaliarl" include substituted or unsubstituted aromatic monocyclic structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, where these ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "hetaliarl" and "hetaliarl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is a heteroaromatic ring, for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Examples of heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0067] As used herein, the term “heteroatom” means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0068] The terms “heterocyclyl,” “heterocyclic,” and “heterocyclic formula” refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, and these ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms “heterocyclyl” and “heterocyclic formula” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of these rings is heterocyclic, for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Examples of heterocyclyl groups include piperidine, piperazine, pyrrolidine, morpholine, lactone, and lactam.

[0069] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclic group.

[0070] As used herein, the term "hydrocarbyl" refers to a group that is linked via a carbon atom without an =O or =S substituent and typically has at least one carbon-hydrogen bond and is primarily a carbon skeleton, but may optionally contain a heteroatom. Therefore, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyls in the spirit of this application, while substituents such as acetyl (which has an =O substituent on the linked carbon) and ethoxy (which is linked via oxygen rather than carbon) are not considered hydrocarbyls. Examples of hydrocarbyl groups, but not limited to, include aryl, heteroaryl, carbocyclic, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

[0071] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group.

[0072] The term “lower” means, when used with a chemical moiety, e.g., acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, a group having 10 or fewer non-hydrogen atoms in the substituent, preferably 6 or fewer. “Lower alkyl” refers to an alkyl group containing, for example, 10 or fewer carbon atoms, preferably 6 or fewer. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents as defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they appear alone or in combination with other substituents, such as in the enumeration of hydroxyalkyl and aralkyl (in which case, for example, atoms in the aryl group are not counted when counting carbon atoms in the alkyl substituent).

[0073] The terms “polycyclyl,” “polycyclic,” and “polycyclic formula” refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings, for example, these rings are “fused rings.” Each of the rings in a polycyclic compound may or may not be substituted. In certain embodiments, each ring in a polycyclic compound contains 3 to 10 atoms, preferably 5 to 7 atoms.

[0074] The term "silyl" refers to the silicon portion formed by the bonding of three hydrocarbyl moieties.

[0075] The term “substituted” refers to a portion of a skeleton having substituents that substitute for hydrogens on one or more carbon atoms. “Substituted” or “substituted with” implies that such substitutions are subject to the permissible valencies of the substituted atom and substituent, and that the substitution results in a stable compound that does not undergo spontaneous transformation, such as rearrangement, cyclization, or elimination. As used herein, the term “substituted” is intended to encompass all permissible substitution values ​​of an organic compound. In a broader embodiment, permissible substituents of an organic compound include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents. For a given organic compound, there may be one or more permissible substituents, and they may be the same or different. In the spirit of the present invention, heteroatoms such as nitrogen may have any permissible substituent of the organic compounds described herein, satisfying the valencies of the hydrogen substituent and / or the heteroatom. Substituents include any substituents described herein, such as halogens, hydroxyls, carbonyls (carboxyls, alkoxycarbonyls, formyls, or acyls, etc.), thiocarbonyls (thioesters, thioacetates, or thioformates, etc.), alkoxyls, phosphoryls, phosphates, phosphonates, phosphinates, aminos, amides, amidines, imines, cyanos, nitros, azides, sulfhydryls, alkylthios, sulfates, sulfonates, sulfamoyls, sulfonamides, sulfonyls, heterocyclyls, aralkyls, or aromatic or heteroaromatic ring moieties. It will be understood by those skilled in the art that substituents may be substituted themselves as needed. Unless specifically stated as “unsubstituted,” references to chemical moieties herein are understood to include substituted forms. For example, references to “aryl” groups or moieties implicitly include both substituted and unsubstituted forms.

[0076] The term "sulfate" is recognized in the art and refers to the group -OSO3H or its pharmaceutically acceptable salts.

[0077] The term "sulfonamide" is recognized in the art and is a general formula [ka] This refers to the group represented by, in the formula, R 11 and R 12 This independently represents a hydrogen or an alkyl hydrocarbyl, or R 11 and R 12 Together with the intervening atoms, it completes a heterocycle having 4 to 8 atoms in its ring structure.

[0078] The term "sulfoxide" is recognized in the art and includes the group -S(O)-R 10 (In the formula, R 10 This refers to hydrocarbyl.

[0079] The term "sulfonate" refers to the group SO3H, or a pharmaceutically acceptable salt thereof, as recognized in the art.

[0080] The term "sulfone" is recognized in the art and includes the group -S(O)2-R 10 (In the formula, R 10 This refers to hydrocarbyl.

[0081] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0082] As used herein, the term "thioester" refers to the group -C(O)SR 10 or -SC(O)R 10 (In the formula, R 10 This refers to hydrocarbyl.

[0083] As used herein, the term "thioether" refers to an ether in which oxygen is replaced by sulfur.

[0084] The term "urea" is recognized in the relevant technical field and is a general formula [ka] It may also be expressed as, in the formula, R 11 and R 12 This independently represents a hydrogen or an alkyl hydrocarbyl, or R 11 The presence of any of the following conditions 12 Together with the intervening atoms, this completes a heterocycle having 4 to 8 atoms in its ring structure.

[0085] The term "protecting group" refers to a group of atoms that, when bonded to a reactive functional group in a molecule, shield, reduce, or prevent the reactivity of that functional group. Typically, protecting groups can be selectively removed during synthesis as needed. Examples of protecting groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry, 3. rd These can be found in Ed., 1999, John Wiley & Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilylethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), and nitroveratryloxycarbonyl ("NVOC"). Typical hydroxyl protecting groups include, but are not limited to, those in which the hydroxyl group is acylated (esterified) or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPS groups), glycol ethers, such as ethylene glycol and propylene glycol derivatives, and allyl ethers.

[0086] The present invention comprises all pharmaceutically acceptable isotope-labeled compounds as described herein, wherein one or more atoms are substituted by atoms having the same atomic number but with an atomic mass or mass number different from that commonly found in nature. In certain embodiments, the compounds of the present invention are rich in such isotope-labeled materials (for example, compounds in which the distribution of isotopes in the compound in the composition differs from the natural or typical distribution of isotopes).

[0087] Examples of isotopes suitable for inclusion in the compound of the present invention include: 2 H and 3 Hydrogen such as H, 11 C, 13 C and 14 Carbon such as C, 36 Chlorine such as Cl 18 Fluorine such as F 123 I and 125 Iodine such as I 13 N and 15 Nitrogen such as N, 15 O, 17 O and 18 Oxygen such as O, 32 Phosphorus such as P, and 35 Examples include sulfur isotopes such as S.

[0088] Certain isotope-labeled compounds, as disclosed herein, for example, those incorporating radioactive isotopes, are useful for drug and / or substrate tissue distribution studies. Radioactive isotope tritium, i.e. 3 H, and carbon-14, that is, 14 C is useful for this purpose considering its ease of implementation and readily available detection means.

[0089] Deuterium, that is, 2 Substitution with heavier isotopes, such as 1H, can be preferable in certain situations because it can result in several therapeutic benefits, such as increased metabolic stability, an increased in vivo half-life, or a reduced required dose.

[0090] 11 C, 18F, 15 O, and 13 Substitution with positron-emitting isotopes such as 16N may be useful in positron emission tomography (PET) studies to investigate substrate receptor occupancy.

[0091] In certain embodiments, the compounds of the present invention may be racemates. In certain embodiments, the compounds of the present invention may be concentrated into one enantiomer. For example, the compounds of the present invention may have more than about 30% ee, about 40% ee, about 50% ee, about 60% ee, about 70% ee, about 80% ee, about 90% ee, or about 95% or more ee. In certain embodiments, the compounds of the present invention may have two or more stereocenters. In certain such embodiments, the compounds of the present invention may be concentrated into one or more diastereomers. For example, the compounds of the present invention may have more than about 30% de, about 40% de, about 50% de, about 60% de, about 70% de, about 80% de, about 90% de, or about 95% or more de.

[0092] In certain embodiments, a therapeutic formulation may be concentrated to primarily provide one enantiomer of the compound (e.g., of formula (I)). A mixture enriched in one enantiomer may, for example, contain at least about 60 mol percent of one enantiomer, or more preferably at least about 75, about 90, about 95, or about 99 mol percent. In certain embodiments, a compound concentrated in one enantiomer is substantially free of the other enantiomer, where substantially free means that the substance constitutes, for example, less than about 10%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than 1% of the other enantiomer in the composition or compound mixture. For example, if the composition or compound mixture contains about 98 grams of the first enantiomer and 2 grams of the second enantiomer, it would be stated that it contains about 98 mol percent of the first enantiomer and only about 2% of the second enantiomer.

[0093] In certain embodiments, the therapeutic formulation may be concentrated to primarily provide one diastereomer of the compound (e.g., of formula (I)). A mixture rich in one diastereomer may, for example, contain at least about 60 mol percent of one diastereomer, or more preferably at least about 75, about 90, about 95, or about 99 mol percent.

[0094] The term “subject” to which administration is intended includes, but is not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, toddlers, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly)) and / or other primates (e.g., crab-eating macaques, rhesus macaques); commercially related mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially related birds such as chickens, ducks, geese, quail, and / or turkeys. The preferred subject is humans.

[0095] As used herein, a therapeutic agent that “prevents” a disorder or condition means a compound that, in a statistical sample, reduces the incidence of the disorder or condition in a treated sample compared to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition compared to an untreated control sample.

[0096] The term “to treat” includes prophylactic and / or therapeutic treatment. The term “prophylactic or therapeutic” treatment includes the administration of one or more subjects of compositions recognized and disclosed in the art. If it is administered before the clinical manifestation of an undesirable condition (e.g., a disease or other undesirable condition of the subject), the treatment is prophylactic (i.e., it protects the subject from the onset of the undesirable condition), whereas if it is administered after the manifestation of the undesirable condition, the treatment is therapeutic (i.e., it is intended to reduce, restore or stabilize an existing undesirable condition or its side effects).

[0097] The term "prodrug" is intended to encompass compounds that, under physiological conditions, are converted into the therapeutically active agents of the present invention (e.g., compounds of formula (I)). A common method for producing a prodrug is to include one or more selected moieties that, under physiological conditions, are hydrolyzed to reveal a desired molecule. In other embodiments, the prodrug is converted by the enzyme activity of the subject. For example, esters or carbonates (e.g., esters or carbonates or carboxylic acids of alcohols) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compounds of formula (I) in the formulations shown above may be replaced with corresponding preferred prodrugs, for example, the hydroxyl in the parent compound is represented as an ester or carbonate or carboxylic acid.

[0098] As used herein, “effective dose” means an amount sufficient to achieve the desired biological effect. As used herein, “therapeutic effective dose” means an amount sufficient to achieve the desired therapeutic effect. For example, a therapeutic effective dose may mean an amount sufficient to improve at least one sign or symptom of cancer.

[0099] The “response” to the treatment method may include, in particular, a reduction or improvement of negative symptoms, a decrease in disease progression or its symptoms, an increase in beneficial symptoms or clinical outcomes, a reduction in side effects, stabilization of the disease, or partial or complete recovery of the disease.

[0100] How to use The PCSK9 gene was identified using genetic mapping techniques on DNA from subjects with autosomal dominant hypercholesterolemia (Abifadel, et al. Nat. Genet. 2003 34:154-6). The encoded protein is a serine protease expressed primarily in the liver, intestines, kidneys, and nervous system. While not bound by any particular theory, studies of mutations in the gene have shown that its putative role is in the degrading of LDL receptors on the cell surface, independently of their catalytic activity (Abifadel, et al. Expert Opin. Ther. Pat. 2010 20:1547-71). Binding of PCSK9 to receptors leads to their lysosomal degradation. This enhanced degradation results in an increase in the amount of circulating low-density lipoprotein LDL (LDL-c). PCSK9 is upregulated by statins, SREBP-1a and SREBP-2, LXR agonists, and insulin, but downregulated by dietary cholesterol, glucagon, ethinylestradiol, chenodeoxycholic acid, and bile acid-activated farnesoid X receptor (FXR) (Maxwell, et al. J. Lipid Res. 2003 44:2109-19; Persson et al. Endocrinology 2009 150:1140-6; Langhi et al. FEBS Lett. 2008 582:949-55). Since elevated PCSK9 levels reduce the abundance of LDL receptors on the cell surface, increasing the dose of statins does not result in a proportional reduction in LDL cholesterol. Therefore, this specification discloses methods for treating a wide range of cardiovascular diseases and conditions that would benefit from lowering LDL-c by inhibiting PCSK9.

[0101] In certain embodiments, methods for inhibiting PCSK9 are performed in subjects requiring it, thereby treating PCSK9-mediated diseases or disorders.

[0102] Furthermore, this specification discloses methods for treating or preventing PCSK9-mediated diseases or disorders, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, this specification discloses methods for treating PCSK9-mediated diseases or disorders, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, this specification discloses methods for preventing PCSK9-mediated diseases or disorders, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. Prevention of cardiovascular events via PCSK9 inhibition is described, for example, in Robinson et al., Artherosclerosis 2015 243:593-597.

[0103] Examples of cardiovascular diseases and conditions include, but are not limited to, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, hypoalphalipoproteinemia, metabolic syndromes, diabetic complications, atherosclerosis, stroke, vascular dementia, chronic kidney disease, coronary heart disease, coronary artery disease, retinopathy, inflammation, thrombosis, peripheral vascular disease, or congestive heart failure.

[0104] In certain embodiments, exemplary cardiovascular diseases and conditions include, but are not limited to, hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, abnormal lipoproteinemia, atherosclerosis, fatty liver, metabolic syndromes, and coronary artery disease. In certain embodiments, the disease is hypercholesterolemia, such as familial hypercholesterolemia or autosomal dominant hypercholesterolemia. In certain embodiments, the disease is hyperlipidemia. In certain embodiments, the disease is coronary artery disease.

[0105] In certain embodiments, the disclosed therapeutic methods can reduce high levels of circulating serum cholesterol, such as LDL-cholesterol and VLDL-cholesterol. In addition, the disclosed methods are useful for reducing circulating serum triglycerides, circulating serum lipoprotein A, circulating serum LDL, and atherogenic lipoproteins. In certain embodiments, diseases or conditions treated with the disclosed compounds and compositions include atherosclerosis and atherosclerotic lesion formation. Subjects with gain-of-function mutations in the PCSK9 gene also benefit from treatment with the disclosed compounds and compositions that suppress the mutations by inhibiting PCSK9.

[0106] PCSK9 inhibition has also shown therapeutic effects in treating sepsis in subjects. Septic shock is often a fatal complication of severe microbial infection (sepsis) that induces an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis is caused by bacterial cell walls containing pathogenic lipid moieties such as lipopolysaccharide (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors [Tall-like receptors (TLRs)] and therefore plays a significant role in septic inflammatory responses (septic shock, or sepsis). PCSK9 reduces the uptake of LPS by hepatic LDL receptors, resulting in free LPS overstimulating the body's immune response to pathogens and causing sepsis. PCSK9 inhibition is beneficial for preserving hepatic LDL receptors, which leads to systemic pathogen clearance and detoxification of the response to sepsis (see, e.g., Walley et al Sci.Translat.Med.2014 6:1-10).

[0107] This specification discloses a method for treating sepsis or septic shock, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, the disclosed therapeutic method is useful for increasing LPS uptake. Certain embodiments provide a method for reducing the inflammatory response induced by sepsis or septic shock.

[0108] Combination therapy The disclosed compounds and compositions may be administered in conjunction with other therapeutic agents, such as other agents suitable for treating high levels of LDL and triglycerides. In certain embodiments, a synergistic effect is achieved by co-administering one or more additional therapeutic agents together with the compounds of the present invention. In certain embodiments, an additive effect is achieved by co-administering one or more additional therapeutic agents together.

[0109] In some embodiments, a method for treating a PCSK9-mediated disease or disorder comprises administering a PCSK9 inhibitor in conjunction with one or more other therapeutic agents. In some embodiments, the PCSK9 inhibitor is a compound of formula (I). The other therapeutic agents may include monoclonal antibodies such as alirocumab, evolocumab, vococizumab, RG7652, LY3015014, and mAb316P.Further exemplary antibodies are found in International Publication Nos. 2015 / 140079, 2015 / 142668, 2015 / 123423, 2015 / 073494, 2015 / 054619, 2014 / 197752, 2014 / 194111, 2014 / 194168, 2014 / 028354, and the country International Publication No. 2013 / 039969, International Publication No. 2013 / 016648, International Publication No. 2012 / 101251, International Publication No. 2012 / 101253, International Publication No. 2012 / 101252, International Publication No. 2014 / 209384, International Publication No. 2014 / 150983, International Publication No. 2014 / 144080, International Publication No. 2013 / 166448, International Publication No. 2012 / 1549 Pamphlet No. 99, International Publication No. 2013 / 008185, International Publication No. 2015 / 200438, International Publication No. 2014 / 107739, International Publication No. 2013 / 188855, International Publication No. 2013 / 169886, International Publication No. 2013 / 148284, International Publication No. 2013 / 091103, International Publication No. 2013 / 039958, International Publication No. 2012 / 177741, International Publication This includes what is described in Pamphlet No. 2012 / 168491, Pamphlet No. 2012 / 170607, Pamphlet No. 2012 / 109530, Pamphlet No. 2012 / 088313, Pamphlet No. 2012 / 054438, Pamphlet No. 2011 / 072263, Pamphlet No. 2011 / 053759, Pamphlet No. 2011 / 053783, and Pamphlet No. 2011 / 037791.Other therapeutic agents that may be used in combination with the disclosed compounds and compositions include plant alkaloids and flavonoids known for their cholesterol-lowering activity, such as berberine and quercetin. In some embodiments, cholesterol-lowering small molecules such as ezetimibe and policosanol may be administered as additional therapeutic agents. In certain embodiments, polypeptides such as BMS-962476 and fibronectin-based skeletal domain proteins may be administered in combination with the disclosed compounds and compositions. Further exemplary compounds include those described in International Publication Nos. 2014 / 150326, 2014 / 150395, 2014 / 139008, 2014 / 140210, 2014 / 127316, 2014 / 005224, 2013 / 177536, 2011 / 152508, 2011 / 130354, 2011 / 006000, and 2015 / 077154.

[0110] In certain embodiments, additional therapeutic agents may be statins such as atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin. In other embodiments, the disclosed compounds may be administered in conjunction with HMG-CoA reductase.

[0111] Any HMG-CoA reductase inhibitor can be used in the combined embodiment of the present invention. The conversion of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) to mevalonic acid is an initial rate-limiting step in the cholesterol biosynthesis pathway. This step is catalyzed by the enzyme HMG-CoA reductase. Statins inhibit HMG-CoA reductase from catalyzing this conversion.

[0112] The term HMG-CoA reductase inhibitor refers to compounds that inhibit the bioconversion of hydroxymethylglutaryl-coenzyme A to mevalonic acid, catalyzed by the enzyme HMG-CoA reductase. Such inhibition is readily determined by those skilled in the art according to standard assays (e.g., Meth. Enzymol. 1981;71:455-509 and the references cited therein). Various of these compounds are described and referenced below, but other HMG-CoA reductase inhibitors will be known to those skilled in the art. U.S. Patent No. 4,231,938 (this disclosure is incorporated herein by reference) discloses certain compounds isolated after culturing microorganisms belonging to the genus Aspergillus, such as lovastatin. Also, U.S. Patent No. 4,444,784 (this disclosure is incorporated herein by reference) discloses synthetic derivatives of the aforementioned compounds, such as simvastatin. Furthermore, U.S. Patent No. 4,739,073 (this disclosure is incorporated by reference) discloses certain substituted indoles, such as fluvastatin. Also, U.S. Patent No. 4,346,227 (this disclosure is incorporated by reference) discloses ML-236B derivatives, such as pravastatin. Also, European Patent Application Publication No. 491226A (this disclosure is incorporated by reference) discloses certain pyridyldihydroxyheptenoic acid, such as cerivastatin. In addition, U.S. Patent No. 5,273,995 (this disclosure is incorporated by reference) discloses certain 6-[2-(substituted pyrrole-1-yl)alkyl]pyran-2-one, such as atorvastatin, and any pharmaceutically acceptable form thereof (i.e., LIPITOR®). Additional HMG-CoA reductase inhibitors include rosuvastatin and pitavastatin.

[0113] Atorvastatin calcium (i.e., atorvastatin hemicalcium), disclosed in U.S. Patent No. 5,273,995, incorporated herein by reference, is currently marketed as Lipitor®.

[0114] Statins also include compounds such as rosuvastatin disclosed in U.S. Patent No. 37,314E, pitivastatin disclosed in European Patent No. 304063B1 and U.S. Patent No. 5,011,930, simvastatin disclosed in U.S. Patent No. 4,444,784 incorporated herein by reference; pravastatin disclosed in U.S. Patent No. 4,346,227 incorporated herein by reference; cerivastatin disclosed in U.S. Patent No. 5,502,199 incorporated herein by reference; mevastatin disclosed in U.S. Patent No. 3,983,140 incorporated herein by reference; both incorporated herein by reference This includes berostatin as disclosed in Japanese Patent No. 4,448,784 and U.S. Patent No. 4,450,171; fluvastatin as disclosed in U.S. Patent No. 4,739,073 incorporated herein by reference; compactin as disclosed in U.S. Patent No. 4,804,770 incorporated herein by reference; lovastatin as disclosed in U.S. Patent No. 4,231,938 incorporated herein by reference; dalvastatin as disclosed in European Patent Application Publication No. 738510A2; fluindostatin as disclosed in European Patent Application Publication No. 363934A1; and dihydrocompactin as disclosed in U.S. Patent No. 4,450,171 incorporated herein by reference.

[0115] Any HMG-CoA synthase inhibitor may be used in the combined embodiments of the present invention. The term HMG-CoA synthase inhibitor refers to a compound that inhibits the bioconversion of acetyl-coenzyme A and acetoacetyl-coenzyme A to hydroxymethylglutaryl-coenzyme A catalyzed by the enzyme HMG-CoA synthase. Such inhibition is readily determined by those skilled in the art according to standard assays (Meth Enzymol. 1975;35:155-160; Meth Enzymol. 1985;110:19-26 and the references cited therein). Various such compounds are described and referenced below, but other HMG-CoA synthase inhibitors will be known to those skilled in the art. U.S. Patent No. 5,120,729 (this disclosure is incorporated herein by reference) discloses certain beta-lactam derivatives. U.S. Patent No. 5,064,856 (this disclosure is incorporated herein by reference) discloses certain spiro-lactone derivatives (MF5253) prepared by culturing microorganisms. U.S. Patent No. 4,847,271 (this disclosure is incorporated herein by reference) discloses certain oxetane compounds, such as 11-(3-hydroxymethyl-4-oxo-2-oxetanyl)-3,5,7-trimethyl-2,4-undeca-dienoic acid derivatives.

[0116] Any compound that reduces HMG-CoA reductase gene expression may be used in the combined embodiments of the present invention. These agents may be HMG-CoA reductase transcription inhibitors that block DNA transcription or translation inhibitors that interfere with or reduce the translation of mRNA encoding HMG-CoA reductase into protein. Such compounds may directly affect transcription or translation, or may be converted in vivo to compounds having the aforementioned activity by one or more enzymes in the cholesterol biosynthesis cascade, or may cause the accumulation of isoprene metabolites having the aforementioned activity. Such compounds may cause this effect by reducing the level of SREBP (sterol regulatory element binding protein) by inhibiting the activity of site-1 protease (SI P), or by activating the oxysterol receptor, or by antagonizing SCAP. Such regulation is readily determined by those skilled in the art according to standard assays (Meth. Enzymol. 1985;110:9-19). While several compounds are described and referenced below, other inhibitors of HMG-CoA reductase gene expression will be known to those skilled in the art. U.S. Patent No. 5,041,432 (this disclosure is incorporated by reference) discloses certain 15-substituted lanosterol derivatives.

[0117] Other oxygenated sterols that inhibit HMG-CoA reductase synthesis are discussed by EI Mercer (Prog. Lip. Res. 1993; 32: 357-416).

[0118] Any compound having activity as a CETP inhibitor may function as a second compound in embodiments of the combination therapy of the present invention. The term CETP inhibitor refers to a compound that inhibits the cholesteryl ester transfer protein (CETP)-mediated transport of various cholesteryl esters and triglycerides from HDL to LDL and VLDL. Such CETP inhibitory activity is readily determined by those skilled in the art according to standard assays (e.g., U.S. Patent No. 6,140,343). Various CETP inhibitors will become known to those skilled in the art, including, for example, those disclosed in U.S. Patent No. 6,140,343 and U.S. Patent No. 6,197,786, both assigned to the assignees of the present invention. CETP inhibitors are also described in U.S. Patent No. 6,723,752 and include several CETP inhibitors, including (2R)-3-{[3-(4-chloro-3-ethyl-phenoxy)-phenyl]-[[3-(1,1,2,2-tetrafluoro-ethoxy)-phenyl]-methyl]-amino}-1,1,1-trifluoro-2-propanol. Furthermore, the CETP inhibitors included herein are also described in U.S. Patent Application No. 10 / 807838, filed on March 23, 2004. U.S. Patent No. 5,512,548 discloses certain polypeptide derivatives having activity as CETP inhibitors, while certain CETP-inhibiting losenonolactone derivatives and phosphate-containing analogs of cholesteryl esters are disclosed in J. Antibiot, 49(8)815-816 (1996) and Bioorg. Med. Chem. Lett.; 6:1951-1954 (1996), respectively.

[0119] Any PPAR modulator may be used in the combined embodiments of the present invention. The term PPAR modulator refers to compounds that modulate peroxisome proliferator-activator receptor (PPAR) activity in mammals, particularly humans. Such modulation is readily determined by those skilled in the art according to standard assays known in the literature. By modulating the PPAR receptor, such compounds regulate the transcription of key genes involved in lipid and glucose metabolism, such as those in fatty acid oxidation, and also involved in high-density lipoprotein (HDL) association (e.g., apolipoprotein A1 gene transcription), thus reducing overall fat and increasing HDL cholesterol. Due to their activity, these compounds also reduce plasma levels of their related components, such as triglycerides, VLDL cholesterol, LDL cholesterol, and apolipoprotein B in mammals, particularly humans, and increase HDL cholesterol and apolipoprotein A1. Therefore, these compounds are useful in the treatment and correction of various dyslipidemias observed to be associated with the development and occurrence of cardiovascular diseases, including atherosclerosis, as well as hypoalphalipoproteinemia and hypertriglyceridemia. Various of these compounds are described and referenced below, but others will be known to those skilled in the art. International Publication Nos. 02 / 064549 and 02 / 064130, and U.S. Patent Application No. 10 / 720942 filed November 24, 2003, and U.S. Patent Application No. 60 / 552114 filed March 10, 2004 (these disclosures are incorporated herein by reference) disclose certain compounds that are PPARa activators.

[0120] Any other PPAR regulator may be used in combination with the present invention. In particular, regulators of PPARp and / or PPARy may be useful in combination with the compounds of the present invention. An example of a PPAR inhibitor is described as {5-methoxy-2-methyl-4-[4-(4-trifluoromethyl-benzyloxy)-benzylsulfanyl]-phenoxy}acetic acid in U.S. Patent Application Publication No. 2003 / 0225158.

[0121] Any MTP / Apo B (microsomal triglyceride transport protein and / or apolipoprotein B) secretion inhibitor may be used in the combined embodiments of the present invention. The term MTP / Apo B secretion inhibitor refers to compounds that inhibit the secretion of triglycerides, cholesteryl esters, and phospholipids. Such inhibition is readily determined by those skilled in the art according to standard assays (e.g., Wetterau, JR 1992; Science 258:999). A variety of MTP / Apo B secretion inhibitors will become known to those skilled in the art and include impritapide (Bayer) and additional compounds disclosed in International Publication No. 96 / 40640 and International Publication No. 98 / 23593 (two exemplary publications).

[0122] Any squalene synthase inhibitor may be used in the combined embodiments of the present invention. The term squalene synthase inhibitor refers to a compound that inhibits the condensation of two molecules of farnesyl pyrophosphate that form squalene, catalyzed by the enzyme squalene synthase. Such inhibition is readily determined by those skilled in the art according to standard assays (Meth. Enzymol. 1969;15:393-454 and Meth. Enzymol. 1985;110:359-373 and the references contained therein). Various such compounds are described and referenced below, but other squalene synthase inhibitors will be known to those skilled in the art. U.S. Patent No. 5,026,554 (this disclosure is incorporated by reference) discloses a fermentation product of the microorganism MF5465 (ATCC 74011) containing zaragodic acid. A summary of other claimed squalene synthase inhibitors is compiled (Curr. Op. Then Patents (1993) 861-4).

[0123] Any squalene epoxidase synthase inhibitor may be used in the combined embodiments of the present invention. The term squalene epoxidase inhibitor refers to a compound that inhibits the bioconversion of squalene and oxygen molecules to squalene-2,3-epoxide, catalyzed by the enzyme squalene epoxidase. Such inhibition is readily determined by those skilled in the art according to standard assays (Biochim. Biophys. Acta 1984;794:466-471). Various such compounds are described and referenced below, but other squalene epoxidase inhibitors will be known to those skilled in the art. U.S. Patent Nos. 5,011,859 and 5,064,864 (this disclosure is incorporated by reference) disclose certain fluoroanalogs of squalene. European Patent Application Publication No. 395,768A (this disclosure is incorporated by reference) discloses certain substituted allylamine derivatives. International Publication No. 9312069A (this disclosure is incorporated herein by reference) discloses certain amino alcohol derivatives. U.S. Patent No. 5,051,534 (this disclosure is incorporated herein by reference) discloses certain cyclopropyloxy-squalene derivatives.

[0124] Any squalene cyclase inhibitor may be used as a second component in the combination of the present invention. The term squalene cyclase inhibitor refers to a compound that inhibits the bioconversion of squalene-2,3-epoxide to lanosterol, which is catalyzed by the enzyme squalene cyclase. Such inhibition is readily determined by those skilled in the art according to standard assays (FEBS Lett. 1989; 244: 347-350). In addition, the compounds described and referenced below are squalene cyclase inhibitors, but other squalene cyclase inhibitors will also be known to those skilled in the art. International Publication No. 09410150 (this disclosure is incorporated herein by reference) discloses certain 1,2,3,5,6,7,8,8a-octahydro-2-allyl-5,5,8(beta)-trimethyl-6(beta)-isoquinolineamine derivatives. French Patent No. 2697250 (this disclosure is incorporated herein by reference) discloses certain beta,beta-dimethyl-4-piperidineethanol derivatives, such as 1-(1,5,9-trimethyldecyl)-beta,beta-dimethyl-4-piperidineethanol.

[0125] Any combined squalene epoxidase / squalene cyclase inhibitor may be used as a second component in the combined embodiments of the present invention. The term combined squalene epoxidase / squalene cyclase inhibitor refers to a compound that inhibits the bioconversion of squalene to lanosterol via a squalene-2,3-epoxide intermediate. In some assays, squalene epoxidase inhibitors and squalene cyclase inhibitors cannot be distinguished; however, these assays are recognized by those skilled in the art. Thus, inhibition by combined squalene epoxidase / squalene cyclase inhibitors is readily determined by those skilled in the art according to the aforementioned standard assays for squalene cyclase or squalene epoxidase inhibitors. Various such compounds are described and referenced below, but other squalene epoxidase / squalene cyclase inhibitors will be known to those skilled in the art. U.S. Patent Nos. 5,084,461 and 5,278,171 (this disclosure is incorporated by reference) disclose certain azadecalin derivatives. European Patent No. 468,434 (this disclosure is incorporated herein by reference) discloses certain piperidyl ethers and thio-ether derivatives, such as 2-(1-piperidyl)pentylisopentyl sulfoxide and 2-(1-piperidyl)ethylethyl sulfide. International Publication No. 9401404 (this disclosure is incorporated herein by reference) discloses certain acyl-piperidines, such as 1-(1-oxopentyl-5-phenylthio)-4-(2-hydroxy-1-methyl)-ethyl)piperidine. U.S. Patent No. 5,102,915 (this disclosure is incorporated herein by reference) discloses certain cyclopropyloxysqualene derivatives.

[0126] The compounds of the present invention may also be administered in combination with naturally occurring compounds that act to lower plasma cholesterol levels. These naturally occurring compounds are commonly referred to as functional foods and include, for example, garlic extract and niacin. A sustained-release form of niacin is available and is known as Niaspan. Niacin may also be combined with other therapeutic agents such as lovastatin or another HMG-CoA reductase inhibitor. This combination therapy with lovastatin is known as ADVICOR® (Kos Pharmaceuticals Inc.).

[0127] Any cholesterol absorption inhibitor may be used as an additional compound in the combined embodiments of the present invention. The term cholesterol absorption inhibitor refers to the ability of a compound to inhibit cholesterol contained in the lumen of the intestine from entering intestinal cells and / or from moving from intestinal cells into the lymphatic system and / or bloodstream. Such cholesterol absorption inhibitory activity is readily determined by those skilled in the art according to standard assays (e.g., J. Lipid Res. (1993) 34:377-395). Cholesterol absorption inhibitors are known to those skilled in the art and are described, for example, in International Publication No. 94 / 00480. An example of a cholesterol absorption inhibitor is ZETIA® (ezetimibe) (Schering-Plough / Merck).

[0128] Any ACAT inhibitor may be used in embodiments of the combination therapy of the present invention. The term ACAT inhibitor refers to a compound that inhibits the intracellular esterification of dietary cholesterol by the enzyme acyl-CoA:cholesterol acyltransferase. Such inhibition can be readily determined by those skilled in the art according to standard assays, such as the method of Heider et al. described in Journal of Lipid Research, 24:1127 (1983). A variety of these compounds are known to those skilled in the art; for example, U.S. Patent No. 5,510,379 discloses certain carboxysulfonates, while both International Publication Nos. 96 / 26948 and International Publication Nos. 96 / 10559 disclose urea derivatives having ACAT inhibitory activity. Examples of ACAT inhibitors include compounds such as abasimib (Pfizer), CS-505 (Sankyo), and eflutimibe (Eli Lilly and Pierre Fabre).

