PCSK9 inhibitors and methods of use thereof

Abstract

The present invention relates to PCSK9 inhibitors and methods of use thereof. The present invention relates to novel heteroaryl compounds and pharmaceutical formulations thereof. The invention further relates to methods of treating or preventing cardiovascular diseases, and methods of treating sepsis or septic shock, using the novel heterocyclic compounds disclosed herein.

Description

[0001] This application is a divisional application of the invention patent application with application date of January 16, 2020, application number 202080009213.4, and invention name “PCSK9 inhibitors and methods of use thereof”.

[0002] Related applications

[0003] This application claims the benefit of priority to U.S. Application No. 62 / 794,234, filed January 18, 2019, which is incorporated herein by reference in its entirety. Background Art

[0004] PCSK9, also known as "proprotein convertase subtilisin / kexin 9," is a member of the secreted proprotein convertase family and plays an important role in cholesterol metabolism. PCSK9 increases circulating LDL cholesterol levels by enhancing 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 internalized into the endosomal / lysosomal compartment. The enhanced binding affinity of PCSK9 to the LDL receptor at the acidic pH of the late endosome / lysosome reduces LDL receptor recycling and targets the LDL receptor for lysosomal degradation. Genetic association studies 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.

[0005] Another biological pathway involving the action of PCSK9 on the LDL receptor is the onset of septic shock. Septic shock is an often fatal complication of severe microbial infection (sepsis) that triggers an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis originates from microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharide (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors (Toll-like receptors (TLRs)) and therefore plays a prominent role in the inflammatory response of sepsis (septic shock or sepsis).

[0006] PCSK9 reduces LPS absorption through the liver's LDL receptors, allowing free LPS to overstimulate the body's immune response to pathogens that cause sepsis. Therefore, inhibiting PCSK9 is beneficial in maintaining liver LDL receptors to achieve systemic pathogen clearance and detoxification in response to sepsis. However, aside from antibiotic therapy, there is currently no effective treatment for sepsis or septic shock.

[0007] For cardiovascular disease, there are few options for inhibiting PCSK9. Statins actually upregulate PCSK9 in HepG2 cells and primary human hepatocytes through increased expression of SREBP-2, a transcription factor that upregulates the LDLR and PCSK9 genes. Because elevated PCSK9 levels reduce the abundance of LDL receptors on the cell surface, increasing statin doses fail to achieve proportional LDL-cholesterol lowering.

[0008] Two monoclonal antibodies (mAbs) that selectively bind to extracellular PCSK9 and prevent it from interacting with the LDL receptor, alizumab and ivocuzumab, have recently received FDA approval for use in lowering LDL-C levels. In clinical trials, alizumab showed a 50% reduction in LDL levels compared to placebo. Elbitar et al., Expert Opin Therapeutic Patents [Expert Review of Therapeutic Patents] 201626:1377-1392. Patients taking ivocuzumab showed 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 can cause allergic reactions or other harmful immune responses in the body.

[0009] Unlike infections, which can occur sporadically, cardiovascular disease often requires management throughout a person's life. Therefore, ease of dosing and administration becomes a key factor in patient adherence to medication. There is a need for PCSK9 inhibitors with the increased efficacy and easier administration that can be achieved with small molecule PCSK9 inhibitors. Summary of the Invention

[0010] Disclosed herein are compounds having formula (I)

[0011]

[0012] in:

[0013] A is selected from H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 ;

[0014] B is selected from H, alkyl and halo, or

[0015] A and B, together with the carbon atoms to which they are attached, form a 5- or 6-membered heteroaryl group;

[0016] X is NR 5 or O;

[0017] R 1 and R 1 ' are each independently selected from H and alkyl; or

[0018] If n is 0, R 1 and R 1 'Together with the atoms to which they are attached, they form a 4-8 membered cycloalkyl or cycloalkenyl ring;

[0019] R 2 is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or

[0020] R 1 and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; or

[0021] R 1 ' and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; R 2 ' is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano, or

[0022] R 2 and R 2 'Together with the carbon atoms to which they are attached, they form a 3- to 8-membered cycloalkyl or heterocyclyl ring;

[0023] Each R 3 and R 4 are independently H or alkyl; or

[0024] R 2 and R 3 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; and

[0025] R 5 is H or alkyl; or

[0026] R 1 and R 5 Together with the atoms to which they are attached, form a 6-8 membered cycloalkyl or heterocyclyl ring; or

[0027] R 2 and R 5 Together with the atoms to which they are attached, they form a 5-8 membered cycloalkyl or heterocyclyl ring; each R 6 and R 7 are independently H or alkyl;

[0028] Y is selected from aryl, heteroaryl and heterocyclyl;

[0029] and

[0030] n is 0 or 1.

[0031] In certain embodiments, the present invention provides a pharmaceutical composition suitable for treating or preventing cardiovascular disease in a subject, comprising an effective amount of any compound described herein (e.g., a compound of the present invention, such as a compound of formula (I)) and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical formulation can be used to treat or prevent the conditions or diseases described herein.

[0032] Disclosed herein are methods for treating diseases and conditions that would benefit from PCSK9 inhibition. These diseases include, but are not limited to, cardiovascular disorders such as hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome, and coronary artery disease.

[0033] Other diseases and conditions that can be treated using the methods described herein include, but are not limited to, sepsis and septic shock.

[0034] Provided herein are combination therapies of compounds of formula (I) with monoclonal antibodies, statins, and other cardiovascular agents that can enhance cardiovascular therapeutic benefits beyond the ability of adjuvant therapy alone. Also provided herein are combination therapies of compounds of formula (I) with antibiotic agents that can reduce the incidence and severity of sepsis and septic shock beyond the ability of adjuvant therapy alone. DETAILED DESCRIPTION

[0035] Disclosed herein are compounds having formula (I)

[0036]

[0037]

[0038] in:

[0039] A is selected from H, halo, hydroxy, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 ;

[0040] B is selected from H, alkyl and halo, or

[0041] A and B, together with the carbon atoms to which they are attached, form a 5- or 6-membered heteroaryl group;

[0042] X is NR 5 or O;

[0043] R 1 and R 1' are each independently selected from H and alkyl; or

[0044] If n is 0, R 1 and R 1 'Together with the atoms to which they are attached, they form a 4-8 membered cycloalkyl or cycloalkenyl ring;

[0045] R 2 is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or

[0046] R 1 and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; or

[0047] R 1 ' and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; R 2 ' is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano, or

[0048] R 2 and R 2 'Together with the carbon atoms to which they are attached, they form a 3- to 8-membered cycloalkyl or heterocyclyl ring;

[0049] Each R 3 and R 4 are independently H or alkyl; or

[0050] R 2 and R 3 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; and

[0051] R 5 is H or alkyl; or

[0052] R 1 and R 5 Together with the atoms to which they are attached, form a 6-8 membered cycloalkyl or heterocyclyl ring; or

[0053] R 2 and R 5 Together with the atoms to which they are attached, they form a 5-8 membered cycloalkyl or heterocyclyl ring; each R 6 and R 7 are independently H or alkyl;

[0054] Y is selected from aryl, heteroaryl and heterocyclyl;

[0055] and

[0056] n is 0 or 1.

[0057] Disclosed herein are compounds having formula (I')

[0058]

[0059] in:

[0060] A is selected from H, halo, alkyl, thioalkyl, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 ;

[0061] B is selected from H, alkyl and halo;

[0062] R 1 and R 1 ' are each independently selected from H and alkyl; or

[0063] If n is 0, R 1 and R 1 'Together with the atoms to which they are attached, they form a 4-8 membered cycloalkyl ring;

[0064] R 2 is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, hydroxyalkyl, alkylamino, cyano, and hydroxy; or

[0065] R 1 and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; or

[0066] R 1 ' and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; R 2 ' is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, and cyano; or

[0067] R 2 and R 2 'Together with the carbon atoms to which they are attached, they form a 3- to 8-membered cycloalkyl or heterocyclyl ring;

[0068] Each R 3 and R 4 are independently H or alkyl; or

[0069] R 2 and R 3 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring; and

[0070] R 5 is H or alkyl; or

[0071] R 1and R 5 Together with the atoms to which they are attached, form a 6-8 membered cycloalkyl or heterocyclyl ring; or

[0072] R 2 and R 5 Together with the atoms to which they are attached, they form a 5-8 membered cycloalkyl or heterocyclyl ring; each R 6 and R 7 are independently H or alkyl;

[0073] Het is heteroaryl or heterocyclyl; and

[0074] n is 0 or 1.

[0075] All examples hereinafter and herein are to be understood as being examples of both formula (I) and formula (I').

[0076] In certain embodiments, the compound of claim 1, wherein A is selected from H, hydroxy, thioalkyl, alkyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 In certain embodiments, A is H, while in other embodiments, A is an alkyl group, such as a thioalkyl group. In certain embodiments, A is selected from -SCH3, -SCHF2, and -OCHF2. In some embodiments, A is an alkoxy group. In other embodiments, A is a cycloalkyl group. In certain embodiments, B is H.

[0077] In certain embodiments, A and B, together with the carbon atoms to which they are attached, form a pyrrolyl or thienyl ring, which is unsubstituted or substituted with one or more alkyl groups.

[0078] In certain embodiments, X is preferably NR 5. In other embodiments, Y is preferably heteroaryl or heterocyclyl.

[0079] In certain embodiments, R 1 and R 1 ' are each H. However, when n is 0, R 1 and R 1 ' together with the atoms to which they are attached may 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 ' Together with the atoms to which they are attached, they can 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.

[0080] In certain embodiments, R 2is selected from H, halo, alkyl, alkoxy, amidoalkyl, aminoalkyl, alkylamino, cyano, and hydroxy. 2 It is C 1-3 Alkyl. R 2 Can be substituted by one or more substituents selected from the group consisting of amino, amide, cyano, hydroxyl, and heterocyclic groups. 2 'It's C 1-3 alkyl, while in other embodiments, R 2 'It's H.

[0081] In certain embodiments, R 1 and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring. 1 ' and R 2 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring. 2 and R 2 Together with the carbon atoms to which they are attached, they form a 3- to 8-membered cycloalkyl or heterocyclyl ring.

[0082] In certain embodiments, R 3 It is C 1-3 alkyl, while in other embodiments, R 3 is H. In certain embodiments, R 4 It’s H.

[0083] In certain embodiments, R 2 and R 3 Together with the atoms to which they are attached, they form a 3-8 membered cycloalkyl or heterocyclyl ring. 1 and R 5 Together with the atoms to which they are attached, they form a 6-8 membered cycloalkyl or heterocyclyl ring. 2 and R 5 Together with the atoms to which they are attached they form a 5-8 membered cycloalkyl or heterocyclyl ring.

[0084] In certain embodiments, Y is a monocyclic heteroaryl, such as but not limited to pyridinyl, pyrazinyl, pyrimidinyl, and thiazolyl. In other embodiments, Y is a monocyclic heteroaryl, such as but not limited to pyridinyl, pyrazinyl, and pyrimidinyl. In some embodiments, Y is selected from triazene, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, and triazolyl. The monocyclic heteroaryl can be unsubstituted or substituted with one or more substituents selected from the following: alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxyl, cyano, halo, heteroaryl, nitro, sulfonamido, and thioalkyl. In another embodiment, Y is a monocyclic heteroaryl, which can be unsubstituted or substituted with one or more substituents selected from the following: alkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxyl, cyano, halo, aryl, heteroaryl, heterocyclic radical, nitro, sulfonamido, and thioalkyl.

[0085] In certain embodiments, the monocyclic heteroaryl is substituted with an aryl, heteroaryl, or heterocyclic group selected from the group consisting of phenyl, pyridyl, 2-hydroxypyridyl, piperidonyl, 2-hydroxy-1-methylpyridyl, triazolyl, imidazolidinonyl, pyrimidonyl, 2-hydroxyisoquinolyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl, and morpholinyl. In certain preferred embodiments, Y is a 6-membered monocyclic heteroaryl. In some embodiments, the monocyclic heteroaryl is substituted with a heteroaryl or heterocyclic group, wherein the heteroaryl or heterocyclic group is substituted with one or more substituents selected from the group consisting of halo, CN, alkyl, alkoxy, hydroxy, carboxyl, -CO2 alkyl, and tetrazolyl. In certain preferred embodiments, the monocyclic heteroaryl is positioned para to A relative to X.

[0086] In other embodiments, Y is a bicyclic heteroaryl, such as but not limited to benzothiazolyl, benzimidazolyl, benzoxazolyl, triazolopyridinyl, thiazolopyridinyl, quinolyl and quinoxalinyl. The bicyclic heteroaryl can be unsubstituted or substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, hydroxyalkyl, thioalkyl, alkoxy, alkoxycarbonyl, amide, carboxyl, cyano, halo, heteroaryl, nitro and sulfonamido. In certain embodiments, the bicyclic heteroaryl is unsubstituted or substituted by one or more substituents selected from the group consisting of thioalkyl, alkoxycarbonyl, amide, carboxyl, halo, and heteroaryl.

[0087] In certain embodiments, Y is a compound having the formula -C(O)NR 8 R 9 or -NR 9 C(O)R 10 is substituted with an amide substituent, wherein

[0088] R 8 and R 9are each independently selected from H, alkyl, heterocyclyl and heteroaryl; or R 8 and R 9 Together with the nitrogen atoms to which they are attached, they form

[0089] a 4-, 5-, 6-, or 7-membered heterocyclic or heteroaryl ring; and

[0090] R 10 It is an alkyl group.

[0091] In certain embodiments, Y is a compound having the formula -S(O)2NR 8 R 9 or -NR 9 S(O)2R 10 substituted with a sulfonylamino substituent; wherein

[0092] R 8 and R 9 are each independently selected from H, alkyl, and heteroaryl; or

[0093] R 8 and R 9 Together with the nitrogen atoms to which they are attached, they form

[0094] a 4-, 5-, 6-, or 7-membered heterocyclic ring; and

[0095] R 10 It is an alkyl group.

[0096] All aforementioned embodiments of the variable Y in formula (I) are to be understood as also embodiments of the variable Het in formula (I′).

[0097] In certain embodiments, R 8 and R 9 Each is independently selected from H, methyl, ethyl, triazolyl, and pyrazolyl. 8 and R 9 In embodiments where one or both of R are alkyl, each alkyl is independently unsubstituted or substituted with one or more substituents selected from the group consisting of methyl, methoxy, carboxyl, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, oxadiazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl, and 2,4-dihydro-3H-1,2,4-triazol-3-onyl. In certain embodiments, R 8 and R 9 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from the group consisting of aziridine, isothiazolidine-1,1-dioxide, azetidine, thiazol-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, 2-oxa-6-azaspiro[3.3]heptane. In some embodiments, R 8and R 9 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from the group consisting of 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]decan-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9 -diazaspiro[5.5]undecan-3-one, tetrahydropyrrolo[3,4-d]imidazole, pyrimidine, 8-oxa-2-azaspiro[4.5]decane, hexahydro-3H-oxazolo[3,4-a]pyrazin-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 -azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, oxadiazole, triazole, pyridone, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3(2H)-one, piperidone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5]nonane, pyrazole, and pyridazin-3(2H)-one.

[0098] In some embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxyl, and heterocyclic. In certain embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from the group consisting of methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxyl, and oxetane.

[0099] In certain embodiments, the present invention provides pharmaceutical formulations suitable for use in human patients, comprising any of the compounds shown above (e.g., compounds of the present invention, such as compounds of formula (I)) and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical formulations can be used to treat or prevent the conditions or diseases described herein.

[0100] Any of the disclosed compounds may be used in the preparation of a medicament for the treatment of any of the diseases or conditions disclosed herein.

[0101] definition

[0102] Unless otherwise indicated, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art of the present disclosure. The following references provide a general definition of the multiple terms used in the present disclosure for technical personnel: Singleton et al., Dictionary of Microbiology and Molecular Biology [microbiology and molecular biology dictionary] (2nd edition 1994);The Cambridge Dictionary of Science and Technology [Cambridge dictionary of science and technology] (Walker writes, 1988);The Glossary of Genetics [genetics vocabulary], 5th edition, R. Rieger et al. (editor), Springer Verlag [Springer Press] (1991);And Hale and Marham, The HarperCollins Dictionary of Biology [HarperCollins biological dictionary] (1991). Unless otherwise indicated, the following terms as used herein have the following meanings given to them.

[0103] In this disclosure, “comprises,” “comprising,” “containing,” and “having,” etc. may have the meanings ascribed to them by U.S. patent law and may mean “includes,” “including,” etc.; “consisting essentially of or consists essentially of” also has the meaning ascribed to it by U.S. patent law and the term is open-ended, allowing for existence beyond what is recited as long as the basic or novel features recited are not altered by existence beyond what is recited, but excluding prior art embodiments.

[0104] Unless expressly stated or obvious from the context, as used herein, the term "or" is understood to be inclusive. Unless expressly stated or obvious from the context, as used herein, the terms "a," "an," and "the" are understood to be singular or plural.

[0105] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0106] The term "amido" is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbyl C(O)NH-.

[0107] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbyl C(O)O-, preferably alkyl C(O)O-.

[0108] 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.

[0109] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula alkyl-O-alkyl.

[0110] As used herein, the term "alkenyl" refers to an aliphatic group containing at least one double bond, and is intended to include "unsubstituted alkenyls" and "substituted alkenyls," the latter of which refers to alkenyl moieties having substituents replacing hydrogen on one or more carbons of the alkenyl group. Such substituents may occur on one or more carbons that are or are not contained in one or more double bonds. Moreover, as discussed below, such substituents include all substituents contemplated for alkyl groups, unless stability does not permit. For example, substitution of an alkenyl group with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0111] "Alkyl" or "alkane" is a fully saturated straight or branched chain non-aromatic hydrocarbon. Typically, unless otherwise defined, a straight or branched chain alkyl group has 1 to about 20 carbon atoms, preferably 1 to about 10 carbon atoms. Examples of straight and branched chain alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, amyl, and octyl. C1-C6 straight or branched chain alkyl groups are also referred to as "lower alkyl groups."

[0112] Furthermore, the term "alkyl" (or "lower alkyl") as used throughout the specification, examples, and claims is intended to include "unsubstituted alkyl" and "substituted alkyl," the latter referring to alkyl moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone. If not otherwise specified, such substituents may include, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that the moieties substituted on the hydrocarbon chain may themselves be substituted, if appropriate. For example, the substituents of the substituted alkyl group may include amino, azido, imino, amido, phosphoryl (including phosphonate and phosphite), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate) and silyl, as well as substituted and unsubstituted forms of ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate and ester), -CF3, -CN and the like. Exemplary substituted alkyl groups are described below. The cycloalkyl group may be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF3, -CN and the like.

[0113] The term "C x-y " when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is intended to include groups containing x to y carbons in the chain. For example, the term "C x-y "Alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight and branched chain alkyl groups containing x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. C0 alkyl represents hydrogen when the group is in the terminal position and a bond if it is internal. The term "C 2-y Alkenyl" and "C 2-y "Alkynyl" refers to a substituted or unsubstituted unsaturated aliphatic group analogous in length and possible substitution to the alkyls described above, but that contains at least one double or triple bond, respectively.

[0114] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0115] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and can be represented by the general formula alkylS-.

[0116] As used herein, the term "alkynyl" refers to an aliphatic group containing at least one triple bond, and is intended to include "unsubstituted alkynyls" and "substituted alkynyls," the latter of which refers to alkynyl moieties having substituents replacing a hydrogen on one or more carbons of the alkynyl group. Such substituents may be present on one or more carbons that are or are not contained in one or more triple bonds. Moreover, as discussed above, such substituents include all substituents contemplated for alkyl groups unless stability does not permit. For example, substitution of an alkynyl group with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0117] As used herein, the term "amide" refers to the following group:

[0118]

[0119] Each R 11 independently represent hydrogen or a hydrocarbon group, or two R 11 Together with the N atom to which they are attached, the heterocycles complete the ring structure having from 4 to 8 atoms.

[0120] The terms "amine" and "amino" are art-recognized and refer to unsubstituted and substituted amines and salts thereof, for example, moieties that can be represented by the formula

[0121]

[0122] Each R 11 independently represent hydrogen or a hydrocarbon group, or two R 11 Together with the N atom to which they are attached, they complete the heterocycle having from 4 to 8 atoms in the ring structure. As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0123] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0124] 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 rings, wherein two or more carbon atoms are shared by two adjacent rings, at least one of which is aromatic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl includes benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0125] The term "carbamate" is art-recognized and refers to the following group:

[0126]

[0127] where R 11and R 12 independently represents hydrogen or a hydrocarbon group, such as an alkyl group, or R 11 and R 12 Together with one or more intermediate atoms, the heterocycle has from 4 to 8 atoms in the ring structure.

[0128] As used herein, the terms "carbocycle" and "carbocyclic" refer to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocycle includes aromatic carbocycles and non-aromatic carbocycles. Non-aromatic carbocycles include cycloalkane rings in which all carbon atoms are saturated, and cycloalkene rings containing at least one double bond.

[0129] The term "carbocycle" includes 5-7 membered monocyclic rings and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with the other ring. Each ring of a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, such as a phenyl ring, can be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings, where valence permits, is included in the definition of carbocycle. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles 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]hept-3-ene. "Carbocycle" may be substituted at any position or positions capable of bearing a hydrogen atom.

[0130] A "cycloalkyl" group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Unless otherwise defined, monocyclic cycloalkyl groups typically have 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms. The second ring of a bicyclic cycloalkyl group can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl groups include bicyclic molecules in which the two rings share one, two, or three or more atoms. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl group in which each ring shares two adjacent atoms with another ring. The second ring of a fused bicyclic cycloalkyl group can be selected from saturated, unsaturated, and aromatic rings. A "cycloalkenyl" group is a cyclic hydrocarbon containing one or more double bonds.

[0131] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0132] The term "carbonate" is art-recognized and refers to the group -OCO2-R 10, where R 10 Represents a hydrocarbon group.

[0133] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0134] As used herein, the term "ester" refers to the group -C(O)OR 10 , where R 10 Represents a hydrocarbon group.

[0135] As used herein, the term "ether" refers to a hydrocarbon group attached to another hydrocarbon group through an oxygen. Thus, an ether substituent of a hydrocarbon group can be hydrocarbon-O-. Ethers can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which can be represented by the general formula alkyl-O-alkyl.

[0136]

[0046] The terms "halo" and "halogen" as used herein mean halogen and include chlorine, fluorine, bromine and iodine.

[0137] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a heteroaryl group.

[0138] As used herein, the term "heteroalkyl" refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, wherein no two heteroatoms are adjacent.

[0139] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic structures, preferably 5 to 7 membered rings, more preferably 5 to 6 membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more rings, wherein two or more carbon atoms are common to two adjacent rings, wherein at least one ring is heteroaromatic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine.

[0140] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0141] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more rings, wherein two or more carbon atoms are shared by two adjacent rings, wherein at least one ring is heterocyclic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0142] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0143] As used herein, the term "hydrocarbyl" refers to a group that is bonded to a carbon atom that does not have a =O or =S substituent and typically has at least one carbon-hydrogen bond and a primary carbon backbone, but may optionally include heteroatoms. Thus, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-nitrophenyl (pyridyl), and trifluoromethyl are considered to be hydrocarbyl groups, but substituents such as acetyl (which has a =O substituent on the connecting carbon) and ethoxy (which is connected through oxygen rather than carbon) are not hydrocarbyl groups. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocycle, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

[0144] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0145] The term "lower" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is intended to include groups in which the substituent has ten or fewer non-hydrogen atoms, preferably six or fewer non-hydrogen atoms. For example, "lower alkyl" refers to an alkyl group containing ten or fewer carbon atoms, preferably six or fewer carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent as defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy group, respectively, whether alone or in combination with other substituents, such as in the description of hydroxyalkyl and aralkyl groups (in which case, for example, when counting the carbon atoms in the alkyl substituent, the atoms in the aryl group are not counted).

[0146] The terms "polycyclic group," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are shared by two adjacent rings, e.g., the rings are "fused rings." Each ring of the polycyclic ring may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring contains 3 to 10 atoms, preferably 5 to 7 atoms, in the ring.

[0147] The term "silyl" refers to a silicon moiety having three hydrocarbyl moieties attached thereto.

[0148] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbon atoms of the main chain. It will be understood that "substituted" or "substituted by" includes implicit conditions, i.e., such substitutions are consistent with the allowed valences of the substituted atom and substituent, and that the substitution produces a stable compound, e.g., a compound that does not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all allowed substituents of an organic compound. In a broad sense, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. For appropriate organic compounds, allowed substituents may be one or more and the same or different. For the purposes of the present invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any allowed substituents of the organic compound described herein that satisfy the heteroatom valences. Substituents may include any substituent described herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl or aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the substituents themselves may be substituted if appropriate. Unless explicitly stated as "unsubstituted", references to chemical moieties herein are understood to include substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0149] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0150] The term "sulfonamide" is art-recognized and refers to a group represented by the following general formula:

[0151]

[0152] where R 11 and R 12independently represents hydrogen or a hydrocarbon group, such as an alkyl group, or R 11 and R 12 Together with the intermediate atoms, the heterocycle has 4 to 8 atoms in the ring structure.

[0153] The term "sulfoxide" is art-recognized and refers to the group -S(O)-R 10 , where R 10 Represents a hydrocarbon group.

[0154] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0155] The term "sulfone" is art-recognized and refers to the group -S(O)2-R 10 , where R 10 Represents a hydrocarbon group.

[0156] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0157] As used herein, the term "thioester" refers to the group -C(O)SR 10 or -SC(O)R 10 , where R 10 Represents a hydrocarbon group.

[0158] As used herein, the term "thioether" is equivalent to an ether in which the oxygen is replaced by sulfur.

[0159] The term "urea" is art-recognized and can be represented by the following general formula:

[0160]

[0161] where R 11 and R 12 independently represents hydrogen or a hydrocarbon group, such as an alkyl group, or any occurrence of R 11 With R 12 Together with one or more intermediate atoms, the heterocycle completes the ring structure having from 4 to 8 atoms.

[0162] The term "protecting group" refers to a reactive atomic group that masks, reduces or prevents the reactive functional group of a molecule when attached to a reactive functional group in the molecule. Typically, protecting groups can be selectively removed as needed during the building-up process. Examples of protecting groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry [protecting groups in organic chemistry], 3rd edition, 1999, John Wiley & Sons, New York, and Harrison et al., Compendium of Synthetic Organic Methods [synthetic organic method compendium], 1-8 volumes, 1971-1996, John Wiley & Sons, New York, NY. Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butyloxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethoxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC"), etc. Representative hydroxy protecting groups include, but are not limited to, those in which the hydroxy 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.

[0163] The present invention includes all pharmaceutically acceptable isotopically labeled compounds as described herein, wherein one or more atoms are replaced by atoms having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. In certain embodiments, the compounds of the present invention are enriched in such isotopically labeled substances (e.g., compounds wherein the isotopic distribution of the compound in the composition is different from the natural or typical distribution of isotopes).

[0164] Examples of suitable isotopes for inclusion in the compounds of the present invention include isotopes of hydrogen, such as 2 H and 3 H; carbon, such as 11 C. 13 C and 14 C; chlorine, such as 36 Cl; fluorine, such as 18 F; iodine, such as 123 I and 125 I; nitrogen, such as 13 N and 15 N; oxygen, such as 15 O. 17 O and 18O; phosphorus, such as 32 P; and sulfur, such as 35 S.

[0165] Certain isotopically labeled compounds disclosed herein, for example those incorporating radioactive isotopes, are useful in drug and / or substrate tissue distribution studies. 3 H) and carbon-14 (i.e. 14 C), are useful for this purpose due to their ease of incorporation and ready means of detection.

[0166] Using heavier isotopes such as deuterium (i.e. 2 H) substitutions may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances.

[0167] Using positron-emitting isotopes such as 11 C. 18 F. 15 O and 13 N) substitutions can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0168] In certain embodiments, the compounds of the present invention may be racemic. In certain embodiments, the compounds of the present invention may be enriched in one enantiomer. For example, the compounds of the present invention may have greater than about 30% ee, about 40% ee, about 50% ee, about 60% ee, about 70% ee, about 80% ee, about 90% ee, or even about 95% or higher ee. In certain embodiments, the compounds of the present invention may have more than one stereocenter. In certain such embodiments, the compounds of the present invention may be enriched in one or more diastereomers. For example, the compounds of the present invention may have greater than about 30% de, about 40% de, about 50% de, about 60% de, about 70% de, about 80% de, about 90% de, or even about 95% or higher de.

[0169] In certain embodiments, therapeutic preparations can be enriched to provide primarily one enantiomer of a compound (e.g., a compound with formula (I)). An enantiomerically enriched mixture can include, for example, at least about 60 mol % of one enantiomer, or more preferably at least about 75 mol %, about 90 mol %, about 95 mol % or even about 99 mol %. In certain embodiments, a compound enriched in one enantiomer is substantially free of another enantiomer, wherein substantially free means that the substance in question accounts for 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 about 1% compared to the amount of another enantiomer in, for example, a composition or compound mixture. For example, if a composition or compound mixture contains about 98 grams of the first enantiomer and about 2 grams of the second enantiomer, it can be said to contain about 98 mol % of the first enantiomer and only about 2% of the second enantiomer.

[0170] In certain embodiments, the therapeutic formulation can be enriched to provide primarily one diastereomer of a compound (e.g., a compound of Formula (I)). A diastereomerically enriched mixture can contain, for example, at least about 60 mole % of one diastereomer, or more preferably at least about 75 mole %, about 90 mole %, about 95 mole % or even about 99 mole %.

[0171] The term "subject" (to which administration is intended) includes, but is not limited to, humans (i.e., male or female of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young, middle-aged, or elderly)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds such as chickens, ducks, geese, quail, and / or turkeys. Preferred subjects are humans.

[0172] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in a treated sample relative to an untreated control sample, or delays the onset of, or reduces the severity of, one or more symptoms of the disorder or condition relative to an untreated control sample.

