PCSK9 inhibitors and methods of use thereof

Abstract

Disclosed herein are PCSK9 inhibitors. Also disclosed herein are methods of using PCSK9 inhibitors to treat certain diseases and disorders. [Formula 1] JPEG2025525415000025.jpg25128

Description

[Technical Field]

[0001] 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 through enhanced degradation of LDL receptors, independent 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 endosomal / lysosomal compartments. The enhanced binding affinity of PCSK9 to LDL receptors at the acidic pH of late endosomes / lysosomes reduces LDL receptor recycling and instead 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.

[0002] Another biological pathway involving the effect of PCSK9 on the LDL receptor is the development of septic shock. Septic shock is an often fatal complication of severe microbial infection (sepsis) that induces an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis is derived from microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharides (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors [Toll-like receptors (TLRs)] and therefore figures prominently in the septic inflammatory response (septic shock or sepsis).

[0003] PCSK9 reduces LPS uptake by the hepatic LDL receptor, resulting in free LPS overstimulating the body's immune response to pathogens and causing sepsis. Therefore, inhibiting PCSK9 is beneficial for preserving hepatic LDL receptors and providing systemic pathogen clearance and detoxification in response to sepsis. However, there is currently no effective treatment for sepsis or septic shock beyond antibiotic therapy.

[0004] For cardiovascular disease, few options exist for inhibiting PCSK9. Statins actually upregulate PCSK9 in HepG2 cells and primary human hepatocytes via increasing expression of SREBP-2, a transcription factor that upregulates both the LDL receptor and PCSK9 genes. Because elevated PCSK9 levels reduce the abundance of LDL receptors on the cell surface, increasing statin doses failed to achieve a proportional LDL-cholesterol-lowering effect.

[0005] Two monoclonal antibodies (mAbs), alirocumab and evolocumab, which selectively bind to extracellular PCSK9 and prevent its interaction with LDL receptors, recently received FDA approval for lowering LDL-C levels. In clinical trials, alirocumab demonstrated an approximately 50% reduction in LDL levels compared to placebo (Elbitar et al., Expert Opin Therapeutic Patents 2016 26:1377-1392). Patients receiving evolocumab demonstrated an approximately 60-75% reduction in LDL levels. The efficacy of these drugs demonstrates the potential for PCSK9 inhibitors to be an effective treatment for individuals with hypercholesterolemia and other cardiovascular diseases. However, both antibody drugs require intravenous administration and may trigger allergic reactions or other adverse immune responses in the body.

[0006] Unlike infections, which may be temporary, cardiovascular diseases often require lifelong management.Therefore, ease of dosing and administration is an important factor for patient compliance with maintenance drug treatment.There is a need for PCSK9 inhibitors with increased efficacy and easier administration, which can be achieved using small molecule PCSK9 inhibitors. Summary of the Invention

[0007] In one aspect, the disclosure provides a compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0008] [ka] [In the ceremony A is H, halo, hydroxy, alkyl, alkylthio, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 , and -C(O)NR 6 R 7 Selected from; B is selected from H, alkyl, and halo; or A and B together with the carbon atom to which they are attached form a 5- or 6-membered heteroaryl; X is NR 5 or O; Z is cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; Each R 6 and R 7 is independently H or alkyl; Y is selected from aryl, heteroaryl, and heterocyclyl; n is 0 or 1].

[0009] In another aspect, the disclosure relates to methods of treating certain diseases and disorders (eg, cardiovascular disease or septic shock) with compounds of formula (I). DETAILED DESCRIPTION OF THE INVENTION

[0010] The present disclosure relates to inhibitors of PCSK9 and uses thereof. In one aspect, the present disclosure provides a compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0011] [ka] [In the ceremony A is H, halo, hydroxy, alkyl, alkylthio, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR6 , and -C(O)NR 6 R 7 Selected from; B is selected from H, alkyl, and halo; or A and B together with the carbon atom to which they are attached form a 5- or 6-membered heteroaryl; X is NR 5 or O; Z is cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; Each R 6 and R 7 is independently H or alkyl; Y is selected from aryl, heteroaryl, and heterocyclyl; n is 0 or 1].

[0012] In certain preferred embodiments of Formula I, Z is cycloalkyl.

[0013] In certain embodiments of Formula (I), the structure of the compound is represented by Formula (Ia), (Ib), (Ic), or a pharmaceutically acceptable salt thereof:

[0014] [ka] During the ceremony, R 1 and R 1’ are each independently selected from alkyl, halo, hydroxyl, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amido, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamido, aryl, heteroaryl, heterocyclyl, and aralkyl; or R 1 and R 1’ combine to form an alkylalkene, cycloalkyl, cycloalkene, or heterocyclyl; Each R 2is independently selected from alkyl, halo, hydroxyl, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amido, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamido, aryl, heteroaryl, heterocyclyl, and aralkyl; n is 1, 2, 3 or 4.