[0129] Lipase inhibitors may be used in embodiments of the combination therapy of the present invention. Lipase inhibitors are compounds that inhibit the metabolic cleavage of dietary triglycerides or plasma phospholipids into free fatty acids and corresponding glycerides (e.g., EL, HL, etc.). Under normal physiological conditions, lipolysis occurs via a two-step process involving acylation of the activated serine portion of the lipase enzyme. This results in the formation of a fatty acid-lipase hemiacetal intermediate, which is subsequently cleaved to release diglycerides. After further deacylation, the lipase-fatty acid intermediate is cleaved to yield free lipase, glycerides, and fatty acids. In the intestinal tract, the resulting free fatty acids and monoglycerides are incorporated into bile acid-phospholipid micelles, which are subsequently absorbed at the brush border level of the small intestine. The micelles eventually enter peripheral circulation as chylomicrons. Such lipase inhibitory activity can be readily determined by those skilled in the art according to standard assays (e.g., Methods Enzymol 286:190-231).

[0130] Pancreatic lipases mediate the metabolic cleavage of fatty acids from triglycerides at the 1- and 3-carbon positions. The primary sites of ingested fat metabolism are the duodenum and proximal jejunum, where pancreatic lipases are secreted in significantly greater quantities than required for fat breakdown in the upper small intestine. Since pancreatic lipases are the key enzymes required for the absorption of dietary triglycerides, inhibitors have potential applications in the treatment of obesity and other related conditions. Such pancreatic lipase inhibitory activity can be readily determined by those skilled in the art according to standard assays (e.g., Methods Enzymol 286:190-231).

[0131] Gastric lipase is an immunologically distinct lipase involved in approximately 10–40% of the digestion of dietary fats. Gastric lipase is secreted in response to mechanical stimulation, food intake, the presence of a high-fat diet, or sympathetic agonists. The breakdown of ingested fats in the stomach is physiologically important for supplying the fatty acids necessary to induce pancreatic lipase activity in the intestines, and is also crucial for fat absorption under various physiological and pathological conditions associated with pancreatic insufficiency. See, for example, CKAbrams, et al., Gastroenterology, 92, 125 (1987). Such gastric lipase inhibitory activity can be readily determined by those skilled in the art according to standard assays (e.g., Methods Enzymol 286:190-231).

[0132] Various gastric and / or pancreatic lipase inhibitors are known to those skilled in the art. Preferred lipase inhibitors are selected from lipustatin, tetrahydrolipustatin (orlistat), valilactone, esteratin, everactone A, and everactone B. The compound tetrahydrolipustatin is particularly preferred. Lipase inhibitors, N-3-trifluoromethylphenyl-N'-3-chloro-4'-trifluoromethylphenylurea, and various related urea derivatives are disclosed in U.S. Patent No. 4,405,644. Lipase inhibitors, esteratin, are disclosed in U.S. Patent Nos. 4,189,438 and 4,242,453. Lipase inhibitors, cyclo-0,0'-[(1,6-hexanediyl)-bis-(iminocarbonyl)]dioximes, and various related bis(iminocarbonyl)dioximes can be prepared as described in Petersen et al., Liebig's Annalen, 562, 205-229 (1949).

[0133] Various pancreatic lipase inhibitors are described below in this specification. The pancreatic lipase inhibitor lipustatin, (2S,3S,5S,7Z,10Z)-5-[(S)-2-formamido-4-methyl-valeryloxy]-2-hexyl-3-hydroxy-7,10-hexadecanoate lactone, and tetrahydrolipustatin (orlistat), (2S,3S,5S)-5-[(S)-2-formamido-4-methyl-valeryloxy]-2-hexyl-3-hydroxy-hexadecane-1,3ate lactone, as well as various substituted N-formylleucine derivatives and their stereoisomers are disclosed in U.S. Patent No. 4,598,089. For example, tetrahydrolipstatin is prepared as described in, for example, U.S. Patent No. 5,274,143; No. 5,420,305; No. 5,540,917; and No. 5,643,874. Pancreatic lipase inhibitors, FL-386, 1-[4-(2-methylpropyl)cyclohexyl]-2-[(phenylsulfonyl)oxy]-ethanone and related various substituted sulfonic acid derivatives are disclosed in U.S. Patent No. 4,452,813. The pancreatic lipase inhibitor, WAY-121898, 4-phenoxyphenyl-4-methylpiperidine-1-yl carboxylate, and various related carbamic esters and pharmaceutically acceptable salts are disclosed in U.S. Patent No. 5,512,565; U.S. Patent No. 5,391,571 and U.S. Patent No. 5,602,151. The pancreatic lipase inhibitor, valilactone, and the process for its preparation by microbial culture of Actinomycetes strain MG147-CF2 are disclosed in Kitahara, et al., J. Antibiotics, 40(11), 1647-1650 (1987). The process for the preparation of the pancreatic lipase inhibitors, everactone A and everactone B, and the Actinomycetes strain MG7-G1 by microbial culture is disclosed in Umezawa, et al., J. Antibiotics, 33, 1594-1596 (1980).The use of everactone A and B in suppressing monoglyceride formation is disclosed in Japanese Patent Publication No. Hei 8-143457, published on June 4, 1996.

[0134] Other compounds marketed for hyperlipidemia, including hypercholesterolemia, and intended to help prevent or treat atherosclerosis include bile acid scavengers such as Welchol®, Colestid®, LoCholest®, and Questran®; and fibric acid derivatives such as Atromid®, Lopid®, and Tricor®.

[0135] Given the association between diabetes and atherosclerosis (e.g., metabolic syndromes), the compound of formula I may be administered in combination with antidiabetic compounds. Diabetes may be treated by administering a therapeutically effective amount of the compound of the present invention in combination with other agents (e.g., insulin) that may be used to treat diabetes to patients with diabetes (particularly type II), insulin resistance, impaired glucose tolerance, metabolic syndromes, or diabetic complications such as neuropathy, nephropathy, retinopathy, or cataracts. This includes the class of antidiabetic agents (and certain agents) described herein.

[0136] Any glycogen phosphorylase inhibitor may be used as a second agent in combination with the compounds of the present invention. The term glycogen phosphorylase inhibitor refers to a compound that inhibits the bioconversion of glycogen to glucose-1-phosphate, which is catalyzed by the enzyme glycogen phosphorylase. Such glycogen phosphorylase inhibitory activity is readily determined by those skilled in the art according to standard assays (e.g., J. MedChem. 41 (1998) 2934-2938). Various glycogen phosphorylase inhibitors are known to those skilled in the art, including those described in International Publication No. 96 / 39384 and International Publication No. 96 / 39385.

[0137] Any aldose reductase inhibitor may be used in combination with the compounds of the present invention. The term aldose reductase inhibitor refers to a compound that inhibits the bioconversion of glucose to sorbitol catalyzed by the enzyme aldose reductase. Aldose reductase inhibition is readily determined by those skilled in the art according to standard assays (e.g., J. Malone, Diabetes, 29:861-864 (1980). “Red Cell Sorbitol, an Indicator of Diabetic Control”). Various aldose reductase inhibitors are known to those skilled in the art, such as 6-(5-chloro-3-methylbenzofuran-2-sulfonyl)-2H-pyridazine-3-one, as described in U.S. Patent No. 6,579,879.

[0138] Any sorbitol dehydrogenase inhibitor may be used in combination with the compounds of the present invention. The term sorbitol dehydrogenase inhibitor refers to a compound that inhibits the bioconversion of sorbitol to fructose catalyzed by the enzyme sorbitol dehydrogenase. Such sorbitol dehydrogenase inhibitor activity is readily determined by those skilled in the art according to standard assays (e.g., Analyt. Biochem (2000) 280:329-331). Various sorbitol dehydrogenase inhibitors are known, and for example, U.S. Patent Nos. 5,728,704 and 5,866,578 disclose compounds and methods for treating or preventing diabetic complications by inhibiting the enzyme sorbitol dehydrogenase.

[0139] Any glucosidase inhibitor can be used in combination with the compounds of the present invention. Glucosidase inhibitors inhibit the enzymatic hydrolysis of complex carbohydrates to bioavailable simple sugars, such as glucose, by glycoside hydrolases, such as amylase or maltase. In particular, the rapid metabolic action of glucosidase after the intake of high levels of carbohydrates causes a state of elevated blood glucose levels due to diet, which in high-fat or diabetic subjects leads to increased insulin secretion, increased fat synthesis, and decreased lipolysis. Following such elevated blood glucose levels, hypoglycemia often occurs due to the increased levels of insulin present. Furthermore, residual digested food in the stomach promotes the production of gastric juice, which is known to induce or facilitate the development of gastritis or duodenal ulcers. Therefore, glucosidase inhibitors are known to be useful in promoting the passage of carbohydrates through the stomach and inhibiting the absorption of glucose from the intestinal tract. Furthermore, carbohydrates are converted into lipids in adipose tissue, and subsequently, the uptake of dietary fats into adipose tissue deposits is reduced or delayed accordingly, thereby reducing or preventing the harmful abnormalities that result. Such glucosidase inhibitory activity can be readily determined by those skilled in the art according to standard assays (e.g., Biochemistry (1969) 8:4214).

[0140] Generally preferred glucosidase inhibitors include amylase inhibitors. Amylase inhibitors are glucosidase inhibitors that inhibit the enzymatic breakdown of starch or glycogen into maltose. Such amylase inhibitory activity can be readily determined by those skilled in the art according to standard assays (e.g., Methods Enzymol (1955) 1:149). Inhibition of such enzymatic breakdown is beneficial in reducing the amount of bioavailable sugars, including glucose and maltose, and the associated adverse conditions that result.

[0141] Various glucosidase inhibitors are known to those skilled in the art, and examples are provided below. Preferred glucosidase inhibitors are selected from acarbose, adiposin, voglibose, miglitol, emiglitate, camiglibose, tendamistate, trestatin, pradymycin-Q, and salvostatin. The glucosidase inhibitor acarbose and various related amino sugar derivatives are disclosed in U.S. Patent No. 4,062,950 and U.S. Patent No. 4,174,439, respectively. The glucosidase inhibitor adiposin is disclosed in U.S. Patent No. 4,254,256. Glucosidase inhibitors, voglibose, 3,4-dideoxy-4-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-2-C-(hydroxymethyl)-D-epi-inositol, and various related N-substituted pseudo-amino sugars are disclosed in U.S. Patent No. 4,701,559. Glucosidase inhibitors, miglitol, (2R,3R,4R,5S)-1-(2-hydroxyethyl)-2-(hydroxymethyl)-3,4,5-piperidinetriol, and various related 3,4,5-trihydroxypiperidines are disclosed in U.S. Patent No. 4,639,436. The glucosidase inhibitor emiglitate, p-[2-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]ethoxy]-ethyl benzoate, various related derivatives and their pharmaceutically acceptable acid addition salts are disclosed in U.S. Patent No. 5,192,772. The glucosidase inhibitor MDL-25637, 2,6-dideoxy-7-0-|3-D-glucopyranosyl-2,6-imino-D-glycero-L-glucoheptitol, various related homodisaccharides and their pharmaceutically acceptable acid addition salts are disclosed in U.S. Patent No. 4,634,765.The glucosidase inhibitor camiglibose, 6-deoxy-6-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]-aD-glucopyranoside methylsesquihydrate, related deoxynojirimycin derivatives, various pharmaceutically acceptable salts thereof, and methods for their synthesis are disclosed in U.S. Patent Nos. 5,157,116 and 5,504,078. The glycosidase inhibitor sarvostatin and various related pseudosaccharides are disclosed in U.S. Patent No. 5,091,524.

[0142] Various amylase inhibitors are known to those skilled in the art. Amylase inhibitors, tendamistat, and various related cyclic peptides are disclosed in U.S. Patent No. 4,451,455. Amylase inhibitor AI-3688 and various related cyclic polypeptides are disclosed in U.S. Patent No. 4,623,714. Amylase inhibitor trestatin, consisting of a mixture of trestatin A, trestatin B, and trestatin C, and various related trehalose-containing amino sugars are disclosed in U.S. Patent No. 4,273,765.

[0143] Additional antidiabetic compounds that can be used as a second agent in combination with the compounds of the present invention include, for example,: biguanides (e.g., metformin), insulin secretagogues (e.g., sulfonylurea and glinides), glitazone, non-glitazone PPARy agonists, PPARp agonists, DPP-IV inhibitors, PDE5 inhibitors, GSK-3 inhibitors, glucagon antagonists, f-1,6-BPase inhibitors (Metabasis / Sankyo), GLP-1 / analogs (AC 2993, also known as exendin-4), insulin and insulin mimetic (Merck natural products). Other examples would include PKC-P inhibitors and AGE degrading agents.

[0144] The compounds of the present invention may also be used in combination with cardiovascular therapeutic agents such as antihypertensive drugs. Any antihypertensive drug may be used as a second agent in such a combination, and examples are provided herein. Such antihypertensive activity can be readily determined by those skilled in the art according to standard assays (e.g., blood pressure measurement).

[0145] Amlodipine and related dihydropyridine compounds are disclosed as potent anti-ischemic and antihypertensive agents in U.S. Patent No. 4,572,909, incorporated herein by reference. U.S. Patent No. 4,879,303, incorporated herein by reference, discloses amlodipine benzenesulfonate (also known as amlodipine besylate). Amlodipine and amlodipine besylate are potent and sustained-release calcium channel blockers. Therefore, amlodipine, amlodipine besylate, amlodipine maleate, and other pharmaceutically acceptable acid addition salts of amlodipine are useful as antihypertensive and anti-ischemic agents. Amlodipine besylate is currently marketed as Norvasc®.

[0146] Calcium channel blockers within the scope of the present invention include bepridil, which can be prepared as disclosed in U.S. Patent No. 3,962,238 or U.S. Reissue Patent No. 30,577; clentiazem, which can be prepared as disclosed in U.S. Patent No. 4,567,175; diltiazem, which can be prepared as disclosed in U.S. Patent No. 3,562; fendiline, which can be prepared as disclosed in U.S. Patent No. 3,262,977; and, as disclosed in U.S. Patent No. 3,261,859. Gallopamil, which can be prepared as disclosed in U.S. Patent No. 4,808,605; mibefuradil, which can be prepared as disclosed in U.S. Patent No. 3,152,173; semothiadil, which can be prepared as disclosed in U.S. Patent No. 4,786,635; telodiline, which can be prepared as disclosed in U.S. Patent No. 3,371,014; verapamil, which can be prepared as disclosed in U.S. Patent No. 3,261,859; Aranipine, which can be prepared as disclosed in National Patent No. 4,572,909; Barnidipine, which can be prepared as disclosed in U.S. Patent No. 4,220,649; Benidipine, which can be prepared as disclosed in European Patent Application Publication No. 106,275; Cilnidipine, which can be prepared as disclosed in U.S. Patent No. 4,672,068; Efonidipine, which can be prepared as disclosed in U.S. Patent No. 4,885,284; In U.S. Patent No. 4,952,592 Ergodipine, which can be prepared as disclosed; ferrodipine, which can be prepared as disclosed in U.S. Patent No. 4,264,611; isradipine, which can be prepared as disclosed in U.S. Patent No. 4,466,972; lasidipine, which can be prepared as disclosed in U.S. Patent No. 4,801,599; relcanidipine, which can be prepared as disclosed in U.S. Patent No. 4,705,797; manidipine, which can be prepared as disclosed in U.S. Patent No. 4,892,875;Nicardipine as can be prepared as disclosed in U.S. Patent No. 3,985,758; Nifedipine as can be prepared as disclosed in U.S. Patent No. 3,485,847; Nilvadipine as can be prepared as disclosed in U.S. Patent No. 4,338,322; Nimodipine as can be prepared as disclosed in U.S. Patent No. 3,799,934; Nisoldipine as can be prepared as disclosed in U.S. Patent No. 4,154,839; Nitrendipine as can be prepared as disclosed in U.S. Patent No. 3,799,934; As disclosed in U.S. Patent No. 2,882,271 Examples of possible preparations include, but are not limited to, cinnarizine; flunarizine, which can be prepared as disclosed in U.S. Patent No. 3,773,939; lidoflazine, which can be prepared as disclosed in U.S. Patent No. 3,267,104; lomerizine, which can be prepared as disclosed in U.S. Patent No. 4,663,325; bencyclan, which can be prepared as disclosed in Hong Kong Patent No. 151,865; etaphenone, which can be prepared as disclosed in German Patent No. 1,265,758; and perhexylin, which can be prepared as disclosed in British Patent No. 1,025,578. All such U.S. patent disclosures are incorporated herein by reference. Examples of currently marketed antihypertensive drugs include calcium channel blockers such as Cardizem®, Adalat®, Calan®, Cardene®, Covera®, Dilacor®, DynaCirc®, Procardia XL®, Sular®, Tiazac®, Vascor®, Verelan®, Isoptin®, Nimotop®, Norvasc®, and Plendil®;Examples of angiotensin-converting enzyme (ACE) inhibitors include Accupril®, Altace®, Captopril®, Lotensin®, Mavik®, Monopril®, Prinivil®, Univasc®, Vasotec®, and Zestril®.

[0147] Angiotensin-converting enzyme inhibitors (ACE inhibitors) within the scope of the present invention include: alacepril, which can be prepared as disclosed in U.S. Patent No. 4,248,883; benazepril, which can be prepared as disclosed in U.S. Patent No. 4,410,520; captopril, which can be prepared as disclosed in U.S. Patents No. 4,046,889 and No. 4,105,776; seronapril, which can be prepared as disclosed in U.S. Patent No. 4,452,790; delapril, which can be prepared as disclosed in U.S. Patent No. 4,385,051; enalapril, which can be prepared as disclosed in U.S. Patent No. 4,374,829; hosinopril, which can be prepared as disclosed in U.S. Patent No. 4,337,201; and the other inhibitors disclosed in U.S. Patent No. 4,508,727. Examples include, but are not limited to, imadapril, which can be prepared as disclosed in U.S. Patent No. 4,555,502; lysinopril, which can be prepared as disclosed in U.S. Patent No. 893,553; modertopril, which can be prepared as disclosed in U.S. Patent No. 4,508,729; perindopril, which can be prepared as disclosed in U.S. Patent No. 4,344,949; quinapril, which can be prepared as disclosed in U.S. Patent No. 4,587,258; ramipril, which can be prepared as disclosed in U.S. Patent No. 4,470,972; temocapril, which can be prepared as disclosed in U.S. Patent No. 4,699,905; and trandolapril, which can be prepared as disclosed in U.S. Patent No. 4,933,361. All such U.S. patent disclosures are incorporated herein by reference.

[0148] Angiotensin-II receptor antagonists (A-II antagonists) within the scope of the present invention include, but are not limited to, candesartan, which can be prepared as disclosed in U.S. Patent No. 5,196,444; eprosartan, which can be prepared as disclosed in U.S. Patent No. 5,185,351; irbesartan, which can be prepared as disclosed in U.S. Patent No. 5,270,317; losartan, which can be prepared as disclosed in U.S. Patent No. 5,138,069; and valsartan, which can be prepared as disclosed in U.S. Patent No. 5,399,578. All such disclosures of U.S. patents are incorporated herein by reference.

[0149] Beta-adrenergic receptor blockers (beta or β-blockers) within the scope of the present invention include acebutolol, which can be prepared as disclosed in U.S. Patent No. 3,857,952; alprenolol, which can be prepared as disclosed in Dutch Patent No. 6,605,692; amosuralol, which can be prepared as disclosed in U.S. Patent No. 4,217,305; arotinolol, which can be prepared as disclosed in U.S. Patent No. 3,932,400; and U.S. Patent No. 3,663,607 or Atenolol, which can be prepared as disclosed in U.S. Patent No. 3,836,671; befnolol, which can be prepared as disclosed in U.S. Patent No. 3,853,923; betaxolol, which can be prepared as disclosed in U.S. Patent No. 4,252,984; bevantrol, which can be prepared as disclosed in U.S. Patent No. 3,857,981; bisoprolol, which can be prepared as disclosed in U.S. Patent No. 4,171,370; disclosed in U.S. Patent No. 4,340,541 Bobindolol which can be prepared as disclosed in U.S. Patent No. 3,663,570; Bufetolol which can be prepared as disclosed in U.S. Patent No. 3,723,476; Buflalol which can be prepared as disclosed in U.S. Patent No. 3,929,836; Bunitrol which can be prepared as disclosed in U.S. Patent No. 3,940,489 and No. 3,961,071; as disclosed in U.S. Patent No. 3,309,406 Buprandrol, which can be prepared in the manner disclosed in French Patent No. 1,390,056; butyrizine hydrochloride, which can be prepared in the manner disclosed in U.S. Patent No. 4,252,825; carazolol, which can be prepared in the manner disclosed in German Patent No. 2,240,599; carteolol, which can be prepared in the manner disclosed in U.S. Patent No. 3,910,924; carvedilol, which can be prepared in the manner disclosed in U.S. Patent No. 4,503,067;Celiprolol, which can be prepared as disclosed in U.S. Patent No. 4,034,009; Cetamolol, which can be prepared as disclosed in U.S. Patent No. 4,059,622; Chloranolol, which can be prepared as disclosed in German Patent No. 2,213,044; Direvalol, which can be prepared as disclosed in Clifton et al., Journal of Medicinal Chemistry, 1982, 25, 670; Epanolol, which can be prepared as disclosed in European Patent Application Publication No. 41,491; Indenolol, which can be prepared as disclosed in U.S. Patent No. 4,045,482; Labetalol, which can be prepared as disclosed in U.S. Patent No. 4,012,444; Levovonolol, which can be prepared as disclosed in U.S. Patent No. 4,463,176; Seeman et al. Mepindolol, which can be prepared as disclosed in al., Helv. Chim. Acta, 1971, 54, 241; Metipranolol, which can be prepared as disclosed in Czechoslovakian Patent Application No. 128,471; Metoprolol, which can be prepared as disclosed in U.S. Patent No. 3,873,600; Moprolol, which can be prepared as disclosed in U.S. Patent No. 3,501,7691; Nadolol, which can be prepared as disclosed in U.S. Patent No. 3,935,267; Nadoxolo, which can be prepared as disclosed in U.S. Patent No. 3,819,702 Nevivalol, which can be prepared as disclosed in U.S. Patent No. 4,654,362; nipradilol, which can be prepared as disclosed in U.S. Patent No. 4,394,382; oxprenolol, which can be prepared as disclosed in British Patent No. 1,077,603; perbutrol, which can be prepared as disclosed in U.S. Patent No. 3,551,493; pindolol, which can be prepared as disclosed in Swiss Patent Nos. 469,002 and 472,404; practrol, which can be prepared as disclosed in U.S. Patent No. 3,408,387;Pronetarol, which can be prepared as disclosed in British Patent No. 909,357; propranolol, which can be prepared as disclosed in U.S. Patents No. 3,337,628 and No. 3,520,919; Uloth et al., Journal of Medicinal Examples include, but are not limited to, sotalol, which can be prepared as disclosed in Chemistry, 1966, 9,88; sfinarol, which can be prepared as disclosed in German Patent No. 2,728,641; talindor, which can be prepared as disclosed in U.S. Patents No. 3,935,259 and No. 4,038,313; tertatrol, which can be prepared as disclosed in U.S. Patent No. 3,960,891; chilisolol, which can be prepared as disclosed in U.S. Patent No. 4,129,565; timolol, which can be prepared as disclosed in U.S. Patent No. 3,655,663; triprolol, which can be prepared as disclosed in U.S. Patent No. 3,432,545; and xybenolol, which can be prepared as disclosed in U.S. Patent No. 4,018,824. All such U.S. patent disclosures are incorporated herein by reference.

[0150] Alpha-adrenergic receptor blockers (alpha or α-blockers) within the scope of the present invention include amosulol, which can be prepared as disclosed in U.S. Patent No. 4,217,307; arotinolol, which can be prepared as disclosed in U.S. Patent No. 3,932,400; dapiprazole, which can be prepared as disclosed in U.S. Patent No. 4,252,721; doxazosin, which can be prepared as disclosed in U.S. Patent No. 4,188,390; fenspiride, which can be prepared as disclosed in U.S. Patent No. 3,399,192; indramin, which can be prepared as disclosed in U.S. Patent No. 3,527,761; and other alpha-adrenergic receptor blockers (alpha or α-blockers) prepared as disclosed above. Examples include, but are not limited to, labetrol; naphtopidil, which can be prepared as disclosed in U.S. Patent No. 3,997,666; nicergoline, which can be prepared as disclosed in U.S. Patent No. 3,228,943; prazosin, which can be prepared as disclosed in U.S. Patent No. 3,511,836; tamsulosin, which can be prepared as disclosed in U.S. Patent No. 4,703,063; trazoline, which can be prepared as disclosed in U.S. Patent No. 2,161,938; trimazosin, which can be prepared as disclosed in U.S. Patent No. 3,669,968; and yohimbine, which can be isolated from natural sources according to methods well known to those skilled in the art. All such U.S. patent disclosures are incorporated herein by reference.

[0151] As used herein, the term “vasodilator” includes cerebral vasodilators, coronary vasodilators, and peripheral vasodilators. Cerebral vasodilators within the scope of the present invention include: bencyclan, which can be prepared as disclosed above; cinnarizine, which can be prepared as disclosed above; citicoline, which can be isolated from natural sources as disclosed in Kennedy et al., Journal of the American Chemical Society, 1955, 77, 250, or synthesized as disclosed in Kennedy, Journal of Biological Chemistry, 1956, 222, 185; cyclanderate, which can be prepared as disclosed in U.S. Patent No. 3,663,597; cyclonicate, which can be prepared as disclosed in German Patent No. 1,910,481; diisopropylamine dichloroacetate, which can be prepared as disclosed in British Patent No. 862,248; Hermann et al., Journal of the American Chemical Society Eblunamonin, which can be prepared as disclosed in Society, 1979, 101, 1540; Fasudil, which can be prepared as disclosed in U.S. Patent No. 4,678,783; Phenoxedil, which can be prepared as disclosed in U.S. Patent No. 3,818,021; Flunarizine, which can be prepared as disclosed in U.S. Patent No. 3,773,939; Ibudilast, which can be prepared as disclosed in U.S. Patent No. 3,850,941; Ifenprodil, which can be prepared as disclosed in U.S. Patent No. 3,509,164; Lomerizine, which can be prepared as disclosed in U.S. Patent No. 4,663,325; Nafronil, which can be prepared as disclosed in U.S. Patent No. 3,334,096; Blicke et al.Examples include, but are not limited to, nicamethate, which can be prepared as disclosed in the Journal of the American Chemical Society, 1942, 64, 1722; nicergoline, which can be prepared as disclosed above; nimodipine, which can be prepared as disclosed in U.S. Patent No. 3,799,934; papaverine, which can be prepared as outlined in Goldberg, Chem. Prod. Chem. News, 1954, 17, 371; pentiphylline, which can be prepared as disclosed in German Patent No. 860,217; thinophedrine, which can be prepared as disclosed in U.S. Patent No. 3,563,997; vincamine, which can be prepared as disclosed in U.S. Patent No. 3,770,724; vinpocetine, which can be prepared as disclosed in U.S. Patent No. 4,035,750; and biquidyl, which can be prepared as disclosed in U.S. Patent No. 2,500,444. All such U.S. patent disclosures are incorporated herein by reference.

[0152] Coronary vasodilators within the scope of the present invention include amotrifen, which can be prepared as disclosed in U.S. Patent No. 3,010,965; J.Chem.Soc.Bendazole, which can be prepared as disclosed in 1958,2426; benphlozyl hemysuccinate, which can be prepared as disclosed in U.S. Patent No. 3,355,463; benziodarone, which can be prepared as disclosed in U.S. Patent No. 3,012,042; chloracidine, which can be prepared as disclosed in British Patent No. 740,932; chromonal, which can be prepared as disclosed in U.S. Patent No. 3,282,938; British Patent No. 1,160,925 Clobenfural, which can be prepared as disclosed in the detailed specification; Chlonitrate, which can be prepared from propanediol according to a method well known to those skilled in the art (see, for example, Annalen, 1870, 155, 165); Chlorichromene, which can be prepared as disclosed in U.S. Patent No. 4,452,811; Dilazep, which can be prepared as disclosed in U.S. Patent No. 3,532,685; Dipyridamole, which can be prepared as disclosed in British Patent No. 807,826; Germany Dropreniramine, which can be prepared as disclosed in Japanese Patent No. 2,521,113; efloroxate, which can be prepared as disclosed in British Patent Nos. 803,372 and 824,547; erythrityl tetranitrate, which can be prepared by nitration of erythritol according to a method well known to those skilled in the art; etaphenone, which can be prepared as disclosed in German Patent No. 1,265,758; fen, which can be prepared as disclosed in U.S. Patent No. 3,262,977 Dilin; Floresyl, which can be prepared as disclosed in German Patent No. 2,020,464; Gangrefen, which can be prepared as disclosed in Soviet Patent No. 115,905; Hexestrol, which can be prepared as disclosed in U.S. Patent No. 2,357,985; Hexobenzine, which can be prepared as disclosed in U.S. Patent No. 3,267,103; Itramintsylate, which can be prepared as disclosed in Japanese Patent No. 168,308; Baxter et al.Kerin, which can be prepared as disclosed in Journal of the Chemical Society, 1949, S 30; Lidoflazin, which can be prepared as disclosed in U.S. Patent No. 3,267,104; Mannitol hexanitrate, which can be prepared by nitration of mannitol according to a method well known to those skilled in the art; Medibazin, which can be prepared as disclosed in U.S. Patent No. 3,119,826; Nitroglycerin; Pentaerythritol tetranitrate, which can be prepared by nitration of pentaerythritol according to a method well known to those skilled in the art; Pentrinitrol, which can be prepared as disclosed in German Patent No. 638,422-3; Perhexylin, which can be prepared as disclosed above; Pimephylline, which can be prepared as disclosed in U.S. Patent No. 3,350,400; U.S. Patent No. 3,152, Examples include, but are not limited to, preniramine, which can be prepared as disclosed in Patent No. 173; propatyl nitrate, which can be prepared as disclosed in French Patent No. 1,103,113; trapidyl, which can be prepared as disclosed in East German Patent No. 55,956; triclomyl, which can be prepared as disclosed in U.S. Patent No. 2,769,015; trimetazidine, which can be prepared as disclosed in U.S. Patent No. 3,262,852; trolnitrate phosphate, which can be prepared by precipitation with phosphoric acid after nitration of triethanolamine according to a method well known to those skilled in the art; and visnadine, which can be prepared as disclosed in U.S. Patents No. 2,816,118 and No. 2,980,699. All such U.S. patent disclosures are incorporated herein by reference.

[0153] Peripheral vasodilators within the scope of the present invention include aluminum nicotinate, which can be prepared as disclosed in U.S. Patent No. 2,970,082; bamethan, which can be prepared as disclosed in Corrigan et al., Journal of the American Chemical Society, 1945, 67, 1894; bencyclan, which can be prepared as disclosed above; betahistine, which can be prepared as disclosed in Walter et al., Journal of the American Chemical Society, 1941, 63, 2771; and Hamburg et al. Bradykinin, which can be prepared as disclosed in al., Arch. Biochem. Biophys., 1958, 76, 252; brobincamine, which can be prepared as disclosed in U.S. Patent No. 4,146,643; bufeniod, which can be prepared as disclosed in U.S. Patent No. 3,542,870; buflomedil, which can be prepared as disclosed in U.S. Patent No. 3,895,030; butaramine, which can be prepared as disclosed in U.S. Patent No. 3,338,899; cethiezyl, which can be prepared as disclosed in French Patent No. 1,460,571; cyclonicate, which can be prepared as disclosed in German Patent No. 1,910,481; White Patent No. Cinepazide, which can be prepared as disclosed in Patent No. 730,345; cinnarizine, which can be prepared as disclosed above; cyclandelate, which can be prepared as disclosed above; diisopropylamine dichloroacetate, which can be prepared as disclosed above; eledoisin, which can be prepared as disclosed in British Patent No. 984,810; phenoxedyl, which can be prepared as disclosed above; flunarizine, which can be prepared as disclosed above; hepronicate, which can be prepared as disclosed in U.S. Patent No. 3,384,642; ifenprodil, which can be prepared as disclosed above; iloprost, which can be prepared as disclosed in U.S. Patent No. 4,692,464; Badgett et al.Inositol niacinate, which can be prepared as disclosed in Journal of the American Chemical Society, 1947, 69, 2907; isoxuprine, which can be prepared as disclosed in U.S. Patent No. 3,056,836; calidine, which can be prepared as disclosed in Biochem. Biophys. Res. Commun., 1961, 6, 210; kallikrein, which can be prepared as disclosed in German Patent No. 1,102,973; moxysilite, which can be prepared as disclosed in German Patent No. 905,738; nafronil, which can be prepared as disclosed above; above Nicomethate, which can be prepared as disclosed above; nicergoline, which can be prepared as disclosed above; nicofuranose, which can be prepared as disclosed in Swiss Patent No. 366,523; niridrine, which can be prepared as disclosed in U.S. Patent Nos. 2,661,372 and 2,661,373; pentifyl, which can be prepared as disclosed above; pentoxifylline, which can be prepared as disclosed in U.S. Patent No. 3,422,107; pyribezil, which can be prepared as disclosed in U.S. Patent No. 3,299,067; Merck Examples of such preparations include, but are not limited to, prostaglandin Ei, which can be prepared by any of the methods referenced in Index, Twelfth Edition, Budaveri, Ed., New Jersey, 1996, p. 1353; throctidyl, which can be prepared as disclosed in German Patent No. 2,334,404; trazoline, which can be prepared as disclosed in U.S. Patent No. 2,161,938; and xanthinol nicotinate, which can be prepared as disclosed in German Patent No. 1,102,750 or Korbonits et al., Acta. Pharm. Hung., 1968, 38, 98. All such U.S. patent disclosures are incorporated herein by reference.