[0173] The term "treatment" includes preventive and / or therapeutic treatments. The term "preventive or therapeutic" treatment is well-known in the art and includes administering one or more disclosed compositions to a subject. If administered before clinically demonstrating an undesirable condition (e.g., a disease or other undesirable state of the subject), the treatment is preventive (i.e., it protects the subject from developing an undesirable condition), while if administered after demonstrating an undesirable condition, the treatment is therapeutic (i.e., it is intended to shorten, improve, or stabilize the undesirable condition or its side effects).

[0174] The term "prodrug" is intended to encompass compounds (e.g., compounds of formula (I)) that are converted under physiological conditions into therapeutic agents of the present invention. A common method for preparing a prodrug is to include one or more selected moieties that are hydrolyzed under physiological conditions to present the desired molecule. In other embodiments, the prodrug is converted by the enzymatic 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 above-mentioned formulations can be replaced with corresponding suitable prodrugs, for example, wherein the hydroxyl group in the parent compound is presented as an ester or carbonate or carboxylic acid.

[0175] As used herein, an "effective amount" refers to an amount sufficient to achieve a desired biological effect. As used herein, a "therapeutically effective amount" refers to an amount sufficient to achieve a desired therapeutic effect. For example, a therapeutically effective amount can refer to an amount sufficient to ameliorate at least one sign or symptom of cancer.

[0176] A "response" to a treatment method can include a decrease or improvement in negative symptoms, a decrease in progression of the disease or its symptoms, an increase in beneficial symptoms or clinical outcomes, a decrease in side effects, stabilization of the disease, partial or complete remission of the disease, etc.

[0177] How to use

[0178] The PCSK9 gene was identified using genetic mapping techniques from DNA of subjects with autosomal dominant hypercholesterolemia (Abifadel et al. Nat. Genet. [Natural Genetics] 2003 34: 154-6). The encoded protein is a serine protease that is primarily expressed in the liver, intestines, kidneys, and nervous system. While not wishing to be bound by any particular theory, studies of mutations in the gene indicate that its putative effect is to reduce LDL receptors on the cell surface without relying on their catalytic activity. (Abifadel et al. Expert Opin. Ther. Pat. [Expert Comments on Therapeutic Patents] 2010 20: 1547-71). The binding of PCSK9 to receptors results in 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 is downregulated by dietary cholesterol, glucagon, ethinyl estradiol, 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). Because elevated levels of PCSK9 reduce the abundance of LDL receptors on the cell surface, increasing statin doses fail to achieve proportional LDL-cholesterol lowering results. Thus, disclosed herein are methods for treating a wide range of cardiovascular diseases and conditions that would benefit from inhibition of PCSK9, thereby lowering LDL-c.

[0179] In certain embodiments, methods of inhibiting PCSK9 are performed in a subject in need thereof, thereby treating a PCSK9-mediated disease or disorder.

[0180] In addition, disclosed herein are methods for treating or preventing a PCSK9-mediated disease or disorder, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, disclosed herein are methods for treating a PCSK9-mediated disease or disorder, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, disclosed herein are methods for preventing a PCSK9-mediated disease or disorder, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. Preventing cardiovascular events by inhibiting PCSK9 has been described, for example, in Robinson et al., Artherosclerosis [arthritis] 2015243:593-597.

[0181] Exemplary cardiovascular diseases and disorders include, but are not limited to, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, hypoalphalipoproteinemia, metabolic syndrome, 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.

[0182] In certain embodiments, exemplary cardiovascular diseases and conditions include, but are not limited to, hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome, 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.

[0183] In certain embodiments, the disclosed methods of treatment can reduce elevated levels of circulating serum cholesterol, such as LDL-cholesterol and VLDL-cholesterol. Furthermore, the disclosed methods can be used to reduce 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 plaque formation. Subjects with gain-of-function mutations in the PCSK9 gene also benefit from treatment with the disclosed compounds and compositions to counteract the mutation by inhibiting PCSK9.

[0184] PCSK9 inhibition has also been shown to have therapeutic benefits in treating sepsis in subjects. Septic shock is a usually fatal complication of severe microbial infection (sepsis), triggering an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis originates from microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharide (LPS; Gram-negative bacteria). LPS is an effective ligand for mammalian innate immune receptors [Toll-like receptors (TLRs)] and therefore plays a prominent role in septic inflammatory responses (septic shock or sepsis). PCSK9 reduces LPS absorption through the liver's LDL receptors, causing free LPS to overstimulate the body's immune response to the pathogens that cause sepsis. Inhibiting PCSK9 is beneficial in maintaining liver LDL receptors to achieve systemic pathogen clearance and detoxification in response to sepsis (see, for example, Walley et al. Sci. Translat. Med. [Science Translational Medicine] 20146: 1-10).

[0185] Disclosed herein are methods for treating sepsis or septic shock comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, the disclosed treatment methods can be used to increase LPS absorption. Certain embodiments provide methods for reducing the inflammatory response induced by sepsis or septic shock.

[0186] Combination therapy

[0187] The disclosed compounds and compositions can be used in combination with other therapeutic agents, such as other agents suitable for treating high levels of LDL and triglycerides. In certain embodiments, one or more additional therapeutic agents are administered in combination with the compounds of the invention to provide a synergistic effect. In certain embodiments, one or more additional therapeutic agents are administered in combination to provide an additive effect.

[0188] In some embodiments, the method for treating a disease or disorder mediated by PCSK9 comprises administering a PCSK9 inhibitor in combination with one or more other therapeutic agents. In some embodiments, the PCSK9 inhibitor is a compound of formula (I). Other therapeutic agents may include monoclonal antibodies such as alizumab, evolocumab, boxilimab, RG7652, LY3015014, and mAb316P. Additional exemplary antibodies include WO / 2015 / 140079, WO / 2015 / 142668, WO / 2015 / 123423, WO / 2015 / 073494, WO / 2015 / 054619, WO / 2014 / 197752, WO / 2014 / 194111, WO / 2014 / 194168, WO / 2014 / 02835 4. WO / 2013 / 039969, WO / 2013 / 016648, WO / 2012 / 101251, WO / 2012 / 101253, WO / 2012 / 1 01252、WO / 2014 / 209384、WO / 2014 / 150983、WO / 2014 / 144080、WO / 2013 / 166448、WO / 201 2 / 154999、WO / 2013 / 008185、WO / 2015 / 200438、WO / 2014 / 107739、WO / 2013 / 188855、WO / 2013 / 169886、WO / 2013 / 148284、WO / 2013 / 091103、WO / 2013 / 039958、WO / 2012 / 177741 , WO / 2012 / 168491, WO / 2012 / 170607, WO / 2012 / 109530, WO / 2012 / 088313, WO / 2012 / 054438, WO / 2011 / 072263, WO / 2011 / 053759, WO / 2011 / 053783, and WO / 2011 / 037791. Other therapeutic agents that can be used in combination with the disclosed compounds and compositions include plant alkaloids and flavonoids known to have cholesterol-lowering activity, such as berberine and quercetin. In some embodiments, cholesterol-lowering small molecules, such as ezetimibe and policosanol, can be administered as additional therapeutic agents. In certain embodiments, polypeptides such as BMS-962476 and fibronectin-based scaffold domain proteins can be administered in combination with the disclosed compounds and compositions.Other exemplary compounds include those described in WO / 2014 / 150326, WO / 2014 / 150395, WO / 2014 / 139008, WO / 2014 / 140210, WO / 2014 / 127316, WO / 2014 / 005224, WO / 2013 / 177536, WO / 2011 / 152508, WO / 2011 / 130354, WO / 2011 / 006000, and WO / 2015 / 077154.

[0189] In certain embodiments, the additional therapeutic agent may be a statin, such as atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin. In other embodiments, the disclosed compounds may be administered in combination with HMG-CoA reductase.

[0190] Any HMG-CoA reductase inhibitor may be used in the combination aspects of the present invention. The conversion of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) to methanol is an early and rate-limiting step in the cholesterol biosynthetic pathway. This step is catalyzed by the enzyme HMG-CoA reductase. Statins inhibit HMG-CoA reductase from catalyzing this conversion.

[0191] The term HMG-CoA reductase inhibitor refers to compounds that inhibit the bioconversion of hydroxymethylglutaryl-CoA to mevalonate catalyzed by HMG-CoA reductase. Such inhibition is readily determined by one skilled in the art according to standard assays (e.g., Meth. Enzymol. 1981; 71:455-509 and references cited therein). A variety of these compounds are described and mentioned below, however, other HMG-CoA reductase inhibitors will be known to one skilled in the art. U.S. Patent No. 4,231,938 (the disclosure of which is hereby incorporated by reference) discloses certain compounds, such as lovastatin, isolated after culturing microorganisms belonging to the genus Aspergillus. In addition, U.S. Patent No. 4,444,784 (the disclosure of which is hereby incorporated by reference) discloses synthetic derivatives of the above compounds, such as simvastatin. In addition, U.S. Patent No. 4,739,073 (the disclosure of which is hereby incorporated by reference) discloses certain substituted indoles, such as fluvastatin. In addition, U.S. Patent No. 4,346,227 (the disclosure of which is incorporated herein by reference) discloses ML-236B derivatives, such as pravastatin. In addition, EP-491226A (the disclosure of which is incorporated herein by reference) discloses certain nitrogen phenyl dihydroxy heptenoic acids, such as cerivastatin. In addition, U.S. Patent No. 5,273,995 (the disclosure of which is incorporated herein by reference) discloses certain 6-[2-(substituted pyrrol-1-yl)alkyl] pyran-2-ones, such as atorvastatin and any pharmaceutically acceptable form thereof (i.e., ). Additional HMG-CoA reductase inhibitors include rosuvastatin and pitavastatin.

[0192] Atorvastatin calcium (i.e., atorvastatin hemi-calcium), disclosed in U.S. Patent No. 5,273,995, incorporated herein by reference, is currently available as sell.

[0193] Statins also include compounds such as rosuvastatin disclosed in USRE 37,314E; pitavastatin disclosed in EP 304063 BI and US5,011,930; simvastatin disclosed in US4,444,784, incorporated herein by reference; pravastatin disclosed in US4,346,227, incorporated herein by reference; cerivastatin disclosed in US5,502,199, incorporated herein by reference; mevastatin disclosed in US3,983,140, ​​incorporated herein by reference; and velostatin disclosed in US4,448,784 and US4,450,171, both incorporated herein by reference. statins; fluvastatin disclosed in U.S. Pat. No. 4,739,073, incorporated herein by reference; compactin disclosed in U.S. Pat. No. 4,804,770, incorporated herein by reference; lovastatin disclosed in U.S. Pat. No. 4,231,938, incorporated herein by reference; darvastatin disclosed in European Patent Application Publication No. 738510A2; fluindostatin disclosed in European Patent Application Publication No. 363934A1; and dihydrocompatin disclosed in U.S. Pat. No. 4,450,171, incorporated herein by reference.

[0194] Any HMG-CoA synthase inhibitor may be used in the combination aspects of the present invention. The term HMG-CoA synthase inhibitor refers to a compound that inhibits the biosynthesis of hydroxymethylglutaryl-CoA from acetyl-CoA and acetoacetyl-CoA catalyzed by the enzyme HMG-CoA synthase. Such inhibition is readily determined by one skilled in the art according to standard assays (Meth. Enzymol. 1975; 35: 155-160; Meth. Enzymol. 1985; 110: 19-26 and references cited therein). A variety of these compounds are described and referenced below, however, other HMG-CoA synthase inhibitors will be known to one skilled in the art. U.S. Patent No. 5,120,729 (the disclosure of which is hereby incorporated by reference) discloses certain β-lactam derivatives. U.S. Patent No. 5,064,856 (the disclosure of which is hereby incorporated by reference) discloses certain spiro-lactone derivatives prepared by culturing a microorganism (MF5253). US Patent No. 4,847,271, the disclosure of which is hereby incorporated by reference, discloses certain oxetane compounds, such as 11-(3-hydroxymethyl-4-oxo-2-oxetanyl)-3,5,7-trimethyl-2,4-undecane-dienoic acid derivatives.

[0195] Any compound that reduces HMG-CoA reductase gene expression can be used in the combination aspects of the present invention. These agents can be HMG-CoA reductase transcription inhibitors that block DNA transcription or translation inhibitors that prevent or reduce the translation of mRNA encoding HMG-CoA reductase into protein. Such compounds can directly affect transcription or translation, or can be bioconverted to compounds with the above-mentioned activities by one or more enzymes in the cholesterol biosynthesis cascade, or can cause the accumulation of isoprene metabolites with the above-mentioned activities. These compounds can cause this effect by inhibiting the activity of site-1 protease (SIP) or by stimulating oxysterol receptors or antagonizing SCAP, thereby reducing the levels of SREBP (sterol regulatory element binding protein). Such modulation is readily determined by those skilled in the art using standard assays (Meth. Enzymol. 1985; 110: 9-19). Several compounds are described and mentioned below, however, those skilled in the art will know other inhibitors of HMG-CoA reductase gene expression. U.S. Patent No. 5,041,432 (the disclosure of which is incorporated by reference) discloses certain 15-substituted lanosterol derivatives.

[0196] El Mercer (Prog. Lip. Res. 1993; 32: 357-416) discusses other oxysterols that inhibit HMG-CoA reductase synthesis.

[0197] Any compound having activity as a CETP inhibitor can be used as the second compound in the combination therapy aspect of the present invention. The term CETP inhibitor refers to a compound that inhibits the cholesteryl ester transfer protein (CETP)-mediated transport of various cholesterol 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). A variety of CETP inhibitors are known to those skilled in the art, for example, those disclosed in co-assigned U.S. Patent No. 6,140,343 and co-assigned U.S. Patent No. 6,197,786. CETP inhibitors are also described in U.S. Patent No. 6,723,752, which include a variety of 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. In addition, CETP inhibitors encompassed herein are also described in U.S. Patent Application No. 10 / 807,838, filed March 23, 2004. U.S. Patent No. 5,512,548 discloses certain polypeptide derivatives having activity as CETP inhibitors, and certain CETP-inhibiting rosenone lactone derivatives and phosphate-containing cholesteryl ester analogs are disclosed in J. Antibiot, 49(8)815-816 (1996) and Bioorg. Med. Chem. Lett., 6:1951-1954 (1996), respectively.

[0198] Any PPAR modulator can be used in the combination aspects of the present invention. The term PPAR modulator refers to a compound that modulates the activity of peroxisome proliferator-activated receptors (PPARs) in mammals, particularly humans. Such modulation is readily determined by those skilled in the art using standard assays known in the literature. It is believed that by modulating PPAR receptors, such compounds modulate the transcription of key genes involved in lipid and glucose metabolism, such as key genes involved in fatty acid oxidation and key genes involved in high-density lipoprotein (HDL) assembly (e.g., apolipoprotein A1 gene transcription), thereby reducing whole body fat and increasing HDL cholesterol. Through their activity, these compounds also reduce plasma levels of triglycerides, VLDL cholesterol, LDL cholesterol, and their related components, such as apolipoprotein B, in mammals, particularly humans, as well as increase HDL cholesterol and apolipoprotein A1. Therefore, these compounds can be used to treat and correct various dyslipidemias observed to be associated with the development and occurrence of atherosclerosis and cardiovascular disease, including hypoalphalipoproteinemia and hypertriglyceridemia. A variety of these compounds are described and mentioned below, but others will be known to those skilled in the art. International Publication Nos. WO 02 / 064549 and 02 / 064130 and U.S. Patent Application Nos. 10 / 720942, filed November 24, 2004, and 60 / 552114, filed March 10, 2004, the disclosures of which are hereby incorporated by reference, disclose certain compounds that are PPARa activators.

[0199] Any other PPAR modulator may be used in the combination aspects of the present invention. In particular, modulators of PPARp and / or PPARy may be used in combination with the compounds of the present invention. An exemplary PPAR inhibitor, {5-methoxy-2-methyl-4-[4-(4-trifluoromethyl-benzyloxy)-benzylsulfanyl]-phenoxy}-acetic acid, is described in US 2003 / 0225158.

[0200] Any MTP / Apo B (microsomal triglyceride transfer protein and or apolipoprotein B) secretion inhibitor can be used in the combination aspects of the present invention. The term MTP / Apo B secretion inhibitor refers to a compound that inhibits the secretion of triglycerides, cholesterol 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 are known to those skilled in the art, including implitapide (Bayer) and other compounds, such as those disclosed in WO 96 / 40640 and WO 98 / 23593 (two exemplary publications).

[0201] Any squalene synthase inhibitor can be used in the combination aspects of the present invention. The term squalene synthase inhibitor refers to a compound that inhibits the condensation of two molecules of farnesyl pyrophosphate to form squalene by the enzyme squalene synthase. Such inhibition is readily determined by one skilled in the art according to standard assays (Meth. Enzymol. 1969; 15: 393-454; and Meth. Enzymol. 1985; 110: 359-373 and references therein). A variety of these compounds are described and mentioned below, but other squalene synthase inhibitors will be known to one skilled in the art. U.S. Patent No. 5,026,554 (the disclosure of which is incorporated by reference) discloses fermentation products of the microorganism MF5465 (ATCC 74011), including squalene synthase. An overview of other patented squalene synthase inhibitors has been compiled (Curr. Op. Then Patents (1993) 861-4).

[0202] Any squalene epoxidase inhibitor can be used in the combination aspects of the present invention. The term squalene epoxidase inhibitor refers to a compound that inhibits the bioconversion of squalene and molecular oxygen 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 [Biochemistry and Biophysics Acta] 1984; 794: 466-471). A variety of these compounds are described and mentioned 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 (the disclosures of which are incorporated by reference) disclose certain fluoro analogs of squalene. EP Publication 395,768A (the disclosures of which are incorporated by reference) discloses certain substituted allylamine derivatives. PCT Publication WO 9312069 A (the disclosures of which are hereby incorporated by reference) discloses certain amino alcohol derivatives. US Patent No. 5,051,534, the disclosure of which is hereby incorporated by reference, discloses certain cyclopropyloxy-squalene derivatives.

[0203] Any squalene cyclase inhibitor may be used as the second component of the combination aspects of the present invention. The term squalene cyclase inhibitor refers to a compound that inhibits the bioconversion of squalene-2,3-epoxide to lanosterol catalyzed by squalene cyclase. Such inhibition is readily determined by one skilled in the art according to a standard assay (FEBS Lett. [Federation of European Biochemical Societies Letters] 1989; 244: 347-350). In addition, the compounds described and referred to below are squalene cyclase inhibitors, but other squalene cyclase inhibitors will also be known to those skilled in the art. Pd publication WO9410150 (the disclosure of which is hereby incorporated by reference) discloses certain 1,2,3,5,6,7,8,8a-octahydro-5,5,8(β)-trimethyl-6-isoquinolinamine derivatives, such as N-trifluoroacetyl-1,2,3,5,6,7,8,8a-octahydro-2-allyl-5,5,8(β)-trimethyl-6(β)-isoquinolinamine. French patent publication 2697250 (the disclosure of which is hereby incorporated by reference) discloses certain β,β-dimethyl-4-piperidinol derivatives, such as 1-(1,5,9-trimethyldecyl)-β,β-dimethyl-4-piperidinol.

[0204] Any combination of squalene epoxidase / squalene cyclase inhibitors can be used as the second component of the combination aspect 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, it is not possible to distinguish between squalene epoxidase inhibitors and squalene cyclase inhibitors; however, those skilled in the art recognize these assays. Therefore, those skilled in the art can easily determine the inhibition of a combined squalene epoxidase / squalene cyclase inhibitor based on the above-mentioned standard assays for squalene cyclase or squalene epoxidase inhibitors. A variety of these compounds are described and mentioned below, however, those skilled in the art will know other squalene epoxidase / squalene cyclase inhibitors. U.S. Patent Nos. 5,084,461 and 5,278,171 (the disclosures of which are incorporated by reference) disclose certain azadecalin derivatives. EP Publication 468,434 (the disclosure of which is hereby incorporated by reference) discloses certain piperidine ether and thioether derivatives, such as 2-(1-piperidinyl)pentyl isopentyl sulfoxide and 2-(1-piperidinyl)ethyl ethyl sulfide. PCT Publication WO 9401404 (the disclosure of which is hereby incorporated 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 (the disclosure of which is hereby incorporated by reference) discloses certain cyclopropyloxy-squalene derivatives.

[0205] The compounds of the present invention can also be administered in combination with naturally occurring compounds for lowering plasma cholesterol levels. These naturally occurring compounds are commonly referred to as nutritional supplements and include, for example, garlic extract and niacin. A slowly released form of niacin is available, called sustained-release niacin (Niaspan). Niacin can 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®. TM (Kos Pharmaceuticals Inc.).

[0206] Any cholesterol absorption inhibitor can be used as another compound of the combination aspect of the present invention. The term cholesterol absorption inhibition refers to the ability of a compound to prevent cholesterol contained in the intestinal lumen from entering the intestinal cells and / or passing through the intestinal cells into the lymphatic system and / or into the bloodstream. Those skilled in the art can easily determine this cholesterol absorption inhibition activity according to standard assays (e.g., J.LipidRes. [Journal of Lipid Research] (1993) 34:377-395). Cholesterol absorption inhibitors are known to those skilled in the art and are described, for example, in PCT WO 94 / 00480. An example of a cholesterol absorption inhibitor is ZETIA TM (ezetimibe) (Schering-Plough / Merck).

[0207] Any ACAT inhibitor can be used in the combination therapy aspects 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 one skilled in the art using standard assays, such as those described in Heider et al., Journal of Lipid Research., 24:1127 (1983). For example, a variety of such compounds are known to those skilled in the art; for example, U.S. Patent No. 5,510,379 discloses certain carboxysulfonates, while WO 96 / 26948 and WO 96 / 10559 both disclose urea derivatives with ACAT inhibitory activity. Examples of ACAT inhibitors include compounds such as avasimib (Pfizer), CS-505 (Sankyo), and eleumimib (Eli Lilly and Pierre Fabre).

[0208] Lipase inhibitors can be used in the combination therapy aspects 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 through a two-step process involving acylation of the activated serine portion of the lipase. This results in the production of a fatty acid-lipase hemiacetal intermediate, which is then cleaved to release diglycerides. After further deacetylation, the lipase-fatty acid intermediate is cleaved to obtain free lipase, glycerides, and fatty acids. In the intestine, the resulting free fatty acids and monoglycerides are incorporated into bile acid-phospholipid micelles and subsequently absorbed at the level of the brush border of the small intestine. The micelles eventually enter the peripheral circulation of chylomicrons. Those skilled in the art can easily determine this lipase inhibitory activity according to standard assays (e.g., Methods Enzymol. [Enzymology Methods] 286: 190-231).

[0209] Pancreatic lipase mediates the metabolic cleavage of fatty acids from the triglyceride at 1- and 3-carbon positions. The original site of intake fat metabolism by pancreatic lipase is the duodenum and proximal jejunum, and the secretion of this pancreatic lipase usually far exceeds the amount required for fat decomposition in the upper part of the small intestine. Because pancreatic lipase is the primary enzyme required for absorbing dietary triglycerides, inhibitors have effectiveness in treating obesity and other related conditions. Those skilled in the art easily determine this pancreatic lipase inhibitory activity according to standard assays (for example, Methods Enzymol. [enzymology method] 286: 190-231).

[0210] Gastric lipase is an immunologically different lipase that is responsible for approximately 10% to 40% of dietary fat digestion. Gastric lipase is secreted in response to mechanical stimulation, food intake, the presence of a fat meal, or sympathetic nerve stimulation factors. The gastric lipolysis of fat intake has physiological importance in providing the fatty acid required for the pancreatic lipase activity in the intestine, and is also important for fat absorption under the various physiological and pathological conditions related to pancreatic insufficiency. For example, referring to C.K. Abrams et al., Gastroenterology [gastroenterology], 92,125 (1987). Those skilled in the art can easily determine that this gastric lipase inhibits activity according to standard assays (for example, Methods Enzymol. [enzymology method] 286:190-231).

[0211] Many gastric and / or pancreatic lipase inhibitors are known to those of ordinary skill in the art. Preferred lipase inhibitors are selected from lipstatin, tetrahydrolipstatin (orlistat), valinolide, esterase inhibitor, erbinolactone A, and erbinolactone B. The compound tetrahydroabscisin is particularly preferred. U.S. Patent No. 4,405,644 discloses the lipase inhibitor N-3-trifluoromethylphenyl-N'-3-chloro-4'-trifluoromethylphenyl urea and various urea derivatives related thereto. U.S. Patent Nos. 4,189,438 and 4,242,453 disclose the lipase inhibitor esteracin. The lipase inhibitor cyclo-0,0'-[(1,6-hexanediyl-1)-bis-(iminocarbonyl)]dioxime and various (iminocarbonyl)dioximes related thereto can be prepared as described by Petersen et al., Liebig's Annalen, 562, 205-229 (1949).

[0212] Various pancreatic lipase inhibitors are described below. U.S. Pat. No. 4,598,089 discloses the pancreatic lipase inhibitor lipstatin, i.e., (2S,3S,5S,7Z,10Z)-5-[(S)-2-formamido-4-methyl-pentanoyloxy]-2-hexyl-3-hydroxy-7,10-hexadecanoic acid lactone, and tetrahydrolipstatin (orlistat), i.e., (2S,3S,5S)-5-[(S)-2-formamido-4-methyl-pentanoyloxy]-2-hexyl-3-hydroxy-hexadecanoic acid lactone, as well as various substituted N-formylleucine derivatives and stereoisomers thereof. For example, tetrahydrolipstatin can be prepared as described in, for example, U.S. Pat. Nos. 5,274,143; 5,420,305; 5,540,917; and 5,643,874. U.S. Patent No. 4,452,813 discloses a pancreatic lipase inhibitor FL-386, 1-[4-(2-methylpropyl)cyclohexyl]-2-[(phenylsulfonyl)oxy]-ethanone and various substituted sulfonated derivatives thereof. U.S. Patent Nos. 5,512,565, 5,391,571 and 5,602,151 disclose a pancreatic lipase inhibitor WAY-121898, 4-phenoxyphenyl-4-methylpiperidin-1-yl-carboxylate and various carbamates and pharmaceutically acceptable salts thereof. Kitahara et al., J. Antibiotics, 40(11), 1647-1650 (1987) discloses an insulin inhibitor valaminolactone and a method for preparing it by microbial cultivation of the actinomycete strain MG147-CF2. Umezawa et al., J. Antibiotics, 33, 1594-1596 (1980) disclose insulin inhibitors erbinolone A and erbinolone B and a method for preparing them by microbial cultivation of actinomycete strain MG7-G1. Japanese Kokai Publication No. 08-143457, published on June 4, 1996, discloses the use of erbinolone A and B for inhibiting monoglyceride formation.

[0213] Other compounds marketed for hyperlipidemia including hypercholesterolemia and intended to help prevent or treat atherosclerosis include bile acid sequestrants, e.g. and and fibric acid derivatives, e.g. and

[0214] In view of the association between diabetes and atherosclerosis (e.g., metabolic syndrome), compounds of Formula I can be used together with antidiabetic compounds. Diabetes can be treated by administering a therapeutically effective amount of a compound of the present invention to a patient suffering from diabetes (especially type II), insulin resistance, impaired glucose tolerance, metabolic syndrome, etc., or any diabetic complications such as neuropathy, nephropathy, retinopathy, or cataracts, in combination with other medicaments (e.g., insulin) that can be used to treat diabetes. This includes the categories of antidiabetic agents (and specific agents) described herein.

[0215] Any glycogen phosphorylase inhibitor can 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 catalyzed by the enzyme glycogen phosphorylase. Such glycogen phosphorylase inhibitory activity is readily determined by one skilled in the art according to standard assays (e.g., J. Med. Chem. 41 (1998) 2934-2938). Various glycogen phosphorylase inhibitors are known to those skilled in the art, including those described in WO 96 / 39384 and WO 96 / 39385.

[0216] Any aldose reductase inhibitor can 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 into sorbitol by the enzyme aldose reductase catalysis. Those skilled in the art can easily determine such aldose reductase inhibition according to standard assays (e.g., J. Malone, Diabetes [diabetes], 29: 861-864 (1980). "Red Cell Sorbitol, an Indicator of Diabetic Control [red cell sorbitol, diabetes control indicator]"). Various aldose reductase inhibitors are known to those skilled in the art, such as those described in U.S. Patent No. 6,579,879, including 6-(5-chloro-3-methyl-benzofuran-2-sulfonyl)-2H-pyridazin-3-one.

[0217] Any sorbitol dehydrogenase inhibitor can 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 by the enzyme sorbitol dehydrogenase. Those skilled in the art can easily determine such sorbitol dehydrogenase inhibitory activity according to standard assays (e.g., Analyt. Biochem [Analytical Biochemistry] (2000) 280: 329-331). A variety of sorbitol dehydrogenase inhibitors are known, 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.

[0218] Any glucosidase inhibitor can be used in combination with the compounds of the present invention. Glucosidase inhibitors inhibit the enzymatic hydrolysis of complex carbohydrates into bioavailable monosaccharides, such as glucose, by glycoside hydrolases (e.g., amylase or maltase). The rapid metabolic action of glucosidases, particularly after high levels of carbohydrate intake, leads to a state of nutritional hyperglycemia, which in obese or diabetic subjects leads to enhanced insulin secretion, increased fat synthesis, and reduced fat degradation. Following this hyperglycemia, hypoglycemia often occurs due to the increased levels of insulin present. In addition, the chyme remaining in the stomach promotes the production of gastric juice, which triggers or promotes the development of gastritis or duodenal ulcers. Therefore, it is known that glucosidase inhibitors are effective in accelerating the passage of carbohydrates through the stomach and inhibiting intestinal glucose absorption. In addition, the conversion of carbohydrates into lipids of adipose tissue and the subsequent incorporation of dietary fat into adipose tissue deposits are correspondingly reduced or delayed, while also having the benefit of reducing or preventing the harmful abnormalities caused thereby. Those skilled in the art can easily determine this glucosidase inhibitory activity according to standard assays (e.g., Biochemistry [biochemistry] (1969) 8: 4214).