[0154] Within the scope of this invention, the term "diuretic" includes diuretic benzothiadiazine derivatives, diuretic organic mercury compounds, diuretic purines, diuretic steroids, diuretic sulfonamide derivatives, diuretic uracil, and amanodine, which can be prepared as disclosed in Australian Patent No. 168,063; amiloride, which can be prepared as disclosed in White Patent No. 639,386; arbutin, which can be prepared as disclosed in Tschitschibabin, Annalen, 1930,479,303; and Australian Patent No. Chlorazanil, which can be prepared as disclosed in U.S. Patent No. 168,063; ethacric acid, which can be prepared as disclosed in U.S. Patent No. 3,255,241; ethozolin, which can be prepared as disclosed in U.S. Patent No. 3,072,653; hydracarbazine, which can be prepared as disclosed in British Patent No. 856,409; isosorbide, which can be prepared as disclosed in U.S. Patent No. 3,160,641; mannitol; Freudenberg This includes methochalcone, which can be prepared as disclosed in et al., Ber., 1957, 90, 957; muzolimin, which can be prepared as disclosed in U.S. Patent No. 4,018,890; perhexylline, which can be prepared as disclosed above; ticlinafene, which can be prepared as disclosed in U.S. Patent No. 3,758,506; triamterene, which can be prepared as disclosed in U.S. Patent No. 3,081,230; and other diuretics such as urea. All disclosures of these U.S. patents are incorporated herein by reference.

[0155] Diuretic benzothiadiazine derivatives within the scope of the present invention include arthiazide, which can be prepared as disclosed in British Patent No. 902,658; bendroflumethiazide, which can be prepared as disclosed in U.S. Patent No. 3,265,573; and benzthiazide. McManus et al., 136th Am. Soc. Meeting (Atlantic City, September 1959), Abstract of Papers, pp. 13-0; Bentilhydrochlorothiazide as prepared as disclosed in U.S. Patent No. 3,108,097; Buthiazide as prepared as disclosed in UK Patent Nos. 861,367 and 885,078; Chlorothiazide as prepared as disclosed in U.S. Patent Nos. 2,809,194 and 2,937,169; Chlorthalidone as prepared as disclosed in U.S. Patent No. 3,055,904; Cyclopenthiazide as prepared as disclosed in Whitehead Patent No. 587,225; Whitehead et al., Journal of Organic Cyclothiazide, which can be prepared as disclosed in Chemistry, 1961, 26, 2814; epithiazide, which can be prepared as disclosed in U.S. Patent No. 3,009,911; ethiazide, which can be prepared as disclosed in British Patent No. 861,367; fenquisone, which can be prepared as disclosed in U.S. Patent No. 3,870,720; indapamide, which can be prepared as disclosed in U.S. Patent No. 3,565,911; hydrochlorothiazide, which can be prepared as disclosed in U.S. Patent No. 3,164,588; hydroflumethiazide, which can be prepared as disclosed in U.S. Patent No. 3,254,076; Close et al.Methiclothiazide, which can be prepared as disclosed in Journal of the American Chemical Society, 1960, 82, 1132; meticran, which can be prepared as disclosed in French Patent No. M2790 and No. 1,365,504; metrazone, which can be prepared as disclosed in U.S. Patent No. 3,360,518; paraflutizide, which can be prepared as disclosed in White Patent No. 620,829; polythiazide, which can be prepared as disclosed in U.S. Patent No. 3,009,911; quinetazone, which can be prepared as disclosed in U.S. Patent No. 2,976,289; teclothiazide, which can be prepared as disclosed in Close et al., Journal of the American Chemical Society, 1960, 82, 1132; and deStevens et al. Examples include, but are not limited to, trichlormethiazide prepared as disclosed in al., Experientia, 1960, 16, 113. All such U.S. patent disclosures are incorporated herein by reference.

[0156] Diuretic sulfonamide derivatives within the scope of the present invention include acetazolamide, which can be prepared as disclosed in U.S. Patent No. 2,980,679; ambuside, which can be prepared as disclosed in U.S. Patent No. 3,188,329; azosemide, which can be prepared as disclosed in U.S. Patent No. 3,665,002; bumetanide, which can be prepared as disclosed in U.S. Patent No. 3,634,583; and British Patent No. 769,757. Butazolamide that can be prepared as disclosed; Chloraminophenamide that can be prepared as disclosed in U.S. Patent Nos. 2,809,194, 2,965,655 and 2,965,656; Clofenamide that can be prepared as disclosed in Olivier, Rec. Trav. Chim., 1918, 37,307; Clopamide that can be prepared as disclosed in U.S. Patent No. 3,459,756; U.S. Patent No. 3,183, Chlorexolone, which can be prepared as disclosed in Patent No. 243; disulfamide, which can be prepared as disclosed in British Patent No. 851,287; etoxolamide, which can be prepared as disclosed in British Patent No. 795,174; furosemide, which can be prepared as disclosed in U.S. Patent No. 3,058,882; mefluside, which can be prepared as disclosed in U.S. Patent No. 3,356,692; U.S. Patent No. 2,783,241 Examples include, but are not limited to, metazolamide, which can be prepared as disclosed in U.S. Patent No. 4,010,273; pyretanide, which can be prepared as disclosed in U.S. Patent No. 4,018,929; tripamide, which can be prepared as disclosed in Japanese Patent No. 7305,585; and xypamide, which can be prepared as disclosed in U.S. Patent No. 3,567,777. All such U.S. patent disclosures are incorporated herein by reference.Other additional agents for combination with the compounds disclosed herein include PCSK9 translation inhibitors, as described in International Publication No. 2016 / 055901, which is incorporated herein by reference.

[0157] In some embodiments, additional therapeutic agents include HMG-CoA reductase inhibitors, HMG-CoA synthase inhibitors, HMG-CoA reductase gene expression inhibitors, HMG-CoA synthase gene expression inhibitors, MTP / Apo B secretion inhibitors, CETP inhibitors, bile acid absorption inhibitors, cholesterol absorption inhibitors, cholesterol synthesis inhibitors, squalene synthase inhibitors, squalene epoxidase inhibitors, squalene cyclase inhibitors, combined squalene epoxidase / squalene cyclase inhibitors, fibrates, niacin, combinations of niacin and lovastatin, ion exchange resins, antioxidants, ACAT inhibitors, bile acid scavengers, and PCSK9 translation inhibitors.

[0158] With regard to the treatment of sepsis or septic shock, the disclosed methods may include additional therapeutic agents such as antibiotics. In certain embodiments, the antibiotics are, but are not limited to, kanamycin, amikacin, tobramycin, dibekacin, gentamicin, shisomecin, netylmycin, streptomycin, and aminoglycosides such as neomycin B, C, and E. Other antibiotics include, but are not limited to, carbapenems such as imipenam, meropenam, ertapenem, doripenam, panipenam, biapenam, raspenem, tebipenam, ranapenam, tomopenam, and chienpenam.

[0159] Further antibiotics include, but are not limited to, ciprofloxacin, garenoxacin, gatifloxacin, gemifloxacin, levofloxacin, cinoxacin, nalidixic acid, moxifloxacin, oxolinic acid, pyromidic acid, pipemidic acid, loxoxacin, enoxacin, freloxacin, lomefloxacin, nadifloxacin, norfloxacin, and Examples of quinolones include floxacin, pefloxacin, rufloxacin, valofloxacin, grepafloxacin, levofloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, clinafloxacin, sitafloxacin, trovafloxacin, prulifloxacin, delafloxacin, JNJ-Q2, nemonoxacin, and zabofloxacin.

[0160] Further antibiotics include, but are not limited to, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, rimecycline, meclocycline, metacycline, minocycline, lolitetracycline, and tetracyclines such as tigecycline.

[0161] Additional antibiotics useful in the methods disclosed herein include, but are not limited to, ampicillin, amoxicillin, augmentin, piperacillin, tazobactam, chloramphenicol, and ticarcillin.

[0162] Pharmaceutical composition The compositions and methods of the present invention may be used to treat subjects that require them. In certain embodiments, the subjects are mammals such as humans or non-human mammals. When administered to subjects such as humans, the compositions or compounds are preferably administered as a pharmaceutical composition comprising, for example, the compounds of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions, such as water or buffered saline or other solvents or vehicles, such as glycols, glycerols, oils such as olive oil, or organic acid esters for injection. In preferred embodiments, when such a pharmaceutical composition is for administration to humans, particularly for administration via an invasive route (i.e., a route such as injection or infusion that avoids transport or diffusion across the epithelial barrier), the aqueous solution is free of or substantially free of pyrogens. Excipients may be selected, for example, to result in delayed release of the drug or to selectively target one or more cells, tissues or organs. The pharmaceutical composition may be in the form of tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilized for reconstitution, powders, liquids, syrups, suppositories, or injections. The composition may also be present in transdermal delivery systems, such as skin patches. The composition may also be present in liquids suitable for topical administration, such as eye drops.

[0163] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent that, for example, stabilizes, increases the solubility of, or increases the absorption of a compound such as the compound of the present invention. Examples of such physiologically acceptable agents include carbohydrates, such as glucose, sucrose, or dextran; antioxidants, such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix, which can, for example, incorporate the compound of the present invention. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively easy to prepare and administer.

[0164] The term "physiologically acceptable" is used herein to mean a compound, material, composition, and / or dosage form that is suitable for use in contact with the target tissue without excessive toxicity, irritation, allergic reaction, or other problems or complications, in proportion to a reasonable risk-benefit ratio, within the bounds of reasonable medical judgment.

[0165] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with other components of the formulation and is not harmful to the subject. Some examples of materials that can function as pharmaceutically acceptable carriers include: (1) sugars, e.g., lactose, glucose, and sucrose; (2) starches, e.g., corn starch and potato starch; (3) cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, e.g., cocoa butter and suppository wax; (9) oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, and corn oil. (10) Glycols, e.g., propylene glycol; (11) Polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; (12) Esters, e.g., ethyl oleate and ethyl laurate; (13) Agar; (14) Buffers, e.g., magnesium hydroxide and aluminum hydroxide; (15) Alginic acid; (16) Water free of pyrogens; (17) Isotonic saline; (18) Ringer's solution; (19) Ethyl alcohol; (20) Phosphate buffer; and (21) Other non-toxic compatible substances used in pharmaceutical formulations.

[0166] Pharmaceutical compositions (formulations) may be administered to a subject by any of several routes of administration, including, for example, orally (e.g., aqueous or non-aqueous liquids or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorbed through the oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., as pessaries, creams, or foams); parenterally (e.g., as sterile liquids or suspensions, including intramuscular, intravenous, subcutaneous, or intrathecal); nasally; intraperitoneally; subcutaneously; percutaneously (e.g., as patches applied to the skin); and topically (e.g., as creams, ointments, or sprays applied to the skin, or as eye drops). Compounds may also be formulated for inhalation. In certain embodiments, compounds may simply be dissolved or suspended in sterile water. Details of suitable administration routes and compositions therefor can be found, for example, in U.S. Patents No. 6,110,973, No. 5,763,493, No. 5,731,000, No. 5,541,231, No. 5,427,798, No. 5,358,970 and No. 4,172,896, and the patents cited herein.

[0167] The formulation may conveniently be provided in unit dosage forms and may be prepared by any method well known in the art of pharmaceuticals. The amount of active ingredient that can be combined with a carrier material to form a single dosage form will vary depending on the subject being treated and the specific method of administration. The amount of active ingredient that can be combined with a carrier material to form a single dosage form is generally the amount of the compound that produces the therapeutic effect. Generally, out of 100 percent, this amount is in the range of about 1 percent to about 99 percent of the active ingredient, preferably about 5 percent to about 70 percent, and most preferably about 10 percent to about 30 percent.

[0168] Methods for preparing these formulations or compositions include the step of combining an active compound, such as the compound of the present invention, with a carrier and, optionally, one or more auxiliary components. Generally, formulations are prepared by uniformly and densely combining the compound of the present invention with a liquid carrier, a pulverized solid carrier, or both, and then, if necessary, shaping the product.

[0169] Formulations of the present invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilized products, powders, granules, or liquids or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions, or as elixirs or syrups, or as lozenges (using an inert base such as gelatin and glycerin, or sucrose and acacia) and / or mouthwashes, each containing a predetermined amount of the compound of the present invention as an active ingredient. The composition or compound may also be administered as a bolus, lick, or paste.

[0170] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, sugar-coated tablets, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable elements, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or bulking agents, e.g., starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, e.g., carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, e.g., glycerol; (4) disintegrants. (1) saturates, e.g., agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (2) solubility retardants, e.g., paraffin; (3) absorption enhancers, e.g., quaternary ammonium compounds; (4) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (5) absorbents, e.g., kaolin and bentonite clay; (6) lubricants, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (7) complexing agents, e.g., modified and unmodified cyclodextrins; and (8) colorants. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also include buffers. Similar types of solid compositions may also be used as fillers in soft and hard-filled gelatin capsules, using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycol, etc.

[0171] Tablets may be prepared by compression or molding with one or more auxiliary components of optional use. Compressed tablets may be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), a surfactant, or a dispersant. Molded tablets may be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0172] Tablets and other solid dosage forms of pharmaceutical compositions, such as sugar-coated tablets, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be slit or prepared with coatings and shells, such as enteric coatings and other coatings well known in the field of pharmaceutical formulation. They may also be formulated to provide sustained or controlled release of the active ingredient therein, for example, using hydroxypropyl methylcellulose in various ratios to provide a desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating a sterilizer in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injection medium immediately before use. These compositions may also optionally contain opacifiers and may be compositions that release the active ingredient in a delayed manner, optionally, only in or preferentially in a specific portion of the gastrointestinal tract. Examples of embedding compositions that may be used include polymer substances and waxes. The active ingredient may also be in a microencapsulated form, using one or more of the excipients described above, as needed.

[0173] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophilized preparations for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrins and their derivatives, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters, as well as mixtures thereof.

[0174] In addition to inert diluents, oral compositions may also contain auxiliary agents such as humectants, emulsifiers and suspending agents, sweeteners, flavoring agents, colorants, fragrances, and preservatives.

[0175] In addition to the active compound, the suspension may contain suspending agents, such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0176] Formulations of pharmaceutical compositions for rectal, vaginal, or urethral administration may be provided as suppositories, which may be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, suppository wax, or salicylate, which are solid at room temperature but liquid at body temperature, and therefore melt in the rectum or vaginal cavity, releasing the active compounds.

[0177] Pharmaceutical compositions for oral administration may be provided as mouthwashes, oral sprays, or oral ointments.

[0178] In addition to or instead of the above, the composition may be formulated for delivery via catheters, stents, wires, or other intracavitary devices. Delivery via such devices may be particularly useful for delivery to the bladder, urethra, ureters, rectum, or intestines.

[0179] Formulations suitable for intravaginal administration also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing a carrier known to be suitable in the art.

[0180] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants as needed.

[0181] In addition to the active compound, ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0182] Powders and sprays may contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures thereof. Sprays may further contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0183] Transdermal patches offer the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0184] Ophthalmic formulations, ophthalmic ointments, powders, and liquids are also assumed to be within the scope of the present invention. Exemplary ophthalmic formulations are described in U.S. Patent Publications 2005 / 0080056, 2005 / 0059744, 2005 / 0031697, and 2005 / 004074, and U.S. Patent No. 6,583,124, the contents of which are incorporated herein by reference. If necessary, liquid ophthalmic formulations have properties similar to or are compatible with such fluids as tears, aqueous humor, or vitreous humor. The preferred route of administration is topical administration (e.g., topical administration such as eye drops, or administration via implant).

[0185] As used herein, the terms “parenteral administration” and “administered parenterally” mean, but are not limited to, methods of administration other than intestinal and local administration (usually by injection), including, but are not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions. Pharmaceutical compositions suitable for parenteral administration include one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into sterile injection solutions or dispersions immediately before use, which may contain antioxidants, buffers, bacteriostatic agents, solutes or suspending agents or thickeners that make the preparation isotonic with the blood of the target recipient.

[0186] Suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, and polyethylene glycol), and suitable mixtures thereof, vegetable oils, such as olive oil, and organic esters for injection, such as ethyl oleate. Appropriate fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0187] These compositions may also contain auxiliary agents such as preservatives, humectants, emulsifiers, and dispersants. Prevention of microbial action can be ensured by the inclusion of various antimicrobial and antifungal agents, such as parabens, chlorobutanol, and phenolsorbic acid. It may also be desirable to include isotonic agents such as sugars and sodium chloride in the composition. Furthermore, long-term absorption of injectable drug forms can be achieved by the inclusion of absorption-delaying agents such as aluminum monostearate and gelatin.

[0188] In some cases, it is desirable to delay the absorption of a drug from subcutaneous or intramuscular injection in order to prolong its effects. This can be achieved by using a liquid suspension of crystalline or amorphous material with low water solubility. In this case, the absorption rate of the drug depends on its dissolution rate, which may further depend on the crystal size and morphology. Alternatively, delayed absorption of a parenterally administered drug form can be achieved by dissolving or suspending the drug in an oil vehicle.

[0189] Depot formulations for injection are prepared by forming a microencapsulation matrix of the compound of the present invention in a biodegradable polymer such as polylactide-polyglycolide. The rate of drug release can be controlled depending on the drug-to-polymer ratio and the properties of the specific polymer used. Examples of other biodegradable polymers include poly(orthoester) and poly(anhydride). Depot injection formulations are also prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

[0190] For use in the method of the present invention, the active compound may be given as itself, or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably 0.5 to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0191] The delivery method may also be provided by refillable or biodegradable devices. Recently, various sustained-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs, including protein-based biologics. Various biocompatible polymers (including hydrogels), including both biodegradable and non-biodegradable polymers, can be used to form implants for the sustained release of compounds at specific target sites.

[0192] The actual dosage level of the active ingredient in a pharmaceutical composition may be varied to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and method of administration without being toxic to the patient.

[0193] The selected dosage level will depend on a variety of factors, including the activity of the specific compound or combination of compounds used, or its ester, salt, or amide; the route of administration; the time of administration; the elimination rate of the specific compound used; the duration of treatment; other drugs, compounds, and / or materials used in combination with the specific compound used; the age, sex, weight, condition, overall health, and medical history of the person being treated; and similar factors well known in the field of medicine.

[0194] A physician or veterinarian with ordinary skill in the art can easily determine and prescribe the therapeutically effective dose of the required pharmaceutical composition. For example, a physician or veterinarian can start the dose of the pharmaceutical composition or compound at a level lower than the level required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. "Therapeutic dose" means a concentration of the compound sufficient to produce the desired therapeutic effect. It is generally understood that the effective dose of a compound varies depending on the subject's weight, sex, age, and medical history. Other factors that may affect the effective dose include, but are not limited to, the severity of the subject's condition, the disorder being treated, the stability of the compound, and, if necessary, another type of therapeutic agent administered with the compound of the present invention. A larger total dose may be delivered by multiple administrations of the drug. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13th ed., 1814-1882, incorporated herein by reference).

[0195] Generally, the preferred daily dose of the active compound used in the compositions and methods of the present invention is the lowest effective dose of the compound that produces a therapeutic effect. Such an effective dose generally depends on the factors described above.

[0196] If necessary, the effective daily dose of the active compound may be administered in one, two, three, four, five, or six or more divided doses, either in unit dosage form, at appropriate intervals throughout the day. In certain embodiments of the present invention, the active compound may be administered two or three times a day. In preferred embodiments, the active compound is administered once a day.

[0197] In certain embodiments, the compounds of the present invention may be used alone or in combination with other types of therapeutic agents. As used herein, the term “combined administration” refers to any form of administration of two or more different therapeutic compounds such that a second compound is administered while a previously administered therapeutic compound is still effective in the body (for example, two compounds are effective simultaneously in the subject, which may include a synergistic effect between the two compounds). For example, different therapeutic compounds may be administered simultaneously or sequentially, either in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds may be administered to each other within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or 1 week. Thus, a subject receiving such treatment may benefit from the combined effect of the different therapeutic compounds.

[0198] In certain embodiments, the combined administration of the compound of the present invention with one or more additional therapeutic agents provides improved efficacy compared to each individual administration of the compound of the present invention (e.g., the compound of formula I or Ia) or one or more additional therapeutic agents. In certain such embodiments, the combined administration provides an additive effect, where the additive effect refers to the sum of the effects of each individual administration of the compound of the present invention and one or more additional therapeutic agents.

[0199] The present invention involves the use of pharmaceutically acceptable salts of the compounds of the present invention in compositions and methods of the present invention. In certain embodiments, the salts envisioned by the present invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. In certain embodiments, the salts envisioned by the present invention include, but are not limited to, L-arginine, benenetamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydravamin, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, the salts envisioned by the present invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

[0200] pharmaceutically acceptable acid addition salts can also exist in a variety of solvents, as well as water, methanol, ethanol, dimethylformamide, and others. Mixtures of such solvates can also be prepared. The source of such solvates may be derived from the crystallization solvent, inherent to the preparation or crystallization solvent, or associated with such solvents.

[0201] Wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavoring agents, and fragrances, preservatives, and antioxidants may also be present in the composition.

[0202] Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, and sodium sulfite; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), lecithin, propyl gallate, and α-tocopherol; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid. [Examples]

[0203] Examples of compounds of formula (I) with useful biological activity, or pharmaceutically acceptable salts thereof, are listed in Tables 1 to 10. 1 ¹H NMR spectra were performed on a Varian MR-400 spectrometer operating at 400 MHz (proton frequency), equipped with a self-shielding Z gradient coil, a 5 mm 1H / nX reverse detection broadband probe head, a deuterium digital lock channel unit, and an orthogonal digital detection unit with transmitter offset frequency shift. Chemical shifts are reported as δ values ​​in ppm relative to trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in Hertz (Hz), and multiplicity is reported using the following abbreviations (s=singlet, d=doublet, t=triplet, q=quadruplet, m=multilet, br=broad, nd=undetermined).

[0204] A. Analysis method Method 1 (Acid FA) UPLC Configuration Solvent: - A 0.1% water containing formic acid (high purity via PureLab optional unit) B. Acetonitrile containing 0.1% (V / V) formic acid (Far UV grade) Column: Acquity UPLC HSS C18 1.8um 100×2.1mm (Plus Guard Cartridge) Flow rate: -0.5mL / min gradient:

[0205] [Table 1]

[0206] Injection 0.5~2uL UV detection via Waters DAD Starting range (nm) 210 Ending range (nm) 400 Resolution (nm) 1.2 MS detection: Waters SQD2, single quadrupole ULC-MS Scan range (m / z) for MS data Start (m / z) 100 End (m / z) 700 or 1500 (if necessary) Includes +ve / -ve switching Ionization is ESI. ESI voltage and temperature are, The power source is 150C 3.5KV, with a 25V capillary cone.

[0207] Method 2 (Basic FA) UPLC Configuration Solvent: Acetonitrile (Far UV grade) Water containing 10 mM ammonium bicarbonate (high purity via PureLab optional unit) Column: Acquity UPLC BEH Shield RP18 1.7um 100×2.1mm (Plus Guard Cartridge) Flow rate: -0.5mL / min Gradient: -A: Water / Basic BMeCN / Basic

[0208] [Table 2]

[0209] Typical injection dose: 0.5–2 μL (concentration approximately 0.2–1 mg / mL). UV detection via Waters DAD Start range (nm) 210 End range (nm) 400 Resolution (nm) 1.2 Other wavelength traces are extracted from the DAD data. MS detection: Waters SQD2, single quadrupole ULC-MS The sample is then sent to mass spectrometry at approximately 300 µl / min using a flow splitter. Scan range (m / z) for MS data Start (m / z)100 End (m / z) 700 or 1500 (if necessary) Includes +ve / -ve switching

[0210] Preparative reverse-phase HPLC conditions Preparative HPLC Waters Micromass ZQ / Sample Manager 2767 Photodiode array detector 2996; Column: XTerra Preparative MS C18 column (5μm, 19×150mm, Waters) Flow rate: 20 mL / min with MS detection UV wavelength: 254nm. Mobile phase: Solvent A (Water:MeCN:HCO2H 95:5:0.05); Solvent B (Water:MeCN:HCO2H 5:95:0.05) gradient:

[0211] [Table 3]

[0212] Flash chromatography is performed using an Isolera MPLC system (manufactured by Biotage) that utilizes pre-filled silica gel or reversed-phase cartridges (supplied by Biotage or Interchim).

[0213] B. Chemical synthesis The general procedure used in the method for preparing the compounds of the present invention is described below. Scheme 1 [ka]

[0214] 2-Chloro-5-(methylthio)pyrimidine (100B) Dimethyl disulfide (13.96 mL, 155.4 mmol, 1.0 equivalent) was added under nitrogen to a -75°C cooled solution of 5-bromo-2-chloropyrimidine (100A) (30 g, 155.4 mmol, 1.0 equivalent) in anhydrous tetrahydrofuran (700 mL). To this mixture, a solution of n-butyllithium (2.5 M, 68.4 mL, 170.9 mmol, 1.0 equivalent) was added dropwise over 1.5 hours, maintaining the internal temperature at -70°C to -75°C throughout the addition. After the addition was complete, the mixture was stirred at -75°C for 4.5 hours, followed by quenching with the slow addition of a saturated solution of ammonium chloride (100 mL). The cold bath was removed, and the reaction mixture was warmed to room temperature under nitrogen for 18 hours. The reaction mixture was diluted with ethyl acetate (500 mL) and washed with water (50 mL), followed by saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The resulting crude pale yellow oil was purified by flash chromatography (eluting at isohexane-ethyl acetate, 0-50%) to obtain the desired product, 2-chloro-5-(methylthio)pyrimidine, as a waxy pale yellow solid (100B). Yield: 7.39g (29%). 1 H NMR(CDCl3)δ 8.48(2H,s),2.54(3H,s);MS(ESI+)m / z 161(M+H) + .

[0215] General method 1 (2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)carbamate tert-butyl(100C) (3-amino-2-methylpropyl)carbamate tert-butyl (4.79 g, 25.52 mmol, 1.05 equivalents) was added to a stirred solution of 2-chloro-5-(methylthio)pyrimidine (100B) (3.89 g, 24.31 mmol, 1.0 equivalent) and cesium carbonate (11.85 g, 36.46 mmol, 1.4 equivalents) in anhydrous dimethylformamide (50 mL). The mixture was stirred at room temperature for 18 hours and then concentrated under reduced pressure to approximately 20 mL. The solution was diluted with ethyl acetate (100 mL), washed with water (75 mL) and brine (50 mL), and then dried over magnesium sulfate. The solvent was removed under vacuum to obtain a crude residue, which was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-50%) to obtain the desired product (2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)carbamate tert-butyl (100C) as a pale yellow oil. Yield: 6.59g (86%) MS(ESI+)m / z 313(M+H) + .

[0216] (S)-(2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl) tert-butyl carbamate (100D) Racemic (2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)carbamate tert-butyl (100C) (5g) was purified by chiral SFC under the following conditions: YMC amylose-C 30 / 70 MeOH / CO2, 100 mL / min, 120 bar, 40°C, GLS 40 psi, System 3900 psi, fall 140 bar, Stacker, DAD 245 nm.

[0217] The first isomer to elute, 1.3 minutes. (R)-(2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl) tert-butyl carbamate Yield: 2.05g. 1H NMR(400MHz,CDCl3)δ 8.34(s,2H),5.79(dd,J=6.7,6.7Hz,1H),5.18(dd,J=5.8,5.8Hz,1H),3.50-3.39(m,1H),3.33-3.14(m, 2H),3.04-2.94(m,1H),2.36(s,3H),1.94-1.85(m,1H),1.45(s,9H),0.95(d,J=6.9Hz,3H).MS(ESI+)m / z 313(M+H) + .

[0218] The second isomer to elute, at 1.7 min: (S)-(2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl) tert-butyl(100D) carbamate Yield: 2.48g. 1 H NMR(400MHz,CDCl3)δ 8.34(s,2H),5.77-5.69(m,1H),5.15(dd,J=6.5,6.5Hz,1H),3.50-3.40(m,1H),3.33-3.16(m,2H), 3.03-2.94(m,1H),2.36(s,3H),1.95-1.85(m,1H),1.47(s,9H),0.95(d,J=6.9Hz,3H);MS(ESI+)m / z 313(M+H) + .

[0219] General method 2 (R)-2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (100E) A solution of hydrogen chloride (15 mL, 4 M in 1,4-dioxane) was added to (S)-(2-ethyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)carbamate tert-butyl (100D) (600 mg, 2.48 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the desired product (R)-2-methyl-N was obtained as a pale yellow semi-solid. 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (100E) was obtained. Yield: 548 mg (100%) HCl salt. 1H NMR(400MHz,DMSO)δ 8.39(s,2H),8.00-7.96(m,3H),7.72-7.72(m,1H),3.32-3.18(m,2H),2.88-2.80(m,1H) ,2.66-2.55(m,1H),2.38(s,3H),2.13-2.00(m,1H),0.96(d,J=6.8Hz,3H);MS(ESI+)m / z 213(M+H) + .

[0220] General method 3 (S)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-carboxylate ethyl(100F) Ethyl-2-chlorobenzothiazole-6-carboxylate (600 mg, 2.48 mmol, 1.0 equivalent) in (R)-2-methyl-N dimethylformamide (20 mL) 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (100E) (548 mg, 2.48 mmol, 1.0 equivalent) and triethylamine (1.73 mL, 12.44 mmol, 5.0 equivalents) were added under nitrogen to a stirred solution. The mixture was stirred at room temperature for 48 hours, and then concentrated under vacuum to approximately 5 mL. Water (25 mL) was added, and the mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic phase was washed with water (20 mL) and brine (25 mL), and then dried over magnesium sulfate. The solvent was removed under vacuum to obtain a crude yellow oil, which was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain the desired (S)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylate ethyl (100F) as a grayish-white solid. Yield: 647 mg (62%). 1H NMR(400MHz,CDCl3)δ 8.37(s,2H),8.27(d,J=1.5Hz,1H),7.99(dd,J=1.8,8.5Hz,1H),7.51(d, J=8.4Hz,1H),7.02-6.98(m,1H),5.83(dd,J=6.7,6.7Hz,1H),4.38(q,J= 7.2Hz,2H),3.67-3.47(m,2H),3.42-3.24(m,2H),2.37(s,3H),2.20-2.1 1(m,1H),1.40(dd,J=7.2,7.2Hz,3H),1.07(d,J=7.0Hz,3H);MS(ESI+)m / z 418(M+H) + .

[0221] Common Method 4 (S)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-carboxylic acid (Example 1) Lithium hydroxide monohydrate (318 mg, 7.75 mmol, 5.0 equivalents) was added to a stirred solution of (S)-2-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid ethyl (100F) (647 mg, 1.55 mmol, 1.0 equivalent) in ethanol (7 mL) and water (5 mL). The mixture was stirred at ambient temperature for 18 hours and then concentrated under reduced pressure. Water (5 mL) was added to the residue, and the mixture was acidified to approximately pH 3 with aqueous hydrochloric acid (2 M). The formed precipitate was collected by filtration, washed with water, and then dried under high vacuum to obtain the desired product (S)-2-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid (Example 1) as a pale yellow solid.

[0222] Common Method 5 (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-carboxamide)methyl propanoate (100G) Hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide (HATU, 146 mg, 0.385 mmol, 1.5 equivalents) was added to a solution of (S)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid (1) (100 mg, 0.257 mmol, 1.0 equivalent), triethylamine (0.36 mL, 2.57 mmol, 10.0 equivalents), and methyl-2-amino-2-methylpropanate hydrochloride (196 mg, 1.28 mmol, 4.9 equivalents) in dimethylformamide (5 mL), and the reaction mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure, and the resulting crude residue was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-75%) to obtain the desired (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamide)methyl propanoate (G) as a grayish-white solid. Yield: 120 mg (95%). 1 1H NMR (400MHz, MeOD) δ 8.34 (s, 2H), 8.10 (d, J=1.8Hz, 1H), 7.77 (dd, J=1.9, 8.5Hz, 1H), 7.47 (d, J=8.5Hz, 1H), 3.74 (s, 3H), 3.45 (dd, J=6.3, 22.4Hz, 4H), 2.34 (s, 3H), 2.29-2.16 (m, 1H), 1.59 (s, 6H), 1.08 (d, J=6.9Hz, 3H) NH exchangeable protons not observed; MS (ESI+) m / z 489 (M+H) + .

[0223] (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamide)propanoic acid (Example 2) A method similar to general method 4 Lithium hydroxide monohydrate (50 mg, 1.22 mmol, 5.0 equivalents) was added to a stirred solution of (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamide)methyl propanoate (100 G) (120 mg, 0.245 mmol, 1.0 equivalent) in ethanol (5 mL) and water (5 mL), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum, the residue was diluted with water (3 mL), and acidified to approximately pH 3 with an aqueous solution of hydrochloric acid (2 M). The formed precipitate was collected by filtration, washed with water, and then dried under high vacuum to obtain the title compound as a grayish-white solid.

[0224] The following examples were synthesized using the procedure described above.

[0225] [Table 4]

[0226] [Table 5]

[0227] [Table 6]

[0228] [Table 7]

[0229] [Table 8]

[0230] [Table 9]

[0231] [Table 10]

[0232] [Table 11]

[0233] [Table 12]

[0234] [Table 13]

[0235] [Table 14]

[0236] Scheme 2 [ka]

[0237] Common Method 6 2-Chloro-N-(4-methoxybenzyl)benzo[d]thiazole-6-sulfonamide (101B) (4-methoxyphenyl)methaneamine (134 mg, 0.97 mmol, 1.05 equivalents) was added dropwise to an ice-cold solution of 2-chlorobenzothiazole-6-sulfonyl chloride (101A) (250 mg, 0.932 mmol, 1.0 equivalent) and triethylamine (0.39 mL, 2.79 mmol, 3.0 equivalents) in tetrahydrofuran (10 mL), and the mixture was stirred at 0°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a solid, washed with ice-cold water (10 mL) and ice-cold tetrahydrofuran (10 mL), and then dried under vacuum to obtain the desired 2-chloro-N-(4-methoxybenzyl)benzo[d]thiazole-6-sulfonamide as a white solid (101B). Yield: 295 mg (85%). 1H NMR(400MHz,DMSO)δ 8.56(d,J=1.6Hz,1H),8.24(s,1H),8.11(d,J=8.7Hz,1H),7.90(dd,J=1.9,8.7Hz,1H) ,7.10(d,J=8.8Hz,2H),6.76(d,J=8.8Hz,2H),3.98(s,2H),3.67(s,3H);MS(ESI+)m / z 369(M+H) + .

[0238] The intermediates in Table 2 were synthesized using the same conditions as those described for intermediate 101B.

[0239] [Table 15]

[0240] [Table 16]

[0241] (S)-N-(4-methoxybenzyl)-2-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-sulfonamide(101C) The methodology applied was the same as that described in General Method 3. Yield: 298 mg (69%). 1 H NMR(400MHz,DMSO)δ 8.56(1H,d,J=1.6Hz),8.24(1H,s),8.11(1H,d,J=8.7Hz),7.90(1H,dd,J=1.9,8.7Hz) ,7.10(2H,d,J=8.8Hz),6.76(2H,d,J=8.8Hz),3.98(2H,s),3.67(3H,s);MS(ESI+)m / z 545(M+H) + .

[0242] (S)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-sulfonamide (Example 87) Trifluoroacetic acid (5 mL) was added dropwise to an ice-cold solution of N-(4-methoxybenzyl)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-sulfonamide (101C) (250 mg, 0.459 mmol, 1.0 equivalent) in anhydrous dichloromethane (5 mL). The mixture was stirred on ice for 30 minutes, followed by heating to ambient temperature over 18 hours. The reaction mixture was diluted with dichloromethane (15 mL), and saturated sodium bicarbonate (15 mL) was slowly added. The organic phase was separated, washed with brine (5 mL), dried over magnesium sulfate, and then concentrated to dryness under reduced pressure. The obtained crude residue was purified by flash chromatography (DCM - methanol, eluting at 0-10%) to obtain the desired (S)-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-sulfonamide (Example 87) as a white solid. Yield: 190 mg (97%). 1 H NMR(400MHz,DMSO)δ 8.40(dd,J=5.5,5.5Hz,1H),8.34(s,2H),8.12(d,J=1.6Hz,1H),7.66(dd,J=2.0,8.4Hz,1H),7.51(dd,J=6.0,6.0Hz,1H),7.44(d,J =8.5Hz,1H),7.21(s,2H),3.49-3.42(m,1H),3.31-3.23(m,3H),2.34(s,3H),2.17-2.07(m,1H),0.96(d,J=6.8Hz,3H);MS(ESI+)m / z 425(M+H) + .

[0243] The following examples were prepared according to General Method 3, using the procedure described in Scheme 2.

[0244] [Table 17]

[0245] [Table 18]

[0246] Scheme 3 [ka]

[0247] 2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (102A) The methodology applied was the same as that of the general method 2. A solution of hydrogen chloride (54 mL, 4 M in 1,4-dioxane) was added to tert-butyl (2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)carbamate (100C) (4.25 g, 13.60 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the desired 2-methyl-N was obtained as a pale yellow semi-solid. 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (102A) was obtained. The semi-crude sample was carried to the next reaction without further purification.

[0248] N 1 -(6-bromobenzo[d]thiazole-2-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine(102B) The methodology applied was the same as that of the general method 3. 6-bromo-2-chlorobenzenethiazole (828 mg, 3.33 mmol, 0.95 equivalents) in 25 mL of anhydrous dimethylformamide (25 mL) 1-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (102A) (1.00 g, 3.51 mmol, 1.0 equivalent) and cesium carbonate (3.43 g, 10.52 mmol, 3.0 equivalents) were added under nitrogen to a stirred suspension. The mixture was stirred at room temperature for 72 hours, then concentrated under vacuum to approximately 3 mL. Water (25 mL) was added, and the mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic phase was washed with water (20 mL) and brine (25 mL), then dried over magnesium sulfate. The solvent was removed under vacuum to obtain a crude yellow oil, which was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain the desired N22A solution as a viscous yellow solid. 1 -(6-bromobenzo[d]thiazole-2-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (102B) was obtained. Yield: 266 mg (18%). 1 H NMR(400MHz,CDCl3)δ 8.37(s,2H),7.66(d,J=0.6Hz,1H),7.37(d,J=1.9Hz,2H),6.58(s,1H),5.69(dd,J=6.3 ,6.3Hz,1H),3.67-3.23(m,4H),2.37(s,3H),2.17-2.09(m,1H),1.06(d,J=6.9Hz,3H).

[0249] Common Method 7 4-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-yl)-1H-pyrazole-1-carboxylate tert-butyl(102C) N 1 -(6-bromobenzo[d]thiazole-2-yl)-2-methyl-N 3A solution of -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (102B) (50 mg, 0.12 mmol, 1.0 equivalent) was added under nitrogen to a solution of (1-(tert-butoxycarbonyl)-1H-pyrazole-4-yl)boronic acid (27.8 mg, 0.13 mmol, 1.0 equivalent), cesium carbonate (58 mg, 0.18 mmol, 1.5 equivalents), and tetrakis(triphenylphosphine)palladium (0) (7 mg, 0.01 mmol, 0.05 equivalents) in water (0.20 mL) and N,N-dimethylformamide (0.80 mL). The reaction mixture was heated to 90°C for 16 hours. An additional fixed amount of (1-(tert-butoxycarbonyl)-1H-pyrazole-4-yl)boronic acid (27.8 mg, 0.13 mmol, 1.0 equivalent) and tetrakis(triphenylphosphine)palladium (0) (7 mg, 0.01 mmol, 0.05 equivalents) were added to the reactants, and the mixture was heated to 90°C under nitrogen for a further 16 hours. The solvent was removed under reduced pressure; water (2 mL) was added, and the mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), passed through a phase separator and dried, and then concentrated under vacuum. The obtained crude residue was purified by flash chromatography (elution at isohexane to ethyl acetate, 0-75%) to obtain the desired 4-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-yl)-1H-pyrazole-1-carboxylate tert-butyl(102C) as a grayish-white solid. The semi-crude product was proceeded to the next reaction without further purification.

[0250] N 1 -(6-(1H-pyrazole-4-yl)benzo[d]thiazole-2-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (Example 97) A solution of hydrogen chloride (2 mL, 4 M in 1,4-dioxane) was added to 4-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-yl)-1H-pyrazole-1-carboxylate tert-butyl (102C) (102C) (100 mg, 0.12 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the desired N2 was obtained as a grayish-white solid. 1 -(6-(1H-pyrazole-4-yl)benzo[d]thiazole-2-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (Example 97) was obtained.

[0251] The following examples were prepared according to General Method 7 using the procedure described in Scheme 3.

[0252] [Table 19]

[0253] General method 8 2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)-N 3 -(6-(oxazol-2-yl)benzo[d]thiazole-2-yl)propan-1,3-diamine (Example 101) N 1 -(6-bromobenzo[d]thiazole-2-yl)-2-methyl-N 3-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (102B) (90 mg, 0.21 mmol, 1.0 equivalent) was added under nitrogen to a solution of 2-(tri-n-butylstannyl)oxazole (0.044 mL, 0.21 mmol, 1.0 equivalent) and tetrakis(triphenylphosphine)palladium (0) (23 mg, 0.02 mmol, 0.1 equivalent) in N,N-dimethylformamide (0.80 mL). The reaction mixture was heated to 90°C for 16 hours. A further fixed amount of 2-(tri-n-butylstannyl)oxazole (0.044 mL, 0.21 mmol, 1.0 equivalent) and tetrakis(triphenylphosphine)palladium (0) (23 mg, 0.02 mmol, 0.1 equivalent) was added to the reaction mixture and heated to 110°C for 18 hours. The reaction mixture was cooled to room temperature, diluted with water (2 mL), and extracted with ethyl acetate (3 × 5 mL). The combined organic phase was washed with water (2 mL) and brine (2 mL), filtered through a Celite pad, dried by passing through a phase separator, and then concentrated to dryness under vacuum. The resulting crude residue was purified twice by flash chromatography (isohexane to ethyl acetate, eluting first at 0-100%, followed by ethyl acetate to methanol, eluting at 0-10%) to obtain a semi-crude residue, which was further purified by reverse-phase preparative HPLC to obtain the desired 2-methyl-N2- 1 -(5-(methylthio)pyrimidine-2-yl)-N 3 -(6-(oxazol-2-yl)benzo[d]thiazole-2-yl)propan-1,3-diamine (101) was obtained.

[0254] The following examples were prepared according to General Method 8 using the procedure described in Scheme 3.

[0255] [Table 20]

[0256] Scheme 4 [ka] The following examples were prepared according to General Method 3, using the procedure described in Scheme 4.

[0257] [Table 21]

[0258] Scheme 7 [ka]

[0259] (2,2-dimethyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl) tert-butyl carbamate (105A) The methodology applied was similar to that described in General Method 1, except that further heating was required. (3-amino-2,2-dimethylpropyl)carbamate tert-butyl (0.13 g, 0.65 mmol, 1.05 equivalents) was added to a stirred suspension of 2-chloro-5-(methylthio)pyrimidine (100B) (0.10 g, 0.62 mmol, 1.0 equivalent) and cesium carbonate (0.24 g, 0.75 mmol, 1.2 equivalents) in anhydrous N,N-dimethylformamide (1.5 mL), and the mixture was stirred at 80°C for 4 hours. The reaction mixture was concentrated under vacuum, diluted with ethyl acetate (20 mL), washed with water (7.5 mL) and brine (5.0 mL), and then passed through a phase separator and dried. The solvent was removed under vacuum to obtain the desired (2,2-dimethyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)carbamate tert-butyl (105A) as a pale yellow oil. Yield: 0.163g (81%). 1 H NMR(400MHz,DMSO)δ 8.37(s,2H),7.24(dd,J=6.6,6.6Hz,1H),6.93(dd,J=6.3,6.3Hz,1H),3.20(d,J=6 .8Hz,2H),2.85(d,J=8.7Hz,2H),2.40(s,3H),1.43(d,J=3.3Hz,9H),0.83(s,6H).

[0260] 2,2-dimethyl-N 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (105B) The methodology applied was the same as that described in General Method 2. A solution of hydrogen chloride (5 mL, 4 M in 1,4-dioxane) was added to tert-butyl (105A) (2,2-dimethyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)carbamate (105A) (0.16 g, 0.50 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the desired 2,2-dimethyl-N was obtained as a pale yellow semi-solid. 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (105B) was obtained. Yield: 0.13g (100%) HCl salt. 1 H NMR(400MHz,DMSO)δ 8.39(s,2H),7.95(s,3H),7.70(s,1H),3.25(d,J=5.5Hz,2H),2.68-2.61(m,2H),2.38(s,3H),0.96(s,6H).

[0261] N1-(benzo[d]oxazol-2-yl)-2,2-dimethyl-N3-(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine (Example 164) The methodology applied was similar to that described in General Method 3, except that cesium carbonate was used as the usual base. 2-chlorobenzoxazole (0.06 mL, 0.54 mmol, 0.1 equivalent) is added to 2,2-dimethyl-N in anhydrous N,N-dimethylformamide (2.0 mL). 1-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (105B) (0.13 g, 0.49 mmol, 1.0 equivalent) and cesium carbonate (0.48 g, 1.48 mmol, 3.0 equivalents) were added under nitrogen to a stirred solution. The mixture was stirred at either 80°C or room temperature for 16 hours, and then concentrated under vacuum. Water (2.5 mL) was added, and the mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL), passed through a phase separator, and dried. The solvent was removed under vacuum to obtain a crude yellow oil, which was purified by preparative HPLC to obtain the desired N2-white solid. 1 -(benzo[d]oxazol-2-yl)-2,2-dimethyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (Example 164) was obtained.

[0262] The following examples were prepared according to General Method 3, using the procedure described in Scheme 7.

[0263] [Table 22]

[0264] [Table 23]

[0265] [Table 24]

[0266] Scheme 8 [ka]

[0267] 3-Chloro-N-(2-chlorobenzo[d]thiazole-6-yl)propan-1-sulfonamide(106B) Sodium hydride (60% dispersion in mineral oil) (326 mg, 8.15 mmol, 3.0 equivalents) was added in small amounts to an ice-cold solution of 2-chlorobenzothiazole-6-amine (500 mg, 2.71 mmol, 1.0 equivalent) in N,N-dimethylformamide (25 mL), and the mixture was stirred under ice for 1 hour. A solution of 3-chloropropane-1-sulfonyl chloride (673 mg, 3.80 mmol, 1.4 equivalents) in N,N-dimethylformamide (3 mL) was added dropwise, and the reaction mixture was then warmed to ambient temperature over 3 hours. The reaction mixture was diluted with saline solution (20 mL) and extracted with ethyl acetate (2 × 25 mL). The combined organic fractions were concentrated under reduced pressure to obtain a pale yellow oil, which was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain the desired product, 3-chloro-N-(2-chlorobenzo[d]thiazole-6-yl)propan-1-sulfonamide 106B, as a grayish-white gummie. Yield: 427 mg (48%). 1 ¹H NMR (400MHz, MeOD) δ 7.87-7.84 (m, 2H), 7.38 (dd, J=2.3, 8.8Hz, 1H), 3.67 (dd, J=6.3, 6.3Hz, 2H), 2.27-2.19 (m, 2H). Note the CH2 protons obscured by MeOD.

[0268] 2-(2-chlorobenzo[d]thiazole-6-yl)isothiazolidine 1,1-dioxide(106C) Sodium hydride (60% dispersion in mineral oil) (98 mg, 2.46 mmol, 2.0 equivalents) was added to an ice-cold solution of 3-chloro-N-(2-chlorobenzo[d]thiazole-6-yl)propan-1-sulfonamide (106B) (400 mg, 1.23 mmol, 1.0 equivalent) in N,N-dimethylformamide (5 mL). The reaction mixture was stirred under ice for 1 hour, followed by careful quenching with the addition of a saturated solution of ammonium chloride (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 20 mL), and the combined organic phase was washed with water (20 mL) and brine (20 mL). The mixture was concentrated under vacuum to obtain a gum-like substance. The crude product was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain the desired product (106C) as a grayish-white gum-like substance. Yield: 220 mg (62%). 1 H NMR(400MHz,CDCl3)δ 7.92(d,J=8.9Hz,1H),7.72(d,J=2.1Hz,1H),7.38(dd,J=2.4,8.9Hz,1H),3 .84(dd,J=6.5,6.5Hz,2H),3.43(dd,J=7.5,7.5Hz,2H),2.63-2.55(m,2H).

[0269] 2-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-yl)isothiazolidine 1,1-dioxide (Example 175) The methodology applied was the same as that described in General Method 3. Yield: 25 mg (15%). 11H NMR (400MHz, DMSO) δ 8.34(s,2H),8.03(dd,J=5.6,5.6Hz,1H),7.57(d,J=2.3Hz,1H),7.51(dd,J=5.9, 5.9Hz,1H),7.35(d,J=8.8Hz,1H),7.14(dd,J=2.4,8.7Hz,1H),3.72(dd,J=6.5,6. 5Hz,2H),3.47(dd,J=7.5,7.5Hz,2H),3.45-3.38(m,1H),3.30-3.22(m,3H),2.44 -2.36(m,2H),2.35(s,3H),2.13-2.06(m,1H),0.95(d,J=6.8Hz,3H);MS(ESI+)m / z 465(M+H) + .

[0270] Scheme 9 [ka]

[0271] N-(2-chlorobenzo[d]thiazole-6-yl)methanesulfonamide (107B) Methanesulfonyl chloride (0.055 mL, 0.706 mmol, 1.3 equivalents) was added dropwise to an ice-cold solution of 2-chlorobenzothiazole-6-amine (100 mg, 0.54 mmol, 1.0 equivalent) and pyridine (0.066 mL, 0.815 mmol, 1.5 equivalents) in anhydrous dichloromethane (5 mL). The mixture was stirred at 0°C for 15 minutes, then warmed to ambient temperature over 1 hour. The reaction mixture was quenched with water (1 mL). The organic phase was removed, and the mixture was concentrated under reduced pressure to obtain a pale yellow oil, which was purified by flash chromatography (eluting at iso-hexane to ethyl acetate, 0-100%) to obtain the desired N-(2-chlorobenzo[d]thiazole-6-yl)methanesulfonamide (107B) as a pale yellow gummie. Yield: 135 mg (94.8%) 11H NMR (400MHz, CDCl3) δ 7.88 (d, J=8.8Hz, 1H), 7.79 (d, J=2.1Hz, 1H), 3.00 (s, 3H). Aromatic H protons are obscured by CDCl3, and no NH exchangeable protons are observed.

[0272] N-(2-chlorobenzo[d]thiazole-6-yl)-N-methylmethanesulfonamide (107C) Sodium hydride (60% dispersion in mineral oil) (31 mg, 0.772 mmol, 1.5 equivalents) was added in small amounts to an ice-cold solution of N-(2-chlorobenzo[d]thiazole-6-yl)methanesulfonamide (107B) (135 mg, 0.515 mmol, 1.0 equivalent) in anhydrous tetrahydrofuran (2 mL). The mixture was stirred at room temperature for 2 hours. Iodomethane (0.048 mL, 0.772 mmol, 1.5 equivalents) was added, and the mixture was stirred at room temperature for a further 2 hours. Water (1 mL) was added, and the solvent was subsequently removed under high vacuum to obtain a pale yellow gummous substance, which was purified by flash chromatography (iso-hexane to ethyl acetate, eluting at 0-100%) to obtain the desired N-(2-chlorobenzo[d]thiazole-6-yl)-N-methylmethanesulfonamide (107C) as a pale yellow gummous substance. Yield: 100 mg (70%) 1 H NMR(400MHz,CDCl3)δ 7.95(d,J=8.8Hz,1H),7.86(d,J=2.1Hz,1H),7.48(dd,J=2.3,8.8Hz,1H),3.39(s,3H),2.88(s,3H).

[0273] N-methyl-N-(2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-yl)methanesulfonamide (Example 176) The methodology applied was the same as that described in General Method 3. Yield: 89 mg (60%) 11H NMR (400MHz, CDCl3) δ 8.38 (s, 2H), 7.62 (d, J=2.0Hz, 1H), 7.51 (d, J=8.7Hz, 1H), 7.24 (d, J=3.2Hz, 1H), 5.74 (dd, J=6.5, 6.5Hz, 1H), 3.34-3.33 (m, 6H), 2.86 (s, 3H), 2.37 (s, 3H), 2.20-2.10 (m, 1H), 1.06 (d, J=6.9Hz, 3H). Not all NH exchangeable protons were observed; MS (ESI+) m / z 453 (M+H) + .

[0274] Scheme 10 [ka]

[0275] 4-(((5-(methylthiopyrimidine-2-yl)amino)methyl)pyrrolidine-2-one(108B) The methodology applied was the same as that described in General Method 1. 5-aminomethyl-pyrrolidine-2-one (108A) (1.0 g, 8.76 mmol, 1.0 equivalent) was added to a stirred solution of 2-chloro-5-(methylthio)pyrimidine (100B) (1.4 g, 8.76 mmol, 1.0 equivalent) and cesium carbonate (8.56 g, 26.28 mmol, 3.0 equivalents) in anhydrous dimethylformamide (10 mL). The mixture was heated to 50°C for 18 hours and then concentrated under reduced pressure. The resulting solution was diluted with ethyl acetate (50 mL), washed with water (10 mL) and brine (10 mL), and then passed through a phase separator and dried. The solvent was removed under reduced pressure to obtain a crude residue, which was purified by trituration in methanol to obtain the desired product 4-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)pyrrolidine-2-one (108B) as a yellow solid. The aqueous phase was concentrated under reduced pressure, combined with the filtrate, and purified by flash chromatography (dichloromethane to methanol, eluting at 0-10%) to obtain the desired product 4-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)pyrrolidine-2-one (108B) as a yellow solid. Both collections were combined and used in the next step. Yield: 0.74g (36%). 1 H NMR(400MHz,CDCl3)δ 8.35(s,2H),5.79-5.76(m,1H),5.48(dd,J=5.5,5.5Hz,1H),3.57-3.50(m,3H),3.20(dd,J=5. 3,9.5Hz,1H),2.89-2.84(m,1H),2.53-2.46(m,1H),2.37(s,3H),2.16(dd,J=6.4,17.1Hz,1H).

[0276] 4-(((5-(methylthiopyrimidine-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate tert-butyl(108C) 4-dimethylaminopyridine (5 mg, 0.04 mmol, 0.1 equivalent) was added to a stirred suspension of 4-(((5-(methylthiopyrimidine-2-yl)amino)methyl)pyrrolidine-2-one (108B) (100 mg, 0.4 mmol, 1.0 equivalent), di-tert-butyl dicarbonate (229 mg, 1.0 mmol, 2.5 equivalents), and triethylamine (0.146 mL, 1.00 mmol, 2.5 equivalents) in dichloromethane (4.2 mL). The mixture was stirred at room temperature for 72 hours, followed by the addition of water (3.0 mL). The mixture was extracted with ethyl acetate (3 × 5.0 mL), the combined organic phase was washed with brine (2.5 mL), and the mixture was passed through a phase separator and dried. The solvent was removed under reduced pressure to obtain a crude residue, which was purified by reverse-phase chromatography (eluting at 0.1% formic acid solution to acetonitrile, 5-100%) to obtain the desired product 4-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate tert-butyl(108C). Yield: 72 mg (53%). 1 H NMR(400MHz,DMSO)δ 8.35(s,2H),7.62(t,J=6.1Hz,1H),3.73(dd,J=7.8,10.4Hz,1H),3.50-3.44(m,1H),3.29( d,J=6.4Hz,2H),2.60-2.55(m,2H),2.36(s,3H),2.28(dd,J=9.6,20.9Hz,1H),1.44(s,9H).

[0277] (4-amino-2-(((5-(methylthiopyrimidine-2-yl)amino)methyl)-4-oxobutyl)carbamate tert-butyl(108D) A solution of ammonium hydroxide (2.2 mL) was added to 4-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate tert-butyl (108C) (72 mg, 0.21 mmol), and the mixture was heated at 80°C for 1.5 hours. The mixture was cooled to room temperature and subsequently extracted with dichloromethane (3 × 5 mL). The organic solvent was passed through a phase separator to dry and then removed under reduced pressure to obtain the desired product (4-amino-2-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)-4-oxobutyl)carbamate tert-butyl (108D). Yield: 61 mg (86%). 1 H NMR(400MHz,DMSO)δ 8.33(s,2H),7.32(s,1H),7.25(dd,J=6.0,6.0Hz,1H),6.81(s,2H),3.24(dd,J=6.2,6.2Hz,2H), 2.97(dd,J=6.0,6.0Hz,2H),2.36(s,3H),2.18-2.08(m,1H),2.04(d,J=6.4Hz,2H),1.38(s,9H).

[0278] 4-Amino-3-(((5-(methylthiopyrimidine-2-yl)amino)methyl)butanamide dihydrochloride (108E) The methodology applied was the same as that described in General Method 2. A solution of hydrogen chloride (0.7 mL, 4 M in 1,4-dioxane) was added to (4-amino-2-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)-4-oxobutyl)carbamate tert-butyl (108D) (61 mg, 0.17 mmol), and the mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure to obtain the desired product, 4-amino-3-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)butanamide dihydrochloride (108E), as a pale yellow solid. Yield: 56 mg (100%) HCl salt.

[0279] 4-((6-(N-(4-methoxybenzyl)sulfamoyl)benzo[d]thiazole-2-yl)amino)-3-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)butanamide(177) The methodology applied was the same as that described in General Method 3. 2-Chloro-N-(4-methoxybenzyl)benzo[d]thiazole-6-sulfonamide (101B) (70 mg, 0.19 mmol, 1.10 equivalents) was added under nitrogen to a stirred solution of 4-amino-3-(((5-(methylthio)pyrimidine-2-yl)amino)methyl)butanamide dihydrochloride (108E) (56 mg, 0.17 mmol, 1.0 equivalent) and triethylamine (0.072 mL, 0.51 mmol, 3.0 equivalents) in anhydrous dimethylformamide (2.0 mL). The mixture was stirred at room temperature for 72 hours and then concentrated under reduced pressure. Water (2.5 mL) was added, and the resulting precipitate was collected by filtration and subsequently washed with methanol. The organic filtrate was concentrated under reduced pressure to obtain a crude residue, which was purified by flash chromatography (eluting at dichloromethane to methanol, 0-25%) to obtain the desired product 4-((6-(N-(4-methoxybenzyl)sulfamoyl)benzo[d]thiazole-2-yl)amino)-3-(((5-(methylthiopyrimidine-2-yl)amino)methyl)butanamide (Example 177) as a grayish-white solid. Yield: 25 mg (25%). 1 H NMR(400MHz,DMSO)δ 8.45(dd,J=5.6,5.6Hz,1H),8.33(s,2H),8.08(d,J=1.8Hz,1H),7.88(dd,J=6.1,6.1Hz,1H ),7.62(dd,J=1.9,8.5Hz,1H),7.49-7.42(m,2H),7.39-7.37(m,1H),7.14(d,J=8.7Hz,2H) ,6.87-6.87(m,1H),6.81(d,J=8.7Hz,2H),3.89(d,J=6.0Hz,2H),3.70(s,3H),3.48(d,J=1 .1Hz,2H),3.37(dd,J=6.1,6.1Hz,2H),2.43-2.37(m,1H),2.34(s,3H),2.22-2.17(m,2H).

[0280] 4-((5-(methylthio)pyrimidine-2-yl)amino)-3-(((6-sulfamoylbenzo[d]thiazole-2-yl)amino)methyl)butanamide(178) Trifluoroacetic acid (0.3 mL) was added dropwise to a solution of 4-((6-(N-(4-methoxybenzyl)sulfamoyl)benzo[d]thiazole-2-yl)amino)-3-(((5-(methylthiopyrimidine-2-yl)amino)methyl)butanamide (177) (20 mg, 0.03 mmol, 1.0 equivalent) cooled to 0°C in anhydrous dichloromethane (0.3 mL). The mixture was stirred for 30 minutes and then warmed to room temperature over 8 hours. A further fixed amount of trifluoroacetic acid (1 mL) was added, and the mixture was stirred for 16 hours. The reaction mixture was concentrated under pressure and then carefully made basic by the addition of saturated aqueous solution of sodium bicarbonate (3 mL). The mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phases were washed with brine (3 mL), dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained crude residue was purified by flash chromatography (dichloromethane to methanol, eluting at 0-20%) to obtain the desired product 4-((5-(methylthio)pyrimidine-2-yl)amino)-3-(((6-sulfamoylbenzo[d]thiazole-2-yl)amino)methyl)butanamide (Example 178) as a white solid. Yield: 12 mg (75%). 1 H NMR(400MHz,DMSO)δ 8.41(dd,J=5.5,5.5Hz,1H),8.33(s,2H),8.13(d,J=1.8Hz,1H),7.67(dd,J=1.9,8.5Hz,1H),7.47-7.43(m,2H),7.38(s,1H), 7.21(s,2H),6.86(s,1H),3.46-3.46(m,2H),3.36(dd,J=6.2,6.2Hz,2H),2.39-2.38(m,1H),2.34(s,3H),2.21-2.17(m,2H).

[0281] The following embodiments were synthesized using the procedure described in Scheme 10.

[0282] [Table 25]

[0283] Scheme 11 [ka]

[0284] 4-((2-chlorobenzo[d]thiazole-6-yl)sulfonyl)morpholine (109B) The methodology applied was similar to that described in General Method 6, except that dichloromethane was used as the solvent instead of tetrahydrofuran. Yield: 686 mg. 1 H NMR(400MHz,CDCl3)δ 8.25(d,J=1.8Hz,1H),8.10(d,J=8.6Hz,1H),7.85(dd,J=1.3,8.6Hz,1H),3.75(dd,J=4.7,4.7Hz,4H),3.04(dd,J=4.7,4.7Hz,4H);MS(ESI+)m / z 319(M+H)+.

[0285] (3-((5-(difluoromethoxy)pyrimidine-2-yl)amino)-2-methylpropyl)carbamate tert-butyl(109D) The methodology applied was the same as that described in General Method 1. Yield: 433 mg. 1 H NMR(400MHz,CDCl3)δ 8.16-8.15(m,2H),5.05-5.05(m,1H),3.47-3.38(m,1H),3.31-3.17(m,2H),3.04- 2.95(m,1H),1.96-1.86(m,1H),1.60-1.58(m,1H),0.96-0.94(m,3H);MS(ESI+)m / z 333(M+H)+.

[0286] N1-(5-(difluoromethoxy)pyrimidine-2-yl)-2-methylpropane-1,3-diamine hydrochloride (109E) A solution of hydrogen chloride (2.7 mL, 4 M in 1,4-dioxane) was added to (3-((5-(difluoromethoxy)pyrimidine-2-yl)amino)-2-methylpropyl)carbamate tert-butyl (109D) (300 mg, 0.903 mmol), and the mixture was stirred at room temperature for 15 minutes. The solvent was removed under vacuum to obtain the crude title compound N1-(5-(difluoromethoxy)pyrimidine-2-yl)-2-methylpropane-1,3-diamine hydrochloride (109E), which was then carried to the next step without further purification. Yield: 225 mg (quantitative). MS(ESI+)m / z 233(M+H)+.

[0287] N1-(5-(difluoromethoxy)pyrimidine-2-yl)-2-methyl-N3-(6-(morpholinosulfonyl)benzo[d]thiazole-2-yl)propan-1,3-diamine(180) The methodology applied was the same as that described in General Method 3. Yield: 225 mg. 1 H NMR(400MHz,CDCl3)δ 8.21(s,2H),7.97(d,J=1.5Hz,1H),7.65(dd,J=1.8,8.5Hz,1H),7.59(d, J=8.5Hz,1H),6.91(s,1H),6.43(t,J=71.6Hz,1H),5.72(dd,J=6.6,6.6H z,1H),3.77-3.73(m,4H),3.62-3.51(m,2H),3.43-3.29(m,2H),3.01(dd ,J=4.6,4.6Hz,4H),2.21-2.12(m,1H),1.08(d,J=6.9Hz,3H);(ESI+)m / z 515(M+H)+.

[0288] The following examples were synthesized according to the procedure described in Scheme 11.

[0289] [Table 26]

[0290] [Table 27]

[0291] Scheme 12 [ka]

[0292] (2-methyl-3-(pyridine-2-ylamino)propyl)carbamate tert-butyl (110B) A solution of tert-butyl 3-amino-2-methylpropylcarbamate (110A) (150 mg, 0.80 mmol, 1.0 equivalent) in 2 mL of 1,4-dioxane was added under nitrogen to a solution of 2-bromopyridine (0.076 mL, 0.80 mmol, 1.0 equivalent), sodium tert-butoxide (383 mg, 3.98 mmol, 5.0 equivalents), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (38 mg, 0.08 mmol, 0.1 equivalent), and tris(dibenzylideneacetone)dipalladium (0) (73 mg, 0.08 mmol, 0.1 equivalent) in 10 mL of 1,4-dioxane. The reaction mixture was heated to 70°C for 72 hours. The solvent was removed under reduced pressure, and the resulting residue was partitioned between water (2 mL) and ethyl acetate (5 mL). The mixture was filtered through Celite, the aqueous phase was removed, and the mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phase was washed with water (2 mL) and brine (2 mL), then dried by passing it through a phase separator, and concentrated to dryness under vacuum. The resulting crude residue was purified by reverse-phase chromatography (eluting at 10 mM aqueous ammonium bicarbonate solution to acetonitrile, 5–95%) to obtain a somewhat pure desired (2-methyl-3-(pyridine-2-ylamino)propyl)carbamate tert-butyl (110B) as a grayish-white solid, which was used in the next step without further purification. Yield: 27 mg (12%). MS(ESI+)m / z 266(M+H) + .

[0293] 2-methyl-N 1 -(pyridine-2-yl)propan-1,3-diamine hydrochloride (110C) The methodology applied was the same as that described in General Method 2. A solution of hydrogen chloride (0.4 mL, 4 M in 1,4-dioxane) was added to tert-butyl (108B) (27 mg, 0.10 mmol) (2-methyl-3-(pyridine-2-ylamino)propyl)carbamate, and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to obtain the desired product, 2-methyl-N1-(pyridine-2-yl)propan-1,3-diamine hydrochloride (110C), as a pale yellow semi-solid. The semi-crude sample was carried to the next reaction without further purification. Yield: 25 mg (assuming quantitative accuracy)

[0294] 2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)-N 3 -(pyridine-2-yl)propan-1,3-diamine (Example 187) The methodology applied was the same as that described in General Method 3. 2-chloro-5-methylsulfanylpyrimidine (100B) (17 mg, 0.11 mmol, 1.05 equivalents) in anhydrous N,N-dimethylformamide (0.5 mL) 1 -(pyridine-2-yl)propan-1,3-diamine hydrochloride (110C) (24 mg, 0.10 mmol, 1.0 equivalent) and cesium carbonate (99 mg, 0.30 mmol, 3.0 equivalents) were added under nitrogen to a stirred solution. The mixture was heated to 50°C for 16 hours and then concentrated under vacuum. Water (2 mL) was added, and the mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL) and dried by passing through a phase separator. The solvent was removed under vacuum to obtain a crude yellow oil, which was purified by reverse-phase chromatography (eluting at 10 mM ammonium bicarbonate aqueous solution ~ acetonitrile, 5-95%) to obtain the desired 2-methyl-N as a viscous yellow solid.1 -(5-(methylthio)pyrimidine-2-yl)-N 3 -(pyridine-2-yl)propan-1,3-diamine (Example 187) was obtained. Yield: 1.5 mg (5%). 1 H NMR(400MHz,CDCl3)δ 8.35(s,2H),8.10(dd,J=0.6,3.6Hz,1H),7.41-7.35(m,1H),6.54(dd,J=5.3,6.8Hz,1H),6.38(d,J=8.4Hz,1H),5.93- 5.93(m,1H),4.96-4.96(m,1H),3.53-3.21(m,4H),2.35(s,3H),2.10-2.01(m,1H),1.03(d,J=6.9Hz,3H);MS(ESI+)m / z 290(M+H) + .