[0219] Generally preferred glucosidase inhibitors include amylase inhibitors. Amylase inhibitors are glucosidase inhibitors that inhibit the enzymatic degradation of starch or glycogen to maltose. Those skilled in the art can readily determine such amylase inhibitory activity according to standard assays (e.g., Methods Enzymol. (1955) 1:149). Inhibiting such enzymatic degradation is beneficial in reducing the amount of bioavailable sugars (including glucose and maltose) and the attendant adverse conditions that result therefrom.

[0220] A variety of glucosidase inhibitors are known to those of ordinary skill in the art, and examples are provided below. Preferred glucosidase inhibitors are selected from acarbose, lipolysin, voglibose, miglitol, emiglitate, canagliflozin, tendamistate, trestatin, pranamicin-Q, and salbostatin. U.S. Patent Nos. 4,062,950 and 4,174,439 disclose the glucosidase inhibitor acarbose and various amino sugar derivatives related thereto, respectively. U.S. Patent No. 4,254,256 discloses the glucosidase inhibitor lipolysin. U.S. Patent No. 4,701,559 discloses the glucosidase inhibitor voglibose, 3,4-dideoxy-4-[[2-hydroxy-1-(hydroxymethyl)ethyl]amino]-2-C-(hydroxymethyl)-D-epinositol, and various N-substituted pseudoamino sugars related thereto. U.S. Patent No. 4,639,436 discloses the glucosidase inhibitors miglitol, (2R,3R,4R,5S)-1-(2-hydroxyethyl)-2-(hydroxymethyl)-3,4,5-piperidinetriol, and various 3,4,5-trihydroxypiperidines related thereto. U.S. Patent No. 5,192,772 discloses the glucosidase inhibitor emiglitate, ethyl p-[2-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]ethoxy]-benzoate, various derivatives thereof, and pharmaceutically acceptable acid addition salts thereof. U.S. Patent No. 4,634,765 discloses the glucosidase inhibitor MDL-25637, 2,6-dideoxy-7-0-|3-D-pyranose-syl-2,6-imino-D-glyceryl-L-glucose-sucrose heptanol, various homodisaccharides related thereto, and pharmaceutically acceptable acid addition salts thereof. U.S. Patent Nos. 5,157,116 and 5,504,078 disclose the glucosidase inhibitor canglibose, methyl 6-deoxy-6-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperido]-aD-pyranose-glucopyranoside sesquihydrate, deoxynojirimycin derivatives related thereto, various pharmaceutically acceptable salts thereof, and synthetic methods for preparing them. U.S. Patent No. 5,091,524 discloses the glucosidase inhibitor salbostatin and various pseudosaccharides related thereto.

[0221] A variety of amylase inhibitors are known to those of ordinary skill in the art. U.S. Patent No. 4,451,455 discloses the amylase inhibitor tendamistat and various cyclic peptides related thereto. U.S. Patent No. 4,623,714 discloses the amylase inhibitor AI-3688 and various cyclic polypeptides related thereto. U.S. Patent No. 4,273,765 discloses the amylase inhibitor tetamistatin (composed of a mixture of tetamistatin A, tetamistatin B, and tetamistatin C) and various amino sugars containing trehalose related thereto.

[0222] Additional antidiabetic compounds that can be used as a second agent in combination with the compounds of the present invention include, for example, the following: biguanides (e.g., metformin), insulin secretagogues (e.g., sulfonylureas and glinides), glitazones, nonglitazones PPARγ agonists, PPARβ agonists, DPP-IV inhibitors, PDE5 inhibitors, GSK-3 inhibitors, glucagon antagonists, inhibitors of β-1,6-BPase (Metabasis / Sankyo), GLP-1 / analogs (AC 2993, also known as exendin-4), insulin and insulin mimetics (Merck Natural Products). Other examples would include PKC-β inhibitors and AGE degraders.

[0223] The compounds of the present invention can also be used in combination with cardiovascular agents such as antihypertensive agents. Any antihypertensive agent can be used as the 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 measurements).

[0224] Amlodipine and related dihydropyridine compounds are disclosed in U.S. Patent No. 4,572,909, which is incorporated herein by reference, as effective anti-ischemic and antihypertensive agents. U.S. Patent No. 4,879,303, which is incorporated herein by reference, discloses amlodipine benzenesulfonate salt (also known as amlodipinebesylate). Amlodipine and amlodipine besylate are effective, long-lasting calcium channel blockers. Therefore, amlodipine, amlodipine besylate, amlodipine maleate and other pharmaceutically acceptable amlodipine acid addition salts have utility as antihypertensive agents and as anti-ischemic agents. Amlodipine besylate is currently used as Sale.

[0225] Calcium channel blockers within the scope of the present invention include, but are not limited to: bepridil, which may be prepared as disclosed in U.S. Pat. No. 3,962,238 or U.S. Pat. No. 30,577; clentiazem, which may be prepared as disclosed in U.S. Pat. No. 4,567,175; diltiazem, which may be prepared as disclosed in U.S. Pat. No. 3,562; fendiline, which may be prepared as disclosed in U.S. Pat. No. 3,262,977; gallopamil, which may be prepared as disclosed in U.S. Pat. No. 3,261,859; mirapridil, which may be prepared as disclosed in U.S. Pat. No. 4,808,605; prenylamine, which may be prepared as disclosed in U.S. Pat. No. 3,152,173; semodil, which may be prepared as disclosed in U.S. Pat. No. 4,786,635; terodiline, which may be prepared as disclosed in U.S. Pat. No. 3,371,014;

[0226] Verapamil, which may be prepared as disclosed in U.S. Patent No. 3,261,859; aranipine, which may be prepared as disclosed in U.S. Patent No. 4,572,909;

[0227] 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; elgodipine, which can be prepared as disclosed in U.S. Patent No. 4,952,592; felodipine, 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; lacidipine, which can be prepared as disclosed in U.S. Patent No. 4,801,599;

[0228] Lercanidipine, which may be prepared as disclosed in U.S. Patent No. 4,705,797; manidipine, which may be prepared as disclosed in U.S. Patent No. 4,892,875; nicardipine, which may be prepared as disclosed in U.S. Patent No. 3,985,758; nifedipine, which may be prepared as disclosed in U.S. Patent No. 3,485,847; nilvadipine, which may be prepared as disclosed in U.S. Patent No. 4,338,322; nimodipine, which may be prepared as disclosed in U.S. Patent No. 3,799,934; nisoldipine, which may be prepared as disclosed in U.S. Patent No. 4,154,839; and nifedipine, which may be prepared as disclosed in U.S. Patent No. 3,799, No. 934; cinnarizine, which can be prepared as disclosed in U.S. Patent No. 2,882,271; 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; benzylcyclane, which can be prepared as disclosed in Hungarian Patent No. 151,865; etabenzone, which can be prepared as disclosed in German Patent No. 1,265,758; and perhexiline, which can be prepared as disclosed in British Patent No. 1,025,578.

[0229] The disclosures of all of these U.S. patents are incorporated herein by reference. Examples of currently marketed products containing antihypertensive agents include calcium channel blockers, such as

[0230]

[0231] and Angiotensin-converting enzyme (ACE) inhibitors, such as

[0232] and

[0233] Angiotensin converting enzyme inhibitors (ACE inhibitors) within the scope of the present invention include, but are not limited to: alacepril, which may be prepared as disclosed in U.S. Pat. No. 4,248,883; benazepril, which may be prepared as disclosed in U.S. Pat. No. 4,410,520; captopril, which may be prepared as disclosed in U.S. Pat. Nos. 4,046,889 and 4,105,776; cilopril, which may be prepared as disclosed in U.S. Pat. No. 4,452,790; delapril, which may be prepared as disclosed in U.S. Pat. No. 4,385,051; enalapril, which may be prepared as disclosed in U.S. Pat. No. 4,374,829; fosinopril, which may be prepared as disclosed in U.S. Pat. No. 4,337,201; fenapryl, which may be prepared as disclosed in U.S. Pat. No. 4,508,727.

[0015] The present invention also includes but is not limited to: imadapril, which can be prepared as disclosed in U.S. Patent No. 4,555,502; lisinopril, which can be prepared as disclosed in U.S. Patent No. 4,555,502; moveltopril, which can be prepared as disclosed in Belgian Patent No. 893,553; perindolapril, which can be prepared as disclosed in U.S. Patent No. 4,508,729; quinapril, which can be prepared as disclosed in U.S. Patent No. 4,344,949; ramipril, which can be prepared as disclosed in U.S. Patent No. 4,587,258; spirapril, 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. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0234] Angiotensin-1L receptor antagonists (A-11 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. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0235] Beta-adrenergic receptor blockers (beta- or beta-blockers) within the scope of the present invention include, but are not limited to: acebutolol, which may be prepared as disclosed in U.S. Patent No. 3,857,952; alprenolol, which may be prepared as disclosed in Netherlands Patent Application No. 6,605,692; amosulalol, which may be prepared as disclosed in U.S. Patent No. 4,217,305; arotinolol, which may be prepared as disclosed in U.S. Patent No. 3,932,400; atenolol, which may be prepared as disclosed in U.S. Patent Nos. 3,663,607 or 3,836,671; benfuranolol, which may be prepared as disclosed in U.S. Patent No. 3,853,923; betaxolol, which may be prepared as disclosed in U.S. Patent No. 4,252,984; and benzyl alcohol, which may be prepared as disclosed in U.S. Patent No. 3,857,981. bevantolol which can be prepared as disclosed in U.S. Patent No. 4,171,370; bisoprolol which can be prepared as disclosed in U.S. Patent No. 4,340,541; buculol 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; bufurolol which can be prepared as disclosed in U.S. Patent No. 3,929,836; bunitolol which can be prepared as disclosed in U.S. Patent Nos. 3,940,489 and 3,961,071; buprandolol which can be prepared as disclosed in U.S. Patent No. 3,309,406; butiridine hydrochloride which can be prepared as disclosed in French Patent No. 1,390,056. hydrochloride); butofilol, which can be prepared as disclosed in U.S. Patent No. 4,252,825; carazolol, which can be prepared as disclosed in German Patent No. 2,240,599; carteolol, which can be prepared as disclosed in U.S. Patent No. 3,910,924; carvedilol, which can be prepared as disclosed in U.S. Patent No. 4,503,067; celiprolol, which can be prepared as disclosed in U.S. Patent No. 4,034,009; cetaphilol, which can be prepared as disclosed in U.S. Patent No. 4,059,622; cloranolol, which can be prepared as disclosed in German Patent No. 2,213,044; chloranolol, which can be prepared as disclosed in Clifton et al., Journal of dilevolol, which may be prepared as disclosed in Journal of Medicinal Chemistry, 1982, 25,670; epanolol, which may be prepared as disclosed in European Patent Publication No. 41,491; innodolol, which may be prepared as disclosed in U.S. Patent No. 4,045,482; labetalol, which may be prepared as disclosed in U.S. Patent No. 4,012,444;Levobunolol, which can be prepared as disclosed in U.S. Patent No. 4,463,176; Mepindolol, which can be prepared as disclosed in Seeman et al., Helv. Chim. Acta [Swiss Chemical Journal], 1971, 54, 241;

[0236] Metipranolol, which may be prepared as disclosed in Czechoslovak Patent Application No. 128,471; metoprolol, which may be prepared as disclosed in U.S. Patent No. 3,873,600; moprolol, which may be prepared as disclosed in U.S. Patent No. 3,501,7691; nadolol, which may be prepared as disclosed in U.S. Patent No. 3,935,267; nadolol, which may be prepared as disclosed in U.S. Patent No. 3,819,702; nebivolol, which may be prepared as disclosed in U.S. Patent No. 4,654,362; neprolol, which may be prepared as disclosed in U.S. Patent No. 4,394,382; and niprolol, which may be prepared as disclosed in British Patent No. 1 ,077,603; perbutolol 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; pralol which can be prepared as disclosed in U.S. Patent No. 3,408,387; propranolol which can be prepared as disclosed in British Patent No. 909,357; propranolol which can be prepared as disclosed in U.S. Patent Nos. 3,337,628 and 3,520,919; and propranolol which can be prepared as disclosed in Uloth et al., Journal of

[0015] The present invention also includes the following: sotalol, which can be prepared as disclosed in the Journal of Medicinal Chemistry, 1966, 9, 88; sufinalol, which can be prepared as disclosed in German Patent No. 2,728,641; talindol, which can be prepared as disclosed in U.S. Patent Nos. 3,935,259 and 4,038,313; tertalol, which can be prepared as disclosed in U.S. Patent No. 3,960,891; tesorolol, 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; toliprolol, which can be prepared as disclosed in U.S. Patent No. 3,432,545; and xibenolol, which can be prepared as disclosed in U.S. Patent No. 4,018,824. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0237] Alpha-adrenergic receptor blockers (α- or α-blockers) within the scope of the present invention include, but are not limited to: amosulalol, which may be prepared as disclosed in U.S. Pat. No. 4,217,307; arotinolol, which may be prepared as disclosed in U.S. Pat. No. 3,932,400; daprazol, which may be prepared as disclosed in U.S. Pat. No. 4,252,721; doxazosin, which may be prepared as disclosed in U.S. Pat. No. 4,188,390; fenspiride, which may be prepared as disclosed in U.S. Pat. No. 3,399,192; indoramine, which may be prepared as disclosed in U.S. Pat. No. 3,527,761; and daprazol, which may be prepared as disclosed in U.S. Pat. No. 4,252,721. Labetalol prepared as disclosed in U.S. Patent No. 3,997,666; Naftopidil prepared as disclosed in U.S. Patent No. 3,997,666; Nicergoline prepared as disclosed in U.S. Patent No. 3,228,943; Prazosin prepared as disclosed in U.S. Patent No. 3,511,836; Tamsulosin prepared as disclosed in U.S. Patent No. 4,703,063; Tolazoline prepared as disclosed in U.S. Patent No. 2,161,938; Trimazosin prepared as disclosed in U.S. Patent No. 3,669,968; and Yohimbine isolated from natural sources according to methods well known to those skilled in the art. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0238] As used herein, the term "vasodilator" is intended to include cerebral vasodilators, coronary vasodilators, and peripheral vasodilators.Cerebral vasodilators within the scope of the present invention include, but are not limited to: benzylcyclane, which may be prepared as disclosed above; cinnarizine, which may be prepared as disclosed above; citicoline, which may 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; cyclomandelate, which may be prepared as disclosed in U.S. Patent No. 3,663,597; cyclonicotinate, which may be prepared as disclosed in German Patent No. 1,910,481; diisopropylamine dichloroethyl ester, which may be prepared as disclosed in British Patent No. 862,248; and hydroxybenzoate, which may be prepared as disclosed in Hermann et al., Journal of the American Chemical Society, 1956, 222, 185. Society, 1979, 101, 1540; enamonin, which can be prepared as disclosed in U.S. Pat. No. 4,678,783; fasudil, which can be prepared as disclosed in U.S. Pat. No. 3,818,021; flunarizine, which can be prepared as disclosed in U.S. Pat. No. 3,773,939; ibudilast, which can be prepared as disclosed in U.S. Pat. No. 3,850,941; ifenprodil, which can be prepared as disclosed in U.S. Pat. No. 3,509,164; lomerizine, which can be prepared as disclosed in U.S. Pat. No. 4,663,325; nafronyl, which can be prepared as disclosed in U.S. Pat. No. 3,334,096; fenoxadil, which can be prepared as disclosed in Blicke et al., Journal of the American Chemical Society, 1989, 101, 1540; nafronyl, which can be prepared as disclosed in U.S. Pat. No. 3,334,096; fenoxadil, which can be prepared as disclosed in U.S. Pat. No. 3,818,021; flunarizine, which can be prepared as disclosed in U.S. Pat. No. 3,773,939; ibudilast, which can be prepared as disclosed in U.S. Pat. No. 3,850,941; Society, 1942, 64, 1722; nicergoline, which can be prepared as disclosed above; nimodipine, which can be prepared as disclosed in U.S. Pat. No. 3,799,934; papaverine, which can be prepared as reviewed in Goldberg, Chem. Prod. Chem. News, 1954, 17, 371; pentifedrine, which can be prepared as disclosed in German Patent No. 860,217; tenofezone, which can be prepared as disclosed in U.S. Pat. No. 3,563,997; vincaine, which can be prepared as disclosed in U.S. Pat. No. 3,770,724; vinpocetine, which can be prepared as disclosed in U.S. Pat. No. 4,035,750; and vequadil, which can be prepared as disclosed in U.S. Pat. No. 2,500,444.The disclosures of all of these US patents are incorporated herein by reference.

[0239] Coronary vasodilators within the scope of the present invention include, but are not limited to, amine oxide triphenyls, which may be prepared as disclosed in U.S. Pat. No. 3,010,965; bendazol, which may be prepared as disclosed in J. Chem. Soc. 1958, 2426; benfuridil hemisuccinate, which may be prepared as disclosed in U.S. Pat. No. 3,355,463; beniodarone, which may be prepared as disclosed in U.S. Pat. No. 3,012,042; chloracizine, which may be prepared as disclosed in British Pat. No. 740,932; carbomer, which may be prepared as disclosed in U.S. Pat. No. 3,282,938; clobenfural, which may be prepared as disclosed in British Pat. No. 1,160,925; chlorohydrin nitrates, which may be prepared from propylene glycol according to methods well known to those skilled in the art, for example, see Annale et al. n,1870,155,165; cloricromen which may be prepared as disclosed in U.S. Patent No. 4,452,811; dilazepoxide which may be prepared as disclosed in U.S. Patent No. 3,532,685; dipyridamole which may be prepared as disclosed in British Patent No. 807,826; hydropramine which may be prepared as disclosed in German Patent No. 2,521,113; efloxate which may be prepared as disclosed in British Patent Nos. 803,372 and 824,547; erythritol tetranitrate which may be prepared by nitrating erythritol according to methods well known to those skilled in the art; etabenzone which may be prepared as disclosed in German Patent No. 1,265,758; fendiline which may be prepared as disclosed in U.S. Patent No. 3,262,977; floxil which may be prepared as disclosed in German Patent No. 2,020,464;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; nitroethylamine tosylate which can be prepared as disclosed in Swedish Patent No. 168,308; and nitroethylamine tosylate which can be prepared as disclosed in Baxter et al., Journal of the Chemical Society, 1949, S. 30; lidoflazine which can be prepared as disclosed in U.S. Pat. No. 3,267,104; mannitol hexanitrate which can be prepared by nitration of mannitol according to methods well known to those skilled in the art; medibazine which can be prepared as disclosed in U.S. Pat. No. 3,119,826; nitroglycerin; pentaerythritol tetranitrate which can be prepared by nitration of pentaerythritol according to methods well known to those skilled in the art; pentanidol which can be prepared as disclosed in German Patent No. 638,422-3; perhexilline which can be prepared as disclosed above; pimephylline which can be prepared as disclosed in U.S. Pat. No. 3,350,400; prenylamine which can be prepared as disclosed in U.S. Pat. No. 3,152,173; propatyl nitrate which can be prepared as disclosed in French Patent No. 1,103,113. nitrate); trapidil, which can be prepared as disclosed in German Patent No. 55,956; tricromyl, 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; triethylnitramine phosphate, which can be prepared by nitration of triethanolamine followed by precipitation with phosphoric acid according to methods well known to those skilled in the art; and visnadine, which can be prepared as disclosed in U.S. Patent Nos. 2,816,118 and 2,980,699. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0240] Peripheral vasodilators within the scope of the present invention include, but are not limited to: aluminum nicotinate, which may be prepared as disclosed in U.S. Patent No. 2,970,082; bamexane, which may be prepared as disclosed in Corrigan et al., Journal of the American Chemical Society, 1945, 67, 1894; benzylcycloalkanes, which may be prepared as disclosed above; and benzophenones, which may be prepared as disclosed in Walter et al., Journal of the American Chemical Society, 1945, 67, 1894. Betahistine, which may be prepared as disclosed in the Journal of the American Chemical Society, 1941, 63, 2771; bradykinin, which may be prepared as disclosed in Hamburg et al., Arch. Biochem. Biophys., 1958, 76, 252; bromovincine, which may be prepared as disclosed in U.S. Pat. No. 4,146,643; buflomedil, which may be prepared as disclosed in U.S. Pat. No. 3,542,870; buflomedil, which may be prepared as disclosed in U.S. Pat. No. 3,895,030; butalamide, which may be prepared as disclosed in U.S. Pat. No. 3,338,899; cetidil, which may be prepared as disclosed in French Patent No. 1,460,571; and dapoxetine, which may be prepared as disclosed in German Patent No. 1,910 ,481; cinepazide which can be prepared as disclosed in Belgian Patent No. 730,345; cinnarizine which can be prepared as disclosed above; cyclomandelate which can be prepared as disclosed above; diisopropylamine dichloroacetate which can be prepared as disclosed above; eledoxin which can be prepared as disclosed in British Patent No. 984,810; fenoxidil which can be prepared as disclosed above; flunarizine which can be prepared as disclosed above; decanenitate 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; and dapoxetine which can be prepared as disclosed in Badgett et al., Journal of Inositol nicotinate prepared as disclosed in the Journal of the American Chemical Society, 1947, 69, 2907; isoxsulin prepared as disclosed in U.S. Patent No. 3,056,836; can be prepared as Biochem.Biophys.Res.Commun.[Biochemical and Biophysical Research Letters], 1961, 6, 210; kallikrein which can be prepared as disclosed in German Patent No. 1,102,973; moxycillin which can be prepared as disclosed in German Patent No. 905,738; naphthofuramide which can be prepared as disclosed above; nicotinamide ethyl which can be prepared as disclosed above; nicergoline which can be prepared as disclosed above; Nicofuroxine, which can be prepared as disclosed in U.S. Patent No. 366,523; nylidrin, which can be prepared as disclosed in U.S. Patent Nos. 2,661,372 and 2,661,373; pentofylline, which can be prepared as disclosed above; pentoxifylline, which can be prepared as disclosed in U.S. Patent No. 3,422,107; piribedil, which can be prepared as disclosed in U.S. Patent No. 3,299,067;

[0241] Prostaglandin Ei, which can be prepared by any method cited in the Merck Index, 12th edition, Budaveri, New Jersey, 1996, page 1353; Sulodip, which can be prepared as disclosed in German Patent No. 2,334,404; Tolazoline, 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 Korbits et al., Acta. Pharm. Hung., 1968, 38, 98. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0242] Within the scope of the present invention, the term "diuretic" refers to diuretic benzothiazine derivatives, diuretic organomercuries, diuretic purines, diuretic steroids, diuretic sulfonamide derivatives, diuretic uracil and other diuretics, such as amanozine, which can be prepared as disclosed in Austrian Patent No. 168,063; amiloride, which can be prepared as disclosed in Belgian Patent No. 639,386; arbutin, which can be prepared as disclosed in Tschitschibabin, Annalen, 1930, 479,303; clorazanide, which can be prepared as disclosed in Austrian Patent No. 168,063; ethacrynic acid, which can be prepared as disclosed in U.S. Patent No. 3,255,241; and phenacylglycerol, which can be prepared as disclosed in U.S. Patent No. 3,255,241. Etoxazoline prepared as disclosed in U.S. Patent No. 3,072,653; hydrazine 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; metochalcone which can be prepared as disclosed in Freudenberg et al., Ber., 1957, 90,957; muzolomide which can be prepared as disclosed in U.S. Patent No. 4,018,890; perhexiline which can be prepared as disclosed above; tinic acid which can be prepared as disclosed in U.S. Patent No. 3,758,506; triaminopterin which can be prepared as disclosed in U.S. Patent No. 3,081,230; and urea. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0243] Diuretic benzothiadiazine derivatives within the scope of the present invention include, but are not limited to: althiazine, which may be prepared as disclosed in British Patent No. 902,658; bendrofluazide, which may be prepared as disclosed in U.S. Patent No. 3,265,573; benzthiazide, McManus et al., 136th Am. Soc. Meeting [136th American Society of Societies] (Atlantic City, September 1959), Abstracts, pp. 13-0; benzyl hydrochlorothiazide, which may be prepared as disclosed in U.S. Pat. No. 3,108,097; buthiazide, which may be prepared as disclosed in British Patent Nos. 861,367 and 885,078; chlorothiazide, which may be prepared as disclosed in U.S. Pat. Nos. 2,809,194 and 2,937,169; chlorthalidone, which may be prepared as disclosed in U.S. Pat. No. 3,055,904; cyclopenthiazide, which may be prepared as disclosed in Belgian Patent No. 587,225; cyclothiazide, which may be prepared as disclosed in Whitehead et al., Journal of Organic Chemistry, 1961, 26, 2814;

[0244] Epitiazide, which may be prepared as disclosed in U.S. Patent No. 3,009,911; ethiazide, which may be prepared as disclosed in British Patent No. 861,367; fenquinazole, which may be prepared as disclosed in U.S. Patent No. 3,870,720; indapamide, which may be prepared as disclosed in U.S. Patent No. 3,565,911; hydrochlorothiazide, which may be prepared as disclosed in U.S. Patent No. 3,164,588; hydroflumethiazide, which may be prepared as disclosed in U.S. Patent No. 3,254,076; and fenquinazole, which may be prepared as disclosed in Close et al., Journal of the American Chemical Society. Society, 1960, 82, 1132; meticyclan, which can be prepared as disclosed in French Patent Nos. M2790 and 1,365,504; metolazone, which can be prepared as disclosed in U.S. Patent No. 3,360,518; paraflumethiazide, which can be prepared as disclosed in Belgian Patent No. 620,829; polythiazide, which can be prepared as disclosed in U.S. Patent No. 3,009,911; quinethazolone, which can be prepared as disclosed in U.S. Patent No. 2,976,289; tetrachlorothiazide, which can be prepared as disclosed in Close et al., Journal of the American Chemical Society, 1960, 82, 1132; and trichloromethiazide, which can be prepared as disclosed in deStevens et al., Experientia, 1960, 16, 113. The disclosures of all of these U.S. patents are incorporated herein by reference.

[0245] Diuretic sulfonamide derivatives within the scope of the present invention include, but are not limited to, acetazolamide, which may be prepared as disclosed in U.S. Pat. No. 2,980,679; ambuside, which may be prepared as disclosed in U.S. Pat. No. 3,188,329; azosemide, which may be prepared as disclosed in U.S. Pat. No. 3,665,002; bumetanide, which may be prepared as disclosed in U.S. Pat. No. 3,634,583; butazolamide, which may be prepared as disclosed in British Patent No. 769,757; clomiphenamide, which may be prepared as disclosed in U.S. Pat. Nos. 2,809,194, 2,965,655, and 2,965,656; chlorphenamide, which may be prepared as disclosed in Olivier, Rec. Trav. Chim., 1918, 37,307; clopamide, which may be prepared as disclosed in U.S. Pat. No. 3,459,756; and clomiphenamide, which may be prepared as disclosed in U.S. Pat. No. 3,634,583. No. 3,183,243; disulfamide, which can be prepared as disclosed in British Patent No. 851,287; ethoxolamide, 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; mefruside, which can be prepared as disclosed in U.S. Patent No. 3,356,692; methazolamide, which can be prepared as disclosed in U.S. Patent No. 2,783,241; piretanide, which can be prepared as disclosed in U.S. Patent No. 4,010,273; torasemide, 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 xipamide, which can be prepared as disclosed in U.S. Patent No. 3,567,777. The disclosures of all of these U.S. patents are incorporated herein by reference. Other additional agents in combination with the compounds disclosed herein include PCSK9 translation inhibitors as described in WO 2016 / 055901, incorporated herein by reference.

[0246] 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, a combination of niacin and lovastatin, ion exchange resins, antioxidants, ACAT inhibitors, bile acid sequestrants, and PCSK9 translation inhibitors.

[0247] To treat sepsis or septic shock, the disclosed methods may include additional therapeutic agents, such as antibiotics. In certain embodiments, the antibiotic is an aminoglycoside, such as, but not limited to, kanamycin, amikacin, tobramycin, dibekacin, gentamicin, sisomicin, netilmicin, streptomycin, and neomycin B, neomycin C, and neomycin E. Other antibiotics include carbapenems, such as, but not limited to, imipenem, meropenem, ertapenem, doripenem, panipenem, biapenem, rozupenem, terbipenem, lenapenem, tomopenem, and cerembenem.

[0248] Additional antibiotics include quinolones such as, but not limited to, ciprofloxacin, garefoxacin, gatifloxacin, gemifloxacin, levofloxacin, cinoxacin, nalidixic acid, moxifloxacin, oxolinic acid, piromidic acid, pipemidic acid, rosofloxacin, enoxacin, fleroxacin, lomefloxacin, naldifloxacin, norfloxacin, ofloxacin, pefloxacin, rufloxacin, balofloxacin, grefloxacin, levofloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, clinafloxacin, sitafloxacin, trovafloxacin, prulifloxacin, delafloxacin, JNJ-Q2, nimofloxacin, and zabofloxacin.

[0249] Additional antibiotics include tetracyclines, such as, but not limited to, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, metacycline, minocycline, rolicycline, and tigecycline.

[0250] Additional antibiotics that can be used in the methods disclosed herein include, but are not limited to, ampicillin, amoxicillin, amoxicillin, piperacillin, tazobactam, chloramphenicol, and ticarcillin.

[0251] Pharmaceutical composition

[0252] The compositions and methods of the present invention can be used to treat subjects in need. In certain embodiments, the subject is a mammal, such as a human or non-human mammal. When applied to a subject such as a human, the composition or compound is preferably administered as a pharmaceutical composition, comprising, for example, a compound 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 physiologically buffered saline or other solvents or vehicles, such as ethylene glycol, glycerol, oil (such as olive oil) or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive routes of administration (i.e., routes of transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free, or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of a medicament or selectively target one or more cells, tissues or organs. Pharmaceutical compositions can be in dosage unit form, such as tablets, capsules (including dispersed capsules and gelatin capsules), granules, lyophiles for reconstruction, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system, such as a skin patch.The composition may also be present in a solution suitable for topical administration, such as eye drops.

[0253] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent that, for example, acts to stabilize a compound (such as a compound of the present invention), increase its solubility, or increase its absorption. Such physiologically acceptable agents include, for example, 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 choice of a pharmaceutically acceptable carrier, including 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 (preparation) may also be a liposome or other polymer matrix, into which, for example, a compound of the present invention may be incorporated. For example, liposomes comprising phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively easy to prepare and administer.

[0254] The phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject without excessive toxicity, irritation, allergic response, or other problems or complications, and with a reasonable benefit / risk ratio commensurate.

[0255] As used herein, the phrase "pharmaceutically acceptable carrier" refers to 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 of being compatible with the other ingredients of the formulation and not injurious to the subject. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other nontoxic compatible substances used in pharmaceutical formulations.

[0256] The pharmaceutical composition (formulation) can be administered to a subject by any of a variety of routes of administration, including, for example, oral administration (e.g., as a drench in the form of an aqueous or non-aqueous solution or suspension, tablet, capsule (including dispersible capsules and gelatin capsules), bolus, powder, granule, paste for application to the tongue); absorption through the oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., in the form of a vaginal suppository, cream, or foam); parenterally (including intramuscular, intravenous, subcutaneous, or intrathecal, e.g., as a sterile solution or suspension); nasally; intraperitoneally; subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment, or spray applied to the skin or as eye drops). The compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable therefor can be found, for example, in US Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0257] The formulation can be conveniently present in unit dosage form and can be prepared by any method well known in the pharmaceutical field. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the subject being treated, the specific mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally the amount of the compound that produces a therapeutic effect. Typically, the range of this amount (in percentage) will be about 1% to about 99% of the active ingredient, preferably about 5% to about 70%, and most preferably about 10% to about 30%.

[0258] The method for preparing these formulations or compositions comprises the step of combining an active compound (such as a compound of the present invention) with a carrier and optionally one or more auxiliary ingredients. In general, the formulation is prepared by uniformly and intimately combining a compound of the present invention with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product.

[0259] Formulations of the present invention suitable for oral administration may be in the form of capsules (including dispersible capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophiles, powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia), and / or as a mouthwash, etc., each containing a predetermined amount of the compound of the present invention as the active ingredient. The composition or compound may also be administered as a bolus, electuary, or paste.

[0260] To prepare solid dosage forms for oral administration (capsules (including dispersible capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic; (3) humectants, Such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retarder, such as paraffin; (6) absorption accelerator, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including dispersible capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Similar types of solid compositions can also be used as fillers in soft-filled and hard-filled gelatin capsules, using excipients such as lactose (lactose or milk sugar) and high molecular weight polyethylene glycols.

[0261] Tablets can be made by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets can 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 carboxymethyl cellulose), a surfactant, or a dispersant. Molded tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0262] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including dispersible capsules and gelatin capsules), pills, and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may also be formulated to provide sustained or controlled release of the active ingredient therein using, for example, hydroxypropylmethylcellulose, other polymer matrices, liposomes, and / or microspheres in varying proportions to provide the desired release characteristics. They may be sterilized, for example, by filtering through a bacteria-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be formulated so that they release one or more active ingredients only or preferentially in a certain portion of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0263] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, lyophiles 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 ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuran methanol, polyethylene glycol, and sorbitan fatty acid esters, and mixtures thereof.

[0264] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0265] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0266] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration may be presented as suppositories, which can 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 salicylates, and which are solid at room temperature but liquid at body temperature and will therefore melt in the rectum or vaginal cavity and release the active compound.

[0267] Formulations of the pharmaceutical composition for administration to the oral cavity may be in the form of a mouthwash or oral spray or oral ointment.

[0268] Alternatively or additionally, the composition can be formulated for delivery via a catheter, stent, wire or other intraluminal device. Delivery via such a device may be particularly useful for delivery to the bladder, urethra, ureter, rectum or intestine.

[0269] Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0270] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.The active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[0271] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0272] In addition to the active compound, powders and sprays can contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays can also contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0273] Transdermal patches have the added advantage of providing controlled delivery of the compound 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 through the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0274] Ophthalmic formulations, eye ointments, powders, solutions etc. are also considered to be within the scope of the present invention. Exemplary ophthalmic formulations are described in U.S. Publication Nos. 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 desired, liquid ophthalmic formulations have a property similar to tears, aqueous humor or vitreous humor, or are compatible with such liquids. Preferred route of administration is localized administration (e.g., topical administration, such as eye drops, or administration via implants).

[0275] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more of the following pharmaceutically acceptable agents: sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions just before use, which may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0276] Examples of 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, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.

[0277] These compositions can also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. Can ensure to prevent the effect of microorganisms by including various antibacterials and antifungals (for example, parabens, chlorobutanol, sorbic acid phenol etc.). It is also possible to wish that isotonic agents, such as sugar, sodium chloride etc., are included in the compositions. In addition, the absorption extension of injectable drug form can be realized by comprising the medicament (such as aluminum monostearate and gelatin) that delays absorption.

[0278] In some cases, in order to prolong the effect of the drug, it is desirable to slow down the absorption of the subcutaneous or intramuscular injection of the drug. This can be achieved by using a liquid suspension of a crystalline or amorphous substance with low water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, the absorption of a parenteral drug form can be delayed by dissolving or suspending the drug in an oily vehicle.

[0279] Injectable depot forms are prepared by forming a microcapsule matrix of the subject compound in a biodegradable polymer (such as polylactide-polyglycolide). According to the ratio of the drug to the polymer and the property of the specific polymer adopted, the drug release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot-type injectable formulations are also prepared by embedding the drug in a liposome or microemulsion compatible with human tissue.

[0280] For use in the methods of the present invention, the active compound can be administered per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) active ingredient in combination with a pharmaceutically acceptable carrier.

[0281] The introduction method can also be provided through rechargeable or biodegradable devices. In recent years, various sustained-release polymer devices have been developed and tested in vivo for the controlled delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers, including hydrogels, including biodegradable and non-degradable polymers, can be used to form implants to continuously release compounds at specific target sites.

[0282] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0283] The selected dosage level will depend upon a variety of factors, including the activity of the specific compound or combination of compounds employed, or their esters, salts, or amides, the route of administration, the time of administration, the rate of excretion of the specific compound or compounds employed, the duration of the treatment, other drugs, compounds, and / or materials used in combination with the specific compound or compounds employed, the age, sex, weight, condition, general health and prior medical history of the subject being treated, and like factors well known in the medical arts.

[0284] A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can start the dosage of the pharmaceutical composition or compound at a dose lower than the level required to obtain the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means the concentration of the compound sufficient to cause the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age, and medical history. Other factors affecting the effective amount may 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 together with the compound of the present invention. A larger total dose can be delivered by multiple administrations of the agent. 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 edition, 1814-1882, incorporated herein by reference).

[0285] In general, a suitable daily dose of the active compound used in the compositions and methods of the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0286] If desired, the effective daily dose of the active compound can be administered as 1, 2, 3, 4, 5, 6 or more sub-doses administered separately at appropriate intervals throughout the day, optionally in unit dosage form. In certain embodiments of the invention, the active compound can be administered twice or three times daily. In preferred embodiments, the active compound will be administered once daily.

[0287] In certain embodiments, the compounds of the present invention can be used alone or in combination with another type of therapeutic agent. As used herein, the phrase "combined administration" refers to the administration of any form of two or more different therapeutic compounds, so that the therapeutic compound previously administered is still effective in vivo while the second compound is administered (for example, two compounds are effective in the subject's body at the same time, which can include the synergistic effect of the two compounds). For example, different therapeutic compounds can be in the form of identical formulations or separate formulations, administered simultaneously or sequentially. In certain embodiments, different therapeutic compounds can be administered within one hour, 12h, 24h, 36h, 48h, 72h or a week from each other. Therefore, the subject receiving this type of treatment can benefit from the combined effects of different therapeutic compounds.

[0288] In certain embodiments, the combined administration of a compound of the invention (e.g., a compound of Formula I or Ia) or one or more additional therapeutic agents provides improved efficacy relative to each administration of the compound of the invention (e.g., a compound of Formula I or Ia) or one or more additional therapeutic agents alone. In certain such embodiments, the combined administration provides an additive effect, wherein the additive effect refers to the sum of the effects of each of the individual administrations of the compound of the invention and the one or more additional therapeutic agents.

[0289] The present invention includes the purposes of the pharmaceutically acceptable salt of the compound of the present invention in the compositions and methods of the present invention.In certain embodiments, the salt of the present invention considered includes but is not limited to alkyl, dialkyl, trialkyl or tetraalkyl ammonium salt.In certain embodiments, the salt of the present invention considered includes but is not limited to, L-arginine, phenethylbenzylamine (benenthamine), benzathine (benzathine), betaine, calcium hydroxide, choline, deanol (deanol), diethanolamine, diethylamine, 2- (diethylamino) ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine (hydrabamine), 1H-imidazole, lithium, L-lysine, magnesium, 4- (2-hydroxyethyl) morpholine, piperazine, potassium, 1- (2-hydroxyethyl) pyrrolidine, sodium, triethanolamine, tromethamine and zinc salt.In certain embodiments, the salt of the present invention considered includes but is not limited to Na, Ca, K, Mg, Zn or other metal salts.

[0290] Pharmaceutically acceptable acid addition salts may also exist as various solvates, such as water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates may also be prepared. The source of such solvates may be derived from the crystallization solvent, inherent in the preparation or crystallization solvent, or adventitious to such solvent.

[0291] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0292] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0293] Examples

[0294] Examples of compounds of formula (I) or pharmaceutically acceptable salts thereof having useful biological activities are listed in Tables 1-10. 1 H NMR spectroscopy was performed on a Varian MR-400 mass spectrometer operating at 400 MHz (proton frequency) and equipped with: a self-shielded z-gradient coil, a 5 mm 1H / nX reverse detection broadband probe, 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 the following abbreviations are used to report multiplicities (s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad, nd = not determined).

[0295] A. Analytical Methods

[0296] Method 1 (acid FA)

[0297] UPLC setup

[0298] Solvent: -A water (high purity obtained by PureLab Option unit) containing 0.1% formic acid

[0299] B Acetonitrile (far UV grade) containing 0.1% (v / v) formic acid

[0300] Column: Acquity UPLC HSS C18 1.8 μm 100x2.1 mm (with guard column)

[0301] Flow rate: -0.5mL / min

[0302] gradient:

[0303] Time (min) ％A ％B 0.00 95 5 1.2 95 5 3.5 0 100 4.9 0 100 5 95 5 6 95 5

[0304] Inject 0.5-2 μL

[0305] UV detection by Waters DAD

[0306] Start range (nm) 210 End range (nm) 400 Resolution (nm) 1.2

[0307] MS detection: Waters SQD2, single quadrupole UPLC-MS

[0308] Scan range of MS data (m / z)

[0309] Start (m / z) 100

[0310] End (m / z) 700 or 1500, as needed

[0311] Perform +ve / -ve switching

[0312] The ionization was ESI.

[0313] The ESI voltage and temperature are:

[0314] Source 150C 3.5KV Capillary 25V Cone

[0315] Method 2 (Alkaline FA)

[0316] UPLC setup

[0317] Solvent: -Acetonitrile (far UV grade)

[0318] Water (high purity obtained using a PureLab Option unit) containing 10 mM ammonium bicarbonate (ammonium hydrogen carbonate)

[0319] Column: -Acquity UPLC BEH Shield RP181.7 um 100x2.1mm.

[0320] (with guard column)

[0321] Flow rate: -0.5mL / min

[0322] Gradient: -A: Water / Basic B: MeCN / Basic

[0323]

[0324]

[0325] A typical injection is 0.5-2 uL (concentration approximately 0.2-1 mg / mL).

[0326] UV detection by Waters DAD

[0327] Start Range (nm) 210 End Range (nm) 400 Resolution (nm) 1.2 Additional wavelength traces are extracted from the DAD data.

[0328] MS detection: Waters SQD2, single quadrupole UPLC-MS

[0329] The splitter provides approximately 300 μl / min of flow to the mass spectrometer.

[0330] Scan range of MS data (m / z)

[0331] Start (m / z) 100

[0332] End (m / z) 700 or 1500, as needed

[0333] Perform +ve / -ve switching

[0334] Preparative reversed-phase HPLC conditions

[0335] Preparative HPLC

[0336] Waters Micromass ZQ / Sample Manager 2767

[0337] Photodiode array detector 2996;

[0338] Column: XTerra Prep MS C18 column (5 μm, 19×150 mm, Waters)

[0339] Flow rate: MS detection 20mL / min

[0340] UV wavelength: 254nm.

[0341] Mobile phase: Solvent A (water: MeCN: HCO2H 95:5:0.05); Solvent B (water: MeCN: HCO2H 5:95:0.05)

[0342] gradient:

[0343] Time (min) ％A ％B 0.00 100.0 0.00 1.00 100 0.00 10.00 0.00 100.0 11.00 0.00 100.0 12.00 100.0 0.00

[0344] Flash chromatography was performed using an Isolera MPLC system (manufactured by Biotage) using pre-packed silica gel or reverse phase cartridges (supplied by Biotage or Interchim).

[0345] B. Chemical synthesis

[0346] The general procedures used in the methods for preparing the compounds of the present invention are as follows: Scheme 1

[0347]

[0348] 2-Chloro-5-(methylthio)pyrimidine (100B)

[0349] Dimethyl disulfide (13.96 mL, 155.4 mmol, 1.0 equiv) was added to a -75°C cooled solution of 5-bromo-2-chloropyrimidine (100A) (30 g, 155.4 mmol, 1.0 equiv) in anhydrous tetrahydrofuran (700 mL) under nitrogen. To this mixture was added dropwise a solution of n-butyllithium (2.5 M, 68.4 mL, 170.9 mmol, 1.0 equiv) over 1.5 h, with the internal temperature maintained at -70°C to -75°C throughout the addition. After the addition was complete, the mixture was stirred at -75°C for 4.5 h and then quenched by slowly adding a saturated solution of ammonium chloride (100 mL). The cooling bath was removed and the reaction was warmed to room temperature under nitrogen over 18 h. The reaction mixture was diluted with ethyl acetate (500 mL), washed with water (50 mL), and then washed with saturated brine (50 mL). The organic phase was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The crude pale yellow oil obtained was purified by flash chromatography (isohexane / ethyl acetate 0-50% elution) to give the desired product 2-chloro-5-(methylthio)pyrimidine (100B) as a waxy pale yellow solid.

[0350] Yield: 7.39 g (29%). 1 H NMR(CDCl3)δ8.48(2H,s),2.54(3H,s); MS(ESI+)m / z 161(M+H) + .

[0351] General Method 1

[0352] tert-Butyl (2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100C)

[0353] Tert-butyl (3-amino-2-methylpropyl) carbamate (4.79 g, 25.52 mmol, 1.05 eq) was added to a stirred suspension of 2-chloro-5-(methylthio)pyrimidine (100B) (3.89 g, 24.31 mmol, 1.0 eq) and cesium carbonate (11.85 g, 36.46 mmol, 1.4 eq) in anhydrous dimethylformamide (50 mL). The mixture was stirred at room temperature for 18 h and then concentrated to approximately 20 mL under reduced pressure. The liquid 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 in vacuo to afford a crude residue which was purified by flash chromatography (isohexane / ethyl acetate 0-50% elution) to give the desired product, tert-butyl (2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100C) as a pale yellow oil.

[0354] Yield: 6.59 g (86%). MS (ESI+) m / z 313 (M+H) + .

[0355] Tert-Butyl (S)-(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100D)

[0356] Racemic tert-butyl (2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100C) (5 g) was purified by chiral SFC using the following conditions: YMC Amylose-C 30 / 70 MeOH / CO, 100 mL / min, 120 bar, 40°C, GLS 40 psi, system 3900 psi, 140 bar drop, Stacker, DAD 245 nm.

[0357] First eluting isomer, 1.3 minutes. tert-Butyl (R)-(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate

[0358] Yield: 2.05 g. 1 H NMR (400MHz, CDCl3) δ8.34(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) + .

[0359] Second eluting isomer, 1.7 minutes. tert-Butyl (S)-(2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100D)

[0360] Yield: 2.48 g. 1 H NMR (400MHz, CDCl3) δ8.34 (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)+ .

[0361] General Method 2

[0362] (R)-2-Methyl-N 1 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E)

[0363] A solution of hydrogen chloride (15 mL, 4 M in 1,4-dioxane) was added to tert-butyl (S)-(2-ethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100D) (600 mg, 2.48 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to provide the desired product (R)-2-methyl-N-(2-ethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100D) as a light yellow semisolid. 1 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E).

[0364] Yield: 548 mg (100%) HCl salt. 1 H NMR(400MHz,DMSO)δ8.39(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) + .

[0365] General Method 3

[0366] Ethyl (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylate (100F)

[0367] Under nitrogen, ethyl-2-chlorobenzothiazole-6-carboxylate (600 mg, 2.48 mmol, 1.0 equiv) was added to (R)-2-methyl-N 1-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) (548 mg, 2.48 mmol, 1.0 equiv) and triethylamine (1.73 mL, 12.44 mmol, 5.0 equiv) in anhydrous dimethylformamide (20 mL) were stirred. The mixture was stirred at room temperature for 48 h and then concentrated under vacuum to approximately 5 mL. Water (25 mL) was added and the mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with water (20 mL) and brine (25 mL) and then dried over magnesium sulfate. The solvent was removed in vacuo to give a crude yellow oil which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired ethyl (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylate (100F) as an off-white solid.

[0368] Yield: 647 mg (62%). 1 H NMR (400MHz, CDCl3) δ8.37(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),2.20-2.11( m,1H),1.40(dd,J=7.2,7.2Hz,3H),1.07(d,J=7.0Hz,3H); MS(ESI+)m / z418(M+H) + .

[0369] General Method 4

[0370] (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid (Example 1)

[0371] Lithium hydroxide monohydrate (318 mg, 7.75 mmol, 5.0 equiv) was added to a stirred solution of ethyl (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylate (100F) (647 mg, 1.55 mmol, 1.0 equiv) in ethanol (7 mL) and water (5 mL). The mixture was stirred at ambient temperature for 18 h and then concentrated under reduced pressure. Water (5 mL) was added to the residue, and the mixture was acidified to a pH of approximately 3 with aqueous hydrochloric acid (2 M). The formed precipitate was collected by filtration, washed with water, and then dried under high vacuum to give the desired product, (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid (Example 1) as a light yellow solid.

[0372] General Method 5

[0373] Methyl (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamido)propionate (100G)

[0374] 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 146 mg, 0.385 mmol, 1.5 equiv) was added to a solution of (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid (1) (100 mg, 0.257 mmol, 1.0 equiv), triethylamine (0.36 mL, 2.57 mmol, 10.0 equiv) and methyl-2-amino-2-methylpropanoate hydrochloride (196 mg, 1.28 mmol, 4.9 equiv) in dimethylformamide (5 mL), and the reaction mixture was stirred at room temperature for 18 h. The solvent was removed under reduced pressure and the crude residue obtained was purified by flash chromatography (isohexane / ethyl acetate 0-75% elution) to give the desired methyl (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamido)propanoate (G) as an off-white solid.

[0375] Yield: 120 mg (95%). 1H NMR (400 MHz, MeOD) δ 8.34 (s, 2H), 8.10 (d, J = 1.8 Hz, 1H), 7.77 (dd, J = 1.9, 8.5 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 3.74 (s, 3H), 3.45 (dd, J = 6.3, 22.4 Hz, 4H), 2.34 (s, 3H), 2.29-2.16 (m, 1H), 1.59 (s, 6H), 1.08 (d, J = 6.9 Hz, 3H). No NH exchangeable protons were observed. MS (ESI+) m / z 489 (M+H). + .

[0376] (S)-2-Methyl-2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamido)propanoic acid (Example 2)

[0377] The method is similar to general method 4

[0378] Lithium hydroxide monohydrate (50 mg, 1.22 mmol, 5.0 equiv) was added to a stirred solution of methyl (S)-2-methyl-2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxamido)propanoate (100G) (120 mg, 0.245 mmol, 1.0 equiv) 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, and the residue was diluted with water (3 mL) and acidified to pH approximately 3 with aqueous hydrochloric acid (2 M). The formed precipitate was collected by filtration, washed with water, and then dried under high vacuum to give the title compound as an off-white solid.

[0379] Using the above procedure, the following examples were synthesized:

[0380] Table 1

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396] Option 2

[0397]

[0398] General Method 6

[0399] 2-Chloro-N-(4-methoxybenzyl)benzo[d]thiazole-6-sulfonamide (101B).

[0400] (4-Methoxyphenyl)methanamine (134 mg, 0.97 mmol, 1.05 eq) was added dropwise to an ice-cold solution of 2-chlorobenzothiazole-6-sulfonyl chloride (101A) (250 mg, 0.932 mmol, 1.0 eq), triethylamine (0.39 mL, 2.79 mmol, 3.0 eq) in tetrahydrofuran (10 mL), and the mixture was stirred at 0° C. for 1 hour. The reaction mixture was concentrated under reduced pressure to give a solid, which was washed with ice-cold water (10 mL), ice-cold tetrahydrofuran (10 mL), and then dried under vacuum to give the desired 2-chloro-N-(4-methoxybenzyl)benzo[d]thiazole-6-sulfonamide (101B) as a white solid.

[0401] Yield: 295 mg (85%). 1 H NMR (400MHz, DMSO) δ8.56(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) + .

[0402] The intermediates in Table 2 were synthesized using conditions similar to those described for Intermediate 101B:

[0403] Table 2

[0404]

[0405]

[0406]

[0407] (S)-N-(4-Methoxybenzyl)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-sulfonamide (101C)

[0408] The method applied was similar to that described in General Method 3.

[0409] Yield: 298 mg (69%). 1 H NMR(400MHz,DMSO)δ8.56(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) + .

[0410] (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-sulfonamide (Example 87)

[0411] Trifluoroacetic acid (5 mL) was added dropwise to an ice-cold solution of N-(4-methoxybenzyl)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-sulfonamide (101C) (250 mg, 0.459 mmol, 1.0 equiv) in anhydrous dichloromethane (5 mL). The mixture was stirred on ice for 30 minutes and then allowed to warm to ambient temperature over 18 h. 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 crude residue obtained was purified by flash chromatography (DCM / methanol 0-10% elution) to give the desired (S)-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-sulfonamide (Example 87) as a white solid.

[0412] Yield: 190 mg (97%). 1 H NMR (400MHz, DMSO) δ8.40(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),2.3 4(s,3H),2.17-2.07(m,1H),0.96(d,J=6.8Hz,3H);MS(ESI+)m / z425(M+H) + .

[0413] Using the procedure described in Scheme 2, following General Method 3, the following examples were prepared:

[0414] Table 3

[0415]

[0416]

[0417]

[0418]

[0419] Option 3

[0420]

[0421] 2-Methyl-N 1 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A)

[0422] The method applied is similar to General Method 2.

[0423] A solution of hydrogen chloride (54 mL, 4 M in 1,4-dioxane) was added to tert-butyl(2-methyl-3-((5-(methylthio)pyrimidin-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 to provide the desired product, 2-methyl-N-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (100C) as a pale yellow semisolid. 1 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A) The semi-crude sample was used in the next reaction without further purification.

[0424] N1 -(6-bromobenzo[d]thiazol-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B)

[0425] The method applied is similar to General Method 3.

[0426] Under nitrogen, 6-bromo-2-chlorobenzothiazole (828 mg, 3.33 mmol, 0.95 equiv) was added to 2-methyl-N 1 -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A) (1.00 g, 3.51 mmol, 1.0 equiv) and cesium carbonate (3.43 g, 10.52 mmol, 3.0 equiv) in a stirred suspension in anhydrous dimethylformamide (25 mL). The mixture was stirred at room temperature for 72 h and then concentrated to about 3 mL under vacuum. Water (25 mL) was added and the mixture was extracted with ethyl acetate (3x50 mL). The combined organic phases were washed with water (20 mL) and brine (25 mL) and then dried over magnesium sulfate. The solvent was removed under vacuum to give a crude yellow oil, which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired N-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A) (1.00 g, 3.51 mmol, 1.0 equiv). 1 -(6-bromobenzo[d]thiazol-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B).

[0427] Yield: 266 mg (18%). 1 H NMR (400MHz, CDCl3) δ8.37(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).

[0428] General Method 7

[0429] Tert-butyl 4-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)-1H-pyrazole-1-carboxylate (102C)

[0430] Under nitrogen, N 1 -(6-bromobenzo[d]thiazol-2-yl)-2-methyl-N 3A solution of -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B) (50 mg, 0.12 mmol, 1.0 equiv) was added to a solution of (1-(tert-butoxycarbonyl)-1H-pyrazol-4-yl)boronic acid (27.8 mg, 0.13 mmol, 1.0 equiv), cesium carbonate (58 mg, 0.18 mmol, 1.5 equiv) and tetrakis(triphenylphosphine)palladium(0) (7 mg, 0.01 mmol, 0.05 equiv) in water (0.20 mL) and N,N-dimethylformamide (0.80 mL). The reaction mixture was heated to 90° C. for 16 h. Additional aliquots of (1-(tert-butoxycarbonyl)-1H-pyrazol-4-yl)boronic acid (27.8 mg, 0.13 mmol, 1.0 equiv) and tetrakis(triphenylphosphine)palladium(0) (7 mg, 0.01 mmol, 0.05 equiv) were added to the reaction, and the mixture was heated to 90° C. under nitrogen for an additional 16 h. 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), dried through a phase separator, and then concentrated under vacuum. The crude residue obtained was purified by flash chromatography (isohexane / ethyl acetate 0-75% elution) to give the desired tert-butyl 4-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)-1H-pyrazole-1-carboxylate (102C) as an off-white solid. The semi-crude product was used in the next reaction without further purification.

[0431] N 1 -(6-(1H-pyrazol-4-yl)benzo[d]thiazol-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 97)

[0432] A solution of hydrogen chloride (2 mL, 4 M in 1,4-dioxane) was added to tert-butyl 4-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)-1H-pyrazole-1-carboxylate (102C) (100 mg, 0.12 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to provide the desired N-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)-1H-pyrazole-1-carboxylate (102C) as an off-white solid. 1 -(6-(1H-pyrazol-4-yl)benzo[d]thiazol-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 97).

[0433] Using the procedure described in Scheme 3, the following examples were prepared according to General Method 7:

[0434] Table 4

[0435]

[0436]

[0437] General Method 8

[0438] 2-Methyl-N 1 -(5-(methylthio)pyrimidin-2-yl)-N 3 -(6-(Oxazol-2-yl)benzo[d]thiazol-2-yl)propane-1,3-diamine (Example 101)

[0439] Under nitrogen, N 1 -(6-bromobenzo[d]thiazol-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (102B) (90 mg, 0.21 mmol, 1.0 equiv) was added to a solution of 2-(tri-n-butylstannyl)oxazole (0.044 mL, 0.21 mmol, 1.0 equiv) and tetrakis(triphenylphosphine)palladium(0) (23 mg, 0.02 mmol, 0.1 equiv) in N,N-dimethylformamide (0.80 mL). The reaction mixture was heated to 90° C. for 16 h. Another aliquot of 2-(tri-n-butylstannyl)oxazole (0.044 mL, 0.21 mmol, 1.0 equiv) and tetrakis(triphenylphosphine)palladium(0) (23 mg, 0.02 mmol, 0.1 equiv) was added to the reaction mixture and heated to 110° C. for 18 h. The reaction is cooled to room temperature, diluted with water (2mL), and extracted with ethyl acetate (3x5mL). The combined organic phase is washed with water (2mL) and brine (2mL), filtered through a diatomaceous earth pad, and dried by a phase separator, and then concentrated to dryness under vacuum. The crude residue obtained is purified twice by flash chromatography (first isohexane / ethyl acetate 0-100% elution, then ethyl acetate / methanol 0-10%) to give a semi-crude residue, which is further purified by reverse phase preparative HPLC to give the desired 2-methyl-N-(2-nitropropene)-1-nitropropene as a pale solid. 1 -(5-(methylthio)pyrimidin-2-yl)-N 3 -(6-(Oxazol-2-yl)benzo[d]thiazol-2-yl)propane-1,3-diamine (101).

[0440] Using the procedure described in Scheme 3, the following examples were prepared according to General Method 8:

[0441] Table 5

[0442]

[0443]

[0444] Option 4

[0445]

[0446] Using the procedure described in Scheme 4, the following examples were prepared according to General Method 3:

[0447] Table 6

[0448]

[0449]

[0450] Option 7

[0451]

[0452] tert-Butyl (2,2-dimethyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (105A)

[0453] The method applied was similar to that described in General Method 1, except that additional heating was required.

[0454] Tert-butyl (3-amino-2,2-dimethylpropyl) carbamate (0.13 g, 0.65 mmol, 1.05 equiv) is added to a stirred suspension of 2-chloro-5-(methylthio) pyrimidine (100B) (0.10 g, 0.62 mmol, 1.0 equiv) and cesium carbonate (0.24 g, 0.75 mmol, 1.2 equiv) in anhydrous N, N-dimethylformamide (1.5 mL), and the mixture is stirred at 80 ° C for 4 h. The reaction mixture is concentrated under vacuum, diluted with ethyl acetate (20 mL), washed with water (7.5 mL) and brine (5.0 mL), and then dried through a phase separator. The solvent is removed under vacuum to give the desired tert-butyl (2,2-dimethyl-3-((5-(methylthio) pyrimidin-2-yl) amino) propyl) carbamate (105A) as a light yellow oil.

[0455] Yield: 0.163 g (81%). 1H NMR (400MHz, DMSO) δ8.37(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).

[0456] 2,2-Dimethyl-N 1 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (105B)

[0457] The method applied was similar to that described in General Method 2.

[0458] A solution of hydrogen chloride (5 mL, 4 M in 1,4-dioxane) was added to tert-butyl(2,2-dimethyl-3-((5-(methylthio)pyrimidin-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 to provide the desired product, 2,2-dimethyl-N-((5-(methylthio)pyrimidin-2-yl)amino)propyl)carbamate (105A) as a light yellow semisolid. 1 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (105B).