[0295] Scheme 13 [ka]

[0296] 5-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)pyrazine-2-carboxylate methyl(188) The methodology applied was the same as that described in General Method 3. 5-bromopyrazine-2-carboxylate methyl (114 mg, 0.53 mmol, 1.0 equivalent) in anhydrous N,N-dimethylformamide (2.0 mL) 1-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (102A) (150 mg, 0.53 mmol, 1.0 equivalent) and cesium carbonate (514 mg, 1.58 mmol, 3.0 equivalents) were added under nitrogen to a stirred solution. The mixture was stirred at room temperature for 16 hours and then concentrated under vacuum. Water (2.5 mL) was added, and the mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic phases were washed with water (2 mL) and brine (2.5 mL), and then passed through a phase separator and dried. The solvent was removed under vacuum to obtain a brown oil, which was purified by flash chromatography (iso-hexane to ethyl acetate, eluting at 0 to 100%). The obtained semi-crude product was further purified by flash chromatography (dichloromethane to methanol, eluting at 0-10%) to obtain the desired 5-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)amino)pyrazine-2-carboxylate methyl (Example 188) as a white solid. Yield: 60 mg (32%). 1 H NMR(400MHz,CDCl3)δ 8.76(s,1H),8.37(s,2H),7.91(d,J=1.4Hz,1H),6.36-6.36(m,1H),5.56(dd,J=6.0,6.0Hz,1H),3.95(s,3H) ),3.60-3.51(m,2H),3.40-3.26(m,2H),2.38(s,3H),2.10-2.03(m,1H),1.04(d,J=6.9Hz,3H).MS(ESI+)m / z 349(M+H) + .

[0297] 5-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)pyrazine-2-carboxylic acid (189) The methodology applied was the same as that described in General Method 4. Lithium hydroxide monohydrate (23 mg, 55 mmol, 5.0 equivalents) was added to a stirred solution of methyl 5-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)amino)pyrazine-2-carboxylate (188) (38 mg, 0.11 mmol, 1.0 equivalent) in ethanol (0.4 mL) and water (0.4 mL). The mixture was stirred at ambient temperature for 72 hours and then concentrated under reduced pressure. Water (0.5 mL) was added to the residue, and the mixture was acidified to approximately pH 3 with aqueous hydrochloric acid (2 M). The viscous precipitate was collected under filtration, subsequently extracted with ethyl acetate (3 × 3 mL), washed with water (1 mL), and then dried by passing through a phase separator. The solvent was removed under vacuum to obtain the desired 5-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)amino)pyrazine-2-carboxylic acid (Example 189) as a pale yellow solid. Yield: 34 mg (94%); 1 ¹H NMR (400MHz, DMSO) δ 8.52 (s, 1H), 8.36 (s, 2H), 7.99 (s, 2H), 7.65-7.64 (m, 1H), 3.39-3.20 (m, 4H), 2.36 (s, 3H), 2.12-2.03 (m, 1H), 0.94 (d, J=6.8Hz, 3H), one NH proton not observed; MS (ESI+) m / z 335 (M+H) + .

[0298] (4-Hydroxypiperidine-1-yl)(5-((2-Methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyrazine-2-yl)methanone (Example 190) The methodology applied was the same as that described in General Method 5. Hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide (HATU, 58 mg, 0.15 mmol, 1.5 equivalents) was added to a solution of 5-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)amino)pyrazine-2-carboxylic acid (189) (34 mg, 0.10 mmol, 1.0 equivalent) and 4-hydroxypiperidine (103 mg, 1.02 mmol, 10 equivalents) in N,N-dimethylformamide (1 mL), and the reaction mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure, and the resulting crude residue was purified by reverse-phase preparative HPLC to obtain the desired (4-hydroxypiperidine-1-yl)(5-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyrazine-2-yl)methanone (Example 190) as a grayish-white solid. Yield: 22 mg (53%). 1 H NMR(400MHz,CDCl3)δ 8.46(s,1H),8.37(s,2H),7.79(s,1H),5.97-5.97(m,1H),5.71-5.71 (m,1H),4.15-4.15(m,2H),4.01-3.94(m,1H),3.58-3.47(m,2H),3.4 2-3.25(m,4H),2.37(s,3H),2.08(ddd,J=11.5,11.5,5.1Hz,1H),2.00-1.94(m,2H),1.62-1.60(m,3H),1.04(d,J=6.8Hz,3H);MS(ESI+)m / z 418(M+H) + .

[0299] Scheme 16 [ka]

[0300] 5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-amine (111B) The methodology applied was the same as that described in General Method 7. 2-bromo-5-methyl-1,3,4-oxadiazole (0.58 g, 3.58 mmol, 1.05 equivalents) was added under nitrogen to a solution of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-amine (111A) (0.75 g, 3.41 mmol, 1.0 equivalent), cesium carbonate (3.33 g, 10.22 mmol, 3.0 equivalents), and tetrakis(triphenylphosphine)palladium (0) (0.39 g, 0.34 mmol, 0.10 equivalents) in water (5.5 mL) and 1,4-dioxane (22.5 mL). The reaction mixture was heated to 100 °C for 16 hours. The solvent was removed under vacuum, water (20 mL) was added, and the mixture was extracted with ethyl acetate (6 × 50 mL). The combined organic phases were washed with water (10 mL) and brine (10 mL), passed through a phase separator and dried, and then concentrated under vacuum. The resulting crude residue was purified by reverse-phase chromatography (elution at 10 mM ammonium bicarbonate solution to acetonitrile, 5-30%) to obtain the desired 5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-amine (111B) as a grayish-white solid. Yield: 153 mg (25%). 1 H NMR(400MHz,DMSO)δ 8.49(d,J=2.0Hz,1H),7.87(dd,J=2.4,8.8Hz,1H),6.75(s,2H),6.57(d,J=8.8Hz,1H),2.53(s,3H).

[0301] (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)amino)propyl) tert-butyl(111C) carbamate N-(2-methyl-3-oxopropyl)carbamate tert-butyl (111B) (225 mg, 1.21 mmol, 1.2 equivalents) was added to a solution of 5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-amine (114B) (153 mg, 1.00 mmol, 1.0 equivalent), acetic acid (0.230 mL, 4.02 mmol, 4.0 equivalents), and molecular sieves (4 Å type, 250 mg) in anhydrous dichloromethane (20 mL). The reaction mixture was stirred at room temperature for 5 minutes, and then sodium triacetoxyborohydride (532 mg, 2.51 mmol, 2.5 equivalents) was added all at once. The mixture was stirred at room temperature for 40 hours. 225 mg, 1.21 mmol, 1.2 equivalents of N-(2-methyl-3-oxopropyl)carbamate tert-butyl was added, and the reaction mixture was stirred at room temperature for a further 72 hours. The reaction was quenched by the careful addition of a saturated aqueous solution of sodium bicarbonate (30 mL). The mixture was vigorously stirred for 30 minutes, and then the dichloromethane layer was isolated and concentrated under reduced pressure to obtain the gum. The crude product was purified by flash chromatography (eluting at isohexane-ethyl acetate, 0-100%) to obtain the desired (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)amino)propyl)carbamate tert-butyl (111C) as a grayish-white gum. The crude sample was moved to the next reaction without further purification. Yield: 101 mg (29%).

[0302] 2-methyl-N 1 -(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)propan-1,3-diamine hydrochloride (111D) The methodology applied was the same as that described in General Method 2. A solution of hydrogen chloride (1.2 mL, 4 M in 1,4-dioxane) was added to tert-butyl (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)aminopropyl)carbamate (110C) (101 mg, 0.29 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the desired 2-methyl-N was obtained as a grayish-white solid. 1 -(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)propan-1,3-diamine hydrochloride (111D) was obtained. The crude sample was carried to the next reaction without further purification. Yield: 93 mg (assuming quantitative accuracy)

[0303] 2-methyl-N 1 -(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (Example 203) The methodology applied was the same as that described in General Method 3. 2-chloro-5-methylsulfanylpyrimidine (100B, 51 mg, 0.32 mmol, 1.1 equivalents) is mixed with 2-methyl-N in anhydrous N,N-dimethylformamide (2.9 mL). 1-(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)propan-1,3-diamine hydrochloride (111D) (93 mg, 0.29 mmol, 1.0 equivalent) and cesium carbonate (283 mg, 0.87 mmol, 3.0 equivalents) were added under nitrogen to a stirred suspension. The mixture was stirred at room temperature for 16 hours, heated to 40°C for a further 16 hours, and then concentrated under vacuum. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic phase was washed with water (10 mL) and brine (10 mL), and then passed through a phase separator and dried. The solvent was removed under vacuum to obtain a yellow oil, which was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain a crude residue, which was further purified by reverse-phase preparative HPLC to obtain the desired 2-methyl-N as a yellow solid. 1 -(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridine-2-yl)-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (Example 203) was obtained. Yield: 3.5 mg (3%). 1 H NMR(400MHz,CDCl3)δ 8.70(d,J=1.8Hz,1H),8.37(s,2H),8.00(dd,J=2.3,8.8Hz,1H),6.47(d,J=8.1Hz,1H),5.79-5.69(m,2 MS(ESI+)m / z 372(M+H) + .

[0304] Scheme 17 [ka]

[0305] 2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carboxylate ethyl(112B) In the reaction tube, (R)-2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (100E) (544 mg, 2.19 mmol), 2-(6-bromopyridine-3-yl)cyclopropane-1-carboxylate ethyl (650 mg, 2.41 mmol), L-proline (101 mg, 0.875 mmol), potassium phosphate (928 mg, 4.37 mmol), and dimethyl sulfoxide (5 mL) were added. The mixture was aerated with nitrogen for 2 minutes, and then copper(I) iodide (83 mg, 0.437 mmol) was added. The tube was sealed under nitrogen and heated overnight at 90°C. The reaction mixture was cooled, diluted with ethyl acetate (20 mL), passed through a Celite pad, and the filtrate was concentrated under vacuum. The resulting residue was diluted with ethyl acetate (30 mL) and water (20 mL), the aqueous phase was separated, and extracted with ethyl acetate (2 × 30 mL). The combined organic matter was washed with water (2 × 40 mL) and brine (2 × 50 mL), dried with magnesium sulfate, and then concentrated under vacuum to obtain the crude title compound 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carboxylate ethyl (112B) as a light brown oil. (Note: Purification via normal-phase chromatography failed to separate some closely spaced impurities.) Yield: 284 mg. 1 H NMR(400MHz,CDCl3)d 8.36(s,2H),8.21(dd,J=2.1,6.2Hz,1H),7.95(s,1H),7.26-7.19(m,1H),7.12(d,J=8.1Hz,1H ),6.33(d,J=8.6Hz,1H),4.22-4.15(m,2H),3.53-3.46(m,1H),3.39-3.31(m,2H),3.23-3.17( m,1H),2.49(ddd,J=11.8,11.8,11.8Hz,1H),2.33(s,3H),2.12-2.02(m,1H),1.79-1.74(m,1H ),1.55-1.48(m,1H),1.32-1.24(m,3H),1.24-1.17(m,1H),1.02(d,J=6.8Hz,3H);MS(ESI+)m / z 402(M+H)+ .

[0306] 2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carboxylic acid (112C) To a solution of ethyl 2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carboxylate (112B, 274 mg, 0.682 mmol) in ethanol (6 mL) and water (4 mL), lithium hydroxide monohydrate (143 mg, 3.41 mmol) was added, and the resulting mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum, and the resulting residue was diluted with water (8 mL). The solution was adjusted to pH approximately 2 with a 2 M aqueous solution of hydrochloric acid, and then extracted with dichloromethane / methanol (20% methanol in dichloromethane, 2 × 10 mL). The combined organic layers were passed through a phase separator cartridge and concentrated under vacuum to obtain the crude title compound 2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carboxylic acid (112C) as a brown oil, which was used directly without further purification. Yield: 280mg. MS(ESI+)m / z 374(M+H) + .

[0307] 7-(2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane-2-carboxylate tert-butyl(112D) 2,7-Diazaspiro[3.5]nonane-2-carboxylate tert-butyl (65 mg, 0.281 mmol) was added to a solution of 2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carboxylic acid (112C, 70 mg, 0.187 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 86 mg, 0.225 mmol), and triethylamine (0.26 mL, 1.87 mmol) in dimethylformamide (2 mL). The resulting mixture was stirred at room temperature for 18 hours. After completion, the reaction mixture was concentrated under vacuum, and the resulting residue was diluted with ethyl acetate (5 mL) and water (3 mL). The aqueous phase was separated and extracted with ethyl acetate (2 × 5 mL). The combined organic matter was washed with water (5 mL) and brine (2 × 10 mL), dried over magnesium sulfate, and then concentrated under vacuum. The resulting crude residue was purified by column chromatography (eluting with 0-10% methanol in dichloromethane) to obtain the title compound 7-(2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl(112D) as a grayish-white solid. Yield: 69 mg. 1H NMR(400MHz,CDCl3)δ 8.35(s,2H),7.92(d,J=1.5Hz,1H),7.15(dd,J=2.1,8.5Hz,1H),6.34(d,J=8.6Hz,1H),5.96 (dd,J=6.2,6.2Hz,1H),5.00(s,1H),3.73-3.63(m,5H),3.49(s,4H),3.38-3.29(m,2H),3.2 6-3.18(m,2H),2.37-2.30(m,4H),2.10-1.98(m,1H),1.89-1.82(m,1H),1.61-1.53(m,2H), 1.44(s,9H),1.29-1.23(m,2H),1.21-1.14(m,1H),1.02(d,J=6.8Hz,3H);MS(ESI+)m / z(M+H) + .

[0308] (2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropyl)(2,7-diazaspiro[3.5]nonane-7-yl)methanone (Example 204) Trifluoroacetic acid (0.2 mL) was added to a stirred solution of 7-(2-(6-(((R)-2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane-2-carboxylate tert-butyl (112D, 69 mg, 0.119 mmol) in dichloromethane (2 mL), and the mixture was stirred at room temperature for 30 minutes. The solvent was removed under vacuum, and then azeotropically mixed with dichloromethane (3 × 5 mL). The resulting crude residue was dissolved in dimethyl sulfoxide (1.5 mL) and purified by preparative HPLC. The resulting liquid was dried under vacuum and subsequently freeze-dried from an acetonitrile / water mixture to obtain the title compound (2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)amino)pyridine-3-yl)cyclopropyl)(2,7-diazaspiro[3.5]nonane-7-yl)methanone (204) as a fluffy white solid. Yield: 15 mg. 1H NMR(400MHz,CDCl3)δ 8.54(s,1H),8.35(s,2H),7.89-7.86(m,1H),7.16(dd,J=2.0,8.6Hz,1H),6.36(d,J=8.6 Hz,1H),5.98(dd,J=6.9,6.9Hz,1H),5.49-5.49(m,1H),3.76(s,4H),3.51-3.43(m,2H),3 .39-3.30(m,3H),3.22(dd,J=6.6,13.4Hz,2H),2.38-2.31(m,4H),2.11-2.01(m,1H),1.8 9-1.79(m,5H),1.59-1.52(m,1H),1.27-1.14(m,1H),1.03(d,J=6.8Hz,3H);MS(ESI+)m / z 482(M+H) + .

[0309] The following examples were synthesized according to the procedure described in Scheme 17.

[0310] [Table 28]

[0311] Scheme 18 [ka]

[0312] N 1 -(5-bromopyrazine-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine(113B) A solution of crude 2-methyl-N-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine hydrochloride (100E) (340 mg, 1.60 mmol, 1.0 equivalent) in dimethylformamide (8 mL) was added to a suspension of 2,5-dibromopyrazine (457 mg, 1.92 mmol, 1.2 equivalents) and cesium carbonate (1.56 g, 4.80 mmol, 3.0 equivalents) in dimethylformamide (2 mL). The reaction mixture was heated to 90°C for 18 hours. The solvent was removed under reduced pressure, and the resulting residue was partitioned between water (20 mL) and dichloromethane (50 mL). The layers were separated, and the aqueous phase was extracted with dichloromethane (3 × 25 mL). The combined organic phase was washed with brine (25 mL), dried by passing through a phase separator, and then concentrated to dryness under vacuum. The crude residue was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain the compound N, which is a pale yellow gum-like substance. 1 -(5-bromopyrazine-2-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (113B) was obtained. Yield: 230 mg (38%). 1 H NMR(400MHz,CDCl3)δ 8.36(s,2H),8.06(d,J=1.3Hz,1H),7.67(d,J=1.4Hz,1H),5.67-5.51(m,2H),3.55 -3.31(m,3H),3.25-3.12(m,1H),2.37(s,3H),2.04(s,1H),1.03(d,J=6.9Hz,3H).

[0313] N 1 -(5-(2-methoxypyridine-3-yl)pyrazine-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine(209) The methodology applied was similar to that described in General Method 7 (note: 1,4-dioxane was used instead of dimethylformamide). N 1A solution of -(5-bromopyrazine-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (113B) (50 mg, 0.135 mmol, 1.0 equivalent) was added under a nitrogen atmosphere to a solution of 2-methoxy-3-phenylboronic acid (23 mg, 0.149 mmol, 1.1 equivalent), cesium carbonate (132 mg, 0.406 mmol, 3 equivalents), and tetrakis(triphenylphosphine)palladium (0) (15.6 mg, 0.0135 mmol, 0.1 equivalent) in water (1 mL) and 1,4-dioxane (2 mL). The reaction mixture was heated to 80°C for 1 hour. The solvent was removed under reduced pressure, water (2 mL) was added, and the mixture was extracted with ethyl acetate (3 × 15 mL). The combined organic phases were washed with brine (10 mL) and concentrated under vacuum. The resulting crude residue was purified by preparative HPLC to obtain the desired nitrogen as a grayish-white solid. 1 -(5-(2-methoxypyridine-3-yl)pyrazine-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (154) was obtained. Yield: 24 mg (44%). 1 H NMR(400MHz,CDCl3)δ 8.77(d,J=1.4Hz,1H),8.37(s,2H),8.18-8.13(m,2H),7.99(d,J=1.5Hz,1H),7.01(dd,J=4.9,7.4Hz,1H),5.69(dd,J=6.5,6.5Hz,1H),5.55( dd,J=6.1,6.1Hz,1H),4.04(s,3H),3.59-3.48(m,2H),3.42-3.25(m,2H),2.37(s,3H),2.19-2.06(m,1H),1.06(d,J=6.9Hz,3H);MS(ESI+)m / z 398(M+H) + .

[0314] Scheme 19 [ka]

[0315] (2-methyl-3-oxopropyl)carbamate tert-butyl(114B) Caution - Exothermic reaction; Dess-Martin periodinane (2.94 g, 6.94 mmol, 1.3 equivalents) was added in small amounts over 20 minutes to a solution of (3-hydroxy-2-methylpropyl)carbamate tert-butyl (1.0 g, 5.34 mmol, 1.0 equivalent) in dichloromethane (50 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with dichloromethane (25 mL) and washed with 1 M sodium dithionite aqueous solution (2 × 10 mL) and saturated sodium bicarbonate aqueous solution (2 × 10 mL). The organic phase was passed through a phase separator and dried, then concentrated under vacuum to obtain crude (2-methyl-3-oxopropyl)carbamate tert-butyl (114B), which was used immediately in the next step without further purification.

[0316] (3-((5-bromopyridine-2-yl)amino)-2-methylpropyl)carbamate tert-butyl(114C) The methodology applied was similar to that described in Scheme 13 (for the generation of 109D). The crude product was purified by flash chromatography (iso-hexane to ethyl acetate, eluting at 0-100%) to obtain the desired (3-((5-bromopyridine-2-yl)amino)-2-methylpropyl)carbamate tert-butyl(114C) as a pale yellow gum. Yield: 875 mg (47%). (MS(ESI+)m / z 345(M+H) + .

[0317] (3-((2'-Methoxy-[3,3'-bipyridine]-6-yl)amino)-2-methylpropyl)carbamate tert-butyl(114D) The methodology applied was the same as that described in General Method 7. Crude (3-((2'-methoxy-[3,3'-bipyridine]-6-yl)amino)-2-methylpropyl)carbamate tert-butyl(114D) was used immediately in the next step without further purification. Yield: 110mg, MS(ESI+)m / z 373(M+H)+ .

[0318] N1-(2'-Methoxy-[3,3'-bipyridine]-6-yl)-2-methylpropane-1,3-diamine(114E) The methodology applied was the same as that described in General Method 2. Crude N 1 -(2'-methoxy-[3,3'-bipyridine]-6-yl)-2-methylpropane-1,3-diamine (114E) was immediately transferred to the next step without further purification. Crude yield: 75mg MS(ESI+)m / z 273(M+H) + .

[0319] N 1 -(2'-Methoxy-[3,3'-bipyridine]-6-yl)-2-methyl-N3-(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine(210) The methodology applied was the same as that described in General Method 1. The obtained crude residue is purified by preparative HPLC to obtain the desired product as a grayish-white solid, and N 1 -(2'-methoxy-[3,3'-bipyridine]-6-yl)-2-methyl-N3-(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine was obtained (155). Yield: 8 mg (7%). 1 H NMR(400MHz,CDCl3)δ 8.36(s,2H),8.30(d,J=2.1Hz,1H),8.11(dd,J=1.9,5.0Hz,1H),7.67(dd,J=2.4,8 .7Hz,1H),7.57(dd,J=1.9,7.3Hz,1H),6.95(dd,J=5.0,7.3Hz,1H),6.45(d,J=8.4 Hz,1H),5.90(dd,J=6.3,6.3Hz,1H),5.11(dd,J=6.0,6.0Hz,1H),3.97(s,3H),3.5 6-3.23(m,4H),2.36(s,3H),2.13-2.04(m,1H),1.05(d,J=6.9Hz,3H);MS(ESI+)m / z 397(M+H) +.

[0320] Scheme 20 [ka]

[0321] (3-([3,3'-bipyridine]-6-ylamino)-2-methylpropyl)carbamate tert-butyl(115A) The methodology applied was similar to that of scheme 19 (Example 115B) for producing the aldehyde (114B), followed by the use of a methodology similar to that of general method 7. Crude N 1 -([3,3'-bipyridine]-6-yl)-2-methylpropane-1,3-diamine (115A) was immediately transferred to the next step. Yield: 253 mg (63%) 1 H NMR(400MHz,DMSO)δ 8.82(d,J=1.8Hz,1H),8.48(dd,J=1.6,4.8Hz,1H),8.36(d,J=2.1Hz,1H), 8.00-7.96(m,1H),7.76(dd,J=2.6,8.7Hz,1H),7.44-7.40(m,1H),6.90-6 .76(m,2H),6.62-6.59(m,1H),4.10(q,J=5.3Hz,1H),3.19-3.18(m,4H),3 .03-2.94(m,1H),2.88-2.80(m,1H),1.93-1.82(m,1H),1.40-1.38(m,9H).

[0322] N 1 -([3,3'-bipyridine]-6-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine(211) The methodology applied was similar to that described in Scheme 13 (for the generation of 109D). The obtained crude residue was purified by reverse-phase chromatography (elution at 10 mM ammonium bicarbonate aqueous solution to acetonitrile, 5-95%) to obtain the desired product as a grayish-white solid. 1-([3,3'-bipyridine]-6-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine (211) was obtained. Yield: 32 mg (14%). 1 1H NMR (400MHz, CDCl3)δ 8.77(1H,d,J=2.1Hz),8.53(1H,dd,J=1.6,4.8Hz),8.37-8.35(3H,m),7.81-7 .77(1H,m),7.63(1H,dd,J=2.5,8.7Hz),7.33(1H,dd,J=4.6,7.8Hz),6.50(1H, d,J=8.7Hz),5.93(1H,dd,J=6.1,6.1Hz),5.25(1H,dd,J=6.1,6.1Hz),3.56-3. 25(4H,m),2.36(3H,s),2.18-2.00(1H,m),1.06(3H,d,J=6.9Hz);MS(ESI+)m / z 367(M+H) + .

[0323] Scheme 21 [ka]

[0324] 2-Bromo-5-((trimethylsilyl)ethynyl)pyridine(116B) Triethylamine (49 mL, 352.25 mmol, 25 equivalents) was added to 2-bromo-5-iodopyridine (4.0 g, 14.09 mmol, 1.0 equivalent), then ethinyltrimethylsilane (2.9 mL, 21.13 mmol, 1.5 equivalents), copper(I) iodide (270 mg, 1.41 mmol, 0.1 equivalent), and bis(triphenylphosphine)palladium(II) dichloride (99 mg, 0.14 mmol, 0.01 equivalent) were added, and the mixture was stirred at room temperature for 17 hours. The mixture was concentrated under vacuum to obtain a crude gum, which was purified by flash chromatography (eluting at iso-hexane to ethyl acetate, 0-40%) to obtain the intermediate 2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B) as a white solid. Yield: 2.5g (69%). MS(ESI+)m / z 254 / 256(M+H) + .

[0325] 2-Bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazole-5-yl)pyridine(116C) To a solution of 2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B) (250 mg, 0.983 mmol, 1.0 equivalent) in ethanol (20 mL), 1-(azidomethyl)-4-methoxybenzene (177 mg, 1.08 mmol, 1.1 equivalent) was added, and the mixture was stirred at room temperature for 1 hour. A solution of 1 M tetrabutylammonium fluoride in tetrahydrofuran (0.54 mL, 1.08 mmol, 1.1 equivalent) was added, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated under vacuum and subsequently purified by flash chromatography (iso-hexane to ethyl acetate, eluting at 0-100%) to obtain 2-bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazole-5-yl)pyridine (116C) as a grayish-white solid. Yield: 0.21g (61%). MS(ESI+)m / z 347(M+H) + .

[0326] (3-((5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridine-2-yl)amino)-2-methylpropyl) tert-butyl carbamate (116D) To a degassed solution of 2-bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazole-5-yl)pyridine (116C) (135 mg, 0.391 mmol, 1.0 equivalent) in dimethyl sulfoxide (5 mL), tert-butyl (3-amino-2-methylpropyl)carbamate (96 mg, 0.508 mmol, 1.3 equivalents), L-proline (18 mg, 0.156 mmol, 0.4 equivalents), potassium phosphate (1.6.5 mg, 0.078 mmol, 0.2 equivalents), and copper(I) iodide (15 mg, 0.782 mmol, 0.2 equivalents) were added. The mixture was stirred at 90°C for 28 hours, diluted with water (20 mL), and extracted with dichloromethane (3 × 20 mL). The organic phases were combined and concentrated under vacuum. The crude residue was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain 2-bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazole-5-yl)pyridine (116D) as a light brown solid. Yield: 0.080g (45%). MS(ESI+)m / z 453(M+H) + .

[0327] (3-((5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridine-2-yl)amino)-2-methylpropyl) tert-butyl carbamate (116E) The methodology applied was the same as that described in General Method 2. Yield: 25 mg (29%). 1 H NMR(400MHz,CDCl3)δ 8.42(d,J=2.0Hz,1H),8.35(s,2H),7.90(dd,J=2.4,8.7Hz,1H),7.52(s, 1H),6.94-6.90(m,2H),6.44(d,J=9.0Hz,1H),5.94(dd,J=6.1,6.1Hz,1H ),5.49(s,2H),5.09(dd,J=6.3,6.3Hz,1H),3.81(s,4H),3.53-3.22(m,4H),2.35(s,3H),2.10-2.01(m,1H),1.03(d,J=6.9Hz,3H),(NH is not observed).

[0328] N 1 -(5-(1H-1,2,3-triazol-5-yl)pyridine-2-yl)-2-methyl-N3-(4-(methylthio)phenyl)propane-1,3-diamine(212) The methodology applied was the same as that described in General Method 3. Yield: 18 mg (96%). 1 H NMR(400MHz,CDCl3)δ 8.53(d,J=2.1Hz,1H),8.38(s,2H),7.84(s,1H),7.82(dd,J=2.3,8.7Hz,1H),6.45(d,J=8.4Hz,1H),6.17(dd,J=6.3,6.3Hz,1H), 5.31(dd,J=6.0,6.0Hz,1H),3.57-3.26(m,4H),2.36(s,3H),2.19-2.06(m,1H),1.06(d,J=6.8Hz,3H),(NH not observed);MS(ESI+)m / z 457(M+H) + .

[0329] Scheme 22 [ka]

[0330] (2-methyl-3-((5-(pyridine-2-yl)pyrazine-2-yl)aminopropyl)carbamate tert-butyl(117B) To a solution of 5-(pyridine-2-yl)pyrazine-2-amine (200 mg, 1.16 mmol, 1.0 equivalent) in dichloromethane (10 mL), tert-butyl (117B) (221 mg, 1.17 mmol, 1.0 equivalent), 3 Å molecular sieve (750 mg), acetic acid (0.266 mL, 4.65 mmol), and sodium triacetoxyborohydride (616 mg, 2.9 mmol) were added, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with dichloromethane (25 mL) and filtered. The organic phase was washed with water (25 mL) and brine (25 mL), and then passed through a phase separator cartridge. The solvent was removed under vacuum to obtain crude (2-methyl-3-((5-(pyridine-2-yl)pyrazine-2-yl)aminopropyl)carbamate tert-butyl (117B), which was used in the next step without further purification. Yield: 310 mg (78%).

[0331] 2-methyl-N 1 -(5-(pyridine-2-yl)pyrazine-2-yl)propane-1,3-diamine(117C) The methodology applied was the same as that described in General Method 2. Yield: 200 mg (100%).

[0332] 2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)-N3-(5-(pyridine-2-yl)pyrazine-2-yl)propane-1,3-diamine(213) The methodology applied was the same as that described in General Method 1. Yield: 20 mg (10%). 1H NMR(400MHz,CDCl3)δ 9.04(d,J=1.4Hz,1H),8.62-8.61(m,1H),8.37(s,2H),8.10(d,J=8.0Hz,1H),7.94(d,J=1.4Hz,1H),7.77-7.72(m,1H),7.20(dd,J=4.8, MS(ESI+)m / z 368(M+H) + .

[0333] The following examples were synthesized according to the procedure described in Scheme 22.

[0334] [Table 29]

[0335] Scheme 23 [ka]

[0336] 5-(1-methyl-1H-1,2,3-triazole-4-yl)pyrazine-2-amine (118B) The methodology applied was similar to that described in General Method 7 (note: 1,4-dioxane / water was used as the solvent instead of dimethylformamide). Yield: 158mg(73%)m / z 176(M+H) + .

[0337] (2-methyl-3-((5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrazine-2-yl)amino)propyl) tert-butyl(118C) carbamate The methodology applied was similar to that described in Scheme 13 (for the generation of 109D). Crude (2-methyl-3-((5-(1-methyl-1H-1,2,3-triazole-4-yl)pyrazine-2-yl)aminopropyl)carbamate tert-butyl(118C) was immediately moved to the next step. Yield: 101mg(34%)MS(ESI+)m / z 347(M+H) + .

[0338] 2-methyl-N1-(5-(1-methyl-1H-1,2,3-triazole-4-yl)pyrazine-2-yl)propane-1,3-diamine(118D) The methodology applied was the same as that described in General Method 2. Crude 2-methyl-N 1 -(5-(1-methyl-1H-1,2,3-triazole-4-yl)pyrazine-2-yl)propan-1,3-diamine (106D) was immediately moved to the next step. Yield: 71 mg (estimated) MS (ESI+) m / z 367 (M+H) + .

[0339] 2-Methyl-N1-(5-(1-methyl-1H-1,2,3-triazole-4-yl)pyrazine-2-yl)-N3-(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine(218) The methodology applied was the same as that described in General Method 1. Yield: 25 mg (23%). 1 H NMR(400MHz,CDCl3)δ 8.35(s,3H),8.30(s,1H),8.05(dd,J=2.3,8.9Hz,1H),7.67(s,1H),6.56(d,J=9.2Hz,1H),6.11-6.06 (m,1H),4.14(s,3H),2.63(s,4H),2.35(s,3H),2.19-2.10(m,1H),1.07(d,J=6.8Hz,3H);MS(ESI+)m / z 371(M+H) + .

[0340] Scheme 24 [ka]

[0341] N 1 -(5-bromopyrazine-2-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine(119a) The methodology applied was the same as that described in General Method 3. Yield: 230 mg (38%). 1 H NMR(400MHz,CDCl3)δ 8.36(s,2H),8.06(d,J=1.3Hz,1H),7.67(d,J=1.4Hz,1H),5.67-5.51(m,2H),5.30(s,2H) ),3.55-3.31(m,3H),3.25-3.12(m,1H),2.37(s,3H),2.04(s,1H),1.03(d,J=6.9Hz,3H).

[0342] 2-methyl-N 1 -(5-(methylthio)pyrimidine-2-yl)-N 3 -(5-(5-(trifluoromethyl)-1H-pyrazole-4-yl)pyrazine-2-yl)propane-1,3-diamine(219) The methodology applied was similar to that described in General Method 7 (note that 1,4-dioxane / water was used, and no additional fixed amounts of reagent were added). Yield: 18 mg (31%). 1 H NMR(400MHz,DMSO)δ 13.70(s,1H),8.35-8.34(m,2H),8.26(d,J=1.0Hz,1H),8.13-8.12(m,1H),8.00(d,J=1.5Hz,1H),7.55-7.44(m ,1H),7.23-7.18(m,1H),3.35-3.18(m,4H),2.35(s,3H),2.11-2.02(m,1H),0.95(d,J=6.8Hz,3H);MS(ESI+)m / z 426(M+H) + .