[0459] Yield: 0.13 g (100%) HCl salt. 1 H NMR (400MHz, DMSO) δ8.39 (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).

[0460] N1-(Benzo[d]oxazol-2-yl)-2,2-dimethyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 164)

[0461] The method applied was similar to that described in General Method 3, except that cesium carbonate was used as the general base.

[0462] Under nitrogen, 2-chlorobenzoxazole (0.06 mL, 0.54 mmol, 0.1 equiv) was added to 2,2-dimethyl-N 1-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (105B) (0.13 g, 0.49 mmol, 1.0 equiv) and cesium carbonate (0.48 g, 1.48 mmol, 3.0 equiv) in a stirred solution in anhydrous N, N-dimethylformamide (2.0 mL). The mixture was stirred at 80 ° C or room temperature for 16 h, and then concentrated under vacuum. Water (2.5 mL) was added, and the mixture was extracted with ethyl acetate (3x5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL) and then dried by a phase separator. The solvent was removed under vacuum to give a thick yellow oil, which was purified by preparative HPLC to give the desired N-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (105B). 1 -(Benzo[d]oxazol-2-yl)-2,2-dimethyl-N 3 -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 164).

[0463] Using the procedure described in Scheme 7, the following examples were prepared according to General Method 3:

[0464] Table 9

[0465]

[0466]

[0467]

[0468] Option 8

[0469]

[0470] 3-Chloro-N-(2-chlorobenzo[d]thiazol-6-yl)propane-1-sulfonamide (106B)

[0471] Sodium hydride (60% dispersion in mineral oil) (326mg, 8.15mmol, 3.0 equivalents) is added portion by portion to the ice-cold solution of 2-chlorobenzothiazole-6-amine (500mg, 2.71mmol, 1.0 equivalents) in DMF (25mL), and the mixture is stirred under ice-cooling for 1 hour. 3-chloropropane-1-sulfonyl chloride (673mg, 3.80mmol, 1.4 equivalents) is added dropwise, and then the reaction mixture is warmed to environment temperature through 3h. Reactant mixture is diluted with saline (20mL) and extracted with ethyl acetate (2x25mL). The combined organic fractions were combined and concentrated under reduced pressure to give a pale yellow oil which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired product 3-chloro-N-(2-chlorobenzo[d]thiazol-6-yl)propane-1-sulfonamide 106B as an off-white gum.

[0472] Yield: 427 mg (48%). 1 H NMR (400 MHz, MeOD) δ 7.87-7.84 (m, 2H), 7.38 (dd, J = 2.3, 8.8 Hz, 1H), 3.67 (dd, J = 6.3, 6.3 Hz, 2H), 2.27-2.19 (m, 2H). Note that the CH2 protons are obscured by MeOD.

[0473] 2-(2-Chlorobenzo[d]thiazol-6-yl)isothiazolidine 1,1-dioxide (106C)

[0474] Sodium hydride (60% dispersion in mineral oil) (98 mg, 2.46 mmol, 2.0 equivalents) is added to an ice-cold solution of 3-chloro-N-(2-chlorobenzo [d] thiazole-6-yl) propane-1-sulfonamide (106B) (400 mg, 1.23 mmol, 1.0 equivalents) in DMF (5 mL). The reaction mixture is stirred for 1 hour under ice cooling and then quenched by carefully adding saturated ammonium chloride solution (20 mL). The resulting mixture is extracted with ethyl acetate (3x20 mL), and the combined organic phase is washed with water (20 mL), saline (20 mL), and then concentrated under vacuum to give glue. The crude product is purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired product (106C) as off-white glue.

[0475] Yield: 220 mg (62%). 1H NMR (400MHz, CDCl3) δ7.92 (d, J = 8.9 Hz, 1H), 7.72 (d, J = 2.1 Hz, 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).

[0476] 2-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)isothiazolidine 1,1-dioxide (Example 175)

[0477] The method applied was similar to that described in General Method 3.

[0478] Yield: 25 mg (15%). 1 H NMR (400MHz, DMSO) δ8.34(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) + .

[0479] Option 9

[0480]

[0481] N-(2-Chlorobenzo[d]thiazol-6-yl)methanesulfonamide (107B)

[0482] Methanesulfonyl chloride (0.055 mL, 0.706 mmol, 1.3 equiv) was added dropwise to an ice-cold solution of 2-chlorobenzothiazole-6-amine (100 mg, 0.54 mmol, 1.0 equiv) and pyridine (0.066 mL, 0.815 mmol, 1.5 equiv) in anhydrous dichloromethane (5 mL). The mixture was stirred at 0 ° C for 15 minutes and then warmed to ambient temperature over 1 hour. The reaction mixture was quenched with water (10 mL). The organic phase was removed and concentrated under reduced pressure to give a light yellow oil, which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired N- (2-chlorobenzo [d] thiazole -6- bases) methanesulfonamide (107B) as a light yellow gum.

[0483] Yield: 135 mg (94.8%) 1 H NMR (400 MHz, CDCl 3 ) δ 7.88 (d, J = 8.8 Hz, 1H), 7.79 (d, J = 2.1 Hz, 1H), 3.00 (s, 3H). The aromatic H protons were obscured by CDCl 3 , and the NH exchangeable protons were not observed.

[0484] N-(2-Chlorobenzo[d]thiazol-6-yl)-N-methylmethanesulfonamide (107C)

[0485] Sodium hydride (60% dispersion in mineral oil) (31 mg, 0.772 mmol, 1.5 eq) was added portionwise to an ice-cold solution of N-(2-chlorobenzo[d]thiazol-6-yl)methanesulfonamide (107B) (135 mg, 0.515 mmol, 1.0 eq) in anhydrous tetrahydrofuran (2 mL). The mixture was stirred at room temperature for 2 h. Iodomethane (0.048 mL, 0.772 mmol, 1.5 eq) was added, and the mixture was stirred at room temperature for another 2 h. Water (10 mL) was added, and the solvent was then removed under high vacuum to give a light yellow gum, which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired N-(2-chlorobenzo[d]thiazol-6-yl)-N-methylmethanesulfonamide (107C) as a light yellow gum.

[0486] Yield: 100 mg (70%) 1 H NMR (400MHz, CDCl3) δ7.95 (d, J = 8.8 Hz, 1H), 7.86 (d, J = 2.1 Hz, 1H), 7.48 (dd, J = 2.3, 8.8 Hz, 1H), 3.39 (s, 3H), 2.88 (s, 3H).

[0487] N-Methyl-N-(2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)methanesulfonamide (Example 176)

[0488] The method applied was similar to that described in General Method 3.

[0489] Yield: 89 mg (60%) 1 H NMR (400 MHz, CDCl3) δ 8.38 (s, 2H), 7.62 (d, J = 2.0 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 7.24 (d, J = 3.2 Hz, 1H), 5.74 (dd, J = 6.5, 6.5 Hz, 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 = 6.9 Hz, 3H). Not all NH exchangeable protons were observed; MS (ESI+) m / z 453 (M+H). + .

[0490] Plan 10

[0491]

[0492] 4-(((5-(Methylthio)pyrimidin-2-yl)amino)methyl)pyrrolidin-2-one (108B)

[0493] The method applied was similar to that described in General Method 1.

[0494] 5-aminomethyl-pyrrolidin-2-one (108A) (1.0 g, 8.76 mmol, 1.0 equivalent) is added to a stirred suspension 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 equivalent) in anhydrous dimethylformamide (10 mL). The mixture is heated to 50 ° C for 18 h and then concentrated under reduced pressure. The liquid obtained is diluted with ethyl acetate (50 mL), washed with water (10 mL) and brine (10 mL), and then dried by a phase separator. The solvent is removed under reduced pressure to provide a crude residue, which is purified by grinding in methanol to give the desired product 4-(((5-(methylthio) pyrimidine-2-yl) amino) methyl) pyrrolidin-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 / methanol 0-10% elution) to give the desired product 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)pyrrolidin-2-one (108B) as a yellow solid. The two batches were combined and used in the next step.

[0495] Yield: 0.74 g (36%). 1 H NMR (400MHz, CDCl3) δ8.35 (s, 2H), 5.79-5.76 (m, 1H), 5.48 (dd, J = 5.5, 5.5Hz, 1H), 3.57-3.50 (m, 3H), 3.2 0(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).

[0496] Tert-butyl 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate (108C)

[0497] 4-Dimethylaminopyridine (5 mg, 0.04 mmol, 0.1 eq) was added to a stirred suspension of 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)pyrrolidin-2-one (108B) (100 mg, 0.4 mmol, 1.0 eq), di-tert-butyl dicarbonate (229 mg, 1.0 mmol, 2.5 eq) and triethylamine (0.146 mL, 1.00 mmol, 2.5 eq) in dichloromethane (4.2 mL). The mixture was stirred at room temperature for 72 h, then water (3.0 mL) was added. The mixture was extracted with ethyl acetate (3 x 5.0 mL), and the combined organic phases were washed with brine (2.5 mL) and then dried through a phase separator. The solvent was removed under reduced pressure to provide a crude residue, which was purified by reverse phase chromatography (0.1% formic acid solution / acetonitrile 5% to 100% elution) to give the desired product, tert-butyl 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate (108C).

[0498] Yield: 72 mg (53%). 1 H NMR (400MHz, DMSO) δ8.35(s,2H),7.62(t,J=6.1Hz,1H),3.73(dd,J=7.8,10.4Hz,1H),3.50-3.44(m,1 H), 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).

[0499] tert-Butyl (4-amino-2-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-4-oxobutyl)carbamate (108D)

[0500] A solution of ammonium hydroxide (2.2 mL) was added to tert-butyl 4-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-2-oxopyrrolidine-1-carboxylate (108C) (72 mg, 0.21 mmol), and the mixture was heated at 80° C. for 1.5 h. The mixture was cooled to room temperature and then extracted with dichloromethane (3×5 mL). The organic solvent was dried through a phase separator and then removed under reduced pressure to provide the desired product, tert-butyl (4-amino-2-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-4-oxobutyl)carbamate (108D).

[0501] Yield: 61 mg (86%). 1H NMR (400MHz, DMSO) δ8.33(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).

[0502] 4-Amino-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide dihydrochloride (108E)

[0503] The method applied was similar to that described in General Method 2.

[0504] A solution of hydrogen chloride (0.7 mL, 4 M in 1,4-dioxane) was added to tert-butyl(4-amino-2-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)-4-oxobutyl)carbamate (108D) (61 mg, 0.17 mmol), and the mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure to afford the desired product, 4-amino-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide dihydrochloride (108E) as a light yellow solid.

[0505] Yield: 56 mg (100%) HCl salt.

[0506] 4-((6-(N-(4-methoxybenzyl)sulfamoyl)benzo[d]thiazol-2-yl)amino)-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide (177)

[0507] The method applied was similar to that described in General Method 3.

[0508] Under nitrogen, 2-chloro-N-(4-methoxybenzyl)benzo[d]thiazole-6-sulfonamide (101B) (70 mg, 0.19 mmol, 1.10 equiv) was added to a stirred solution of 4-amino-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide dihydrochloride (108E) (56 mg, 0.17 mmol, 1.0 equiv) and triethylamine (0.072 mL, 0.51 mmol, 3.0 equiv) in anhydrous dimethylformamide (2.0 mL). The mixture was stirred at room temperature for 72 h and then concentrated under reduced pressure. Water (2.5 mL) was added and the resulting precipitate was collected by filtration and then washed with methanol. The organic filtrate was concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography (dichloromethane / methanol 0-25% elution) to give the desired product 4-((6-(N-(4-methoxybenzyl)sulfamoyl)benzo[d]thiazol-2-yl)amino)-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide (Example 177) as an off-white solid.

[0509] Yield: 25 mg (25%). 1 H NMR (400MHz, DMSO) δ8.45(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).

[0510] 4-((5-(Methylthio)pyrimidin-2-yl)amino)-3-(((6-sulfamoylbenzo[d]thiazol-2-yl)amino)methyl)butanamide (178)

[0511] Trifluoroacetic acid (0.3 mL) was added dropwise to a 0 ° C cooled solution of 4-((6-(N-(4-methoxybenzyl)sulfamoyl)benzo[d]thiazol-2-yl)amino)-3-(((5-(methylthio)pyrimidin-2-yl)amino)methyl)butanamide (177) (20 mg, 0.03 mmol, 1.0 equiv) in anhydrous dichloromethane (0.3 mL). The mixture was stirred for 30 minutes and then warmed to room temperature over 8 hours. Another aliquot of trifluoroacetic acid (1 mL) was added, and the mixture was stirred for 16 hours. The reaction mixture was concentrated under pressure and carefully basified by adding saturated aqueous sodium bicarbonate solution (3 mL). The mixture was extracted with ethyl acetate (3x5 mL). The combined organic phases were washed with brine (3 mL), dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The crude residue obtained was purified by flash chromatography (dichloromethane / methanol 0-20% elution) to give the desired product 4-((5-(methylthio)pyrimidin-2-yl)amino)-3-(((6-sulfamoylbenzo[d]thiazol-2-yl)amino)methyl)butanamide (Example 178) as a white solid.

[0512] Yield: 12 mg (75%). 1 H NMR (400MHz, DMSO) δ8.41(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).

[0513] Using the procedure described in Scheme 10, the following examples were synthesized:

[0514] Table 10

[0515]

[0516]

[0517] Plan 11

[0518]

[0519] 4-((2-Chlorobenzo[d]thiazol-6-yl)sulfonyl)morpholine (109B)

[0520] The method applied was similar to that described in General Method 6, using dichloromethane as solvent instead of tetrahydrofuran.

[0521] Yield: 686 mg. 1 H NMR (400MHz, CDCl3) δ8.25(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)+.

[0522] tert-Butyl (3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)-2-methylpropyl)carbamate (109D)

[0523] The method applied was similar to that described in General Method 1.

[0524] Yield: 433 mg. 1 H NMR (400MHz, CDCl3) δ8.16-8.15(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)+.

[0525] N1-(5-(Difluoromethoxy)pyrimidin-2-yl)-2-methylpropane-1,3-diamine hydrochloride (109E)

[0526] A solution of hydrogen chloride (2.7 mL, 4 M in 1,4-dioxane) was added to tert-butyl (3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)-2-methylpropyl)carbamate (109D) (300 mg, 0.903 mmol) and stirred at room temperature for 15 minutes. The solvent was removed under vacuum to give the crude title compound N1-(5-(difluoromethoxy)pyrimidin-2-yl)-2-methylpropane-1,3-diamine hydrochloride (109E), which was used in the next step without further purification.

[0527] Yield: 225 mg (quantitative). MS (ESI+) m / z 233 (M+H)+.

[0528] N1-(5-(difluoromethoxy)pyrimidin-2-yl)-2-methyl-N3-(6-(morpholinosulfonyl)benzo[d]thiazol-2-yl)propane-1,3-diamine (180)

[0529] The method applied was similar to that described in General Method 3.

[0530] Yield: 225 mg. 1 H NMR (400MHz, CDCl3) δ8.21(s,2H),7.97(d,J=1.5Hz,1H),7.65(dd,J=1.8,8.5 Hz,1H),7.59(d,J=8.5Hz,1H),6.91(s,1H),6.43(t,J=71.6Hz,1H),5.72(dd,J =6.6,6.6Hz,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)+.

[0531] Following the procedure described in Scheme 11, the following examples were synthesized:

[0532] Table 11

[0533]

[0534]

[0535]

[0536] Plan 12

[0537]

[0538] tert-Butyl (2-methyl-3-(pyridin-2-ylamino)propyl)carbamate (110B)

[0539] Under nitrogen, a solution of tert-butyl 3-amino-2-methylpropylcarbamate (110A) (150 mg, 0.80 mmol, 1.0 equiv) in 1,4-dioxane (2 mL) was added to 2-bromopyridine (0.076 mL, 0.80 mmol, 1.0 equiv), sodium tert-butoxide (383 mg, 3.98 mmol, 5.0 equiv), 2-dicyclohexylphosphino-2', 4', 6'-triisopropylbiphenyl (38 mg, 0.08 mmol, 0.1 equiv) and tris(dibenzylideneacetone)dipalladium(0) (73 mg, 0.08 mmol, 0.1 equiv) in 1,4-dioxane (10 mL). The reaction mixture was heated to 70 ° C for 72 h. The solvent was removed under reduced pressure, and the obtained residue was partitioned between water (2 mL) and ethyl acetate (5 mL). In 40 DEG C of 40 DEG C, 80 DEG C of 30 DEG C, 100 DEG C of 48 DEG C, 200 DEG C of 84 DEG C, 200 DEG C of 145 DEG C, 200 DEG C of 180 DEG C, 200 DEG C of 185 DEG C, 200 DEG C of 127 DEG C, 200 DEG C of 187 DEG C, 200 DEG C of 188 DEG C, 200 DEG C of 189 DEG C, 210 DEG C of 201 DEG C, 220 DEG C of 202 DEG C, 230 DEG C of 210 DEG C, 240 DEG C of 227 DEG C, 250 DEG C of 189 DEG C, 260 DEG C of 201 DEG C, 270 DEG C of 227 DEG C, 280 DEG C of 228 DEG C, 290 DEG C of 301 DEG C, 300 DEG C of 229 DEG C, 310 DEG C of 230 DEG C, 320 DEG C of 230 DEG C, 330 DEG C of 231 DEG C, 340 DEG C of 231 DEG C, 350 DEG C of 232 DEG C, 360 DEG C of 233 DEG C, 370 DEG C of 234 DEG C, 380 DEG C of 235 DEG C, 390 DEG C of 245 DEG C, 400 DEG C of 245 DEG C, 410 DEG C of 245 DEG C, 420

[0540] Yield: 27 mg (12%). MS (ESI+) m / z 266 (M+H) + .

[0541] 2-Methyl-N 1 -(Pyridin-2-yl)propane-1,3-diamine hydrochloride (110C)

[0542] The method applied was similar to that described in General Method 2.

[0543] A solution of hydrogen chloride (0.4 mL, 4 M in 1,4-dioxane) was added to tert-butyl (2-methyl-3-(pyridin-2-ylamino)propyl)carbamate (108B) (27 mg, 0.10 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to provide the desired product, 2-methyl-N1-(pyridin-2-yl)propane-1,3-diamine hydrochloride (110C), as a light yellow semisolid. The semi-crude sample was used in the next reaction without further purification.

[0544] Yield: 25 mg (assuming % quantitative).

[0545] 2-Methyl-N 1 -(5-(methylthio)pyrimidin-2-yl)-N 3-(Pyridin-2-yl)propane-1,3-diamine (Example 187)

[0546] The method applied was similar to that described in General Method 3.

[0547] Under nitrogen, 2-chloro-5-methylsulfanyl-pyrimidine (100B) (17 mg, 0.11 mmol, 1.05 equiv) was added to 2-methyl-N 1 -(pyridin-2-yl)propane-1,3-diamine hydrochloride (110C) (24 mg, 0.10 mmol, 1.0 equivalent) and cesium carbonate (99 mg, 0.30 mmol, 3.0 equivalent) in anhydrous N, N-dimethylformamide (0.5 mL) in a stirred solution. The mixture was heated to 50 ° C for 16 h and then concentrated under vacuum. Water (2 mL) was added, and the mixture was extracted with ethyl acetate (3x5 mL). The combined organic phases were washed with water (2 mL) and brine (2 mL) and then dried by a phase separator. The solvent was removed under vacuum to give a thick yellow oil, which was purified by reverse phase chromatography (10 mM aqueous ammonium bicarbonate solution / acetonitrile 5-95% elution) to give the desired 2-methyl-N-(pyridin-2-yl)propane-1,3-diamine hydrochloride (110C) as a sticky yellow solid. 1 -(5-(methylthio)pyrimidin-2-yl)-N 3 -(pyridin-2-yl)propane-1,3-diamine (Example 187).

[0548] Yield: 1.5 mg (5%). 1 H NMR (400MHz, CDCl3) δ8.35(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) + .

[0549] Plan 13

[0550]

[0551] Methyl 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylate (188)

[0552] The method applied was similar to that described in General Method 3.

[0553] Under nitrogen, methyl 5-bromopyrazine-2-carboxylate (114 mg, 0.53 mmol, 1.0 equiv) was added to 2-methyl-N 1 -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (102A) (150 mg, 0.53 mmol, 1.0 equiv) and cesium carbonate (514 mg, 1.58 mmol, 3.0 equiv) in anhydrous N,N-dimethylformamide (2.0 mL) in a stirred solution. The mixture was stirred at room temperature for 16 h and then concentrated under vacuum. Water (2.5 mL) was added and the mixture was extracted with ethyl acetate (3x5 mL). The combined organic phases were washed with water (2 mL) and brine (2.5 mL) and then dried through a phase separator. The solvent was removed under vacuum to give a brown oil, which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution). The semi-crude product obtained was further purified by flash chromatography (dichloromethane / methanol 0-10% elution) to give the desired methyl 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylate (Example 188) as a white solid.

[0554] Yield: 60 mg (32%). 1 H NMR (400MHz, CDCl3) δ8.76(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) + .

[0555] 5-((2-Methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylic acid (189)

[0556] The method applied was similar to that described in General Method 4.

[0557] Lithium hydroxide monohydrate (23 mg, 55 mmol, 5.0 equiv) was added to a stirred solution of methyl 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylate (188) (38 mg, 0.11 mmol, 1.0 equiv) in ethanol (0.4 mL) and water (0.4 mL). The mixture was stirred at ambient temperature for 72 h and then concentrated under reduced pressure. Water (0.5 mL) was added to the residue, and the mixture was acidified to pH approximately 3 with aqueous hydrochloric acid (2 M). The sticky precipitate was collected upon filtration, then extracted with ethyl acetate (3 x 3 mL), washed with water (1 mL), and then dried through a phase separator. The solvent was removed in vacuo to give the desired 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylic acid (Example 189) as a light yellow solid.

[0558] Yield: 34 mg (94%); 1 H NMR (400 MHz, 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 = 6.8 Hz, 3H), one NH proton not observed; MS (ESI+) m / z 335 (M+H) + .

[0559] (4-Hydroxypiperidin-1-yl)(5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazin-2-yl)methanone (Example 190)

[0560] The method applied was similar to that described in General Method 5.

[0561] 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 58 mg, 0.15 mmol, 1.5 equiv) was added to a solution of 5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazine-2-carboxylic acid (189) (34 mg, 0.10 mmol, 1.0 equiv) and 4-hydroxypiperidine (103 mg, 1.02 mmol, 10 equiv) in N,N-dimethylformamide (1 mL), and the reaction mixture was stirred at room temperature for 18 h. The solvent was removed under reduced pressure and the crude residue obtained was purified by reverse phase preparative HPLC to give the desired (4-hydroxypiperidin-1-yl)(5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazin-2-yl)methanone (Example 190) as an off-white solid.

[0562] Yield: 22 mg (53%). 1 H NMR(400MHz, CDCl3)δ8.46(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),3.42-3.25(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) + .

[0563] Plan 16

[0564]

[0565] 5-(5-Methyl-1,3,4-oxadiazol-2-yl)pyridin-2-amine (111B)

[0566] The method applied was similar to that described in General Method 7.

[0567] Under nitrogen, 2-bromo-5-methyl-1,3,4-oxadiazole (0.58 g, 3.58 mmol, 1.05 equiv) was added to a solution of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (111A) (0.75 g, 3.41 mmol, 1.0 equiv), cesium carbonate (3.33 g, 10.22 mmol, 3.0 equiv) and tetrakis(triphenylphosphine)palladium(0) (0.39 g, 0.34 mmol, 0.10 equiv) in water (5.5 mL) and 1,4-dioxane (22.5 mL). The reaction mixture was heated to 100 ° C. for 16 h. The solvent was removed under vacuum, water (20 mL) was added and the mixture was extracted with ethyl acetate (6 x 50 mL). The combined organic phases were washed with water (10 mL) and brine (10 mL), dried over a phase separator, and then concentrated under vacuum. The crude residue obtained was purified by reverse phase chromatography (eluting with 10 mM ammonium bicarbonate solution / acetonitrile 5%-30%) to give 5- (5- methyl -1,3,4- oxadiazole -2- bases) pyridine -2- amine (111B) as a desired off-white solid.

[0568] Yield: 153 mg (25%). 1 H NMR (400MHz, DMSO) δ8.49 (d, J = 2.0 Hz, 1H), 7.87 (dd, J = 2.4, 8.8 Hz, 1H), 6.75 (s, 2H), 6.57 (d, J = 8.8 Hz, 1H), 2.53 (s, 3H).

[0569] tert-Butyl (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)amino)propyl)carbamate (111C)

[0570] Tert-butyl N-(2-methyl-3-oxopropyl)carbamate (111B) (225 mg, 1.21 mmol, 1.2 equiv) was added to 5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-amine (114B) (153 mg, 1.00 mmol, 1.0 equiv), acetic acid (0.230 mL, 4.02 mmol, 4.0 equiv) and molecular sieves ( The mixture was stirred at room temperature for 5 minutes, and then sodium triacetoxyborohydride (532 mg, 2.51 mmol, 2.5 equivalents) was added in one portion. The mixture was stirred at room temperature for 40 h. Tert-butyl N-(2-methyl-3-oxopropyl)carbamate (225 mg, 1.21 mmol, 1.2 equivalents) was added, and the reaction mixture was stirred at room temperature for another 72 h. The reaction was quenched by carefully adding saturated aqueous sodium bicarbonate solution (30 mL). The mixture was vigorously stirred for 30 minutes, and the dichloromethane layer was then separated and concentrated under reduced pressure to give a gum. The crude product was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give the desired tert-butyl (2-methyl-3-((5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)amino)propyl)carbamate (111C) as an off-white gum. The crude sample was used in the next reaction without further purification.

[0571] Yield: 101 mg (29%).

[0572] 2-Methyl-N 1 -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride (111D)

[0573] The method applied was similar to that described in General Method 2.

[0574] 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-oxadiazol-2-yl)pyridin-2-yl)amino)propyl)carbamate (110C) (101 mg, 0.29 mmol), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to provide the desired 2-methyl-N-((5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)amino)propyl)carbamate (110C) as an off-white solid. 1 -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride (111D) The crude sample was used in the next reaction without further purification.

[0575] Yield: 93 mg (assuming quantitative %).

[0576] 2-Methyl-N 1 -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 203)

[0577] The method applied was similar to that described in General Method 3.

[0578] Under nitrogen, 2-chloro-5-methylsulfanyl-pyrimidine (100B, 51 mg, 0.32 mmol, 1.1 equiv) was added to 2-methyl-N 1 -(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride (111D) (93 mg, 0.29 mmol, 1.0 equiv) and cesium carbonate (283 mg, 0.87 mmol, 3.0 equiv) in a stirred suspension in anhydrous N,N-dimethylformamide (2.9 mL). The mixture was stirred at room temperature for 16 h, heated to 40 ° C for another 16 h, and then concentrated under vacuum. Water (10 mL) was added and the mixture was extracted with ethyl acetate (3x50 mL). The combined organic phases were washed with water (10 mL) and brine (10 mL) and then dried through a phase separator. The solvent was removed under vacuum to give a yellow oil, which was purified by flash chromatography (isohexane / ethyl acetate 0-100% elution) to give a crude residue, which was further purified by reverse phase preparative HPLC to give the desired 2-methyl-N-(5-(5-methyl-1,3,4-oxadiazole-2-yl)pyridin-2-yl)propane-1,3-diamine hydrochloride (111D) (93 mg, 0.29 mmol, 1.0 equiv). 1 -(5-(5-methyl-1,3,4-oxadiazol-2-yl)pyridin-2-yl)-N 3 -(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (Example 203).

[0579] Yield: 3.5 mg (3%). 1 H NMR (400MHz, CDCl3) δ8.70(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,2H),3.55-3.25(m,4H),2.59(s,3H),2.37(s,3H),2.12-2.04(m,1H),1.05(d,J=6.8Hz,3H); MS(ESI+)m / z 372(M+H) + .

[0580] Plan 17

[0581]

[0582] Ethyl 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylate (112B)

[0583] Add (R)-2-methyl-N 1-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) (544 mg, 2.19 mmol), ethyl 2-(6-bromopyridin-3-yl)cyclopropane-1-carboxylate (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 and the mixture was sparged with nitrogen for 2 minutes, after which copper (I) iodide (83 mg, 0.437 mmol) was added. The test tube was sealed under nitrogen and heated at 90 ° C overnight. The reaction 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 (2x30 mL). The combined organics were washed with water (2 x 40 mL) and then with brine (2 x 50 mL), dried over magnesium sulfate, and then concentrated under vacuum to give the crude title compound, ethyl 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylate (112B) as a light brown oil.

[0584] (Note: Several following impurities could not be separated by purification by normal phase chromatography).

[0585] 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) + .

[0586] 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylic acid (112C)

[0587] To a solution of ethyl 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylate (112B, 274 mg, 0.682 mmol) in ethanol (6 mL) and water (4 mL) was added lithium hydroxide monohydrate (143 mg, 3.41 mmol), and the resulting mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum, and the obtained residue was diluted with water (8 mL). The solution was adjusted to pH approximately 2 with 2M aqueous hydrochloric acid solution and then extracted with dichloromethane / methanol (20% methanol in dichloromethane, 2x10 mL). The combined organic layers were passed through a phase separator column and then concentrated under vacuum to give the crude title compound 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carboxylic acid (112C) as a brown oil which was used directly without further purification.

[0588] Yield: 280 mg. MS (ESI+) m / z 374 (M+H) + .

[0589] Tert-butyl 7-(2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane-2-carboxylate (112D)

[0590] Tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (65 mg, 0.281 mmol) was added to a solution of 2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-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), and the resulting mixture was stirred at room temperature for 18 h. Upon completion, the reaction mixture was concentrated under vacuum, and the obtained residue was diluted with ethyl acetate (5 mL) and water (3 mL). The aqueous phase is separated and extracted with ethyl acetate (2x5mL).The organic matter merged is washed with water (5mL), brine (2x10mL), dried over magnesium sulfate, then concentrated under vacuum.The crude residue obtained is purified using column chromatography (0-10% methanol elution in dichloromethane) to give the title compound, tert-butyl 7- (2- (6- (((R) -2- methyl -3- ((5- (methylthio) pyrimidin-2-yl) amino) propyl) amino) pyridine-3-yl) cyclopropane -1- carbonyl) -2,7- diazaspiro [3.5] nonane -2- formate (112D) in off-white solid.

[0591] Yield: 69 mg. 1 H NMR (400MHz, CDCl3) δ8.35(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),3.26-3.18(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) + .

[0592] (2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropyl)(2,7-diazaspiro[3.5]nonan-7-yl)methanone (Example 204)

[0593] Trifluoroacetic acid (0.2 mL) was added to a stirred solution of tert-butyl 7-(2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropane-1-carbonyl)-2,7-diazaspiro[3.5]nonane-2-carboxylate (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 azeotroped with dichloromethane (3x5 mL). The crude residue obtained was dissolved in dimethyl sulfoxide (1.5 mL) and purified by preparative HPLC. The obtained liquid was dried under vacuum and then freeze-dried from acetonitrile / water mixture to provide the title compound (2-(6-(((R)-2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyridin-3-yl)cyclopropyl)(2,7-diazaspiro[3.5]nonan-7-yl)methanone (204) as a fluffy white solid.

[0594] Yield: 15 mg. 1 H NMR (400MHz, CDCl3) δ8.54 (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=8.6Hz,1H),5.98(dd,J=6.9,6.9Hz,1H),5.49-5.49(m,1H),3.76(s,4H),3.51-3.4 3(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 ),1.89-1.79(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) + .

[0595] Following the procedure described in Scheme 17, the following examples were synthesized:

[0596] Table 14

[0597]

[0598]

[0599] Plan 18

[0600]

[0601] N 1 -(5-Bromopyrazin-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (113B)

[0602] A solution of crude 2-methyl-N-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine hydrochloride (100E) (340 mg, 1.60 mmol, 1.0 equivalent) in dimethylformamide (8 mL) is 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 is heated to 90 ° C for 18 h. The solvent is removed under reduced pressure, and the obtained residue is distributed between water (20 mL) and dichloromethane (50 mL). Each layer is separated, and the aqueous phase is extracted with dichloromethane (3x25 mL). The combined organic phase is washed with brine (25 mL), dried by a phase separator, and then concentrated to dryness under vacuum. The crude residue was purified by flash chromatography (eluting with isohexane / ethyl acetate 0-100%) to afford the title compound as a pale yellow gum. 1 -(5-bromopyrazin-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (113B).

[0603] Yield: 230 mg (38%). 1 H NMR (400MHz, CDCl3) δ8.36(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).

[0604] N 1 -(5-(2-methoxypyridin-3-yl)pyrazin-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (209)

[0605] The method applied was similar to that described in General Method 7 (note: 1,4-dioxane was used instead of dimethylformamide).

[0606] Under nitrogen atmosphere, N 1 -(5-bromopyrazin-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (113B) (50 mg, 0.135 mmol, 1.0 equiv) was added to a solution of 2-methoxy-3-phenylboronic acid (23 mg, 0.149 mmol, 1.1 equiv), cesium carbonate (132 mg, 0.406 mmol, 3 equiv) and tetrakis(triphenylphosphine)palladium(0) (15.6 mg, 0.0135 mmol, 0.1 equiv) 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 (3x15 mL). The combined organic phases were washed with brine (10 mL) and concentrated under vacuum. The crude residue obtained was purified by preparative HPLC to afford the desired N-(2-hydroxy-4-oxo-1-yl)-1-nitropropene as an off-white solid. 1 -(5-(2-methoxypyridin-3-yl)pyrazin-2-yl)-2-methyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (154).

[0607] Yield: 24 mg (44%). 1 H NMR (400MHz, CDCl3) δ8.77(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) + .

[0608] Plan 19

[0609]

[0610] tert-Butyl (2-methyl-3-oxopropyl) carbamate (114B)

[0611] Warning-exothermic reaction; Over 20 minutes, Dess-Martin periodinane (2.94g, 6.94mmol, 1.3 equivalents) was added portionwise to a solution of tert-butyl (3-hydroxy-2-methylpropyl) carbamate (1.0g, 5.34mmol, 1.0 equivalent) in dichloromethane (50mL), and the mixture was stirred at room temperature for 2h. The mixture was diluted with dichloromethane (25mL), and washed with 1M sodium bisulfite aqueous solution (2x10mL) and saturated sodium bicarbonate aqueous solution (2x10mL). The organic phase was dried by a phase separator, and then concentrated under vacuum to give thick tert-butyl (2-methyl-3-oxopropyl) carbamate (114B), which was immediately used in the next step without further purification.

[0612] tert-Butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carbamate (114C)

[0613] The method used was similar to that described in Scheme 13 (for the generation of 109D).

[0614] The crude product was purified by flash chromatography (isohexane / ethyl acetate 0-100%) to give 875 mg (47%) of the desired tert-butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carbamate (114C) as a pale yellow gum.

[0615] Yield: (MS (ESI+) m / z 345 (M+H) + .

[0616] tert-Butyl (3-((2'-methoxy-[3,3'-bipyridyl]-6-yl)amino)-2-methylpropyl)carbamate (114D)

[0617] The method applied was similar to that described in General Method 7.

[0618] The crude tert-butyl (3-((2'-methoxy-[3,3'-bipyridinyl]-6-yl)amino)-2-methylpropyl)carbamate (114D) was used immediately in the next step without further purification.

[0619] Yield: 110 mg, MS (ESI+) m / z 373 (M+H) + .

[0620] N1-(2'-methoxy-[3,3'-bipyridyl]-6-yl)-2-methylpropane-1,3-diamine (114E)

[0621] The method applied was similar to that described in General Method 2.

[0622] The coarse N1 -(2'-Methoxy-[3,3'-bipyridyl]-6-yl)-2-methylpropane-1,3-diamine (114E) was used immediately in the next step without further purification.

[0623] Crude yield: 75 mg MS (ESI+) m / z 273 (M+H) + .

[0624] N 1 -(2'-methoxy-[3,3'-bipyridyl]-6-yl)-2-methyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (210)

[0625] The method applied was similar to that described in General Method 1.

[0626] The crude residue obtained was purified by preparative HPLC to afford the desired product N as an off-white solid. 1 -(2'-methoxy-[3,3'-bipyridyl]-6-yl)-2-methyl-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (155)

[0627] Yield: 8 mg (7%). 1 H NMR (400MHz, CDCl3) δ8.36 (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),6.4 5(d,J=8.4Hz,1H),5.90(dd,J=6.3,6.3Hz,1H),5.11(dd,J=6.0,6.0Hz,1H),3.97(s,3H) ,3.56-3.23(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) + .

[0628] Plan 20

[0629]

[0630] tert-Butyl (3-([3,3'-bipyridyl]-6-ylamino)-2-methylpropyl)carbamate (115A)

[0631] A method similar to Scheme 19 (Example 115B) was applied to generate the aldehyde (114B), followed by a method similar to General Method 7.

[0632] The coarse N 1 -([3,3'-Bipyridyl]-6-yl)-2-methylpropane-1,3-diamine (115A) was used immediately in the next step.

[0633] Yield: 253 mg (63%) 1 H NMR (400MHz, DMSO) δ8.82(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,4 H),3.03-2.94(m,1H),2.88-2.80(m,1H),1.93-1.82(m,1H),1.40-1.38(m,9H).

[0634] N 1 -([3,3'-bipyridyl]-6-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine(211)

[0635] The method used was similar to that described in Scheme 13 (for the generation of 109D).

[0636] The crude residue obtained was purified by reverse phase chromatography (10 mM aqueous ammonium bicarbonate / acetonitrile 5%-95% elution) to give the desired product N as an off-white solid. 1 -([3,3'-bipyridyl]-6-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (211).

[0637] Yield: 32 mg (14%). 1H NMR(400MHz, CDCl3)δ8.77(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) + .

[0638] Plan 21

[0639]

[0640] 2-Bromo-5-((trimethylsilyl)ethynyl)pyridine (116B)

[0641] Triethylamine (49 mL, 352.25 mmol, 25 eq) was added to 2-bromo-5-iodopyridine (4.0 g, 14.09 mmol, 1.0 eq) followed by ethynyltrimethylsilane (2.9 mL, 21.13 mmol, 1.5 eq), copper(I) iodide (270 mg, 1.41 mmol, 0.1 eq), bis(triphenylphosphine)palladium(II) dichloride (99 mg, 0.14 mmol, 0.01 eq) and the mixture was stirred at room temperature for 17 h. The mixture was concentrated under vacuum to give a crude gum which was purified by flash chromatography (eluting with isohexane / ethyl acetate 0-40%) to give the intermediate, 2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B) as a white solid.

[0642] Yield: 2.5 g (69%). MS (ESI+) m / z 254 / 256 (M+H) + .

[0643] 2-Bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridine (116C)

[0644] To a solution of 2-bromo-5-((trimethylsilyl)ethynyl)pyridine (116B) (250 mg, 0.983 mmol, 1.0 eq) in ethanol (20 mL) was added 1-(azidomethyl)-4-methoxybenzene (177 mg, 1.08 mmol, 1.1 eq), and the mixture was stirred at room temperature for 1 hour. A 1M solution of tetrabutylammonium fluoride in tetrahydrofuran (0.54 mL, 1.08 mmol, 1.1 eq) was added, and the mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated under vacuum and then purified by flash chromatography (eluting with isohexane / ethyl acetate 0-100%) to give 2-bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridine (116C) as an off-white solid.

[0645] Yield: 0.21 g (61%). MS (ESI+) m / z 347 (M+H) + .

[0646] tert-Butyl (3-((5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridin-2-yl)amino)-2-methylpropyl)carbamate (116D)

[0647] To a degassed solution of 2-bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridine (116C) (135 mg, 0.391, 1.0 equiv) in dimethyl sulfoxide (5 mL) was added tert-butyl(3-amino-2-methylpropyl)carbamate (96 mg, 0.508 mmol, 1.3 equiv), L-proline (18 mg, 0.156 mmol, 0.4 equiv), potassium phosphate (1.6.5 mg, 0.078 mmol, 0.2 equiv) and copper(I) iodide (15 mg, 0.782 mmol, 0.2 equiv). The mixture was stirred at 90° C. for 28 h, diluted with water (20 mL), and extracted with dichloromethane (3×20 mL). The organic phases were combined and concentrated in vacuo, and the crude residue was purified by flash chromatography (eluting with isohexane / ethyl acetate 0-100%) to give 2-bromo-5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridine (116D) as a light brown solid.

[0648] Yield: 0.080 g (45%). MS (ESI+) m / z 453 (M+H) + .

[0649] tert-Butyl (3-((5-(1-(4-methoxybenzyl)-1H-1,2,3-triazol-5-yl)pyridin-2-yl)amino)-2-methylpropyl)carbamate (116E)

[0650] The method applied was similar to that described in General Method 2.

[0651] Yield: 25 mg (29%). 1 H NMR (400MHz, CDCl3) δ8.42(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 not observed).

[0652] N 1 -(5-(1H-1,2,3-triazol-5-yl)pyridin-2-yl)-2-methyl-N3-(4-(methylthio)phenyl)propane-1,3-diamine (212)

[0653] The method applied was similar to that described in General Method 3.

[0654] Yield: 18 mg (96%). 1 H NMR (400MHz, CDCl3) δ8.53(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) + .

[0655] Plan 22

[0656]

[0657] tert-Butyl (2-methyl-3-((5-(pyridin-2-yl)pyrazin-2-yl)amino)propyl)carbamate (117B)

[0658] To a solution of 5-(pyridin-2-yl)pyrazin-2-amine (200 mg, 1.16 mmol, 1.0 equiv) in dichloromethane (10 mL) were added tert-butyl(2-methyl-3-oxopropyl)carbamate (117B) (221 mg, 1.17 mmol, 1.0 equiv), To the product of 4-(5-(pyridin-2-yl)pyrazine-2-yl)amino)propyl group)carbamate (117B) was added to the 4-thiazolinone solution.The mixture was stirred at room temperature for 18 hours.Molecular sieve (750mg), acetic acid (0.266mL, 4.65mmol) and sodium triacetoxyborohydride (616mg, 2.9mmol), and the mixture was stirred at room temperature for 18h.Reaction mixture was diluted and filtered with dichloromethane (25mL).Organic phase was washed with water (25mL) and salt solution (25mL), then passed through phase separator post.Solvent was removed under vacuum, to provide thick tert-butyl (2-methyl-3-((5-(pyridin-2-yl)pyrazine-2-yl)amino)propyl group)carbamate (117B), which was not further purified and used for next step.

[0659] Yield: 310 mg (78%).

[0660] 2-Methyl-N 1 -(5-(Pyridin-2-yl)pyrazin-2-yl)propane-1,3-diamine (117C)

[0661] The method applied was similar to that described in General Method 2.

[0662] Yield: 200 mg (100%).

[0663] 2-Methyl-N 1 -(5-(Methylthio)pyrimidin-2-yl)-N3-(5-(pyridin-2-yl)pyrazin-2-yl)propane-1,3-diamine (213)

[0664] The method applied was similar to that described in General Method 1.

[0665] Yield: 20 mg (10%). 1 H NMR (400MHz, CDCl3) δ9.04(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=4.8,6.5Hz,1H),5.75-5.63(m,2H),3.58-3.50(m,2H),3.42-3.28(m ,2H),2.37(s,3H),2.19-2.06(m,1H),1.06(d,J=6.9Hz,3H); MS(ESI+)m / z 368(M+H) + .

[0666] Following the procedure described in Scheme 22, the following examples were synthesized:

[0667] Table 15

[0668]

[0669]

[0670] Plan 23

[0671]

[0672] 5-(1-Methyl-1H-1,2,3-triazol-4-yl)pyrazin-2-amine (118B)

[0673] The method used was similar to that described in General Method 7 (note: 1,4-dioxane / water was used as solvent instead of dimethylformamide).

[0674] Yield: 158 mg (73%) m / z 176 (M+H) + .

[0675] tert-Butyl (2-methyl-3-((5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrazin-2-yl)amino)propyl)carbamate (118C)

[0676] The method used was similar to that described in Scheme 13 (for the generation of 109D).

[0677] The crude tert-butyl(2-methyl-3-((5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrazin-2-yl)amino)propyl)carbamate (118C) was used immediately in the next step.

[0678] Yield: 101 mg (34%) MS (ESI+) m / z 347 (M+H) + .

[0679] 2-Methyl-N1-(5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrazin-2-yl)propane-1,3-diamine (118D)

[0680] The method applied was similar to that described in General Method 2.

[0681] The crude 2-methyl-N 1 -(5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrazin-2-yl)propane-1,3-diamine (106D) was used immediately in the next step.

[0682] Yield: 71 mg (assumed) MS (ESI+) m / z 367 (M+H) + .

[0683] 2-Methyl-N1-(5-(1-methyl-1H-1,2,3-triazol-4-yl)pyrazin-2-yl)-N3-(5-(methylthio)pyrimidin-2-yl)propane-1,3-diamine (218)

[0684] The method applied was similar to that described in General Method 1.

[0685] Yield: 25 mg (23%). 1 H NMR (400MHz, CDCl3) δ8.35(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) + .

[0686] Plan 24

[0687]

[0688] N 1 -(5-bromopyrazin-2-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine (119a)

[0689] The method applied was similar to that described in General Method 3.

[0690] Yield: 230 mg (38%). 1 H NMR (400MHz, CDCl3) δ8.36 (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).

[0691] 2-Methyl-N 1 -(5-(methylthio)pyrimidin-2-yl)-N 3-(5-(5-(Trifluoromethyl)-1H-pyrazol-4-yl)pyrazin-2-yl)propane-1,3-diamine (219)

[0692] The method applied was similar to that described in General Method 7 (note: 1,4-dioxane / water was used and no additional aliquots of reagents were added)

[0693] Yield: 18 mg (31%). 1 H NMR (400MHz, DMSO) δ13.70(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),7.5 5-7.44(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) + .

[0694] Plan 25

[0695]

[0696] tert-Butyl (2-methyl-3-(pyrazolo[1,5-a]pyrimidin-5-ylamino)propyl)carbamate (120A).

[0697] A microwave vial containing 5-chloropyrazolo[1,5-c]pyrimidine (500 mg, 3.26 mmol, 1 eq) and tert-butyl(3-amino-2-methylpropyl)carbamate (100E) (6.13 g, 32.56 mmol, 10 eq) was heated to 140°C under microwave irradiation for 30 minutes. The crude reaction mixture was purified by flash chromatography (eluting with isohexane / ethyl acetate 0-100%) to give tert-butyl(2-methyl-3-(pyrazolo[1,5-a]pyrimidin-5-ylamino)propyl)carbamate (120A).

[0698] Yield: 700 mg (70%). MS (ESI+) m / z 306 (M+H) + .

[0699] 2-Methyl-N 1 -(Pyrazolo[1,5-a]pyrimidin-5-yl)propane-1,3-diamine (120B)

[0700] The method applied was similar to that described in General Method 2.

[0701] MS (ESI+) m / z 229 (M+H) + .

[0702] Tert-butyl (2-methyl-3-(pyrazolo[1,5-a]pyrimidin-5-ylamino)propyl)carbamate (220)

[0703] The method applied was similar to that described in General Method 3.

[0704] Yield: 20 mg (14%). 1 H NMR (400MHz, DMSO) δ7.34(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),(2 x NH not observed); MS(ESI+)m / z 330(M+H) + .

[0705] Plan 26

[0706]

[0707] Methyl 6-bromonicotinimidate (121B)

[0708] Sodium methoxide (0.71 g, 13.22 mmol, 1.1 equiv) was added to an ice-cold solution of 6-bromonicotinonitrile (121A) (2.2 g, 12.02 mmol, 1.0 equiv) in dioxane / water (20 mL / 20 mL). The mixture was stirred under ice-cooling for 30 minutes and 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 organics were combined and the solvent was removed under vacuum to provide crude methyl 6-bromonicotinoimidate (121B), which was used in the next step without further purification.

[0709] Yield: 2.5 g (96%).

[0710] 6-Bromo-N'-methylnicotinimide hydrazide (121C)

[0711] Methylhydrazine (0.73 mL, 13.96 mmol, 1.2 eq) was added to a solution of 6-bromonicotinoimidate (121B) (2.5 g, 11.63 mmol, 1.0 eq), and the mixture was stirred at room temperature for 1 hour. The solvent was removed under vacuum to provide crude 6-bromo-N'-methylnicotinoimidyl hydrazide (121C), which was used in the next step without further purification.

[0712] Yield: 2 g (75%).

[0713] 2-Bromo-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridine (121D)

[0714] Formic acid (10 mL, 265 mmol, 30.4 equiv) was added to 6-bromo-N'-methylnicotinimide hydrazide (121C) (2.0 g, 8.73 mmol, 1.0 equiv), and the mixture was heated to reflux for 1 hour. The reaction mixture was diluted with water (100 mL) and then extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed with saturated aqueous sodium bicarbonate solution (50 mL), dried over sodium sulfate, and filtered. The solvent was removed under vacuum to provide a crude brown residue, which was purified by flash chromatography (eluting with 0-10% methanol in dichloromethane / dichloromethane) to give 2-bromo-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridine (121D) as an off-white solid.

[0715] Yield: 500 mg (23%). MS (ESI+) m / z 240 (M+H) + .

[0716] tert-Butyl (2-methyl-3-((5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-yl)amino)propyl)carbamate (121E)

[0717] The method applied was similar to that described in General Method 7 (note: 1,4-dioxane / water was used and no additional aliquots of reagents were added)

[0718] Yield: 125 mg 2-methyl-N 1 -(5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-yl)-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine(221)

[0719] The applied method was similar to that described in General Method 2 for deprotection followed by General Method 1.

[0720] Yield: 25 mg (18%).1 H NMR (400MHz, DMSO) δ8.42(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) + .

[0721] Plan 27

[0722]

[0723] tert-Butyl (3-((5-bromopyridin-2-yl)amino)-2-methylpropyl)carbamate (122B)

[0724] Tert-butyl N-(2-methyl-3-oxopropyl)carbamate (100E) (1.0 g, 5.34 mmol, 1.0 equiv) was added to 5-bromopyridin-2-amine (122A) (920 mg, 5.34 mmol, 1.0 equiv), acetic acid (1.2 mL, 21.36 mmol, 4.0 equiv) and molecular sieves ( To the mixture was added 1-[[(5-bromopyridin-2-yl)amino]-2-methylpropyl]-1-[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[

[0725] Yield: 875 mg (47%).

[0726] tert-Butyl (3-((2'-methoxy-[3,3'-bipyridyl]-6-yl)amino)-2-methylpropyl)carbamate (122C)

[0727] The method applied was similar to that described in General Method 7.

[0728] Yield: 138 mg.

[0729] N 1 -(2'-methoxy-[3,3'-bipyridyl]-6-yl)-2-methyl-N 3 -(5-(Methylthio)pyrimidin-2-yl)propane-1,3-diamine(222)

[0730] The applied method was similar to that described in General Method 2 for deprotection followed by General Method 1.

[0731] Yield: 57 mg. 1 H NMR (400MHz, CDCl3) δ8.36 (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,2 H),3.82(s,3H),3.56-3.47(m,1H),3.44-3.35(m,2H),3.28(dd,J=6.6,13.4Hz,1 H),2.35(s,3H),2.18-2.07(m,1H),1.06(d,J=6.9Hz,3H); MS(ESI+)m / z396(M+H) + .

[0732] Plan 28

[0733]

[0734] 6'-Chloro-2H-[1,3'-bipyridyl]-2-one (123B)

[0735] 2-Amino-5-iodopyridine (123A) (1.12 g, 5.00 mmol, 1.0 equiv) was combined with 2-hydroxypyridine (582 mg, 6.00 mmol, 1.2 equiv), potassium carbonate (760 mg, 5.50 mmol, 1.1 equiv), copper (I) iodide (143 mg, 0.75 mmol, 0.15 equiv) and 8-hydroxyquinoline (110 mg, 0.75 mmol, 0.15 equiv) in anhydrous dimethylsulfonamide (5 mL). The mixture was degassed under a stream of nitrogen and then heated at 130 ° C for 21 h. The reaction mixture was cooled to room temperature and then poured into a mixture of 10% aqueous ammonium hydroxide solution (100 mL) and ethyl acetate (50 mL). Activated carbon (1 g) was added and the mixture was filtered through a pad of celite and washed with ethyl acetate (2x50 mL). The layers were separated and the aqueous phase was extracted with ethyl acetate (2x50 mL). The combined organics were 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 (dichloromethane / methanol 0-10% elution) to give the desired product 6'-chloro-2H-[1,3'-bipyridinyl]-2-one (123B) as an off-white solid.

[0736] Yield: 245 mg (26%). 1 H NMR (400MHz, DMSO) δ7.88(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) + .

[0737] tert-Butyl (2-methyl-3-((2-oxo-2H-[1,3'-bipyridyl]-6'-yl)amino)propyl)carbamate (123C)

[0738] The method applied was similar to that described in Scheme 19.

[0739] 6'-Chloro-2H-[1,3'-bipyridinyl]-2-one (123B) was used in excess (245 mg, 1.31 mmol, 1.1 eq). The crude product was purified by flash chromatography (dichloromethane / methanol 0-7% elution) to give the desired product tert-butyl(2-methyl-3-((2-oxo-2H-[1,3'-bipyridyl]-6'-yl)amino)propyl)carbamate (123C) as a light brown solid.

[0740] Yield: 249 mg (58%). MS (ESI+) m / z 359 (M+H) + .

[0741] 6'-((2-Methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)-2H-[1,3'-bipyridyl]-2-one (223)

[0742] The method applied was similar to that described in General Method 2 using 4M HCl-water in 1,4-dioxane in a (4:1) ratio, followed by General Method 3 using a combination of cesium carbonate (3.0 eq.) and triethylamine (2.0 eq.).

[0743] Yield: 152 mg (57% over 2 steps).

[0744] 1 H NMR (400MHz, DMSO) δ8.35(2H,s),7.91(1H,d,J=2.6Hz),7.61(1H,ddd,J=0.6,2.1,6.8H z),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) ,3.33-3.14(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) + .

[0745] Using the procedure described in Scheme 28, the following examples were synthesized:

[0746] Table 16

[0747]

[0748]

[0749]

[0750]

[0751] Plan 29

[0752]

[0753] tert-Butyl (2-methyl-3-((5-(2-oxopyrrolidin-1-yl)pyrazin-2-yl)amino)propyl)carbamate (124B)

[0754] A solution of tert-butyl N-(3-amino-2-methylpropyl)carbamate (100E) (436 mg, 2.20 mmol, 1.1 equiv) in anhydrous dimethylsulfonamide (10 mL) was added to 2-bromo-5-(pyrrolidinone-1-yl)pyrazine (124A) (510 mg, 2.00 mmol, 1.0 equiv), tripotassium phosphate (866 mg, 4.00 mmol, 2 equiv), L-proline (94 mg, 0.80 mmol, 0.4 equiv) and copper (I) iodide (76 mg, 0.40 mmol, 0.2 equiv). The mixture was degassed and maintained under a stream of nitrogen, then heated at 90° C. for 20 h with stirring. The reaction mixture was cooled 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 organics were 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 (dichloromethane / methanol 0-6% elution) to give the desired product, tert-butyl (2-methyl-3-((5-(2-oxopyrrolidin-1-yl)pyrazin-2-yl)amino)propyl)carbamate (124B), as a yellow solid.

[0755] Yield: 375 mg (54%). MS (ESI+) m / z 350 (M+H) + .

[0756] 1-(5-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)pyrazin-2-yl)pyrrolidin-2-one (233)

[0757] The method applied was similar to that described in General Method 2 using 4M HCl-water in 1,4-dioxane in a [4:1] ratio followed by General Method 3.

[0758] Yield: 71 mg (18% over 2 steps).

[0759] 1H NMR(400MHz, CDCl3)δ9.02(1H,d,J=1.5Hz),8.35(2H,s),7.64(1H,d,J=1.5H z), 5.63 (1H, dd, J = 6.2, 6.2 Hz), 5.24 (1H, dd, J = 5.7, 5.7 Hz), 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) + .

[0760] Plan 30

[0761]

[0762] Tert-butyl 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (125B).

[0763] 1,1-difluoro-2-iodo-ethane (128 mg, 0.67 mmol, 1.2 equivalents) is added to a suspension of tert-butyl 2,7-diazaspiro [3.5] nonane-7-formate (125A) (126 mg, 0.56 mmol, 1.0 equivalents), potassium carbonate (231 mg, 1.67 mmol, 3.0 equivalents) in dimethylformamide (1.0 mL), and the mixture is stirred at 60 ° C for 2 h. The reaction mixture is cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase is washed with brine, dried by a phase separator, and then concentrated under reduced pressure to provide the desired tert-butyl 2- (2,2-difluoroethyl) -2,7-diazaspiro [3.5] nonane-7-formate (125B) as a light yellow oil.

[0764] Yield: 73 mg (45%). 1 H NMR (400MHz, CDCl3) δ5.87-5.57(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).

[0765] 2-(2,2-Difluoroethyl)-2,7-diazaspiro[3.5]nonane (125C)

[0766] Trifluoroacetic acid (1.3 mL) was added dropwise to a solution of tert-butyl 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (125B) (73 mg, 0.25 mmol, 1.0 equiv) in dichloromethane (1.3 mL). The mixture was stirred for 30 minutes and then concentrated to dryness under reduced pressure to provide the desired product, 2-(2,2-difluoroethyl)-2,7-diazaspiro[3.5]nonane (125C), as a light yellow oil, which was used immediately in the next reaction without further purification.

[0767] The intermediates in Table 17 were synthesized using conditions similar to those described for Intermediate 125C:

[0768] Table 17

[0769]

[0770] Plan 31

[0771]

[0772] Tert-butyl 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (126B).

[0773] 2,2,2- trifluoroethyl trifluoromethanesulfonate (153mL, 1.06mmol, 1.2 equivalents) is added to a suspension of tert-butyl 2,7- diazaspiro [3.5] nonane -7- formate (126A) (200mg, 0.88mmol, 1.0 equivalents), cesium carbonate (862mg, 2.65mmol, 3.0 equivalents) in acetonitrile (2.0mL), and the mixture is stirred at 80 ° C for 16h. The reaction mixture is cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase merged is washed with salt water, dried by a phase separator, and then concentrated under reduced pressure to provide tert-butyl 2- (2,2,2- trifluoroethyl) -2,7- diazaspiro [3.5] nonane -7- formate (126B) as a sticky white solid. The sample is used in the next step without further purification.

[0774] Yield: 341 mg (quantitative). 1 H NMR (400MHz, CDCl3) δ3.38-3.30 (m, 4H), 3.19 (s, 4H), 3.01 (q, J = 9.4Hz, 2H), 1.73-1.69 (m, 4H), 1.45 (s, 9H).

[0775] 2-(2,2,2-Trifluoroethyl)-2,7-diazaspiro[3.5]nonane (126C)

[0776] Trifluoroacetic acid (1.3 mL) was added dropwise to a solution of tert-butyl 2-(2,2,2-trifluoroethyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (126B) (110 mg, 0.357 mmol, 1.0 equiv) in dichloromethane (1.0 mL). The mixture was stirred for 30 minutes and then concentrated to dryness under reduced pressure to provide 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, assuming 100% yield.

[0777] Plan 32

[0778]

[0779] 2-Methyl-5-oxa-2,8-diazaspiro[3.5]nonane (127B)

[0780] Under nitrogen, lithium aluminum hydride (1M solution in tetrahydrofuran, 1.97 mL, 1.97 mmol, 3.0 equivalents) was added dropwise 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 equivalents) in tetrahydrofuran (4.0 mL). The reaction was stirred at 0°C for 5 minutes, the cooling bath was removed, and the reaction was then heated to 70°C for 16 hours (effervescence was observed at 0°C and 35°C). The reaction mixture was cooled to 0°C and quenched by slowly adding water (0.075 mL), aqueous sodium hydroxide solution (15%, 0.075 mL), and then water (0.22 mL). The cooling bath was removed, the reaction mixture was stirred for 15 minutes, magnesium sulfate was added and stirred for an additional 40 minutes. The mixture was dried through a phase separator, washed with tetrahydrofuran and diethyl ether, and then concentrated to dryness under reduced pressure to provide 2-methyl-5-oxa-2,8-diazaspiro[3.5]nonane (127B) as a light brown oil. The sample was used in the next reaction without further purification.