[0343] Scheme 25 [ka]

[0344] (2-methyl-3-(pyrazolo[1,5-a]pyrimidine-5-ylamino)propyl)carbamate tert-butyl(120A) A microwave vial containing 5-chloropyrazolo[1,5-c]pyrimidine (500 mg, 3.26 mmol, 1 equivalent) and (3-amino-2-methylpropyl)carbamate tert-butyl (100E) (6.13 g, 32.56 mmol, 10 equivalents) was heated to 140°C for 30 minutes under microwave irradiation. The crude reaction mixture was purified by flash chromatography (isohexane to ethyl acetate, eluting at 0-100%) to obtain (2-methyl-3-(pyrazolo[1,5-a]pyrimidine-5-ylamino)propyl)carbamate tert-butyl (120A). Yield: 700 mg (70%). MS(ESI+)m / z 306(M+H) + .

[0345] 2-methyl-N 1 -(pyrazolo[1,5-a]pyrimidine-5-yl)propane-1,3-diamine(120B) The methodology applied was the same as that described in General Method 2. MS(ESI+)m / z 229(M+H) + .

[0346] (2-methyl-3-(pyrazolo[1,5-a]pyrimidine-5-ylamino)propyl) tert-butyl carbamate (220) The methodology applied was the same as that described in General Method 3. Yield: 20 mg (14%). 1¹H NMR (400MHz, DMSO) δ 7.34 (s, 2H), 7.27 (d, J=7.6Hz, 1H), 6.79 (d, J=1.8Hz, 1H), 5.29 (d, J=7.6Hz, 1H), 5.08 (d, J=1.5Hz, 1H), 2.44-2.37 (m, 4H), 1.36 (s, 3H), 1.20-1.11 (m, 1H), (2xNH not observed); MS (ESI+) m / z 330 (M+H) + .

[0347] Scheme 26 [ka]

[0348] 6-Methyl bromonicotinimide (121B) Sodium methoxide (0.71 g, 13.22 mmol, 1.1 equivalents) was added to an ice-cold solution of 6-bromonicotinonitrile (121A) (2.2 g, 12.02 mmol, 1.0 equivalent) in dioxane / water (20 mL / 20 mL). The mixture was stirred on ice for 30 minutes, then warmed to room temperature. After 1 hour, the mixture was diluted with ethyl acetate (200 mL) and water (200 mL). The organic phase was separated, and the aqueous phase was further extracted with ethyl acetate (50 mL). The organic compounds were combined, and the solvent was removed under vacuum to obtain crude methyl 6-bromonicotinimide (121B), which was used in the next step without further purification. Yield: 2.5g (96%).

[0349] 6-Bromo-N'-methylnicotinimidohydrazide (121C) Methylhydrazine (0.73 mL, 13.96 mmol, 1.2 equivalents) was added to a solution of 6-bromonicotinimidate (121B) (2.5 g, 11.63 mmol, 1.0 equivalent), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to obtain 6-bromo-N'-methylnicotinimidohydrazide (121C), and this was then used. It was used in the next process without further purification. Yield: 2g (75%).

[0350] 2-Bromo-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridine (121D) Formic acid (10 mL, 265 mmol, 30.4 equivalents) was added to 6-bromo-N'-methylnicotinimide hydrazide (121C) (2.0 g, 8.73 mmol, 1.0 equivalent), and the mixture was heated under reflux for 1 hour. The reaction mixture was diluted with water (100 mL) and subsequently extracted with ethyl acetate (3 × 50 mL). The combined organic phase was washed with a saturated aqueous solution of sodium bicarbonate (50 mL), dried over sodium sulfate, and filtered. The solvent was removed under vacuum to obtain a crude brown residue, which was purified by flash chromatography (eluting with 0-10% methanol in dichloromethane / dichloromethane) to obtain 2-bromo-5-(1-methyl-1H-1,2,4-triazole-3-yl)pyridine (121D) as a grayish-white solid. Yield: 500 mg (23%). MS(ESI+)m / z 240(M+H) + .

[0351] (2-methyl-3-((5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridine-2-yl)amino)propyl)carbamate tert-butyl(121E) The methodology applied was similar to that described in General Method 7 (note that 1,4-dioxane / water was used, and no additional fixed amounts of reagent were added). Yield: 125 mg

[0352] 2-methyl-N 1 -(5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridine-2-yl)-N 3 -(5-(methylthio)pyrimidine-2-yl)propan-1,3-diamine(221) The methodology applied was similar to that described in General Method 2 and subsequent General Method 1 for deprotection. Yield: 25 mg (18%). 1H NMR(400MHz,DMSO)δ 8.42(d,J=2.0Hz,1H),8.34(s,2H),8.04(s,1H),7.76(dd,J=2.4,8.8Hz,1H),7.48(dd,J=6.0,6.0Hz,1H),6.82(dd,J=5.8,5.8Hz ,1H),6.57(d,J=8.4Hz,1H),4.16(s,3H),3.32-3.17(m,4H),2.35(s,3H),2.09-1.99(m,1H),0.93(d,J=6.7Hz,3H);MS(ESI+)m / z 371(M+H) + .

[0353] Scheme 27 [ka]

[0354] (3-((5-bromopyridine-2-yl)amino)-2-methylpropyl)carbamate tert-butyl(122B) N-(2-methyl-3-oxopropyl)carbamate tert-butyl (100E) (1.0 g, 5.34 mmol, 1.0 equivalent) was added to a solution of 5-bromopyridine-2-amine (122A) (920 mg, 5.34 mmol, 1.0 equivalent), acetic acid (1.2 mL, 21.36 mmol, 4.0 equivalents), and molecular sieve (4 Å type, 1.0 g) in anhydrous dichloromethane (50 mL). The reaction mixture was stirred at room temperature for 5 minutes, followed by the addition of sodium triacetoxyborohydride (2.83 g, 13.35 mmol, 2.5 equivalents) all at once. The mixture was stirred at room temperature for 5 hours. The reaction was quenched by the careful addition of saturated aqueous solution of sodium bicarbonate (30 mL). The mixture was vigorously stirred for 30 minutes, after which the dichloromethane layer was separated and washed with an aqueous solution of sodium thiosulfate (1 M, 15 mL). The organic phase was isolated and concentrated under reduced pressure to obtain a semi-crude (3-((5-bromopyridine-2-yl)amino)-2-methylpropyl)carbamate tert-butyl (122B) as a light brown gummie, which was used in the next reaction without further purification. Yield: 875 mg (47%).

[0355] (3-((2'-Methoxy-[3,3'-bipyridine]-6-yl)amino)-2-methylpropyl)carbamate tert-butyl(122C) The methodology applied was the same as that described in General Method 7. Yield: 138 mg.

[0356] N 1 -(2'-methoxy-[3,3'-bipyridine]-6-yl)-2-methyl-N 3 -(5-(methylthio)pyrimidine-2-yl)propane-1,3-diamine(222) The methodology applied was similar to that described for General Method 2 and subsequent General Method 1 for deprotection. Yield: 57 mg. 1 H NMR(400MHz,CDCl3)δ 8.36(s,2H),8.23(d,J=2.0Hz,1H),7.68(dd,J=2.3,8.7Hz,1H),7.32-7. 27(m,2H),7.03-6.95(m,2H),6.48(d,J=8.7Hz,1H),5.94-5.90(m,2H),3. 82(s,3H),3.56-3.47(m,1H),3.44-3.35(m,2H),3.28(dd,J=6.6,13.4Hz ,1H),2.35(s,3H),2.18-2.07(m,1H),1.06(d,J=6.9Hz,3H);MS(ESI+)m / z 396(M+H) + .

[0357] Scheme 28 [ka]

[0358] 6'-Chloro-2H-[1,3'-bipyridine]-2-one (123B) 2-amino-5-iodopyridine (123A) (1.12 g, 5.00 mmol, 1.0 equivalent) was combined with 2-hydroxypyridine (582 mg, 6.00 mmol, 1.2 equivalents), potassium carbonate (760 mg, 5.50 mmol, 1.1 equivalents), copper(I) iodide (143 mg, 0.75 mmol, 0.15 equivalents), and 8-hydroxyquinoline (110 mg, 0.75 mmol, 0.15 equivalents) in anhydrous dimethyl sulfonamide (5 mL). The mixture was degassed under a nitrogen stream and heated at 130°C for 21 hours. The reaction mixture was cooled to room temperature and then poured into a mixture of 10% aqueous ammonium hydroxide (100 mL) and ethyl acetate (50 mL). Activated carbon (1 g) was added, the mixture was filtered through a Celite pad, and washed with ethyl acetate (2 × 50 mL). The layers were separated, and the aqueous phase was extracted with ethyl acetate (2 × 50 mL). The combined organic extract was washed with saturated brine (50 mL), dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The crude pale yellow solid was purified by flash chromatography (eluting at dichloromethane to methanol, 0-10%) to obtain the desired product 6'-chloro-2H-[1,3'-bipyridine]-2-one as a grayish-white solid (123B). Yield: 245 mg (26%). 1 H NMR(400MHz,DMSO)δ 7.88(1H,d,J=2.5Hz),7.60(1H,ddd,J=0.7,2.1,6.8Hz),7.49(1H,ddd,J=2.2,6.7,9.1Hz),7.41(1H,dd,J=2.7,8.7Hz),6 .52(1H,dd,J=0.4,8.8Hz),6.45(1H,ddd,J=0.7,1.3,9.2Hz),6.28(1H,ddd,J=6.7,6.7,1.3Hz),6.23(2H,s);MS(ESI+)m / z 188(M+H) + .

[0359] (2-methyl-3-((2-oxo-2H-[1,3'-bipyridine]-6'-yl)aminopropyl)carbamate tert-butyl(123C) The methodology applied was the same as that described in Scheme 19. 6'-chloro-2H-[1,3'-bipyridine]-2-one (123B) was used in excess (245 mg, 1.31 mmol, 1.1 equivalents). A 4 Å molecular sieve was used in the reaction. The crude product was purified by flash chromatography (eluting at dichloromethane-methanol, 0-7%) to obtain the desired product (2-methyl-3-((2-oxo-2H-[1,3'-bipyridine]-6'-yl)amino)propyl)carbamate tert-butyl (123C) as a light brown solid. Yield: 249mg, (58%). MS(ESI+)m / z 359(M+H) + .

[0360] 6'-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)aminopropyl)amino)-2H-[1,3'-bipyridine]-2-one(223) The methodology applied was similar to that described in General Method 2, which uses a (4:1) ratio of 1,4-dioxane to 4M HCl in water, followed by General Method 3, which uses cesium carbonate (3.0 equivalents) in combination with triethylamine (2.0 equivalents). Yield: 152 mg (57% after two processes). 1 H NMR(400MHz,DMSO)δ 8.35(2H,s),7.91(1H,d,J=2.6Hz),7.61(1H,ddd,J=0.6,2.1,6.8Hz),7.52-7.45( 2H,m),7.40(1H,dd,J=2.7,8.9Hz),6.90(1H,dd,J=5.8,5.8Hz),6.57(1H,dd,J=0.5 ,8.9Hz),6.45(1H,ddd,J=0.7,1.2,9.2Hz),6.28(1H,ddd,J=6.7,6.7,1.4Hz),3.3 3-3.14(4H,m),2.35(3H,s),2.10-2.00(1H,m),0.94(3H,d,J=6.8Hz);MS(ESI+)m / z 383(M+H) + .

[0361] The following examples were synthesized using the procedure described in Scheme 28.

[0362] [Table 30]

[0363] [Table 31]

[0364] [Table 32]

[0365] Scheme 29 [ka]

[0366] (2-methyl-3-((5-(2-oxopyrrolidine-1-yl)pyrazine-2-yl)amino)propyl) tert-butyl carbamate (124B) A solution of tert-butyl N-(3-amino-2-methylpropyl)carbamate (100E) (436 mg, 2.20 mmol, 1.1 equivalents) in dimethyl sulfonamide anhydrous (10 mL) was added to 2-bromo-5-(pyrrolidinon-1-yl)pyrazine (124A) (510 mg, 2.00 mmol, 1.0 equivalent), tripotassium phosphate (866 mg, 4.00 mmol, 2 equivalents), L-proline (94 mg, 0.80 mmol, 0.4 equivalents), and copper(I) iodide (76 mg, 0.40 mmol, 0.2 equivalents). The mixture was degassed and maintained under a nitrogen stream and heated at 90°C for 20 hours with stirring. The reaction mixture was allowed to cool to room temperature and then poured into a mixture of water (50 mL) and ethyl acetate (50 mL). The layers were separated, and the aqueous phase was extracted with ethyl acetate (2 × 50 mL). The combined organic extract was washed with saturated brine (50 mL), dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The crude product was purified by flash chromatography (eluting at dichloromethane to methanol, 0-6%) to obtain the desired product (2-methyl-3-((5-(2-oxopyrrolidine-1-yl)pyrazine-2-yl)aminopropyl)carbamate tert-butyl (124B) as a yellow solid. Yield: 375mg, (54%). MS(ESI+)m / z 350(M+H) + .

[0367] 1-(5-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)pyrazine-2-yl)pyrroridine-2-one(233) The methodology applied was similar to that described in General Method 2, followed by General Method 3, which uses a [4:1] ratio of 1,4-dioxane to 4M HCl in water. Yield: 71 mg (18% after two processes). 1H NMR(400MHz,CDCl3)δ 9.02(1H,d,J=1.5Hz),8.35(2H,s),7.64(1H,d,J=1.5Hz),5.63(1H,dd,J=6.2,6.2Hz),5.24(1H,dd,J=5.7,5.7Hz),3.97(2H,t,J=7.1Hz),3.55 -3.46(1H,m),3.45-3.32(2H,m),3.28-3.19(1H,m),2.62(2H,t,J=8.1Hz),2.36(3H,s),2.20-2.02(3H,m),1.03(3H,d,J=6.8Hz);MS(ESI+)m / z 374(M+H) + .

[0368] Scheme 30 [ka]

[0369] 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl(125B) 1,1-Difluoro-2-iodoethane (128 mg, 0.67 mmol, 1.2 equivalents) was added to a suspension of 2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl (125A) (126 mg, 0.56 mmol, 1.0 equivalent) and potassium carbonate (231 mg, 1.67 mmol, 3.0 equivalents) in dimethylformamide (1.0 mL), and the mixture was stirred at 60°C for 2 hours. The reaction mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase was washed with brine, dried by passing it through a phase separator, and then concentrated under reduced pressure to obtain the desired 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl (125B) as a pale yellow oil. Yield: 73 mg (45%). 1 H NMR(400MHz,CDCl3)δ 5.87-5.57(m,1H),3.41-3.30(m,4H),3.14(s,4H),2.89-2.77(m,2H),1.76-1.67(m,4H),1.46(s,9H).

[0370] 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane(125C) Trifluoroacetic acid (1.3 mL) was added dropwise to a solution of 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl (125B) (73 mg, 0.25 mmol, 1.0 equivalent) in dichloromethane (1.3 mL). The mixture was stirred for 30 minutes, then concentrated to dryness under reduced pressure to obtain the desired product 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane (125C) as a pale yellow oil, which was used in the next reaction without further purification.

[0371] The intermediates in Table 17 were synthesized using the same conditions as those described for intermediate 125C.

[0372] [Table 33]

[0373] Scheme 31 [ka]

[0374] 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl(126B) 2,2,2-trifluoroethyl trifluoromethanesulfonic acid (153 mL, 1.06 mmol, 1.2 equivalents) was added to a suspension of 2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl (126A) (200 mg, 0.88 mmol, 1.0 equivalent) and cesium carbonate (862 mg, 2.65 mmol, 3.0 equivalents) in acetonitrile (2.0 mL), and the mixture was stirred at 80°C for 16 hours. The reaction mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phase was washed with brine, passed through a phase separator and dried, and then concentrated under reduced pressure to obtain 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl (126B) as a viscous white solid. The sample was used in the next step without further purification. Yield: 341 mg (quantitative). 1 H NMR(400MHz,CDCl3)δ 3.38-3.30(m,4H),3.19(s,4H),3.01(q,J=9.4Hz,2H),1.73-1.69(m,4H),1.45(s,9H).

[0375] 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane(126C) Trifluoroacetic acid (1.3 mL) was added dropwise to a solution of 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate tert-butyl (126B) (110 mg, 0.357 mmol, 1.0 equivalent) in dichloromethane (1.0 mL). The mixture was stirred for 30 minutes, followed by concentration under reduced pressure to dryness to obtain 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane (126C) as a light brown oil, which was used directly in the next step without further purification, and was assumed to be in 100% yield.

[0376] Scheme 32 [ka]

[0377] 2-methyl-5-oxa-2,8-diazaspiro[3.5]nonane(127B) Lithium aluminum hydride (1.97 mL, 1.97 mmol, 3.0 equivalents, 1 M solution in tetrahydrofuran) was added dropwise under nitrogen to a 0°C cooled solution of tert-butyl 5-oxa-2,8-diazaspiro[3.5]nonane-2-carboxylate (127A) (150 mg, 0.657 mmol, 1.0 equivalent) in tetrahydrofuran (4.0 mL). The reaction mixture was stirred at 0°C for 5 minutes, the cold bath was removed, and the reaction mixture was subsequently heated to 70°C for 16 hours (boiling was observed at 0°C and 35°C). The reaction mixture was cooled to 0°C and quenched by the slow addition of water (0.075 mL), aqueous sodium hydroxide solution (15%, 0.075 mL), and then water (0.22 mL). The cold bath was removed, the reaction mixture was stirred for 15 minutes, magnesium sulfate was added, and the mixture was stirred for a further 40 minutes. The mixture was passed through a phase separator and dried, washed with tetrahydrofuran and diethyl ether, and then concentrated to dryness under reduced pressure to obtain 2-methyl-5-oxa-2,8-diazaspiro[3.5]nonane (127B) as a light brown oil. The sample was then carried to the next reaction without further purification. Yield: 100% is assumed. 1 1H NMR (400MHz, CDCl3): δ 3.63-3.56 (m,2H), 3.47-3.43 (m,2H), 3.00 (s,2H), 2.94-2.86 (m,2H), 2.83-2.79 (m,2H), 2.40 (s,3H). No NH was observed.

[0378] The intermediates in Table 18 were synthesized using the same conditions as those described for intermediate 127B.

[0379] [Table 34]

[0380] Scheme 33 [ka]

[0381] 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate tert-butyl(128B) The method used was similar to that described in General Method 5 for obtaining the desired product 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-6-carboxylic acid tert-butyl (128B). Yield: 111 mg (quantitative). 1 H NMR(400MHz,DMSO)δ 3.87(s,2H),3.70(s,2H),3.23-3.14(m,1H),2.86(s,3H),2.79(s,3H),2.32-2.24(m,4H),1.37-1.36(m,9H).

[0382] N,N-dimethyl-2-azaspiro[3.3]heptane-6-carboxamide(128C) Trifluoroacetic acid (2.1 mL) was added dropwise to a solution of 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate tert-butyl (128B) (110 mg, 0.357 mmol, 1.0 equivalent) in dichloromethane (2.1 mL). The mixture was stirred for 30 minutes, followed by concentration under reduced pressure to dryness to obtain the desired product, N,N-dimethyl-2-azaspiro[3.3]heptane-6-carboxamide (128C), as a brown oil. The sample was then carried to the next reaction without further purification, and a 100% yield was assumed.

[0383] Scheme 34 [ka]

[0384] (S)-5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-carboxylate methyl(129A) The methodology applied was the same as that described in General Method 3. Yield: 134 mg (20%). 1 H NMR(400MHz,CDCl3)δ 8.38(s,2H),8.13(d,J=6.9Hz,1H),7.21(d,J=12.2Hz,1H),7.12(s,1H),5.62(t,J=6.7Hz,1H),3.93(s,3H),3 .64-3.46(m,2H),3.42-3.22(m,2H),2.38-2.38(m,3H),2.18-2.09(m,1H),1.07(d,J=6.9Hz,3H);MS(ESI+)m / z 422(M+H) + .

[0385] (S)-5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)aminopropyl)amino)benzo[d]thiazole-6-carboxylic acid (129B) The methodology applied was the same as that described in General Method 4. Yield: 98 mg (quantitative). 1 1H NMR (400MHz, DMSO) δ 12.82 (s, 1H), 8.57-8.54 (m, 1H), 8.33-8.32 (m, 2H), 8.18 (d, J=7.4Hz, 1H), 7.50 (t, J=6.1Hz, 1H), 7.16 (d, J=12.5Hz, 1H), 2.34 (s, 3H), 2.16-2.06 (m, 1H), 0.97-0.94 (m, 3H). No NH exchangeable protons were observed; MS (ESI+) m / z 408 (M+H) + .

[0386] (S)-6-(5-fluoro-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carbonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate tert-butyl(129C) The methodology applied was the same as that described in General Method 5. Yield: 20 mg (54%). 1H NMR(400MHz,CDCl3)δ 8.27(d,J=1.8Hz,1H),8.14(s,1H),7.99(dd,J=1.7,8.5Hz,1H),7.51(d, J=8.5Hz,1H),7.00-6.97(m,1H),5.47(dd,J=6.7,6.7Hz,1H),4.37(q,J= 7.2Hz,2H),3.63-3.49(m,2H),3.40-3.23(m,2H),2.47(s,3H),2.17-2.0 9(m,1H),1.40(dd,J=7.2,7.2Hz,3H),1.06(d,J=6.9Hz,3H);MS(ESI+)m / z 489(M+H) + .

[0387] (S)-(5-fluoro-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-yl)(2,6-diazaspiro[3.4]octan-6-yl)methanone formate (Example 240) Trifluoroacetic acid (1 mL) was added dropwise to a solution of (S)-6-(5-fluoro-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carbonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate tert-butyl (149) (17 mg, 0.0166 mmol, 1.0 equivalent) cooled to 0°C in anhydrous dichloromethane (1 mL). The mixture was stirred for 30 minutes and then concentrated under reduced pressure. The obtained crude residue was purified by preparative HPLC (with formic acid additive) to obtain the desired product (S)-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-yl)(2,6-diazaspiro[3.4]octan-6-yl)methanone formate (Example 189) as a grayish-white solid. Yield: 9 mg (100%). 11H NMR (400MHz, CDCl3) δ 8.38-8.36 (m,3H), 7.60-7.52 (m,1H), 5.84-5.72 (m,1H), 3.98 (d,J=9.0Hz,1H), 3.84-3.57 (m,6H), 3.50-3.24 (m,4H), 2.86 (s,12H), 2.38-2.37 (m,3H), 2.29-2.13 (m,3H), 1.09-1.04 (m,3H). No NH exchangeable protons were observed; MS (ESI+) m / z 502 (M+H) + .

[0388] The following examples were prepared using the procedure described in Scheme 34.

[0389] [Table 35]

[0390] Scheme 35 [ka]

[0391] 2-((3-((5-mercaptopyrimidine-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide(130A) N,N-dimethyl-2-((2-methyl-3-((5-(methylthiopyrimidine-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamide (compound 10, 75 mg, 0.18 mmol), sodium methanethiolate (126 mg, 1.80 mmol, 10.0 equivalents), and N-methylpyrrolidine (1.5 mL) were added to a microwave vessel under nitrogen, sealed, and subsequently heated to 160°C for 1 hour. LC-MS analysis showed complete conversion to the desired thiol 2-((3-((5-((difluoromethyl)thiopyrimidine-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (130A), which was used directly in the next step without purification.

[0392] 2-((3-((5-((difluoromethyl)thiopyrimidine-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide(244) Potassium hydroxide (201 mg, 3.58 mmol, 20.0 equivalents) and water (0.5 mL) were added to a crude thiol solution of 2-((3-((5-((difluoromethyl)thio)pyrimidine-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide in N-methylpyrrolidine (1.5 mL). The mixture was cooled to -70°C and frozen to solid. Bromodifluoromethyldiethylphosphonate (40 μL, 0.215 mmol, 1.2 equivalents) was added all at once, and the reaction mixture was heated to ambient temperature over 1 hour, followed by a cooling to -70°C. A further fixed amount of bromodifluoromethyldiethylphosphonate (40 μL, 0.215 mmol, 1.2 equivalents) was added, and the mixture was heated to ambient temperature over 1 hour. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic phase was washed with water (20 mL) and brine (25 mL), and then dried over magnesium sulfate. The solvent was removed under reduced pressure to obtain a crude brown oil, which was purified by flash chromatography (eluting at dichloromethane to methanol, 0-10%), followed by preparative HPLC, to obtain the desired 2-((3-((5-((difluoromethyl)thio)pyrimidine-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (244) as a grayish-white solid. Yield: 15 mg (19%). 1 H NMR(400MHz,CDCl3)δ 8.42-8.39(m,2H),7.69(d,J=1.4Hz,1H),7.55-7.52(m,1H),7.36(dd,J=1.8,8.3Hz,1H),6.33(s,1H),6.15(t,J= MS(ESI+)m / z 453(M+H)+ .

[0393] The following examples were prepared according to the methodology described in Scheme 35.

[0394] [Table 36]

[0395] [Table 37]

[0396] Scheme 36 [ka]

[0397] ((1R,3R)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)carbamate rac-tert-butyl(131B) The methodology applied was the same as that described in General Method 1. Yield: 220 mg (44%). 1 H NMR(400MHz,CDCl3)δδ 8.35-8.34(m,4H),5.35(s,1H),5.18-5.15(m,1H),4.84-4.82(m,1H),4.56-4.53(m,1H),4.36(dd,J=6.9,13.8Hz,1H),4.26-4.18(m,1H) ),4.02-3.94(m,2H),2.55-2.47(m,1H),2.36(d,J=1.0Hz,6H),2.30-2.16(m,2H),2.10-1.89(m,4H),1.46-1.43(m,24H).;MS(ESI+)m / z 325(M+H) + .

[0398] rac-(1R,3R)-N 1 -(5-(methylthio)pyrimidine-2-yl)cyclopentan-1,3-diamine hydrochloride (131C) The methodology applied was the same as that described in General Method 2. Yield: 250 mg (quantitative %). MS(ESI+)m / z 225(M+H) + . It was used in the next process without further purification.

[0399] rac-N,N-bis(4-methoxybenzyl)-2-(((trans)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide(131D) The methodology applied was the same as that described in General Method 3. Yield: 52 mg (23%). 1 H NMR(400MHz,CDCl3)δ 8.36(s,2H),8.00(d,J=1.6Hz,1H),7.74(dd,J=1.9,8.5Hz,1H),7.58- 7.55(m,1H),7.01-6.96(m,4H),6.76-6.73(m,4H),5.59-5.54(m,1H),5 .22(d,J=6.9Hz,1H),4.50-4.44(m,1H),4.35-4.30(m,1H),4.25(s,4H) ,3.77-3.76(m,6H),2.37-2.37(m,6H),2.23-2.07(m,2H);MS(ESI+)m / z 677(M+H) + .

[0400] rac-2-(((trans)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide(250) Trifluoroacetic acid (1 mL) was added dropwise to an ice-cold solution of rac-N,N-bis(4-methoxybenzyl)-2-(((trans)-3-((5-(methylthio)pyrimidine-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide (131D) (45 mg, 66.48 mmol, 1.0 equivalent) in anhydrous dichloromethane (1 mL). The mixture was warmed to ambient temperature over 18 hours, followed by concentration to dryness under reduced pressure. The resulting crude residue was purified by preparative HPLC, and the resulting clear fraction was freeze-dried to obtain the desired 2-(((trans)-3-((5-(methylthio)pyrimidine-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide (Example 250) as a white powder. Yield: 23 mg (79%). 1 H NMR(CDCl3)δ 8.36(s,2H),8.17(d,J=1.8Hz,1H),7.84(dd,J=2.0,8.5Hz,1H),7.59-7.52(m,1H),5.34(d,J=6.5Hz,1H) ,4.79(s,2H),4.50-4.43(m,1H),4.34-4.27(m,1H),2.37-2.37(m,6H),2.22-2.07(m,2H).;MS(ESI+)m / z 437(M+H) + .

[0401] Scheme 41 [ka]

[0402] Step 1: Example 301C To a solution of Example 301A (1 g, 3.14 mmol) in DMF (30 mL), Cs2CO3 (2.05 g, 6.28 mmol) and Example 301B (5.6 g, 62.8 mmol) were added. The mixture was heated to 25°C for 1 hour. TLC detected that the starting material had been consumed. The reaction product was filtered, and the filtrate (crude Example 301C) was used in the next step without any further purification.

[0403] Step 2: Example 301D The solution from Example 301C was treated with Boc2O (1.03 g, 4.72 mmol) and stirred at room temperature for 2 hours. Water (100 mL) was added, followed by extraction with EA (50 mL x 2), washing with water and brine, drying over Na2SO4, filtering, and the filtrate was concentrated under reduced pressure. The filtrate was then purified by silica gel chromatography (eluting at petroleum ether / dimethyl = 1 / 5 to 1 / 1) to obtain the desired product (Example 301D, 1.2 g, yield 81%) as a yellow solid. LCMS[M+H] + =573

[0404] Step 3: Example 301E To a solution of Example 301D (1.2 g, 2.54 mmol) in DCM (3 mL), TFA (1 mL) was added at room temperature. After the addition, the reaction mixture was stirred at room temperature for 1 hour. TLC detected that the starting material had been consumed, and the mixture was concentrated to obtain the desired product (Example 301E, 945 mg, yield: 100%) as yellow oil, which was used in the next step without further purification.

[0405] Step 4: Example 301 To a solution of Example 301E (945 mg, 2.54 mmol) in ACN (10 mL), TEA (514 mg, 5.08 mmol) and Example 301F (408 mg, 2.54 mmol) were added at room temperature, followed by incubation at 70°C for 18 hours. TLC detected the consumption of the starting material. The reaction product was concentrated and purified by silica gel chromatography (eluting at petroleum ether / Ã = 3 / 1 to 5 / 3) to obtain the desired product Example 301 (780 mg, yield: 61%) as a white solid. LCMS[M+H] + =497. 1 H NMR(400MHz,DMSO-d6)δ 8.78(s,1H),8.34(s,2H),8.12(d,J=1.6Hz,1H),7.54(dd,J=8.5,1.8Hz,1H),7.48(d,J=8.5Hz,1H),3.94- 3.86(m,1H),3.59(dd,J=16.6,11.9Hz,6H),3.39(dd,J=19.4,5.9Hz,4H),2.85-2.81(m,4H),2.33(s,3H).

[0406] Scheme 42 [ka]

[0407] Step 1: Example 302B A solution of Example 302A (6.77 g, 21 mmol) in THF (200 mL) under an N2 atmosphere was cooled to -65°C. MeMgBr (21 mL, 63.1 mmol, 3 M in THF) was added dropwise, and the mixture was stirred at -65°C for 0.5 hours. The reaction mixture was warmed to room temperature for 2 hours and quenched by adding water (200 mL). After extraction with siRNA (200 mL x 2), the combined organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography (elution at petroleum ether / siRNA = 3 / 1) to obtain the desired product Example 302B (4 g, yield 56%) as a yellow oil. LCMS[M+H] + =339

[0408] Step 2: Example 302C A suspension of Pd / C (400 mg) catalyst was introduced into a reactor to a solution of Example 302B (4 g, 0.012 mmol) in MeOH (40 mL). The vessel was purged with nitrogen, followed by hydrogen, and the reaction mixture was stirred at room temperature for 18 hours. TLC and LCMS detected that the starting material had been consumed. Example 302C (4 g, yield: 100%) was obtained by filtration, concentrated, and used in the next step without any further purification. LCMS[M+H] + =205

[0409] Step 3: Example 302D To a solution of Example 301A (154 mg, 0.48 mmol) in DMF (30 mL), Cs2CO3 (313 mg, 0.96 mmol) and Example 302C (982 mg, 4.8 mmol) were added, and the mixture was stirred at 25°C for 2 hours. TLC detected that the starting material had been consumed. The reaction product was filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel chromatography (eluting at petroleum ether / Â1 = 1 / 1 to 1 / 4) to obtain the desired product Example 302D (500 mg, yield: 100%) as a white solid. LCMS[M+H] + =487

[0410] Step 4: Example 302E To a solution of Example 302D (486 mg, 2 mmol) in DCM (2 mL), TFA (1 mL) was added at room temperature. After the addition, the reaction mixture was stirred at room temperature for 1 hour. TLC detected that the starting material had been consumed, and the mixture was concentrated to obtain the desired product Example 302E (532 mg, yield: 100%) as yellow oil, which was used in the next step without further purification.

[0411] Step 5: Example 302 Example 302E (193 mg, 0.5 mmol), TEA (101 mg, 1 mmol), and Example 301F (81 mg, 0.5 mmol) were dissolved in ACN (5 mL), and the mixture was heated to 60°C for 18 hours. LC-MS detected the formation of TM. Purification by preparative HPLC yielded the desired product Example 302 (15 mg, yield: 6%) as a white solid. LC-MS[M+H] + =511 1 H NMR(400MHz,CDCl3)δ 8.41(s,2H),7.98(s,1H),7.65(s,2H),6.10(s,1H),3.77-3.72(m,4H),3.67-3.52(m,4H),3.04-2.98(m,4H),2.38(s,3H),1.33(s,3H).