[0781] Yield: Assume 100%. 1 H NMR (400 MHz, 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), NH not observed.

[0782] The intermediates in Table 18 were synthesized using conditions similar to those described for Intermediate 127B:

[0783] Table 18

[0784]

[0785] Plan 33

[0786]

[0787] tert-Butyl 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (128B)

[0788] The procedure used was similar to that described in General Procedure 5 using 2-(tert-butoxycarbonyl)-2-azaspiro[3.3]heptane-6-carboxylic acid to provide the desired product tert-butyl 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (128B).

[0789] Yield: 111 mg (quantitative). 1 H NMR (400MHz, DMSO) δ3.87(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).

[0790] N,N-Dimethyl-2-azaspiro[3.3]heptane-6-carboxamide (128C)

[0791] Trifluoroacetic acid (2.1 mL) was added dropwise to a solution of tert-butyl 6-(dimethylcarbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (128B) (110 mg, 0.357 mmol, 1.0 equivalent) in dichloromethane (2.1 mL). The mixture was stirred for 30 minutes and then concentrated to dryness under reduced pressure to provide the desired product, N,N-dimethyl-2-azaspiro[3.3]heptane-6-carboxamide (128C), as a light brown oil. The sample was used in the next reaction without further purification, assuming a 100% yield.

[0792] Plan 34

[0793]

[0794] Methyl (S)-5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylate (129A)

[0795] The method applied was similar to that described in General Method 3.

[0796] Yield: 134 mg (20%). 1H NMR (400MHz, CDCl3) δ8.38(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),3.9 3(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) + .

[0797] (S)-5-Fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carboxylic acid (129B)

[0798] The method applied was similar to that described in General Method 4.

[0799] Yield: 98 mg (quant.). 1 H NMR (400 MHz, DMSO) δ 12.82 (s, 1H), 8.57-8.54 (m, 1H), 8.33-8.32 (m, 2H), 8.18 (d, J = 7.4 Hz, 1H), 7.50 (t, J = 6.1 Hz, 1H), 7.16 (d, J = 12.5 Hz, 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) + .

[0800] Tert-Butyl (S)-6-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carbonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate (129C)

[0801] The method applied was similar to that described in General Method 5.

[0802] Yield: 20 mg (54%). 1H NMR (400MHz, CDCl3) δ8.27(d,J=1.8Hz,1H),8.14(s,1H),7.99(dd,J=1.7,8.5 Hz,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 -2.09(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) + .

[0803] (S)-(5-Fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)(2,6-diazaspiro[3.4]octan-6-yl)methanone formate (Example 240)

[0804] Trifluoroacetic acid (1 mL) was added dropwise to a 0° C. cooled solution of tert-butyl (S)-6-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazole-6-carbonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate (149) (17 mg, 0.0166 mmol, 1.0 equiv) in anhydrous dichloromethane (1 mL). The mixture was stirred for 30 minutes and then concentrated under reduced pressure. The crude residue obtained was purified by preparative HPLC (formic acid additive) to give the desired product (S)-(5-fluoro-2-((2-methyl-3-((5-(methylthio)pyrimidin-2-yl)amino)propyl)amino)benzo[d]thiazol-6-yl)(2,6-diazaspiro[3.4]octan-6-yl)methanone formate (Example 189) as an off-white solid.

[0805] Yield: 9 mg (100%). 1 H NMR (400 MHz, CDCl3) δ 8.38-8.36 (m, 3H), 7.60-7.52 (m, 1H), 5.84-5.72 (m, 1H), 3.98 (d, J = 9.0 Hz, 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)+ .

[0806] Using the procedure described in Scheme 34, the following examples were prepared:

[0807] Table 19

[0808]

[0809] Plan 35

[0810]

[0811] 2-((3-((5-Mercaptopyrimidin-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (130A)

[0812]

[0149] Under nitrogen, N,N-dimethyl-2-((2-methyl-3-((5-(methylthio)pyrimidin-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 equiv) and N-methylpyrrolidine (1.5 mL) were added to a microwave vessel, sealed, and then heated to 160°C for 1 hour. LCMS analysis indicated complete conversion to the desired thiol, 2-((3-((5-((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (130A), which was used directly in the next step without purification.

[0813] 2-((3-((5-((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (244)

[0814] Potassium hydroxide (201 mg, 3.58 mmol, 20.0 equiv) and water (0.5 mL) were added to a solution of the crude thiol of 2-((3-((5-((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide in N-methylpyrrolidine (1.5 mL), and the mixture was cooled to -70°C and freeze-dried to a solid. Bromodifluoromethyl diethylphosphonate (40 uL, 0.215 mmol, 1.2 equiv) was added in one portion, and the reaction was allowed to warm to ambient temperature over 1 hour, then re-cooled to -70°C. Another aliquot of bromodifluoromethyl diethylphosphonate (40 uL, 0.215 mmol, 1.2 equiv) was added, and the mixture was allowed to warm to ambient temperature over 1 hour. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic phases were washed with water (20 mL) and brine (25 mL), then dried over magnesium sulfate. The solvent was removed under vacuum to give a crude brown oil, which was purified by flash chromatography (dichloromethane / methanol 0-10% elution) and then by preparative HPLC to give the desired 2-((3-((5-((difluoromethyl)thio)pyrimidin-2-yl)amino)-2-methylpropyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (244) as an off-white solid.

[0815] Yield: 15 mg (19%). 1 H NMR (400MHz, CDCl3) δ8.42-8.39(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=6.7Hz,1H),3.65-3.51(m,2H),3.43-3.29(m,2H),3.08(s,6H),2.20-2.10(m,1H),1.09-1.06(m,3H); MS(ESI+)m / z 453(M+H) + .

[0816] Following the procedure described in Scheme 35, the following examples were prepared:

[0817] Table 20

[0818]

[0819]

[0820] Plan 36

[0821]

[0822] rac-tert-Butyl ((1R,3R)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)carbamate (131B)

[0823] The method applied was similar to that described in General Method 1.

[0824] Yield: 220 mg (44%). 1 H NMR (400MHz, CDCl3) δ 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) + .

[0825] Racemic-(1R,3R)-N 1 -(5-(Methylthio)pyrimidin-2-yl)cyclopentane-1,3-diamine hydrochloride (131C)

[0826] The method applied was similar to that described in General Method 2.

[0827] Yield: 250 mg (quantitative %). MS (ESI+) m / z 225 (M+H) + .

[0828] Used in the next step without further purification.

[0829] rac-N,N-bis(4-methoxybenzyl)-2-(((trans)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide (131D)

[0830] The method applied was similar to that described in General Method 3.

[0831] Yield: 52 mg (23%). 1H NMR(400MHz, CDCl3) δ8.36(s,2H),8.00(d,J=1.6Hz,1H),7.74(dd,J=1.9,8.5H z,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 / z677(M+H) + .

[0832] rac-2-(((trans)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide (250)

[0833] Trifluoroacetic acid (10 mL) was added dropwise to an ice-cold solution of rac-N,N-bis(4-methoxybenzyl)-2-(((trans)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide (131D) (45 mg, 66.48 mmol, 1.0 equiv) in anhydrous dichloromethane (10 mL). The mixture was warmed to ambient temperature over 18 h and then concentrated to dryness under reduced pressure. The crude residue obtained was purified by preparative HPLC, and the clear fractions obtained were freeze-dried to give the desired 2-(((trans)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)benzo[d]thiazole-6-sulfonamide (Example 250) as a white powder.

[0834] Yield: 23 mg (79%). 1 H NMR(CDCl3)δ8.36(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) + .

[0835] Plan 41

[0836]

[0837] Step 1: Example 301C

[0838] To a solution of Example 301A (1 g, 3.14 mmol) in DMF (30 mL) was added CsCO (2.05 g, 6.28 mmol) and Example 301B (5.6 g, 62.8 mmol). The mixture was heated to 25° C. for 1 h. TLC indicated that the starting material was consumed. The reaction was filtered, and the filtrate (crude Example 301C) was used in the next step without further purification.

[0839] Step 2: Example 301D

[0840] The solution of Example 301C was treated with Boc2O (1.03 g, 4.72 mmol) and stirred at room temperature for 2 h. Water (100 mL) was added, followed by extraction with EA (50 mL x 2), washed with water and brine, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure and then purified by silica gel chromatography (eluted with petroleum ether / EtOAc=1 / 5 to 1 / 1) to give the desired product (Example 301D, 1.2 g, 81% yield) as a yellow solid. LCMS [M+H] + =573

[0841] Step 3: Instance 301E

[0842] To a solution of Example 301D (1.2 g, 2.54 mmol) in DCM (3 mL) was added TFA (1 mL) at room temperature. After addition, the reaction mixture was stirred at room temperature for 1 h. TLC detected that the starting material was consumed, and the mixture was concentrated to give the desired product Example 301E (945 mg, yield: 100%) as a yellow oil, which was used in the next step without further purification.

[0843] Step 4: Instance 301

[0844] To a solution of Example 301E (945 mg, 2.54 mmol) in ACN (10 mL) was added TEA (514 mg, 5.08 mmol) and Example 301F (408 mg, 2.54 mmol) at room temperature, and then to 70 ° C for 18 h. TLC detected that the starting material was consumed. The reaction was concentrated and purified by silica gel chromatography (eluted with petroleum ether / EtOAc=3 / 1 to 5 / 3) to give the desired product Example 301 (780 mg, yield: 61%) as a white solid. LCMS [M+H] + =497. 1H NMR (400MHz, DMSO-d6) δ8.78(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).

[0845] Plan 42

[0846]

[0847] Step 1: Instance 302B

[0848] A solution of Example 302A (6.77 g, 21 mmol) in THF (200 mL) was cooled to -65 ° C under an N2 atmosphere. MeMgBr (21 mL, 63.1 mmol, 3 M in THF) was added dropwise and then stirred at -65 ° C for 0.5 h. The reaction was warmed to room temperature for 2 h and quenched by adding water (200 mL). After extraction with EtOAc (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 (eluted with petroleum ether / EtOAc=3 / 1) to give the desired product Example 302B (4 g, 56% yield) as a yellow oil. LCMS[M+H] + =339

[0849] Step 2: Example 302C

[0850] To a solution of Example 302B (4 g, 0.012 mmol) in MeOH (40 mL) was introduced a suspension of Pd / C (400 mg) catalyst into a reactor. The vessel was purged with nitrogen and then hydrogen, and the reaction mixture was stirred at room temperature for 18 h. TLC and LCMS showed that the starting material was consumed. Example 302C (4 g, yield: 100%) was obtained by filtration and concentrated and used in the next step without any purification. LCMS [M+H] + =205

[0851] Step 3: Instance 302D

[0852] To a solution of Example 301A (154 mg, 0.48 mmol) in DMF (30 mL) was added Cs2CO3 (313 mg, 0.96 mmol) and Example 302C (982 mg, 4.8 mmol), and the mixture was heated to 25 ° C for 2 h. TLC detected that the starting material was consumed. The reaction was filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel chromatography (eluted with petroleum ether / EtOAc=1 / 1 to 1 / 4) to give the desired product Example 302D (500 mg, yield: 100%) as a white solid. LCMS [M+H] + =487

[0853] Step 4: Instance 302E

[0854] To a solution of Example 302D (486 mg, 2 mmol) in DCM (2 mL) was added TFA (1 mL) at room temperature. After addition, the reaction mixture was stirred at room temperature for 1 h. TLC detected that the starting material was consumed, and the mixture was concentrated to give the desired product Example 302E (532 mg, yield: 100%) as a yellow oil, which was used in the next step without further purification.

[0855] Step 5: Instance 302

[0856] 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 h. TM formation was detected by LCMS. Purification by preparative HPLC gave the desired product Example 302 (15 mg, yield: 6%) as a white solid. LCMS [M+H] + =511

[0857] 1 H NMR(400MHz, CDCl3)δ8.41(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).

[0858] Plan 43

[0859]

[0860] Step 1: Example 303B

[0861] A solution of Example 303A (33 g, 500 mmol) and methyl chloroformate (49.5 g, 520 mmol) in THF (75 mL) was added over a period of 30 min. KOH (56.1 g, 1000 mmol) in HO (50 mL) was added (maintaining the internal temperature below 40° C.). After the addition, the suspension was stirred at room temperature for 16 h. The suspension was filtered, the filter cake was washed with EtOH (50 mL x 3), and dried under vacuum to provide the desired product Example 303B (67 g, 82.7% yield) as a white solid. LCMS [M+H] + =163

[0862] Step 2: Example 303C

[0863] To a solution of Example 303B (4 g, 24.52 mmol) in MeOH (300 mL) and HCl (5 mL, 4.0 M in MeOH) was added Pd / C (8 g) at room temperature. The suspension was stirred at room temperature for 44 h under 15 psi of H2. LCMS showed that the starting material was completely consumed and the desired product was detected. The mixture was filtered and the filter cake was washed with MeOH (30 mL x 3). The filtrate was concentrated under reduced pressure to provide the product Example 303C (3.7 g, yield: 54.4%) as a yellow solid. LCMS [M+H] + =133.2

[0864] Step 3: Example 303D

[0865] A solution 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) was heated to 80° C. and stirred for 4 h. LCMS showed complete consumption of the starting material and the desired product was detected. The reaction mixture Example 303D was used in the next step without further purification. LCMS [M+H] + =415.5

[0866] Step 4: Instance 303E

[0867] Boc2O (5 g, 22.9 mmol) was added to the solution of Example 303D at room temperature and stirred for 1 h. LCMS showed that the starting material was consumed. The reaction was diluted with DCM (200 mL), washed with water (150 mL x 2), brine (200 mL x 3), dried over Na2SO4, filtered and concentrated under reduced pressure, which was purified by silica gel chromatography (eluted with petroleum ether / EtOAc=2 / 1 to 1 / 9) to provide the product Example 303E (680 mg, yield: 76.7% over two steps) as a white solid. LCMS [M+H] + =515.5

[0868] Step 5: Example 303F

[0869] To a solution of Example 303E (840 mg, 1.63 mmol) in THF (20 mL) was added CaCl2 (363 mg, 3.27 mmol) and NaBH4 (124 mg, 3.27 mmol) at room temperature, and the reaction was stirred for 16 h. The reaction was poured into saturated aqueous NH4Cl (100 mL) and diluted with EtOAc (30 mL) and separated. The aqueous layer was extracted with EtOAc (30 mL x 2), and the combined organic layers were concentrated and purified by silica gel chromatography (eluted with petroleum ether / EtOAc=3 / 1 to 1 / 9) to provide the product Example 303F (383 mg, yield: 48.3%) as a white solid. LCMS[M+H] + =487.6

[0870] Step 6: Instance 303G

[0871] To a solution of Example 303F (553 mg, 1.14 mmol) dissolved in DCM (10 mL) was added HCl / dioxane (4.0 M, 10 mL) at room temperature and the reaction was stirred for 0.5 h. TLC showed that the starting material was consumed. The reaction mixture was concentrated under reduced pressure to provide the product Example 303G (563 mg, yield: 100%) as a white solid. LCMS [M+H] + =387.5

[0872] Step 7: Example 303

[0873] To a solution of Example 303G (563 mg, 1.14 mmol) and KCO (940 mg, 6.82 mmol) in DMF (10 mL) was added Example 301A (183 mg, 1.14 mmol) at room temperature. The reaction was then heated to 60 ° C and stirred for 16 h. TLC showed that the starting material was consumed. The reaction mixture was cooled to room temperature and diluted with EtOAc (30 mL) and washed with water (20 mL). The aqueous layer was extracted with EtOAc (20 mL x 4), and the combined organic layers were washed with brine (20 mL x 2), dried over NaSO, filtered and concentrated. The residue was purified by silica gel chromatography (eluted with petroleum ether / EtOAc=3 / 1 to 1 / 9) to provide a crude product, which was purified by preparative HPLC to provide the product Example 303 (75.2 mg, yield: 12.93%) as a light yellow solid. LCMS[M+H] + =511

[0874] 1 H NMR(400MHz, CDCl3)δ8.37(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).

[0875] Plan 44

[0876]

[0877] Step 1: Example 304B

[0878] Example 304A (23.6 g, 0.12 mol), NaN (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 h. After TLC detected that the reaction was complete, EtOAc (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 with petroleum ether / EtOAc=5 / 1) to give the desired product Example 304B (29 g, yield 100%) as a white solid. LCMS [M+H] + =247

[0879] Step 2: Example 304C

[0880] To a solution of Example 304B (29 g, 0.12 mol) in EtOH (400 mL) was added HCl (22 mL) and Pd / C (1 g), and the heterogeneous mixture was stirred under H at room temperature for 18 h. TLC detected that most of the starting material was consumed. The mixture was filtered and the filtrate was concentrated to give the desired product Example 304C (21 g, yield: 100%) as a white solid. LCMS [M+H] + =221

[0881] Step 3: Example 304D

[0882] To a solution of Example 304C (6.6 g, 3 mol) in DMF (50 mL) was added 2-chloro-5-(methylthio)pyrimidine (2.4 g, 15 mmol) and TEA (7.6 g, 75 mmol). The resulting mixture was stirred at 100 ° C for 1 h. TLC detected that the starting material was consumed. The reaction mixture was concentrated and purified by silica gel chromatography (eluted with petroleum ether / EtOAc=1 / 1 to 1 / 4) to give the desired product Example 304D (450 mg, yield: 8%) as a white solid. LCMS [M+H] + =345

[0883] Step 4: Instance 304E

[0884] To a solution of Example 304D (450 mg, 1.3 mmol) in EtOH (5 mL) was added hydrazine hydrate (131 mg, 2.6 mmol) at room temperature. After 3 h, TLC detected consumption of the starting material, at which point the reaction was concentrated and purified by silica gel chromatography (eluted with DCM / MeOH=10 / 1) to give the desired product Example 304E (120 mg, 43% yield) as a white solid.

[0885] Step 5: Instance 304

[0886] 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) was added DBU (40 mg, 0.26 mmol). The resulting mixture was stirred at 50 ° C for 1 h. The starting material was consumed as detected by TLC, and the reaction mixture was concentrated and purified by preparative TLC to give the desired product Example 304 (30 mg, yield: 51%) as a white solid. LCMS [M+H] + =454.9. 1H NMR (400MHz, CDCl3) δ8.35(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).

[0887] Plan 45

[0888]

[0889] Step 1: Example 305

[0890] 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) was added DBU (47 mg, 0.18 mmol). After addition, the mixture was stirred at 50 ° C for 1 h. LCMS determined that the starting material was consumed. The reaction mixture was cooled to room temperature, diluted with EtOAc (5 mL) and washed with brine (5 mL). The organic layer was concentrated and purified by preparative TLC to give the desired product Example 305 (23.8 mg, yield: 60%) as a white solid. LCMS [M+H] + =440.9

[0891] 1 H NMR (400MHz, MeOD-d4) δ8.35 (s, 2H), 8.07 (s, 1H), 7.69 (dd, J = 8.0, 1H), 7.5 0(d,J=8.0Hz,1H),4.06(m,0.46H),3.57(m,4H),2.51(s,3H),2.32(s,3H).

[0892] Plan 46

[0893]

[0894] In a similar manner, starting from Example 306A, Example 306 (30 mg, yield: 50%) was prepared as a white solid: LCMS [M+H] + =454.9. 1H NMR (400MHz, CDCl3) δ8.35(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),6.0 4(t,J=5.6Hz,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).

[0895] Plan 47

[0896]

[0897] Step 1: Example 307A

[0898] To a solution of Example 306E (100 mg, 0.47 mmol) in DMF (0.5 mL) was added DBU (89 mg, 0.58 mmol) and methyl 2-chlorobenzo [d] thiazole-6-carboxylate (89 mg, 0.39 mmol). The mixture was heated to 50 ° C for 1 h. After TLC determined that the starting material was consumed, the reaction mixture was concentrated and purified by silica gel chromatography (eluted with petroleum ether / EtOAc=1 / 1 to 1 / 4) to give the desired product Example 307A (50 mg, yield: 26%) as a white solid. LCMS [M+H] + =406

[0899] Step 2: Example 307B

[0900] To a solution of Example 307A (50 mg, 0.12 mmol) in THF (1 mL) was added LiOH (0.4 mL, 1 M in water), and the resulting mixture was stirred at room temperature for 18 h. After TLC confirmed the consumption of the starting material, the reaction mixture was concentrated and purified by silica gel chromatography (eluted with DCM / MeOH=10 / 1) to give the desired product Example 307B (5 mg, yield: 11%) as a white solid. LCMS [M+H] + =392

[0901] Step 3: Example 307

[0902] A solution of Example 307B (10 mg, 0.025 mmol) was filled 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 h. After TLC detected the consumption of the starting material, the mixture was concentrated and purified by preparative TLC (eluted with EtOAc) to give the desired product Example 307 (2 mg, yield: 19%) as a white solid. LCMS [M+H] + =419. 1 H NMR (400MHz, CDCl3) δ8.36(s,2H),7.67(s,1H),7.53(d,J=8.2Hz,1H),7.35(dd,J=8.3,1.5Hz,1H),5.9 1(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).

[0903] Plan 48

[0904]

[0905] Step 2: Example 308A

[0906] A Schlenk tube equipped with a magnetic stirring bar is filled with TBAI (5 g, 13.6 mmol), cyclopentene (9.25 g, 136 mmol) and phthalimide (10 g, 68 mmol) in 250 mL of benzene. A solution of 65% TBHP (18.8 g, 136 mmol) is added before sealing the vial, and the reaction mixture is stirred at 80 ° C for 12 h. After cooling to room temperature, the reaction mixture is diluted in ethyl acetate and washed with brine. The aqueous phase is extracted with ethyl acetate. The combined organic layers are concentrated under reduced pressure. The residue is purified by silica gel column chromatography (eluted with PE / ethyl acetate=5 / 1) to provide the desired compound example 308A (6.5 g) as a white solid. LCMS[M+H] + =214.

[0907] 1H NMR (400MHz, CDCl3) δ2.08-2.17(m,1H),2.32-2.47(m,1H),2.41-2.53(m,1H),2.80-2.88(m,1H) ,5.33-5.46(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).

[0908] Step 2: Example 308B

[0909] To a solution of Example 308A (5 g, 23.5 mmol) in THF (25 mL) was added 50% hydrazine hydrate (3.52 g, 35.2 mmol) in H2O. The mixture was stirred at 70 ° C for 2 h. The mixture was then filtered and concentrated under reduced pressure. Di-tert-butyl dicarbonate (10.2 g, 47 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluted with PE / ethyl acetate=5 / 1) to provide the desired compound example 308B (550 mg) as a white solid. LCMS[M+H] + =184.

[0910] Step 3: Example 308C

[0911] To a solution of Example 308B (700 mg, 3.825 mmol) in DCM (5 mL) was added portionwise m-CPBA (790 mg, 4.59 mmol) at 0 ° C. After addition, the mixture was stirred at room temperature overnight. The resulting mixture was cooled to 0 ° C., and m-chlorobenzoic acid was filtered out and washed with additional cold DCM. The combined filtrate and washings were stirred for 30 min with 20% NaHSO3. The DCM layer was separated and extracted with 3N NaOH (3x30 mL), saturated NaCl (30 mL), and then dried over Na2SO4. The evaporation left a white solid, which was purified by silica gel column chromatography (eluted with PE / ethyl acetate=5 / 1) to provide the desired compound example 308C (455 mg) as a white solid. LCMS[M+H] + =144. 1 H NMR (400MHz, CDCl3) δ1.07-1.17(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).

[0912] Step 4: Example 308D

[0913] A mixture of Example 308C (445 mg, 2.28 mmol), NaN (297 mg, 4.57 mmol), NH4Cl (61 mg, 1.14 mmol), 2-methoxyethanol (5 mL) and H2O (1 mL) was stirred in a bath, maintained at 80 ° C for 16 hr. The resulting solution was evaporated to dryness and the residue was dissolved in H2O (5 mL). The solution was saturated with NaCl and then extracted with DCM (4x5 mL). The DCM solution was evaporated and the residue was purified by silica gel column chromatography (eluted with MeOH / DCM=3%-5%) to provide the desired compound example 308D (420 mg) as a colorless oil. LCMS [M+H] + =188.

[0914] Step 5: Instance 308E

[0915] Under H2 atmosphere, a suspension of Example 308D (420 mg, 1.74 mmol) and Pd / C (catalyst) in EtOH (5 mL) was stirred at room temperature for 16 h. The mixture was filtered and concentrated under vacuum. The residue was dried and used directly in the next step without further purification. LCMS [M+H] + =217.

[0916] Step 6: Example 308F

[0917] A mixture of Example 308E (150 mg, 0.694 mmol), 2-chloro-5-(methylthio)pyrimidine (111 mg, 0.694 mmol), DIPEA (180 mg, 1.4 mmol) in DMSO (5 mL) was stirred at 130 ° C for 3 hr. The resulting solution was cooled to room temperature, poured into water, and extracted with EtOAc (3x10 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA=2 / 1) to provide the desired compound example 308F (100 mg) as a colorless oil. LCMS[M+H] + =341.

[0918] 1 H NMR (400MHz, CDCl3) δ1.45(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).

[0919] Step 7: Instance 308G

[0920] To a solution of Example 308F (100 mg, 0.294 mmol) in DCM (3 mL) was added 4 M HCl in dioxane (3 mL). The mixture was stirred at room temperature for 2 h. 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.

[0921] Step 8: Example 308H

[0922] A mixture of Example 308G (70.6 mg, 0.256 mmol), tert-butyl 6-((2-chlorobenzo[d]thiazol-6-yl)sulfonyl)-2,6-diazaspiro[3.4]octane-2-carboxylate (130 mg, 0.294 mmol), 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 EtOAc (3x10 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with MeOH / DCM=5%) to provide the desired compound example 308H (70 mg) as a white solid. LCMS[M+H] + =648.

[0923] Step 9: Example 308

[0924] To a solution of Example 308H (70 mg, 0.108 mmol) in DCM (3 mL) was added 4M HCl in dioxane (3 mL). The mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated and purified by preparative HPLC to provide the title compound Example 308 (20 mg) as a white solid. LCMS [M+H] + =548.

[0925] 1H NMR (400MHz, DMSO-d6) δ1.47-1.56(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=8.3 3,1.88Hz,2H),8.16(d,J=1.88Hz,1H),8.37(s,2H),8.49(d,J=7.79Hz,2H),8.66(br.s.,1H).

[0926] Plan 49

[0927]

[0928] Step 1: Example 309A

[0929] Cyclopentadienyl sodium (2M solution 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 min at -40 ° C, the reaction mixture was poured into a 2: 1 mixture of pentane / ice-cold water (900 mL). After shaking and separating 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 isomerization of the double bond. After removing the drying agent by filtration, pentane was removed under vacuum at 0 ° C to provide (benzyloxymethyl)cyclopent-2,4-ene 1 as a light orange oil. Under argon, the obtained crude material is maintained at 0 ℃ and diluted with THF (160mL), cooled to -78 ℃, and at -78 ℃, is added dropwise to a suspension of (-)-Ipc2BH (1M solution in THF, 100mL, 100mmol, 1 equivalent) in THF (400mL) via a cannula. The mixture is slowly warmed to -10 ℃ and stirred at this temperature for 3 days. The reaction is then quenched by adding MeOH (40mL), followed by adding a 3M NaOH aqueous solution (40mL) and 30% H2O2 (40mL). After vigorous stirring for 24h at room temperature, THF is removed under reduced pressure and the remaining aqueous suspension is distributed between EtOAc (400mL) and salt solution (200mL). After extraction, the organic layer is dried over Na2SO4 and concentrated in vacuo. The crude orange oil was purified by column chromatography (eluent: 9:1 to 8:2 heptane:EtOAc) to give Example 309A (4.8 g, 23.53 mmol, 23%) as a light yellow oil. Rf = 0.29 (eluent: 7:3 heptane: EtOAc); 1H NMR (400 MHz, 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 = 8.9 Hz, 1H), 3.53 (dd, J = 5.4, 9.1 Hz, 1H), 4.29 (td, J = 4.1, 7.0 Hz, 1H), 4.52 (s, 2H), 5.53-5.58 (m, 1H), 5.70-5.74 (m, 1H), 7.24-7.37 (m, 5H).

[0930] Step 2: Example 309B

[0931] To a solution of Example 309A (4.8 g, 23.53 mmol, 1 eq) in anhydrous THF (100 mL) was added NaH (50% in mineral oil, 1.13 g, 28.2 mmol, 1.2 eq) at 0°C, and the mixture was stirred at temperature for 20 min. Benzyl bromide (BnBr, 3.6 mL, 30.5 mmol, 1.3 eq) and tetrabutylammonium iodide (TBAI, 100 mg, 0.3 mmol, 0.01 eq) were then added at 0°C, and the reaction mixture was stirred at room temperature. After 15 h, crushed ice was carefully added and the mixture was stirred for 30 min. After extraction with EtOAc (150 mL), the organic layer was washed with H2O (150 mL), brine (150 mL), dried over Na2SO4, and concentrated under vacuum. Purification by column chromatography (eluent: 98:2 to 95:5 heptane:EtOAc) gave Example 309B (6.3 g, 21.4 mmol, 80%) as a colorless syrup. Rf = 0.41 (eluent: 9:1 heptane:EtOAc). LCMS [M+H] + =295.1HNMR(500MHz, CDCl3)δ2.42(d,J=17.4Hz,1H),2.65-2.70(m,1H),3.07(brs,1H),3.33and 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).

[0932] Step 3: Example 309C

[0933] Under nitrogen, a 0.5 M solution of 9-BBN in THF (88 mL, 44 mmol) was added dropwise to a solution of Example 309B (6.50 g, 22.0 mmol) in anhydrous THF (10 mL) at 0°C. The reaction was slowly warmed to room temperature overnight. The reaction was cooled to 0°C and treated with EtOH (7 mL), 3NNaOH solution (20 mL) and H2O2 (33%, 20 mL) in sequence. The resulting mixture was stirred at room temperature overnight. The resulting residue was filtered and washed with EtOAc (200 mL). Water (150 mL) was added to the suspension and after phase separation, the aqueous layer was extracted with EtOAc (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:EtOAc) to give Example 309C (6.0 g, 19.3 mmol, 87%) as a yellow oil. LCMS [M+H] + =313.1H NMR (400MHz, CDCl3) δ7.35-7.25(m,10H,CH-arom.),4.52(s,2H,CH2-benzyl),4.49(d,1H,J=11.8Hz,CHH- benzyl),4.44(d,1H,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(dd,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).

[0934] Step 4: Example 309D

[0935] Compound Example 309C (6.0 g, 19.3 mmol) was dissolved in anhydrous pyridine (30 mL) and cooled to 0 ° C. TosCl (5.5 g, 28.9 mmol) was added in portions over a period of 30 min. After addition, the reaction was stirred at room temperature for 18 h. 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), brine (100 mL), and dried over MgSO4. The solvent was evaporated, and the residue was purified by flash silica gel chromatography (hexane / ethyl acetate 5: 1) to give (1R, 3S, 4R)-3-(benzyloxy)-4-((benzyloxy)methyl)cyclopentyl 4-methylbenzenesulfonate (1.4 g, 3.0 mmol, 15%) as a colorless oil. Rf = 0.26 (20% ethyl acetate in hexane). LCMS [M+H] + =467. 1HNMR(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 -3.92(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).

[0936] Compound (1R,3S,4R)-3-(benzyloxy)-4-((benzyloxy)methyl)cyclopentyl 4-methylbenzenesulfonate (1.4 g, 3.0 mmol) was dissolved in anhydrous DMF (20 mL) and NaN3 (2.1 g, 15.4 mmol) was added. The mixture was stirred at 60 ° C for 14 h. After adding 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 silica gel chromatography (hexane / ethyl acetate 10:1) to give 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) δ7.39-7.25(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).

[0937] Step 5: Example 309E

[0938] Compound example 309D (1.0 g, 2.95 mmol) was dissolved in anhydrous CH2Cl2 (20 mL) and cooled to -78 ° C. Over a period of 45 min, a solution of 1 M Cl3 (40 mL) in CH2Cl2 was added through a dropping funnel, and the mixture was stirred at -78 ° C for 3 h and then warmed to room temperature. The reaction was quenched with anhydrous MeOH (20 mL) at -78 ° C and warmed to room temperature overnight. The solvent was evaporated, and the residue was purified on silica gel (hexane / ethyl acetate 1: 1) to give (1S, 2R, 4S) -4- azido -2- (hydroxymethyl) cyclopentanol (420 mg, 2.67 mmol, 90%) obtained as a yellow oil. LCMS [M + H] + =158. 1 HNMR(400MHz, CDCl3) δ4.07(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(dddd,J=13.9,9.8,6.8Hz,1H).

[0939] Compound (1S, 2R, 4S) -4- azido -2- (hydroxymethyl) cyclopentanol (0.42 g, 2.67 mmol) was dissolved in anhydrous DMF (10 mL) and imidazole (198 mg, 2.94 mmol) was added at room temperature. After adding TBDPSCl (808 mg, 2.94 mmol) portionwise at 0 ° C, the mixture was stirred at room temperature for 16 h. The reaction 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 by silica gel (hexane / ethyl acetate 10: 1) to give Example 309E (630 mg, 1.6 mmol, 60%) as a yellow oil. LCMS [M + H] + =360. 1 H NMR (400MHz, CDCl3) δ7.69-7.66(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).

[0940] Step 6: Example 309F

[0941] 10% Pd / C (63 mg) was added to a suspension of compound example 309E (630 g, 1.6 mmol) in anhydrous EtOH (100 mL). The reaction was evacuated twice to exchange the inert gas atmosphere and then connected to two balloons filled with H2. The suspension was vigorously stirred at room temperature for 15 h. The palladium catalyst was removed by 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 used without further purification. LCMS [M + H] + =370.

[0942] To a solution of the amine (588 mg, 1.6 mmol) in DCM (50 mL) was added Boc O (700 mg, 3.2 mmol) in DCM (10 mL) at 0° C. and stirred at 0° C., then at room temperature for 2 h. The reaction mixture was concentrated, and the crude product was purified on silica gel (hexane / ethyl acetate 5:1) to give Example 309F (650 mg, 1.38 mmol, 87%) obtained as a colorless oil. LCMS [M+H]+ =470. 1 H NMR (400MHz, CDCl3) δ7.68-7.65(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).

[0943] Step 7: Instance 309G

[0944] 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 added to TosCl (384 mg, 2.0 mmol) at 0 ° C. The mixture was stirred at room temperature for 14 h and then concentrated to dryness. The crude product was purified by silica gel (hexane / ethyl acetate 10: 1) to give (1S, 2R, 4S) -4- ((tert- butoxycarbonyl) amino) -2- (((tert- butyldiphenylsilyl) oxy) methyl) cyclopentyl 4- methylbenzenesulfonate (590 mg, 0.94 mmol, 70%) obtained as a colorless oil. LCMS [M + H] + =625.

[0945] (1S,2R,4S)-4-((tert-butoxycarbonyl)amino)-2-(((tert-butyldiphenylsilyl)oxy)methyl)cyclopentyl 4-methylbenzenesulfonate (590 g, 0.94 mmol) was dissolved in anhydrous DMF (10 mL) and NaN (92 mg, 1.4 mmol) was added. The mixture was stirred at 60 ° C for 14 h. After adding ethyl acetate (100 mL), the organic layer was washed with saturated NaHCO solution (2×50 mL) and brine (50 mL) and dried over MgSO. The solvent was removed under reduced pressure. The residue was purified by flash silica gel chromatography (hexane / ethyl acetate 5:1) to give 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. 1H NMR (400MHz, CDCl3) δ7.72-7.67(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).

[0946] 10% Pd / C (46 mg) was added to a suspension of tert-butyl ((1S, 3R, 4S) -3-azido-4- (((tert-butyldiphenylsilyl) oxy) methyl) cyclopentyl) carbamate (460 g, 0.93 mmol) in anhydrous EtOH (50 mL). The reaction was evacuated twice to exchange the inert gas atmosphere and then connected to two H2-filled balloons. The suspension was vigorously stirred at room temperature for 15 h. The palladium catalyst was removed by 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 used without further purification. LCMS [M+H] + =470

[0947] Step 8: Example 309H

[0948] To a solution of Example 309G (70 mg, 0.17 mmol) in THF (5 mL) was added 1M TBAF (1.7 mL, 0.17 mmol) under N2 at 0°C, followed by stirring at room temperature for 2 h. The reaction mixture was concentrated to give the crude product 13, which was used in the next step. LCMS [M+H] + =231.

[0949] Step 9: Example 309I

[0950] 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 at 60° C. overnight. The mixture was purified by preparative HPLC to give compound Example 309I (38 mg, 47%) as a white solid. LCMS [M+H] + =472.

[0951] Step 10: Example 309J

[0952] 4M HCl / dioxane solution (5 L) was added to Example 309I (38 mg, 0.08 mmol) in DCM (1 mL), and the mixture was stirred at room temperature for 1 h. The mixture was concentrated to give Example 309J (30 mg, 100%) as a white solid. LCMS [M+H] + =372.

[0953] Step 16: Example 309

[0954] 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 h. The mixture was purified by preparative HPLC to give compound example 309 (5 mg, 12%) as a white solid. LCMS [M+H] + =496.

[0955] 1 H NMR (400MHz, CD3OD) δ8.36(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).

[0956] Plan 50

[0957]

[0958] Step 1: Example 310A

[0959] To a mixture of tert-butyl ((1S, 3S) -3-aminocyclopentyl) carbamate (1.5 g, 7.5 mmol), 2-chloro-5- (methylthio) pyrimidine (1.3 g, 8.3 mmol) in dimethyl sulfoxide (28 mL) was added N, N-diisopropylethylamine (3.7 mL, 22.5 mmol) at room temperature. Under an N2 atmosphere, the resulting mixture was stirred at 130 ° C for 6.5 h. Water (90 mL) and ethyl acetate (160 mL) were then added to the mixture. The combined organic layers were 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 to PE: EA = 6: 1) to give Example 310A (1.8 g, 76% yield) as a yellow oil. LCMS [M + H] + =325.

[0960] Step 2: Example 310B

[0961] To a mixture of Example 310A (1.8 g, 5.7 mmol) in methanol (2 mL) was added HCl / dioxane (8.0 mL, 4 mol / L) at room temperature. The resulting mixture was stirred at room temperature under a N atmosphere for 2.5 h. The mixture was then evaporated to give Example 310B (1.5 g, 100% yield) as a brown solid. LCMS [M+H] + =225.

[0962] Step 3: Example 310C

[0963] To a mixture of Example 310B (1.5 g, 5.7 mmol), ethyl 2-chlorobenzo [d] thiazole-6-carboxylate (1.4 g, 5.7 mmol) in DMSO (28 mL) was added DIEA (2.9 mL, 17.3 mmol) at room temperature. Under an N2 atmosphere, the resulting mixture was stirred at 80 ° C for 18 h. Water (80 mL) and ethyl acetate (150 mL) were then added to the mixture. The combined organic layers were 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 to 5: 1) to give Example 310C (1.9 g, 77% yield) as a yellow solid. LCMS [M + H] + =430.

[0964] Step 4: Example 310D

[0965] To a mixture of Example 310C (1.9 g, 4.4 mmol) in ethanol (20 mL) and water (10 mL) was added lithium hydroxide monohydrate (372 mg, 8.9 mmol) at room temperature. Under a nitrogen atmosphere, the resulting mixture was stirred at room temperature for 18 h. The mixture was then concentrated. Water (60 mL) and HCl (3 mol / L) were added to the residue until pH = 5. The precipitate was filtered and the filter cake was dried to give Example 310D (1.5 g, 84% yield) as a brown solid. LCMS [M+H] + =402. 1 H NMR (400MHz, DMSO-d6) δ8.49-8.47(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=7.2H z,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).

[0966] Step E: Example 310

[0967] A mixture of Example 310D (30 mg, 0.075 mmol), dimethylamine (6.75 mg, 0.15 mmol), HATU (28.5 mg, 0.075 mmol) in DCM was stirred at room temperature for 2 h. The resulting solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to provide the title compound Example 310 (20 mg) as a yellow solid. LCMS [M+H] + =429. 1 H NMR (600MHz, DMSO-d6) δ1.54-1.64(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).

[0968] Using the above procedure, the following examples were synthesized:

[0969] Table 31

[0970]

[0971]

[0972]

[0973]

[0974]

[0975]

[0976]

[0977]

[0978]

[0979]

[0980]

[0981]

[0982]

[0983]

[0984]

[0985]

[0986]

[0987]

[0988]

[0989]

[0990]

[0991]

[0992]

[0993]

[0994] Plan 51

[0995]

[0996] Step 1: Example 371A

[0997] Under N2 atmosphere, 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:HO=4:1 (50 mL) was stirred at 110 ° C overnight. The mixture was cooled to room temperature and water (50 mL) was added. The mixture was extracted with EtOAc (20 mL x 3), and the combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated; The residue was purified by silica gel chromatography eluted with petroleum ether:EtOAc=5:1 to provide Example 371A (660 mg, 72%) as a white solid.

[0998] Step 2: Example 371

[0999] A mixture of (1S, 3S)-N1-(5-(methylthio)pyrimidin-2-yl)cyclopentane-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 EtOAc (10 mL x5), and the combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC to provide Example 371 (10 mg, 6.4%). LCMS [M+H] + =409.

[1000] 1 H NMR (400MHz, CD3OD) δ8.41(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).

[1001] Plan 52

[1002]

[1003] Step 1: Example 372A

[1004] Under N2 atmosphere, a mixture of 2-fluoro-4-iodopyridine (200mg, 0.9mmol), pyridin-2 (1H) -one (102mg, 1.1mmol), CuI (17mg, 0.09mmol), N, N'-dimethyl-1,2-cyclohexanediamine (19mg, 0.17mmol) and K3PO4 (381mg, 1.8mmol) in DMSO (5mL) was stirred at 100 ° C for 3h. The mixture was cooled and water (20mL) was added. The mixture was extracted with EtOAc (20mL x 3), and the combined organic layers were washed with brine (50mL), dried over Na2SO4, filtered, and concentrated to give a crude product, which was purified by silica gel chromatography using petroleum ether: EtOAc = 5: 1-1: 1 elution to provide Example 372A (97mg, 57%) as an off-white solid.

[1005] Step 2: Example 372

[1006] A mixture of Example 372A (50 mg, 0.19 mmol), (1S,3S)-N1-(5-(methylthio)pyrimidin-2-yl)cyclopentane-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 EtOAc (10 mL x 5) and the combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC to provide Example 372 (10 mg, 13%). LCMS [M+H] + =395. 1 H NMR (400MHz, CD3OD) δ8.39(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).

[1007] Using the above procedure, the following examples were synthesized:

[1008]

[1009] Plan 54

[1010]

[1011] General Method 1

[1012] 6'-Chloro-5-methoxy-2H-[1,3'-bipyridyl]-2-one (Example 400A)

[1013] Under N2 atmosphere, a mixture of compound 2-chloro-5-iodopyridine (191 mg, 0.8 mmol, 1.0 eq), 5-methoxypyridin-2(1H)-one (100 mg, 0.8 mmol, 1.0 eq), CuI (15 mg, 0.08 mmol, 0.1 eq), N,N'-dimethyl-1,2-cyclohexanediamine (23 mg, 0.16 mmol, 0.2 eq) and K3PO4 (339 mg, 1.6 mmol, 0.2 eq) in DMSO (5 mL) was stirred overnight at 120° C. The mixture was cooled to room temperature and water (20 mL) was added. The mixture was extracted with EtOAc (20 mL x 2), the combined organic layers were washed with brine (30 mL), dried over Na 2 SO 4 , filtered, concentrated and purified by silica gel chromatography eluting with PE:EA=1:1 to afford 400A (60 mg, 0.23 mmol, 32%) as an off-white solid.

[1014] General Program 2

[1015] 5-Methoxy-6'-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridyl]-2-one (Example 400)

[1016] Under N2 atmosphere, a mixture of 400A (53 mg, 0.22 mmol, 1.0 equiv), 131C (50 mg, 0.22 mmol, 1.0 equiv), tBuXphos Pd G3 (18 mg, 0.022 mmol, 0.1 equiv) and tBuOK (49 mg, 0.44 mmol, 0.2 equiv) in dioxane (5 mL) was stirred at 110 ° C overnight. The mixture was cooled to room temperature and filtered. The filtrate was concentrated and purified by preparative TLC and then by preparative HPLC to provide compound 400 (50 mg, 53%, TFA salt) as an off-white solid. LCMS [M+H] + =425.

[1017] 1H NMR (400MHz, CD3OD) δ8.36(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).

[1018] Using the above procedure, the following examples were synthesized:

[1019]

[1020] Plan 55

[1021]

[1022] 5-Hydroxy-6'-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,3'-bipyridyl]-2-one (Example 402)

[1023] To a solution of 400 (50 mg, 0.12 mmol, 1.0 equiv) in DCM (10 mL) was added BBr (1 mL) at 0°C. The mixture was stirred at room temperature overnight, quenched with MeOH, and then concentrated. The residue was purified by preparative TLC and then by preparative HPLC to provide compound example 402 (50 mg, 53%, TFA salt) as an off-white solid: 1 H NMR (400MHz, CD3OD) δ8.37(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.

[1024] Plan 56

[1025]

[1026] 4-Hydroxy-5'-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2H-[1,2'-bipyridyl]-2-one (Example 403)

[1027] Analogously to the procedures described in General Method 1, General Method 2 and General Method 3, the starting material 4-(benzyloxy)pyridin-2(1H)-one provided the title compound: 1 H NMR (400MHz, CD3OD) δ8.32(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(d d,J=7.5,2.6Hz,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.

[1028] Plan 57

[1029]

[1030] General Program 4

[1031] 6'-Chloro-4-methoxy-3,3'-bipyridine (404)

[1032] Under N2 atmosphere, a mixture of (6-chloropyridin-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 at 105 ° C overnight. The mixture was cooled to room temperature and quenched with water (10 mL). The mixture was extracted with EtOAc (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 silica gel chromatography eluted with petroleum ether: EtOAc = 5: 1 to provide compound 404 (200 mg, 14%) as a brown solid.

[1033] Using the above procedure, the following examples were synthesized:

[1034]

[1035] Plan 58

[1036]

[1037] 2-Chloro-5-(2,2,2-trifluoroethoxy)pyrimidine(408)

[1038] To a solution of 2-chloropyrimidin-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) was added 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.97 g, 4.2 mmol, 1.1 equivalents). The resulting suspension was stirred at room temperature for 16 h and then distributed between EtOAc (30 mL) and water (80 mL). The separated aqueous layer was extracted with EtOAc (20 mL x 3). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated to provide 408 (0.74 g), which was used directly in the next step. LCMS [M+H] + =213.

[1039] Plan 59

[1040]

[1041] 2-Chloro-5-(difluoromethoxy)pyrimidine (409)

[1042] A solution of 2-chloropyrimidin-5-ol (101.6 g, 0.76 mol, 1.0 equivalent) in DMF (2000 mL) was filled with CsCO (300 g, 0.92 mol, 1.2 equivalents) and then stirred at room temperature for 1.5 h. 2-chloro-2,2-difluoroacetic acid sodium (340 g, 2.3 mol, 3.0 equivalents) was added and the reaction mixture was stirred at 100 ° C for 3.5 h. The reaction mixture was poured into water (5 L) and extracted with EA (3x1 L). The combined organic phases were dried over anhydrous NaSO and concentrated. The residue was purified by flash silica gel chromatography (PE: EA = 10: 1) to provide 409 (70 g).

[1043] Plan 60

[1044]

[1045] 2-Chloro-5-cyclopropylpyrimidine (410A)

[1046] To a solution of 5-bromo-2-chloropyrimidine (100 g, 518 mmol, 1.0 equiv), cyclopropylboronic acid (53 g, 616 mmol, 1.2 equiv) and Pd(dppf)Cl2 (10 g, 13.7 mmol, 0.03 equiv) in dioxane (1.5 L) was added Cs2CO3 (250 g, 769 mmol, 1.5 equiv) and stirred at 110 ° C under N2 for 12 h. 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 (eluted with hexane: ethyl acetate = 15: 1) to provide 410A (55 g) as a yellow solid. LCMS [M+H] + =155.

[1047] General Program 5

[1048] tert-Butyl ((1S,3S)-3-((5-cyclopropylpyrimidin-2-yl)amino)cyclopentyl)carbamate (410B)

[1049] Under N2, a mixture of 410A (40 g, 260 mmol, 1 equiv), tert-butyl ((1S, 3S) -3-aminocyclopentyl) carbamate (55 g, 275 mmol, 1.05 equiv) and DIPEA (105 g, 814 mmol, 3.13 equiv) in DMSO (400 mL) was stirred at 110 ° C for 12 h. The mixture was then cooled to room temperature and diluted with water (1000 mL). The resulting mixture was extracted with EtOAc (100 mL x 3), and the combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography (eluted with hexane: ethyl acetate = 5: 1 to 4: 1) to provide compound 410B (55 g) as a light solid. LCMS [M+H] + =319.

[1050] General Program 6

[1051] (1S,3S)-N 1 -(5-Cyclopropylpyrimidin-2-yl)cyclopentane-1,3-diamine (410C)

[1052] To a solution of 410B (44 g, 13.8 mmol, 1.0 equiv) in MeOH (250 mL) was added HCl (4 M in dioxane, 250 mL) dropwise and the resulting solution was stirred at room temperature for 2.5 h. After the reaction was complete, the mixture was concentrated to dryness under vacuum. The residue was redissolved in MeOH (500 mL) and an ion exchange resin ( 900OH -The mixture was filtered off and the filtrate was concentrated to provide 410C (44.5 g) as a yellow oil. LCMS [M+H] + =219.

[1053] Using the above procedure, the following examples were synthesized:

[1054]

[1055] Plan 61

[1056]

[1057] 2-Bromo-5-(methylthio)pyridine(416)

[1058] Under N2, a solution of 2,5-dibromopyridine (1 g, 4.22 mmol, 1 equivalent) in THF (20 mL) was cooled to -78 ° C. Then n-BuLi (2.5 M, 1.77 mL, 4.43 mmol, 1.05 equivalents) was added dropwise at -78 ° C. The reaction mixture was stirred for 20 minutes, followed by the slow addition of 1,2-dimethyldisulfane (0.411 mL, 4.64 mmol, 1.1 equivalents). The reaction mixture was stirred for another hour and then quenched by saturated NH4Cl. The reaction mixture was extracted with EA (100 mL) and water (100 mL), and then washed with brine. The organic phase was washed with anhydrous Na s The mixture was dried over SO4. The mixture was filtered and concentrated under reduced pressure. The crude residue was purified by flash silica gel chromatography (PE:EA=10:1) to give 416 (86.8 mg): ESI [M+H] + =204.1 1 H NMR (400MHz, CDCl3) δ8.25 (t, J = 6.2 Hz, 1H), 7.41 (ddd, J = 8.9, 8.3, 1.6 Hz, 2H), 2.58-2.40 (m, 3H).

[1059] Plan 62

[1060]

[1061] (1S,3S)-N1-(Thieno[3,2-b]pyridin-5-yl)cyclopentane-1,3-diamine (417)

[1062] The title compound was synthesized from the starting material 5-chlorothieno[3,2-b]pyridine in analogy to the procedures described in General Method 2 and General Method 6.

[1063] Using the above procedure, the following examples were synthesized:

[1064]

[1065] Plan 63

[1066]

[1067] General Program 7

[1068] 3-(6-Chloropyridin-3-yl)pyrimidin-4(3H)-one (419)

[1069] Pyrimidin-4(3H)-one (1.5 g, 15.61 mmol, 1.0 equiv), (6-chloropyridin-3-yl)boronic acid (2.9 g, 18.73 mmol, 1.2 equiv), Cu(OAc)2 (7.9 g, 43.71 mmol, 2.8 equiv), pyridine (2.5 mL, 31.22 mmol, 2.0 equiv) and A solution of molecular sieves (8 g) in DCM (60 mL) was stirred at room temperature for 48 h. The mixture was filtered through celite, and the filtrate was concentrated and purified by flash column (petroleum ether: EtOAc: 1:1) to afford Example 419 (170 mg, 5.3% yield) as a yellow solid.

[1070] 1 H NMR (400MHz, CDCl3) δ8.43(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) +

[1071] Using the procedure described in Scheme 63, the following examples were prepared:

[1072]

[1073] Plan 64

[1074]

[1075] Ethyl (E)-3-(2-fluoropyridin-4-yl)acrylate (423A)

[1076] Under N2 atmosphere, to a suspension of 60% NaH (1.54 g, 38.4 mmol, 1.2 equiv) in THF (15 mL) was added dropwise a solution of ethyl 2-(ethoxy(propoxy)phosphoryl)acetate (8.6 g, 38.4 mmol, 1.2 equiv) in THF at 0 ° C. The mixture was stirred at 0 ° C for 25 min, followed by the addition of a solution of 2-fluoroisonicotinaldehyde (4.0 g, 32.0 mmol, 1.0 equiv) in DMF (15 mL). The resulting mixture was stirred at room temperature for 16 h and then quenched by saturated aqueous NH4Cl 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 to PE:EA=1:15) to give 423A (3.0 g, 50% yield) as a white solid. LCMS: m / z 196 [M+H] + ,rt 2.890min. 1 H-NMR(400MHz, CDCl3)δ8.26(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).

[1077] Ethyl 3-(2-fluoropyridin-4-yl) propionate (423B)

[1078] To a solution of 423A (3.0 g, 20.5 mmol) in EtOH (20 mL) was added 10% Pd / C (400 mg). The reaction was purged with H2 and the mixture was stirred under H2 atmosphere overnight. The Pd / C was filtered off and the filtrate was concentrated under vacuum to give 423B (2.3 g, 76.7% yield), which was used directly: LCMS: m / z 198.1 [M+H] + ,rt 2.801min. 1 H-NMR (400MHz, CDCl3) δ8.12(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).

[1079] 3-(2-Oxo-1,2-dihydropyridin-4-yl)propanoic acid (423C)

[1080] To a 100 mL flask containing 423B (1.53 g, 7.77 mmol) was added concentrated HCl (5 mL). The mixture was heated to 100 ° C for 16 h. The mixture was then cooled to room temperature and 2 mL of water was added. Solid NaHCO 3 was added portionwise to adjust the pH to 6. The mixture was extracted with 30% i-PrOH in CHCl 3, and the combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by column chromatography (MeOH / DCM) to give 423C (1.6 g, yield 91%), which was used directly in the next step. LCMS: m / z 167.8 [M + H] + ; 1 H NMR (400MHz, CDCl3) δ8.14 (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).

[1081] Methyl 3-(2-oxo-1,2-dihydropyridin-4-yl)propanoate (423D)

[1082] To a 100 mL flask containing a solution of crude 423C (1.6 g) in MeOH (28 mL) was added 98% H2SO4 (0.15 mL). The reaction was heated to 90 ° C for 16 h and then cooled to room temperature. Saturated aqueous NaCl was added to the above mixture, which was extracted with 30% i-PrOH in CHCl3. The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by column chromatography (MeOH:DCM=1:15) to give the product 423D (702 mg, 55% yield). LCMS: m / z 182.1 [M+H] + .

[1083] General Program 8

[1084] Methyl 3-(6'-fluoro-2-oxo-2H-[1,3'-bipyridyl]-4-yl) propionate (423)

[1085] To a sealed tube was added 423D (100.0 mg, 0.55 mmol, 1.0 equiv), 2-fluoro-5-iodopyridine (147 mg, 0.66 mmol, 1.2 equiv), CuI (21.0 mg, 0.10 mmol, 0.2 equiv), N,N'-dimethyl-1,2-cyclohexanediamine (15.5 mg, 0.10 mmol, 0.2 equiv) and K2CO3 (151.0 mg, 1.10 mmol, 2.0 equiv). Dioxane (3 mL) was added, the resulting mixture was purged with N2 and stirred at 110°C for 16 h. The mixture was then 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 give 423 (42 mg, 27.6% yield). LCMS: m / z 278.08 [M+H] +

[1086] Plan 65

[1087]

[1088] Methyl 6'-fluoro-2-oxo-2H-[1,3'-bipyridyl]-5-carboxylate (424A)

[1089] A mixture of 6-fluoropyridin-3-amine (1.1 g, 10 mmol, 1.0 equivalent) and methyl 2-oxo-2H-pyran-5-carboxylate (1.5 g, 10 mmol, 1.0 equivalent) in EtOH (10 mL) was stirred overnight at reflux and then cooled to room temperature. The resulting precipitate was collected by filtration and purified by silica gel chromatography using petroleum ether: EtOAc = 2: 1 elution to provide compound 424A (440 mg, 18%) as an off-white solid.

[1090] Methyl 6'-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2-oxo-2H-[1,3'-bipyridine]-5-carboxylate (424B)

[1091] Similar to the method described above, 424B is provided from starting materials 131C and 424A.

[1092] General Program 9

[1093] 6'-(((1S,3S)-3-((5-(methylthio)pyrimidin-2-yl)amino)cyclopentyl)amino)-2-oxo-2H-[1,3'-bipyridine]-5-carboxylic acid (Example 424)

[1094] To a solution of compound 424B (100 mg, 0.22 mmol, 1.0 equiv) in MeOH:HO=5:1 (10 mL) was added NaOH (35 mg, 0.88 mmol, 4.0 equiv). The mixture was stirred at 50 ° C for 3 h and then acidified to pH = 5-6 after cooling to room temperature. The resulting mixture was concentrated and redissolved in THF (50 mL). The solid was filtered off, and the filtrate was concentrated and triturated with EA:MeOH=5:1 to provide Example 424 (90 mg, 93%) as a light yellow solid: 1 H NMR (400MHz, DMSO-d6) δ: 8.35 (s, 2H), 8.25s, 1H), 8.101H), 7.87 (d, J = 9.2Hz, 1H), 7.56 (s, 1H), 6.87 (s, 1H), 6 .51(d,J=9.2Hz,1H),4.36-...

Claims

1. Use of a compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cardiovascular diseases, wherein the compound is:

2. The method of claim 1, wherein the compound is:

3. The use according to claim 1, wherein the compound is a pharmaceutically acceptable salt of the following compound:

4. The method of claim 1 , wherein the cardiovascular disease is selected from the group consisting of hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome, and coronary artery disease.

5. The use according to any one of claims 1 to 4, wherein the cardiovascular disease is a. Familial hypercholesterolemia; b. Autosomal dominant hypercholesterolemia; or c. Atherosclerosis.

6. The use according to any one of claims 1 to 4, wherein the drug is used to reduce a. circulating serum cholesterol levels in the subjects; b. circulating serum LDL-cholesterol levels in the subjects; c. circulating serum VLDL-cholesterol levels in the subjects; d. circulating serum triglyceride levels in the subject; or e. Circulating serum lipoprotein A levels in subjects.

7. The use according to any one of claims 1 to 4, wherein the medicament is further administered in combination with one or more additional therapeutic agents.

8. The use of claim 7, wherein the one or more additional therapeutic agents are selected from alizumab, evolocumab, bocillinam, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimibe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin.

9. The use of claim 7, 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, a combination of niacin and lovastatin, ion exchange resins, antioxidants, ACAT inhibitors, bile acid sequestrants and PCSK9 translation inhibitors.

10. The use of claim 7, wherein the one or more additional therapeutic agents is a statin.

11. The use of claim 7, wherein the one or more additional therapeutic agents are selected from atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin.

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