[0412] Scheme 43 [ka]

[0413] Step 1: Example 303B To a solution of Example 303A (33 g, 500 mmol) and methyl carbonochloride (49.5 g, 520 mmol) in THF (75 mL), KOH (56.1 g, 1000 mmol) in H2O (50 mL) was added over 30 minutes (maintaining the internal temperature below 40°C). After the addition, the suspension was stirred at room temperature for 16 hours. The suspension was filtered, the filtrate was washed with EtOH (50 mL x 3), and dried under vacuum to obtain the desired product Example 303B (67 g, yield 82.7%) as a white solid. LCMS[M+H] + =163

[0414] Step 2: Example 303C To a solution of Example 303B (4 g, 24.52 mmol) in MeOH (300 mL) and HCl (5 mL, 4.0 M in MeOH), Pd / C (8 g) was added at room temperature. The suspension was stirred at room temperature for 44 hours under 15 psi of H2. LC-MS showed that the starting material was completely consumed and the desired product was detected. The mixture was filtered, and the filtrate was washed with MeOH (30 mL x 3). The filtrate was concentrated under reduced pressure to obtain product Example 303C (3.7 g, yield: 54.4%) as a yellow solid. LC-MS[M+H] + =133.2

[0415] Step 3: Example 303D Solutions of Example 303C (2.18 g, 7.84 mmol), Example 301A (0.5 g, 1.57 mmol), and DIEA (13.35 g, 103.5 mmol) in MeCN (100 mL) were heated to 80°C and stirred for 4 hours. LC-MS showed that the starting materials were completely consumed and the desired product was detected. The reaction mixture Example 303D was used for the next step without further purification. LC-MS[M+H] + =415.5

[0416] Step 4: Example 303E Boc2O (5 g, 22.9 mmol) was added to the solution of Example 303D at room temperature and stirred for 1 hour. LC-MS showed that the starting material had been consumed. The reaction mixture was diluted with DCM (200 mL), washed with water (150 mL x 2) and brine (200 mL x 3), dried over Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel chromatography (eluted at petroleum ether / siRNA = 2 / 1 to 1 / 9) to obtain product Example 303E (680 mg, yield: 76.7% in 2 steps) as a white solid. LC-MS[M+H] + =515.5

[0417] Step 5: Example 303F To a solution of Example 303E (840 mg, 1.63 mmol) in THF (20 mL), CaCl2 (363 mg, 3.27 mmol) and NaBH4 (124 mg, 3.27 mmol) were added at room temperature, and the reaction mixture was stirred for 16 hours. The reaction mixture was poured into a saturated aqueous solution of NH4Cl (100 mL), diluted with HCl (30 mL), and separated. The aqueous layer was extracted with HCl (30 mL x 2), and the combined organic layers were concentrated and purified by silica gel chromatography (eluting at petroleum ether / HCl = 3 / 1 to 1 / 9) to obtain product Example 303F (383 mg, yield: 48.3%) as a white solid. LCMS[M+H] + =487.6

[0418] Step 6: Example 303G To a solution of Example 303F (553 mg, 1.14 mmol) dissolved in DCM (10 mL), HCl / dioxane (4.0 M, 10 mL) was added at room temperature, and the reaction mixture was stirred for 0.5 hours. TLC showed that the starting material had been consumed. The reaction mixture was concentrated under reduced pressure to obtain product Example 303G (563 mg, yield: 100%) as a white solid. LCMS[M+H] + =387.5

[0419] Step 7: Example 303 To a solution of Example 303G (563 mg, 1.14 mmol) and K2CO3 (940 mg, 6.82 mmol) in DMF (10 mL), Example 301A (183 mg, 1.14 mmol) was added at room temperature. The reaction mixture was then heated to 60°C and stirred for 16 hours. TLC showed that the starting materials had been consumed. The mixture was cooled to room temperature, diluted with siRNA (30 mL), and washed with water (20 mL). The aqueous layer was extracted with siRNA (20 mL x 4), the combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (eluted at petroleum ether / siRNA = 3 / 1 to 1 / 9) to obtain the crude product, which was then purified by preparative HPLC to obtain Product Example 303 (75.2 mg, yield: 12.93%) as a pale yellow solid. LCMS[M+H] + =511 1 H NMR(400MHz,CDCl3)δ 8.37(s,2H),7.96(s,1H),7.66-7.58(m,2H),6.82(s,1H),5.87(s,1H),3.83- 3.73(m,5H),3.59-3.44(m,5H),3.01(s,4H),2.38(s,3H),2.14-2.08(m,1H).

[0420] Scheme 44 [ka]

[0421] Step 1: Example 304B Example 304A (23.6 g, 0.12 mol), NaN3 (15.1 g, 0.23 mol), and NH4Cl (7.5 g, 0.14 mol) were suspended in DMF (250 mL), and the resulting mixture was heated to 80°C for 1 hour. After detecting completion of the reaction by TLC, HCl (1 L) was added, and the organic extract was washed with water (200 mL x 5), dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluted at petroleum ether / HCl = 5 / 1) to obtain the desired product Example 304B (29 g, 100% yield) as a white solid. LCMS[M+H] + =247

[0422] Step 2: Example 304C To a solution of Example 304B (29 g, 0.12 mol) in EtOH (400 mL), HCl (22 mL) and Pd / C (1 g) were added, and the heterogeneous mixture was stirred under H2 at room temperature for 18 hours. TLC detected that most of the starting material had been consumed. The mixture was filtered, and the filtrate was concentrated to obtain the desired product Example 304C (21 g, yield: 100%) as a white solid. LCMS[M+H] + =221

[0423] Step 3: Example 304D To a solution of Example 304C (6.6 g, 3 mol) in DMF (50 mL), 2-chloro-5-(methylthio)pyrimidine (2.4 g, 15 mmol) and TEA (7.6 g, 75 mmol) were added. The resulting mixture was stirred at 100°C for 1 hour. TLC detected that the starting material had been consumed. The reaction mixture was concentrated and purified by silica gel chromatography (eluting at petroleum ether / siRNA = 1 / 1 to 1 / 4) to obtain the desired product Example 304D (450 mg, yield: 8%) as a white solid. LCMS[M+H] + =345

[0424] Step 4: Example 304E To a solution of Example 304D (450 mg, 1.3 mmol) in EtOH (5 mL), hydrazine hydrate (131 mg, 2.6 mmol) was added at room temperature. After 3 hours, TLC detected that the starting material had been consumed, at which point the reaction product was concentrated and purified by silica gel chromatography (eluting at DCM / MeOH = 10 / 1) to obtain the desired product Example 304E (120 mg, yield: 43%) as a white solid.

[0425] Step 5: Example 304 To a solution of Example 304E (43 mg, 0.20 mmol) and 2-chloro-N,N-dimethylbenzo[d]thiazole-6-sulfonamide (37 mg, 0.13 mmol) in DMF (1 mL), DBU (40 mg, 0.26 mmol) was added. The resulting mixture was stirred at 50°C for 1 hour. TLC detected that the starting material had been consumed. The reaction mixture was concentrated and purified by preparative TLC to obtain the desired product Example 304 (30 mg, yield: 51%) as a white solid. LCMS[M+H] + =454.9. 1 H NMR(400MHz,CDCl3)δ 8.35(s,2H),7.99(d,J=1.5Hz,1H),7.67(dd,J=8.5,1.7Hz,1H),7.59(d,J=8.5Hz,1H),5.97(s,1H),4.11(dd,J=9 .3,4.8Hz,1H),3.74(dd,J=13.7,4.3Hz,1H),3.66(s,1H),3.58(dd,J=13.6,6.3Hz,2H),2.70(s,6H),2.37(s,3H).

[0426] Scheme 45 [ka]

[0427] Step 1: Example 305 To a solution of Example 304E (30 mg, 0.14 mmol) and 2-chloro-N-methylbenzo[d]thiazole-6-sulfonamide (24.5 mg, 0.09 mmol) in DMF (2 mL), DBU (47 mg, 0.18 mmol) was added. After the addition, the mixture was stirred at 50°C for 1 hour. LC-MS confirmed that the starting material had been consumed. The mixture was cooled to room temperature, diluted with  (5 mL), and washed with brine (5 mL). The organic layer was concentrated and purified by preparative TLC to obtain the desired product Example 305 (23.8 mg, yield: 60%) as a white solid. LC-MS[M+H] + =440.9 1 H NMR(400MHz,MeOD-d4)δ 8.35(s,2H),8.07(s,1H),7.69(dd,J=8.0,1H),7.50(d,J=8.0Hz,1H),4.06(m,0.46H),3.57(m,4H),2.51(s,3H),2.32(s,3H).

[0428] Scheme 46 [ka] Example 306 (30 mg, yield: 50%) was prepared as a white solid in a similar manner starting from Example 306A. LCMS[M+H] + =454.9. 1 H NMR(400MHz,CDCl3)δ 8.35(s,2H),7.98(d,J=1.5Hz,1H),7.66(dd,J=8.5,1.7Hz,1H),7.58(d,J=8.5Hz,1H),6.04(t,J=5.6 Hz,1H),4.11(dd,J=9.3,5.1Hz,1H),3.74-3.64(m,2H),3.61-3.54(m,2H),2.70(s,6H),2.36(s,3H).

[0429] Scheme 47 [ka]

[0430] Step 1: Example 307A To a solution of Example 306E (100 mg, 0.47 mmol) in DMF (0.5 mL), DBU (89 mg, 0.58 mmol) and 2-chlorobenzo[d]thiazole-6-carboxylate methyl (89 mg, 0.39 mmol) were added. The mixture was heated to 50°C for 1 hour. After determining that the starting material had been consumed by TLC, the mixture was concentrated and purified by silica gel chromatography (eluting at petroleum ether / Â5 = 1 / 1 to 1 / 4) to obtain the desired product Example 307A (50 mg, yield: 26%) as a white solid. LCMS[M+H] + =406

[0431] Step 2: Example 307B To a solution of Example 307A (50 mg, 0.12 mmol) in THF (1 mL), LiOH (0.4 mL, 1 M in water) was added, and the resulting mixture was stirred at room temperature for 18 hours. After determining that the starting material had been consumed by TLC, the mixture was concentrated and purified by silica gel chromatography (eluting at DCM / MeOH = 10 / 1) to obtain the desired product Example 307B (5 mg, yield: 11%) as a white solid. LCMS[M+H] + =392

[0432] Step 3: Example 307 A solution of Example 307B (10 mg, 0.025 mmol) was packed with dimethylamine hydrochloride (2.5 mg, 0.031 mmol), TEA (8 mg, 0.075 mmol), and HBTU (14 mg, 0.038 mmol). The resulting mixture was stirred at room temperature for 1 hour. After detecting the consumption of the starting material by TLC, the mixture was concentrated and purified by preparative TLC to obtain the desired product Example 307 (2 mg, yield: 19%) as a white solid. LCMS[M+H] + =419. 1H NMR(400MHz,CDCl3)δ 8.36(s,2H),7.67(s,1H),7.53(d,J=8.2Hz,1H),7.35(dd,J=8.3,1.5Hz,1H),5.91(s,1H),4 .14-4.00(m,1H),3.72-3.64(m,2H),3.53(dd,J=13.8,6.3Hz,2H),3.07(s,6H),2.37(s,3H).

[0433] Scheme 48 [ka]

[0434] Step 2: Example 308A A Schlenk tube equipped with a magnetic stirring bar was packed with TBAI (5 g, 13.6 mmol), cyclopentene (9.25 g, 136 mmol), and O-phthalimide (10 g, 68 mmol) in 250 mL of benzene. After adding a solution of 65% TBHP (18.8 g, 136 mmol), the vial was sealed and the reaction mixture was stirred at 80°C for 12 hours. After cooling to room temperature, the reaction mixture was diluted in ethyl acetate and washed with brine. The aqueous phase was extracted with ethyl acetate. The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (elution at PE / ethyl acetate = 5 / 1) to obtain the desired compound Example 308A (6.5 g) as a white solid. LCMS[M+H] + =214. 1 H NMR(400MHz,CDCl3)δ 2.08-2.17(m,1H),2.32-2.47(m,1H),2.41-2.53(m,1H),2.80-2.88(m,1H),5.33-5.4 6(m,1H),5.61-5.70(m,1H),6.07-6.16(m,1H),7.67-7.75(m,2H),7.80-7.87(m,2H).

[0435] Step 2: Example 308B To a solution of Example 308A (5 g, 23.5 mmol) in THF (25 mL), 50% hydrazine hydrate (3.52 g, 35.2 mmol) in H2O was added. The mixture was stirred at 70°C for 2 hours. Next, the mixture was filtered and concentrated under reduced pressure. Di-tert-butyl dicarbonate (10.2 g, 47 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with PE / ethyl acetate = 5 / 1) to obtain the desired compound Example 308B (550 mg) as a white solid. LCMS[M+H] + =184.

[0436] Step 3: Example 308C To a solution of Example 308B (700 mg, 3.825 mmol) in DCM (5 mL), m-CPBA (790 mg, 4.59 mmol) was added in small increments at 0°C. After addition, the mixture was stirred overnight at room temperature. The resulting mixture was cooled to 0°C, m-chlorobenzoic acid was filtered off, and the mixture was washed with additional cold DCM. The combined filtrate and washings were stirred with 20% NaHSO3 for 30 minutes. The DCM layer was separated, extracted with 3N NaOH (3 × 30 mL) and saturated NaCl (30 mL), and dried over Na2SO4. A white solid remained after evaporation, and it was purified by silica gel column chromatography (eluting with PE / ethyl acetate = 5 / 1) to obtain the desired compound Example 308C (455 mg) as a white solid. LCMS[M+H] + =144. 1 H NMR(400MHz,CDCl3)δ 1.07-1.17(m,1H),1.48(s,9H),1.66-1.76(m,1H),1.89-1.96(m,1H),2.05-2.16(m,1H),3.41-3.50(m,1H),3.53(br s,1H),4.07-4.26(m,1H),4.63-4.77(m,1H).

[0437] Step 4: Example 308D A mixture of Example 308C (445 mg, 2.28 mmol), NaN3 (297 mg, 4.57 mmol), NH4Cl (61 mg, 1.14 mmol), 2-methoxyethanol (5 mL), and H2O (1 mL) was stirred for 16 hours in a bath maintained at 80°C. The resulting solution was evaporated to dryness, and the residue was dissolved in H2O (5 mL). This solution was saturated with NaCl and subsequently extracted with DCM (4 × 5 mL). The DCM solution was evaporated, and the residue was purified by silica gel column chromatography (elution occurs at MeOH / DCM = 3%~5%) to obtain the desired compound Example 308D (420 mg) as a colorless oil. LCMS[M+H] + = 188.

[0438] Step 5: Example 308E A suspension of Example 308D (420 mg, 1.74 mmol) and Pd / C (catalyst amount) in EtOH (5 mL) was stirred at room temperature under an H2 atmosphere for 16 hours. The mixture was filtered and concentrated under vacuum. The residue was dried and used directly in the next step without further purification. Example 308E (320 mg). LCMS[M+H] + =217.

[0439] Step 6: Example 308F A mixture of Example 308E (150 mg, 0.694 mmol), 2-chloro-5-(methylthio)pyrimidine (111 mg, 0.694 mmol), and DIPEA (180 mg, 1.4 mmol) in DMSO (5 mL) was stirred at 130°C for 3 hours. The resulting solution was cooled to room temperature, poured into water, and extracted with RINKAN (3 × 10 mL). The combined organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting at PE / EA = 2 / 1) to obtain the desired compound Example 308F (100 mg) as a colorless oil. LCMS[M+H] + =341. 1H NMR(400MHz,CDCl3)δ 1.45(s,9H),1.75-1.85(m,1H),2.13-2.21(m,1H),2.23-2.30(m,1H),2.38(s,3H),3.83-4.05(m,3H),5.33(br s,1H),5.57(br s,1H),8.35(s,2H).

[0440] Step 7: Example 308G To a solution of Example 308F (100 mg, 0.294 mmol) in DCM (3 mL), 4 M HCl (3 mL) in dioxane was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, dried, and used directly in the next step without further purification. Example 308G (70.6 mg). LCMS[M+H] + =241.

[0441] Step 8: Example 308H A mixture of Example 308G (70.6 mg, 0.256 mmol), 6-((2-chlorobenzo[d]thiazole-6-yl)sulfonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate tert-butyl (130 mg, 0.294 mmol), and DIPEA (99 mg, 0.768 mmol) in DMF (4 mL) was stirred at 40°C for 2 days. The resulting solution was cooled to room temperature, poured into water, and extracted with siRNA (3 × 10 mL). The combined organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting at MeOH / DCM = 5%) to obtain the desired compound Example 308H (70 mg) as a white solid. LCMS[M+H] + =648.

[0442] Step 9: Example 308 To a solution of Example 308H (70 mg, 0.108 mmol) in DCM (3 mL), 4 M HCl (3 mL) in dioxane was added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and purified by preparative HPLC to obtain the title compound Example 308 (20 mg) as a white solid. LCMS[M+H] + = 548. 1H NMR(400MHz,DMSO-d6)δ 1.47-1.56(m,1H),1.63-1.73(m,1H),1.97(t,J=6.98Hz,2H),2.06-2.23(m,2H),2.31- 2.41(m,3H),3.19(t,J=6.98Hz,2H),3.35(s,2H),3.68(t,J=6.04Hz,4H),3.95-4.02(m ,1H),4.15(br.s.,1H),4.38(br.s.,1H),7.48(d,J=8.33Hz,1H),7.59(dd,J=8.33,1.8 8Hz,2H),8.16(d,J=1.88Hz,1H),8.37(s,2H),8.49(d,J=7.79Hz,2H),8.66(br.s.,1H).

[0443] スキーム49

change

[0444] Project 1: Implementation Example 309A Sodium cyclopentadienyl (2M in THF, 50 mL, 100 mmol, 1 equivalent) was added dropwise to a solution of benzyl chloromethyl ether (90%, 23 g, 130 mmol, 1.3 equivalents) in DMF (200 mL) at -40°C. After vigorous stirring for 20 minutes at -40°C, the reaction mixture was poured into a 2:1 mixture of pentane / ice-cold water (900 mL). After shaking to separate the phases, the organic layer was washed twice with 150 mL of cold water and dried over Na₂SO₄ with stirring, maintaining the temperature below 0°C to avoid double bond isomerization. After removal of the drying agent by filtration, pentane was removed under vacuum at 0°C to obtain (benzyloxymethyl)cyclopenta-2,4-ene 1 as a pale orange oil. The obtained crude material was maintained at 0°C under argon, diluted with THF (160 mL), cooled to -78°C, and added dropwise via cannula at -78°C to a suspension of (-)-Ipc2BH (100 mL, 100 mmol, 1 equivalent, 1 M solution in THF) in THF (400 mL). The mixture was slowly warmed to -10°C and stirred at that temperature for 3 days. Next, the reaction was quenched by adding MeOH (40 mL), followed by the addition of a 3 M aqueous solution of NaOH (40 mL) and 30% H2O2 (40 mL). After vigorous stirring at room temperature for 24 hours, the THF was removed under reduced pressure, and the remaining aqueous suspension was partitioned between SiO (400 mL) and brine (200 mL). After extraction, the organic layer was dried over Na2SO4 and concentrated under vacuum. The crude orange oil was purified by column chromatography (eluent: 9:1-8:2 heptane: siRNA) to obtain Example 309A (4.8 g, 23.53 mmol, 23%) as a pale yellow oil. Rf=0.29 (eluent: 7:3 heptane:EtOAc); 1H NMR (400MHz, CDCl3)δ 2.23-2.32(m,1H),2.35(s,1H),2.63-2.73(m,1H),2.81-2.88(m,1H),3.28(t,J=8.9Hz,1H),3.53(dd,J=5.4, 9.1Hz,1H),4.29(td,J=4.1,7.0Hz,1H),4.52(s,2H),5.53-5.58(m,1H),5.70-5.74(m,1H),7.24-7.37(m,5H).

[0445] Step 2: Example 309B To a solution of Example 309A (4.8 g, 23.53 mmol, 1 equivalent) in anhydrous THF (100 mL), NaH (50% of mineral oil, 1.13 g, 28.2 mmol, 1.2 equivalents) was added at 0°C, and the mixture was stirred at room temperature for 20 minutes. Next, benzyl bromide (BnBr, 3.6 mL, 30.5 mmol, 1.3 equivalents) and tetrabutylammonium iodide (TBAI, 100 mg, 0.3 mmol, 0.01 equivalents) were added at 0°C, and the reaction mixture was stirred at room temperature. After 15 hours, crushed ice was carefully added, and the mixture was stirred for 30 minutes. After extraction with RINKAN (150 mL), the organic layer was washed with H2O (150 mL) and brine (150 mL), dried over Na2SO4, and concentrated under vacuum. Purification by column chromatography (eluent: 98:2-95:5 heptane:¼) yielded Example 309B (6.3 g, 21.4 mmol, 80%) as a colorless syrup. Rf = 0.41 (eluent: 9:1 heptane:¼). LCMS[M+H] + =295.1H NMR(500MHz,CDCl3)δ 2.42(d,J=17.4Hz,1H),2.65-2.70(m,1H),3.07(brs,1H),3.33 and 3.44(ABX,JAB=9.2Hz,JAX=5.7Hz,JBX=7.3Hz,2H),4.08(ddd,J=3.0,3.3,7.0Hz,1H),4.5 1(d,J=3.4Hz,2H),4.54(s,2H),5.64-5.66(m,1H),5.74-5.75(m,1H),7.22-7.34(m,10H).

[0446] Step 3: Example 309C A 0.5 M solution of 9-BBN (88 mL, 44 mmol) in THF was added dropwise to a solution of Example 309B (6.50 g, 22.0 mmol) in anhydrous THF (10 mL) under nitrogen at 0°C. The reaction mixture was slowly warmed to room temperature overnight. The reaction mixture was cooled to 0°C and sequentially treated with EtOH (7 mL), 3N NaOH solution (20 mL), and H2O2 (33%, 20 mL). The resulting mixture was stirred overnight at room temperature. The resulting residue was filtered and washed with RINKAN (200 mL). Water (150 mL) was added to this suspension, and after phase separation, the aqueous layer was extracted with RINKAN (3 × 50 mL). The combined organic layers were dried (Na2SO4) and concentrated to dryness. The crude product was purified on silica gel (eluent:1:1 heptane:Â) to obtain Example 309C (6.0 g, 19.3 mmol, 87%) as a yellow oil. LCMS[M+H] + =313.1H NMR(400MHz,CDCl3)δ 7.35-7.25(m,10H,CH-arom.),4.52(s,2H,CH2-benzyl),4.49(d,1H,J=11.8Hz,CHH-benzyl),4.44(d,1 H,J=11.8Hz,CHH-benzyl),4.33-4.28(m,1H,H-1),4.07(ddd,1H,J=6.6Hz,6.6Hz,4.1Hz,H-3),3.53(d d,1H,J=9.0Hz,4.2Hz,OCHH),3.49(d,1H,J=9.0Hz,4.3Hz,OCHH),2.35-2.25(m,2H,H-4,H-5a),2.05 (dddd,1H,J=13.5Hz,6.7Hz,3.5Hz,1.7Hz,H-2a),1.89-1.82(m,1H,H-2b),1.52-1.46(m,1H,H-5b).

[0447] Step 4: Example 309D Compound Example 309C (6.0 g, 19.3 mmol) was dissolved in dry pyridine (30 mL) and cooled to 0°C. TosCl (5.5 g, 28.9 mmol) was added in small amounts over 30 minutes. After the addition, the reaction mixture was stirred at room temperature for 18 hours. The suspension was diluted with ethyl acetate (300 mL) and H2O (200 mL). The organic phase was separated, washed with saturated NH4Cl solution (3 × 200 mL) and brine (100 mL), and dried over MgSO4. The solvent was evaporated, and the residue was purified by flash chromatography on silica gel (hexane / ethyl acetate 5:1) to obtain 4-methylbenzenesulfonic acid (1R,3S,4R)-3-(benzyloxy)-4-((benzyloxy)methyl)cyclopentyl (1.4 g, 3.0 mmol, 15%) as a colorless oil. Rf = 0.26 (20% ethyl acetate in hexane). LCMS[M+H] + =467.1H NMR(400MHz,CDCl3)δ7.79(m,2H),7.38-7.27(m,12H),5.05-4.99(m,1H),4.50(s,2H),4.45(s,2H),3.96-3.9 2(m,1H),3.48-3.40(td,2H,J=6.3Hz),2.45(s,3H),2.33-2.20(m,2H),2.14-2.01(m,2H),1.69-1.62(m,1H).

[0448] Compound 4-methylbenzenesulfonic acid (1R,3S,4R)-3-(benzyloxy)-4-((benzyloxy)methyl)cyclopentyl (1.4 g, 3.0 mmol) was dissolved in dry DMF (20 mL), and NaN3 (2.1 g, 15.4 mmol) was added. The mixture was stirred at 60°C for 14 hours. After the addition of ethyl acetate (300 mL), the organic layer was washed with saturated NaHCO3 solution (2 × 100 mL) and brine (100 mL), and dried over MgSO4. The solvent was removed under reduced pressure. The residue was purified by flash chromatography on silica gel (hexane / ethyl acetate 10:1) to obtain Example 309D (1.0 g, 2.95 mmol, 95%) as a colorless oil. Rf = 0.54 (20% ethyl acetate in hexane). LCMS[M+H] + =338. 1H NMR(400MHz,CDCl3)δ 7.39-7.25(m,10H),4.52(dd,J=26.0,18.6Hz,2H),4.49(s,2H),3.97-3.89(m,1H),3.86(td,J=7.0,5.1Hz,1H),3.44(d,J=5.5Hz,2H) ,2.54-2.43(m,1H),2.24(td,J=13.8,6.8Hz,1H),1.99(dddd,J=13.4,8.7,4.7,1.3Hz,1H),1.84(dddd,J=28.2,20.8,9.5,4.2Hz,2H).

[0449] Step 5: Example 309E Compound Example 309D (1.0 g, 2.95 mmol) was dissolved in dry CH2Cl2 (20 mL) and cooled to -78°C. A solution of 1 M BCl3 (40 mL) in CH2Cl2 was added over 45 minutes using a dropping funnel, and the mixture was stirred at -78°C for 3 hours, followed by warming to room temperature. The reaction product was quenched with dry MeOH (20 mL) at -78°C and warmed overnight to room temperature. The solvent was evaporated, and the residue was purified on silica gel (hexane / ethyl acetate 1:1) to obtain (1S,2R,4S)-4-azido-2-(hydroxymethyl)cyclopentanol (420 mg, 2.67 mmol, 90%) as a yellow oil. LCMS[M+H] + = 158. 1 H NMR(400MHz,CDCl3)δ 4.07(dd,J=13.3,6.0Hz,1H),4.02-3.95(m,1H),3.79(ddd,J=10.4,5.2,3.0Hz,1H),3.56(dd,J=10.4,8.0Hz,1H), 2.36-2.20(m,4H),1.99-1.92(m,1H),1.77(dddd,J=14.0,6.0,4.5,1.6Hz,1H),1.58(ddd,J=13.9,9.8,6.8Hz,1H).

[0450] The compound (1S,2R,4S)-4-azido-2-(hydroxymethyl)cyclopentanol (0.42 g, 2.67 mmol) was dissolved in dry DMF (10 mL), and imidazole (198 mg, 2.94 mmol) was added at room temperature. After the addition of small amounts of TBDPSCl (808 mg, 2.94 mmol) at 0 °C, the mixture was stirred at room temperature for 16 hours. The reaction product was diluted with CH2Cl2 (200 mL), washed once with saturated NH4Cl (70 mL) and brine (50 mL), and dried over MgSO4. The solvent was removed under vacuum, and the crude product was purified with silica gel (hexane / ethyl acetate 10:1) to obtain Example 309E (630 mg, 1.6 mmol, 60%) as a yellow oil. LCMS[M+H] + =360. 1 H NMR(400MHz,CDCl3)δ 7.69-7.66(m,4H),7.49-7.40(m,6H),4.15(dd,J=7.0,7.0Hz,1H),4.02-3.99(m,1H),3.81(dd,J=4.8,4.8Hz,1H),3 .59(dd,J=7.2,7.2Hz,1H),2.35-2.28(m,2H),1.91-1.88(m,1H),1.83-1.78(m,1H),1.70-1.62(m,1H),1.08(s,9H).

[0451] Step 6: Example 309F 10% Pd / C (63 mg) was added to a suspension of compound Example 309E (630 g, 1.6 mmol) in dry EtOH (100 mL). The reactants were evaporated twice to change the inert gas atmosphere, and then connected to two balloons filled with H2. The suspension was vigorously stirred at room temperature for 15 hours. The palladium catalyst was removed using a PTFE filter (Whatman Puradisc), and the solvent was removed under reduced pressure. The amine (588 g, 100%) was obtained as a colorless liquid and was used without further purification. LCMS[M+H] + =370.

[0452] To a solution of amine (588 mg, 1.6 mmol) in DCM (50 mL) at 0°C, Boc2O (700 mg, 3.2 mmol) in DCM (10 mL) was added and stirred at 0°C, followed by stirring at room temperature for 2 hours. The reaction mixture was concentrated, and the crude product was purified using silica gel (hexane / ethyl acetate 5:1) to obtain Example 309F (650 mg, 1.38 mmol, 87%) as a colorless oil. LCMS[M+H] + =470. 1 H NMR(400MHz,CDCl3)δ 7.68-7.65(m,4H),7.48-7.40(m,6H),4.97(br s,1H),4.20(m,1H),4.02(br s,1H),3.74(dd,J=4.2,4.2Hz,1H),3.51(dd,J=8.4,8.4Hz,1H),2.29-2.22(m,2H),1.78-1.57(m,3H),1.45(s,9H),1.07(s,9H).

[0453] Step 7: Example 309G Compound Example 309F (630 mg, 1.34 mmol), DIEA (300 mg, 2.28 mmol), and DMAP (278 mg, 2.28 mmol) in DCM (50 mL) were mixed with TosCl (384 mg, 2.0 mmol) at 0°C. The mixture was stirred at room temperature for 14 hours, followed by concentration to dryness. The crude product was purified using silica gel (hexane / ethyl acetate 10:1) to obtain 4-methylbenzenesulfonic acid (1S,2R,4S)-4-((tert-butoxycarbonyl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopentyl (590 mg, 0.94 mmol, 70%) as a colorless oil. LCMS[M+H] + = 625.

[0454] 4-methylbenzenesulfonic acid (1S,2R,4S)-4-((tert-butoxycarbonyl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopentyl (590 g, 0.94 mmol) was dissolved in dry DMF (10 mL), and NaN3 (92 mg, 1.4 mmol) was added. The mixture was stirred at 60°C for 14 hours. After the addition of ethyl acetate (100 mL), the organic layer was washed with saturated NaHCO3 solution (2 × 50 mL) and brine (50 mL), and dried over MgSO4. The solvent was removed under reduced pressure. The residue was purified by flash chromatography on silica gel (hexane / ethyl acetate 5:1) to obtain tert-butyl ((1S,3R,4S)-3-azido-4-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopentyl)carbamate (460 mg, 0.93 mmol, 99%) as a colorless oil. Rf = 0.54 (20% ethyl acetate in hexane). LCMS[M+H] + =496. 1 H NMR(400MHz,CDCl3)δ 7.72-7.67(m,4H),7.46-7.40(m,6H),4.45(br s,1H),4.20(m,1H),4.08(br s,1H),3.79-3.72(m,1H),3.68-3.62(m,1H),2.41-2.35(m,2H),1.80-1.47(m,5H),1.46(s,9H),1.08(s,9H).

[0455] 10% Pd / C (46 mg) was added to a suspension of ((1S,3R,4S)-3-azido-4-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopentyl)carbamate tert-butyl (460 g, 0.93 mmol) in dry EtOH (50 mL). The reactants were evaporated twice to change the inert gas atmosphere, and then connected to two balloons filled with H2. The suspension was vigorously stirred at room temperature for 15 hours. The palladium catalyst was removed using a PTFE filter (Whatman Puradisc), and the solvent was removed under reduced pressure. Amine Example 309G (460 mg, 0.93 mmol, quantitative) was obtained as a colorless liquid and was used without further purification. LCMS[M+H]+ =470

[0456] Step 8: Example 309H To a solution of compound Example 309G (70 mg, 0.17 mmol) in THF (5 mL), 1 M TBAF (1.7 mL, 0.17 mmol) was added under N2 at 0°C, followed by stirring at room temperature for 2 hours. The reaction mixture was concentrated to obtain crude product 13, and the crude product was processed in the following step: LCMS[M+H] + =231.

[0457] Step 9: Example 309I A mixture of compound Example 309H (crude, 0.17 mmol), 2-chloro-N,N-dimethylbenzo[d]thiazole-6-sulfonamide (46 mg, 0.17 mmol), and DIEA (65 mg, 0.51 mmol) in DMSO was stirred overnight at 60°C. The mixture was purified by preparative HPLC to obtain compound Example 309I (38 mg, 47%) as a white solid. LCMS[M+H] + = 472.

[0458] Step 10: Example 309J A 4M HCl / dioxane (5L) solution was added to Compound Example 309I (38 mg, 0.08 mmol) in DCM (1 mL), and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated to obtain Compound Example 309J (30 mg, 100%) as a white solid. LCMS[M+H] + =372.

[0459] Step 16: Example 309 A mixture of Compound Example 309J (30 mg, 0.08 mmol), 2-chloro-5-(methylthio)pyrimidine (15 mg, 0.09 mmol), and DIEA (34 mg, 0.25 mmol) in DMSO (2.5 mL) was stirred at 120°C for 4 hours. The mixture was purified by preparative HPLC to obtain Compound Example 309 (5 mg, 12%) as a white solid. LCMS[M+H] + =496. 1H NMR(400MHz,CD3OD)δ 8.36(s,2H),8.09(d,J=1.6Hz,1H),7.68(dd,J=2.0,1.6Hz,1H),7.56(d,J=8.4Hz,1H),4.62(br s,5H),4.54(br m,1H),3.63-3.58(m,2H),2.70(s,3H),2.69-2.65(m,1H),2.37(s,3H),2. 36-2.28(m,1H),2.16-2.10(m,1H),2.05-1.99(m,1H),1.86-1.82(m,1H).

[0460] Scheme 50 [ka]

[0461] Step 1: Example 310A To a mixture of tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (1.5 g, 7.5 mmol) and 2-chloro-5-(methylthio)pyrimidine (1.3 g, 8.3 mmol) in dimethyl sulfoxide (28 mL), N,N-diisopropylethylamine (3.7 mL, 22.5 mmol) was added at room temperature. The resulting mixture was stirred at 130 °C for 6.5 hours under an N2 atmosphere. Next, water (90 mL) and ethyl acetate (160 mL) were added to the mixture. The combined organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated. The residue was purified by silica gel column chromatography (PE:EA=20:1~PE:EA=6:1) to obtain Example 310A (1.8 g, yield 76%) as yellow oil. LCMS[M+H] + =325.

[0462] Step 2: Example 310B To a mixture of Example 310A (1.8 g, 5.7 mmol) in methanol (2 mL), HCl / dioxane (8.0 mL, 4 mol / L) was added at room temperature. The resulting mixture was stirred at room temperature under an N2 atmosphere for 2.5 hours. Next, the mixture was evaporated to obtain Example 310B (1.5 g, 100% yield) as a brown solid. LCMS[M+H] + =225.

[0463] Step 3: Example 310C A mixture of Example 310B (1.5 g, 5.7 mmol) and ethyl 2-chlorobenzo[d]thiazole-6-carboxylate (1.4 g, 5.7 mmol) in DMSO (28 mL) was mixed with DIEA (2.9 mL, 17.3 mmol) at room temperature. The resulting mixture was stirred at 80°C for 18 hours under an N2 atmosphere. Next, water (80 mL) and ethyl acetate (150 mL) were added to the mixture. The combined organic layer was washed with brine (90 mL), dried over anhydrous sodium sulfate, filtered, and evaporated. The residue was purified by silica gel column chromatography (PE:EA = 30:1~5:1) to obtain Example 310C (1.9 g, yield 77%) as a yellow solid. LCMS[M+H] + =430.

[0464] Step 4: Example 310D To a mixture of Example 310C (1.9 g, 4.4 mmol) in ethanol (20 mL) and water (10 mL), lithium hydroxide monohydrate (372 mg, 8.9 mmol) was added at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. Next, the mixture was concentrated. Water (60 mL) and HCl (3 mol / L) were added to the residue until the pH reached 5. The precipitate was filtered, and the filtrate was dried to obtain Example 310D (1.5 g, yield 84%) as a brown solid. LCMS[M+H] + =402. 1H NMR(400MHz,DMSO-d6)δ 8.49-8.47(d,J=6.8Hz,1H),8.34(s,2H),8.26-8.25(d,J=1.6Hz,1H),7.81-7.79(m,1H),7.55-7.53(d,J=7.2Hz,1H),7.40 -7.38(d,J=8.4Hz,1H),4.39-4.33(m,2H),2.35(s,3H),2.21-2.19(m,2H),2.12-2.10(m,2H),1.98-1.94(t,J=13.6Hz,1H).

[0465] Process E: Example 310 A mixture of Example 310D (30 mg, 0.075 mmol), dimethylamine (6.75 mg, 0.15 mmol), and HATU (28.5 mg, 0.075 mmol) in DCM was stirred at room temperature for 2 hours. The resulting solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain the title compound Example 310 (20 mg) as a yellow solid. LCMS[M+H] + = 429. 1 H NMR(600MHz,DMSO-d6)δ 1.54-1.64(m,2H),1.98(t,J=6.75Hz,2H),2.09-2.16(m,1H),2.18-2.27(m,1H),2.36(s,3H),2.97(s,6H),4.35(br s,2H),7.30(dd,J=8.35,1.49Hz,1H),7.40(d,J=8.24Hz,1H),7.57(br s,1H),7.74-7.81(m,1H),8.35(s,2H),8.58(br s,1H).

[0466] The following examples were synthesized using the procedure described above.

[0467] [Table 38]

[0468] [Table 39]

[0469] Table 40

[0470] Table 41

[0471] Table 42

[0472] Table 43

[0473] Table 44

[0474] Table 45

[0475] Table 46

[0476] Table 47

[0477] Table 48

[0478] Table 49

[0479] Table 50

[0480] [Table 51]

[0481] [Table 52]

[0482] [Table 53]

[0483] [Table 54]

[0484] [Table 55]

[0485] [Table 56]

[0486] Scheme 51 [ka]

[0487] Step 1: Example 371A A mixture of 2-fluoro-5-4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (1.0 g, 4.5 mmol), 3-bromo-2-methoxypyridine (930 mg, 4.9 mmol), Pd(dppf)Cl2 (320 mg, 0.45 mmol), and Na2CO3 (950 mg, 9.0 mmol) in dioxane:H2O=4:1 (50 mL) was stirred overnight at 110°C under an N2 atmosphere. The mixture was cooled to room temperature and water (50 mL) was added. The mixture was extracted with  (20 mL × 3), the combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated; the residue was purified by chromatography on silica gel eluted with petroleum ether:Â=5:1 to obtain Example 371A (660 mg, 72%) as a white solid.

[0488] Step 2: Example 371 A mixture of (1S,3S)-N1-(5-(methylthiopyrimidine-2-yl)cyclopentan-1,3-diamine hydrochloride (100 mg, 0.38 mmol), Example 371A (86 mg, 0.42 mmol), and Cs2CO3 (376 mg, 1.15 mmol) in DMSO (5 mL) was stirred at 130 °C for 2 days. The mixture was cooled to room temperature and water (10 mL) was added. The mixture was extracted with RINKAN (10 mL x 5), the combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC to obtain Example 371 (10 mg, 6.4%). LCMS[M+H] += 409. 1 H NMR(400MHz,CD3OD)δ 8.41(s,2H),8.20-8.17(m,2H),8.09(s,1H),7.82(d,J=7.2Hz,1H),7.13-7.07(m,2H),4.51-4.49(m,1H) ),4.29-4.27(m,1H),3.99(s,1H),2.41-2.33(m,2H),2.38(s,3H),2.19-2.15(m,2H),1.78-1.75(m,1H).

[0489] Scheme 52 [ka]

[0490] Step 1: Example 372A A mixture of 2-fluoro-4-iodopyridine (200 mg, 0.9 mmol), pyridine-2(1H)-one (102 mg, 1.1 mmol), CuI (17 mg, 0.09 mmol), N,N'-dimethyl-1,2-cyclohexanediamine (19 mg, 0.17 mmol), and K3PO4 (381 mg, 1.8 mmol) in DMSO (5 mL) was stirred at 100 °C for 3 hours under an N2 atmosphere. The mixture was cooled and water (20 mL) was added. The mixture was extracted with siRNA (20 mL x 3), the combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. This crude product was purified by chromatography on silica gel eluting with petroleum ether:siRNA = 5:1 to 1:1 to obtain Example 372A (97 mg, 57%) as a grayish-white solid.

[0491] Step 2: Example 372 A mixture of Example 372A (50 mg, 0.19 mmol), (1S,3S)-N1-(5-(methylthiopyrimidine-2-yl)cyclopentan-1,3-diamine hydrochloride (40 mg, 0.21 mmol), and Cs2CO3 (185 mg, 0.57 mmol) in DMSO (3 mL) was stirred at 130 °C for 2 days. The mixture was cooled and water (10 mL) was added. The mixture was extracted with RINKAN (10 mL x 5), the combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC to obtain Example 372 (10 mg, 13%). LCMS[M+H] += 395. 1H NMR(400MHz,CD3OD)δ 8.39(s,2H),7.99(d,J=7.2Hz,1H),7.70-7.64(m,2H),7.20(s,1H),7.05-7.03(m,1H),7.67(d,J=9.2Hz,1H),6.58-6. 54(m,1H),4.51-4.47(m,1H),4.29-4.27(m,1H),2.42-2.33(m,2H),2.38(s,3H),2.20-2.14(m,2H),1.78-1.74(m,1H).

[0492] The following examples were synthesized using the procedure described above.

[0493] [Table 57]

[0494] Scheme 54 [ka]

[0495] General method 1 6'-Chloro-5-methoxy-2H-[1,3'-bipyridine]-2-one (Example 400A) The compounds 2-chloro-5-iodopyridine (191 mg, 0.8 mmol, 1.0 equivalent), 5-methoxypyridine-2(1H)-one (100 mg, 0.8 mmol, 1.0 equivalent), CuI (15 mg, 0.08 mmol, 0.1 equivalent), N,N'-dimethyl-1,2-cyclohexanediamine (23 mg, 0.16 mmol, 0.2 equivalents), and K3PO4 (339 mg, 1.6 mmol, 0.2 equivalents) in DMSO (5 mL) were stirred overnight at 120 °C under an N2 atmosphere. The mixture was cooled to room temperature and water (20 mL) was added. The mixture was extracted with  (20 mL x 2). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, concentrated, and purified by chromatography on silica gel eluting PE:EA=1:1 to obtain 400A (60 mg, 0.23 mmol, 32%) as a grayish-white solid.

[0496] General procedure 2 5-Methoxy-6'-(((1S,3S)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridine]-2-one (Example 400) A mixture of 400A (53 mg, 0.22 mmol, 1.0 equivalent), 131C (50 mg, 0.22 mmol, 1.0 equivalent), tBuXphos Pd G3 (18 mg, 0.022 mmol, 0.1 equivalent), and tBuOK (49 mg, 0.44 mmol, 0.2 equivalents) in 5 mL of dioxane was stirred overnight at 110°C under an N2 atmosphere. The mixture was cooled to room temperature and filtered. The filtrate was concentrated and purified by preparative TLC and then preparative HPLC to obtain compound 400 (50 mg, 53%, TFA salt) as a grayish-white solid. LCMS[M+H] + = 425. 1 H NMR(400MHz,CD3OD)δ 8.36(s,2H),8.09(s,1H),7.95(d,J=9.6Hz,1H),7.50(d,J=9.6Hz,1H),7.23(s,1H),7.10(d,J=9.6Hz,1H),6.61(d,J=9 .6Hz,1H),4.48-4.46(m,1H),4.25-4.23(m,1H),3.73(s,3H),2.40-2.35(m,5H),2.16-2.13(m,2H),1.76-1.74(m,2H).

[0497] The following examples were synthesized using the procedure described above.

[0498] [Table 58]

[0499] Scheme 55 [ka]

[0500] 5-Hydroxy-6'-(((1S,3S)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridine]-2-one (Example 402) To a solution of 400 (50 mg, 0.12 mmol, 1.0 equivalent) in DCM (10 mL), BBr3 (1 mL) was added at 0°C. The mixture was stirred overnight at room temperature, quenched with MeOH, and then concentrated. The residue was purified by preparative TLC and then preparative HPLC to obtain compound Example 402 (50 mg, 53%, TFA salt) as a grayish-white solid. 1 H NMR(400MHz,CD3OD)δ 8.37(s,2H),8.06(s,1H),7.94(d,J=9.6Hz,1H),7.45(d,J=9.6Hz,1H),7.12-7.09(m,2H),6.58(d,J=9.6Hz ,1H),4.49-4.46(m,1H),4.31-4.28(m,1H),2.42-2.30(m,5H),2.17-2.13(m,2H),1.79-1.73(m,2H).[M+H] + =411.

[0501] Scheme 56 [ka]

[0502] 4-Hydroxy-5'-(((1S,3S)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)-2H-[1,2'-bipyridine]-2-one (Example 403) The title compound was obtained from the starting material 4-(benzyloxy)pyridine-2(1H)-one by the same method as described in General Method 1, General Method 2, and General Method 3. 1H NMR(400MHz,CD3OD)δ 8.32(s,2H),7.89(d,J=2.2Hz,1H),7.45(d,J=7.5Hz,2H),7.39(dd,J=9.0,2.7Hz,1H),6.60(d,J=8.6Hz,1H),6.13(dd,J=7.5,2.6 Hz,1H),5.88(d,J=8.6,2.5Hz,1H),4.48-4.25(m,2H),2.35(s,3H),2.32-2.19(m,2H),2.06-1.92(m,2H),1.70-1.52(m,2H).[M+H] + =411.49.

[0503] Scheme 57 [ka]

[0504] General procedure 4 6'-Chloro-4-methoxy-3,3'-bipyridine(404) A mixture of (6-chloropyridine-3-yl)boronic acid (1.0 g, 6.4 mmol, 1 equivalent), 3-bromo-4-methoxypyridine (1.2 g, 6.4 mmol, 1 equivalent), Pd(dppf)Cl2 (468 mg, 0.64 mmol, 0.1 equivalent), and Na2CO3 (1.3 g, 12.8 mmol, 2 equivalents) in dioxane (8 mL) and water (2 mL) was stirred overnight at 105 °C under an N2 atmosphere. The mixture was cooled to room temperature and quenched with water (10 mL). The mixture was extracted with HCl (20 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by chromatography on silica gel eluting with petroleum ether:HCl = 5:1 to obtain compound 404 (200 mg, 14%) as a brown solid.

[0505] The following examples were synthesized using the procedure described above.

[0506] [Table 59]

[0507] Scheme 58 [ka]

[0508] 2-Chloro-5-(2,2,2-trifluoroethoxy)pyrimidine (408) To a solution of 2-chloropyrimidine-5-ol (0.5 g, 3.8 mmol, 1.0 equivalent) and CS2CO3 (1.49 g, 4.6 mmol, 1.2 equivalents) in DMF (20 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonic acid (0.97 g, 4.2 mmol, 1.1 equivalents) was added. The resulting suspension was stirred at room temperature for 16 hours, then partitioned between SiO (30 mL) and water (80 mL). The separated aqueous layer was extracted with SiO (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated to obtain 408 (0.74 g), which was used directly in the next step. LCMS[M+H] + =213.

[0509] Scheme 59 [ka]

[0510] 2-Chloro-5-(difluoromethoxy)pyridine (409) A solution of 2-chloropyrimidine-5-ol (101.6 g, 0.76 mol, 1.0 equivalent) in DMF (2000 mL) was packed with Cs2CO3 (300 g, 0.92 mol, 1.2 equivalents) and stirred at room temperature for 1.5 hours. Sodium 2-chloro-2,2-difluoroacetate (340 g, 2.3 mol, 3.0 equivalents) was added, and the reaction mixture was stirred at 100°C for 3.5 hours. The reaction mixture was poured into water (5 L) and extracted with EA (3 × 1 L). The combined organic phase was dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA = 10:1) to obtain 409 (70 g).

[0511] Scheme 60 [ka]

[0512] 2-Chloro-5-cyclopropylpyrimidine (410A) To a solution of 5-bromo-2-chloropyrimidine (100 g, 518 mmol, 1.0 equivalent), cyclopropylboronic acid (53 g, 616 mmol, 1.2 equivalents), and Pd(dppf)Cl2 (10 g, 13.7 mmol, 0.03 equivalents) in dioxane (1.5 L), Cs2CO3 (250 g, 769 mmol, 1.5 equivalents) was added and the mixture was stirred at 110°C for 12 hours under N2. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with hexane:ethyl acetate = 15:1) to obtain 410A (55 g) as a yellow solid. LCMS[M+H] + = 155.

[0513] General procedure 5 ((1S,3S)-3-((5-Cyclopropylpyrimidine-2-yl)amino)cyclopentyl)carbamate tert-butyl(410B) A mixture of 410A (40 g, 260 mmol, 1 equivalent), ((1S,3S)-3-aminocyclopentyl)carbamate tert-butyl (55 g, 275 mmol, 1.05 equivalents), and DIPEA (105 g, 814 mmol, 3.13 equivalents) in DMSO (400 mL) was stirred at 110 °C for 12 hours under N2. The mixture was then cooled to room temperature and diluted with water (1000 mL). The resulting mixture was extracted with SiO2 (100 mL x 3), the combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by chromatography on silica gel (eluting with hexane:ethyl acetate = 5:1 to 4:1) to obtain compound 410B (55 g) as a pale blue solid. LCMS[M+H] + =319.

[0514] General procedure 6 (1S,3S)-N 1 -(5-Cyclopropylpyrimidine-2-yl)cyclopentan-1,3-diamine(410C) To a solution of 410B (44 g, 13.8 mmol, 1.0 equivalent) in MeOH (250 mL), HCl (4 M in dioxane, 250 mL) was added dropwise, and the resulting solution was stirred at room temperature for 2.5 hours. After the reaction was complete, the mixture was concentrated to dryness under vacuum. The residue was redissolved in MeOH (500 mL) and ion-exchange resin (Ambersep® 900 OH) was used. - The mixture was filtered, and the pH was adjusted to approximately 8. The filtrate was concentrated to obtain 410C (44.5) as yellow oil. LCMS[M+H] += 219.

[0515] The following examples were synthesized using the procedure described above.

[0516] [Table 60]

[0517] Scheme 61 [ka]

[0518] 2-Bromo-5-(methylthio)pyridine (416) A solution of 2,5-dibromopyridine (1 g, 4.22 mmol, 1 equivalent) in THF (20 mL) was cooled to -78°C under N2. Next, n-BuLi (2.5 M, 1.77 mL, 4.43 mmol, 1.05 equivalents) was added dropwise at -78°C. After stirring the reaction mixture for 20 minutes, 1,2-dimethyldisulfan (0.411 mL, 4.64 mmol, 1.1 equivalents) was slowly added. After stirring the reaction mixture for a further 1 hour, it was quenched with saturated NH4Cl. The reaction mixture was extracted with EA (100 mL) and water (100 mL), followed by washing with brine. The organic phase was then treated with anhydrous sodium sThe mixture was dried with SO4. The mixture was filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography (PE:EA=10:1) on silica gel to obtain 416 (86.8 mg). ESI[M+H] + =204.1 1 H NMR(400MHz,CDCl3)δ 8.25(t,J=6.2Hz,1H),7.41(ddd,J=8.9,8.3,1.6Hz,2H),2.58-2.40(m,3H).

[0519] Scheme 62 [ka]

[0520] (1S,3S)-N1-(thieno[3,2-b]pyridine-5-yl)cyclopentan-1,3-diamine(417) The compound of the title can be synthesized from the starting material 5-chlorothieno[3,2-b]pyridine by the same method as described in General Method 2 and General Method 6.

[0521] The following examples were synthesized using the procedure described above.

[0522] [Table 61]

[0523] Scheme 63 [ka]

[0524] General procedure 7 3-(6-chloropyridine-3-yl)pyrimidine-4(3H)-one(419) A solution of pyrimidine-4(3H)-one (1.5 g, 15.61 mmol, 1.0 equivalent), (6-chloropyridine-3-yl)boronic acid (2.9 g, 18.73 mmol, 1.2 equivalents), Cu(OAc)2 (7.9 g, 43.71 mmol, 2.8 equivalents), pyridine (2.5 mL, 31.22 mmol, 2.0 equivalents), and 4 Å molecular sieve (8 g) in DCM (60 mL) was stirred at room temperature for 48 hours. The mixture was filtered through Celite, the filtrate was concentrated, and purified by flash column (petroleum ether: siRNA: 1:1) to obtain Example 419 as a yellow solid (170 mg, yield 5.3%). 1 H NMR(400MHz,CDCl3)δ 8.43(d,J=2.6Hz,1H),8.15(s,1H),7.97(d,J=6.8Hz,1H),7.80(dd,J=8.4, 2.8Hz,1H),7.53(d,J=8.5Hz,1H),6.59(dd,J=6.8,0.8Hz,1H);MS(ESI+)m / z 208.0(M+H) +

[0525] The following examples were prepared using the procedure described in Scheme 63.

[0526] [Table 62]

[0527] Scheme 64 [ka]

[0528] (E)-3-(2-fluoropyridine-4-yl)ethyl acrylate (423A) To a suspension of 60% NaH (1.54 g, 38.4 mmol, 1.2 equivalents) in THF (15 mL), a solution of 2-(ethoxy(propoxy)phosphoryl)ethyl acetate (8.6 g, 38.4 mmol, 1.2 equivalents) in THF was added dropwise at 0°C under an N2 atmosphere. The mixture was stirred at 0°C for 25 minutes, and then a solution of 2-fluoroisonicotinaldehyde (4.0 g, 32.0 mmol, 1.0 equivalent) in DMF (15 mL) was added. The resulting mixture was stirred at room temperature for 16 hours, followed by quenching with saturated aqueous NH4Cl solution at 0°C. The aqueous phase was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under vacuum. The residue was purified by column chromatography (PE~PE:EA = 1:15) to obtain 423A as a white solid (3.0 g, 50% yield). LCMS: m / z 196[M+H] + rt 2.890 min. 1 H-NMR(400MHz,CDCl3)δ 8.26(d,J=5.2Hz,1H),7.59(d,J=16.1Hz,1H),7.29-7.26(m,1H),7.00(d,J=1. 7Hz,1H),6.60(d,J=16.0Hz,1H),4.30(q,J=7.1Hz,2H),1.35(t,J=7.1Hz,3H).

[0529] 3-(2-fluoropyridine-4-yl)propanoate ethyl (423B) To a solution of 423A (3.0 g, 20.5 mmol) in EtOH (20 mL), 10% Pd / C (400 mg) was added. The reaction system was purged with H2, and the mixture was stirred overnight under an H2 atmosphere. Pd / C was filtered off, and the filtrate was concentrated under vacuum to obtain 423B (2.3 g, yield 76.7%), which was used directly. LCMS: m / z 198.1 [M+H] + rt 2.801 min. 1 H-NMR(400MHz,CDCl3)δ 8.12(d,J=5.5Hz,1H),7.04(d,J=4.8Hz,1H),6.78(d,J=4.0Hz,1H),4.13(p,J =6.8Hz,2H),3.03-2.97(m,2H),2.66(t,J=7.6Hz,2H),1.24(t,J=6.7Hz,3H).

[0530] 3-(2-oxo-1,2-dihydropyridine-4-yl)propanoic acid (423C) A 100 mL flask containing 423B (1.53 g, 7.77 mmol) was mixed with concentrated HCl (5 mL). The mixture was heated to 100 °C for 16 hours. The mixture was then cooled to room temperature, and 2 mL of water was added. Solid NaHCO3 was added in small increments to adjust the pH to 6. The mixture was extracted with 30% i-PrOH in CHCl3, and the combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by column chromatography (MeOH / DCM) to obtain 423C (1.6 g, 91% yield), which was used directly in the next step. LCMS: m / z 167.8 [M+H] + ; 1 H NMR(400MHz,CDCl3)δ 8.14(d,J=5.1Hz,1H),7.05(dt,J=5.3,1.7Hz,1H),6.82-6.76(m,1H),3.01(t,J=7.5Hz,2H),2.73(t,J=7.5Hz,2H).

[0531] 3-(2-oxo-1,2-dihydropyridine-4-yl)propanoate methyl(423D) A 100 mL flask containing a solution of crude 423C (1.6 g) in MeOH (28 mL) was mixed with 98% H2SO4 (0.15 mL). The reaction mixture was heated to 90 °C for 16 hours and then cooled to room temperature. A saturated NaCl aqueous solution was added to the mixture and extracted with 30% i-PrOH in CHCl3. The combined organic layer was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (MeOH:DCM = 1:15) to obtain product 423D (702 mg, yield 55%). LCMS: m / z 182.1 [M+H] + .

[0532] General procedure 8 3-(6'-fluoro-2-oxo-2H-[1,3'-bipyridine]-4-yl)methyl propanoate (423) In a sealed tube, 423D (100.0 mg, 0.55 mmol, 1.0 equivalent), 2-fluoro-5-iodopyridine (147 mg, 0.66 mmol, 1.2 equivalents), CuI (21.0 mg, 0.10 mmol, 0.2 equivalents), N,N'-dimethyl-1,2-cyclohexanediamine (15.5 mg, 0.10 mmol, 0.2 equivalents), and K2CO3 (151.0 mg, 1.10 mmol, 2.0 equivalents) were added. Dioxane (3 mL) was added, the resulting mixture was purged with N2, and stirred at 110°C for 16 hours. Next, the mixture was diluted with dichloromethane and filtered. The filtrate was washed with water and separated. The aqueous phase was extracted three times with dichloromethane. The combined organic layers were dried over Na2SO4, concentrated, and purified by silica gel chromatography to obtain 423 (42 mg, yield 27.6%). LCMS: m / z 278.08 [M+H] +

[0533] Scheme 65 [ka]

[0534] 6'-Fluoro-2-oxo-2H-[1,3'-bipyridine]-5-methylcarboxylate (424A) A mixture of 6-fluoropyridine-3-amine (1.1 g, 10 mmol, 1.0 equivalent) and 2-oxo-2H-pyran-5-carboxylate methyl (1.5 g, 10 mmol, 1.0 equivalent) in EtOH (10 mL) was stirred overnight under reflux and then cooled to room temperature. The resulting precipitate was collected by filtration and purified by chromatography on silica gel eluted with petroleum ether:siRNA = 2:1 to obtain compound 424A (440 mg, 18%) as a grayish-white solid.

[0535] 6'-(((1S,3S)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)-2-oxo-2H-[1,3'-bipyridine]-5-methylcarboxylate(424B) 424B was obtained from the starting materials 131C and 424A by the same method as described above.

[0536] General procedure 9 6'-(((1S,3S)-3-((5-(methylthiopyrimidine-2-yl)amino)cyclopentyl)amino)-2-oxo-2H-[1,3'-bipyridine]-5-carboxylic acid (Example 424) To a solution of compound 424B (100 mg, 0.22 mmol, 1.0 equivalent) in MeOH:H2O=5:1 (10 mL), NaOH (35 mg, 0.88 mmol, 4.0 equivalents) was added. The mixture was stirred at 50°C for 3 hours, then cooled to room temperature and acidified to pH=5-6. The resulting mixture was concentrated and redissolved in THF (50 mL). The solid was filtered off, the filtrate was concentrated, and triturated with EA:MeOH=5:1 to obtain Example 424 (90 mg, 93%) as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6)δ:8.35(s,2H),8.25s,1H),8.10 1H),7.87(d,J=9.2Hz,1H),7.56(s,1H),6.87(s,1H),6.51(d,J=9.2Hz,1H),...

Claims

1. Compound of formula (I) 【Chemistry 1】 (In the formula, A stands for H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR ６ , and -C(O)NR ６ R ７ Selected from; B is selected from H, alkyl, and halo, or A and B, together with the carbon atoms to which they are bonded, form a 5- or 6-membered heteroaryl ring, or an unsubstituted or one or more alkyl-substituted pyrrolyl or thienyl ring; X is NR ５ And; n is 0; R １ and R １’ These, together with the atoms to which they are bonded, form a cyclopentyl ring, where the cyclopentyl ring may be substituted with hydroxyl or hydroxyalkyl atoms; R ２ is selected from H, hydroxyalkyl, and hydroxy; R ２’ H is; R ５ is H or alkyl; Each R ６ and R ７ is independently H or alkyl; Y is selected from aryl and heteroaryl groups; Here A) Y is given by formula -C(O)NR ８ R ９ or -NR ９ C(O)R １０ It can be substituted with an amide substituent. Here i) R ８ and R ９ Each of these is independently selected from H, alkyl, heterocyclyl, and heteroaryl; or ii) R ８ and R ９ They, together with the nitrogen atom to which they are bonded, form a 4, 5, 6, or 7-membered heterocyclic or heteroaryl ring; or iii) R8 and R9, together with the nitrogen atom to which they are bonded, form a heterocycle selected from azirazine, isothiazolidine-1,1-dioxide, azetidine, thiazole-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, and 2-oxa-6-azaspiro[3.3]heptane; or iv) R8 and R9, together with the nitrogen atom to which they are bound, form 2,8-diazaspiro[5,5]undecene, tetrahydroimidazo[1,2-a]pyrazine, octahydropyrazino[2,1-c][1,4]oxazine, tetrahydropyrido[3,4-d]pyrimidine, 2-oxa-8-azaspiro[4.5]decane, tetrahydropyrrolo[3,4-c]pyrazole, thiomorpholine, 2-oxa-7-azaspiro[3.5]nonane, 2,8-diazaspiro[4.5]decane-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9-diaza Spiro[5.5]undecane-3-one, tetrahydropyrrolo[3,4-d]imidazole, pyrimidine, 8-oxa-2-azaspiro[4.5]decane, hexahydro-3H-oxazolo[3,4-a]pyrazine-3-one, 1-oxa-7-azaspiro[3.5]nonane, octahydrocyclopenta[c]pyrrole, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine, 2,7-diazaspiro[4.4]nonane, 2,6-diazaspiro[3.4]octane, 7-oxa-2-azaspiro[3.5]nonane, 1-oxa-8λ Forms a heterocycle selected from 2-azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, oxadiazole, triazole, pyridinone, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3(2H)-one, piperidinone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5]nonane, pyrazole, and pyridazine-3(2H)-one; and R １０ is alkyl; or B)Y is given by equation -S(O) ２ NR ８ R ９ or -NR ９ S(O) ２ R １０ It may be substituted with a sulfonamide substituent; Here i) R ８ and R ９ Each of these is independently selected from H, alkyl, and heteroaryl; or ii) R ８ and R ９ They, together with the nitrogen atoms to which they are bonded, form a 4, 5, 6, or 7-membered heterocycle; and R １０ It is alkyl; If Y is a monocyclic heteroaryl, the monocyclic heteroaryl is either unsubstituted or substituted with one or more substituents selected from alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxy, cyano, halo, aryl, heteroaryl, heterocyclyl, nitro, sulfonamide, and thioalkyl, or the monocyclic heteroaryl is substituted with a heteroaryl or heterocyclyl substituted with one or more substituents selected from halo, CN, alkyl, alkoxy, hydroxy, carboxy, -CO2 alkyl, and tetrazolyl; If Y is a bicyclic heteroaryl, the bicyclic heteroaryl is either unsubstituted or substituted with one or more substituents selected from alkyl, haloalkyl, hydroxyalkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxy, cyano, halo, heteroaryl, nitro, and sulfonamide; Here, the heterocycle is either unsubstituted or substituted with one or more substituents selected from alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxy, and heterocyclyl; Herein, "alkyl," "alkenyl," and "alkoxy" may be substituted with one or more substituents selected from halogens, hydroxyl, carbonyl, carboxyl, alkoxycarbonyl, formyl, acyl, thiocarbonyl, thioester, thioacetate, thioformate, alkoxy, phosphoryl, phosphate, phosphonate, phosphine, amino, amide, amidine, imine, cyano, nitro, azide, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic ring moieties; The aforementioned "aryl," "heterocyclyl," "heteroaryl," and "heteroaryl ring" may be substituted with one or more substituents selected from halogens, hydroxyl, carbonyl, carboxyl, alkoxycarbonyl, formyl, acyl, thiocarbonyl, thioester, thioacetate, thioformate, alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amide, amidine, imine, cyano, nitro, azide, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic ring moieties. (However, the aforementioned compounds may be substituted with one or more substituents selected from halogens, hydroxyl, carbonyl, carboxyl, alkoxycarbonyl, formyl, acyl, thiocarbonyl, form 【change】 (not) or its pharmaceutically acceptable salt.

2. A is H, hydroxy, thioalkyl, alkyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR ６ , and -C(O)NR ６ R ７ A compound according to claim 1 or a pharmaceutically acceptable salt thereof, selected from the above.

3. A is -SCH ３ , -SCHF ２ , and -OCHF ２ A compound according to claim 1 or a pharmaceutically acceptable salt thereof, selected from the above.

4. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein B is H.

5. A compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein Y is a heteroaryl compound.

6. R ２ The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein the compound is hydroxyl.

7. A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein Y is a monocyclic heteroaryl compound.

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein Y is selected from pyridyl, pyrazinyl, pyrimidinyl, and thiazolyl.

9. The compound according to any one of claims 7 to 8 or a pharmaceutically acceptable salt thereof, wherein the monocyclic heteroaryl is either unsubstituted or substituted with one or more substituents selected from alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxy, cyano, halo, aryl, heteroaryl, heterocyclyl, nitro, sulfonamide, and thioalkyl.

10. 【Chemistry 2】 【Transformation 3】 【Chemistry 4】 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 A compound selected from or a pharmaceutically acceptable salt thereof.

11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

12. A pharmaceutical product for treating cardiovascular disease in a subject, comprising a compound described in any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof.

13. The pharmaceutical product according to claim 12, wherein the cardiovascular disease is selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, abnormal lipoproteinemia, atherosclerosis, and coronary artery disease.

14. The aforementioned cardiovascular disease, a) Familial hypercholesterolemia, b) Autosomal dominant hypercholesterolemia, or c) Atherosclerosis, The pharmaceutical product according to claim 12 or 13.

15. a) The level of circulating serum LDL cholesterol is reduced in the subject. b) The level of circulating serum VLDL-cholesterol is reduced in the subject. c) The level of circulating serum triglycerides is reduced in the subject, d) The level of circulating serum lipoprotein A is reduced in the subject. The pharmaceutical product according to any one of claims 12 to 14.

16. The pharmaceutical product according to any one of claims 12 to 15, further comprising co-administering one or more additional therapeutic agents.

17. The pharmaceutical product according to claim 16, wherein the one or more additional therapeutic agents are selected from alirocumab, evolocumab, vococizumab, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimibe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

18. The pharmaceutically acceptable agent according to claim 16, wherein the one or more additional therapeutic agents are selected from HMG-CoA reductase inhibitors, HMG-CoA synthase inhibitors, HMG-CoA reductase gene expression inhibitors, HMG-CoA synthase gene expression inhibitors, MTP / Apo B secretion inhibitors, CETP inhibitors, bile acid absorption inhibitors, cholesterol absorption inhibitors, cholesterol synthesis inhibitors, squalene synthase inhibitors, squalene epoxidase inhibitors, squalene cyclase inhibitors, combined squalene epoxidase / squalene cyclase inhibitors, fibrates, niacin, combinations of niacin and lovastatin, ACAT inhibitors, bile acid scavengers, and PCSK9 translation inhibitors.

19. The pharmaceutical product according to claim 16, wherein one or more additional therapeutic agents are statins.

20. The pharmacopoeia according to claim 16, wherein one or more additional therapeutic agents are selected from atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

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