[0015] In certain embodiments of Formula (I), (Ia), (Ib), or (Ic), the structure of the compound is represented by Formula (IIa), (IIb), (IIc), or a pharmaceutically acceptable salt thereof:

[0016] [ka]

[0017] In other embodiments of Formula (I), (Ia), (Ib), or (Ic), the structure of the compound is represented by Formula (IIIa), (IIIb), (IIIc), or a pharmaceutically acceptable salt thereof:

[0018] [ka]

[0019] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc), A is H, halo, hydroxy, alkylthio, alkyl, alkoxy, acyloxy, cyano, cycloalkyl, —C(O)OR 6 and -C(O)NR 6 R 7In certain embodiments, A is H. In other embodiments, A is alkyl. In other embodiments, A is halo (e.g., chloro). In still other embodiments, A is alkylthio. In still other embodiments, A is alkoxy. In still other embodiments, A is cycloalkyl. In certain preferred embodiments, A is selected from -SCH3, -SCHF2, and -OCHF2. In still other embodiments, A and B, together with the carbon atoms to which they are attached, form a pyrrolyl or thienyl ring that is unsubstituted or substituted with one or more alkyls.

[0020] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc), B is H.

[0021] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc), X is NR 5 is.

[0022] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc), Y is heteroaryl or heterocyclyl, preferably a monocyclic heteroaryl, such as pyridyl, pyridinyl, pyrazinyl, pyrimidinyl, or thiazolyl. In other embodiments, Y is a monocyclic heteroaryl selected from triazenyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, and triazolyl.

[0023] In certain such embodiments, the monocyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from alkyl, alkylthio, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, aryl, heteroaryl, heterocyclyl, nitro, sulfonamido, and alkylthio. In certain embodiments, the monocyclic heteroaryl is substituted with a 6-membered aryl, heteroaryl, or heterocyclyl selected from phenyl, pyridinyl, 2-hydroxypyridinyl, piperidinonyl, 2-hydroxy-1-methylpyridinyl, triazolyl, imidazolidinonyl, pyrimidonyl, 2-hydroxyisoquinolinyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl, and morpholinonyl. In certain embodiments, the monocyclic heteroaryl is substituted with a heteroaryl or heterocyclyl substituted with one or more substituents selected from halo, CN, alkyl, alkoxy, hydroxy, carboxy, -CO2 alkyl, and tetrazolyl. In certain preferred embodiments, the substituent is positioned para to Y relative to X.

[0024] In other embodiments, Y is a bicyclic heteroaryl such as benzothiazolyl, benzoxazolyl, benzimidazolyl, triazolopyridinyl, thiazolopyrindinyl, quinolinyl, or quinoxalinyl. In certain such embodiments, the bicyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from alkyl, haloalkyl, hydroxyalkyl, alkylthio, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, heteroaryl, nitro, and sulfonamido. In certain embodiments, the bicyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from alkylthio, alkoxycarbonyl, amido, carboxy, halo, and heteroaryl. In certain preferred embodiments, Y is of the formula -C(O)NR 8 R 9 or -NR 9 C(O)R 10wherein R 8 and R 9 are each independently selected from H, alkyl, heterocyclyl, and heteroaryl; or R 8 and R 9 together with the nitrogen atom to which they are attached form a 4-, 5-, 6-, or 7-membered heterocyclic or heteroaryl ring; R 10 is alkyl.

[0025] In another preferred embodiment, Y is a group of formula -S(O)NR 8 R 9 or -NR 9 S(O)2R 10 and substituted with a sulfonamide substituent of the formula: R 8 and R 9 are each independently selected from H, alkyl, and heteroaryl; or R 8 and R 9 together with the nitrogen atom to which they are attached form a 4-, 5-, 6-, or 7-membered heterocyclic ring; R 10 is alkyl.

[0026] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc), R 8 and R 9 are each independently selected from H, methyl, ethyl, triazolyl, and pyrazolyl. 8 and R 9is alkyl, and each alkyl is independently unsubstituted or substituted with one or more substituents selected from methyl, methoxy, carboxy, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, oxadiazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl, and 2,4-dihydro-3H-1,2,4-triazol-3-onyl. 8 and R 9 are taken together with the nitrogen atom to which they are attached to form a heterocyclic ring selected from aziradine, isothiazolidine-1,1-dioxide, azetindine, thiazol-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, and 2-oxa-6-azaspiro[3.3]heptane. In certain embodiments, R 8 and R 9 together with the nitrogen atom to which they are attached, form 2,8-diazaspiro[5,5]undecene, tetrahydroimidazo[1,2-a]pyrazine, octahydropyrazino[2,1-c][1,4]oxazine, tetrahydropyrido[3,4-d]pyrimidine, 2-oxa-8-azaspiro[4.5]decane, tetrahydropyrrolo[3,4-c]pyrazole, thiomorpholine, 2-oxa-7-azaspiro[3.5]nonane, 2,8-diazaspiro[4.5]decan-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9 ... Zaspiro[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, pyridinone, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3(2H)-one, piperidinone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5]nonane, pyrazole, and pyridazin-3(2H)-one. In certain embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxy, and heterocyclyl. In certain embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxy, and oxetanyl.

[0027] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc), R 1 and R 1’ combine to form a heterocyclyl (e.g., azetidinyl). In other embodiments, R 1 and R 1’ combine to form an alkylalkene (e.g., aminoallyl).

[0028] In certain embodiments, the compound is

[0029] [ka] or a pharmaceutically acceptable salt thereof.

[0030] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure (e.g., a compound of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc)) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0031] In yet another aspect, the present disclosure provides a method of treating a cardiovascular disease or disorder in a subject, the method comprising administering to the subject a compound of the present disclosure (e.g., a compound of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc)) or a pharmaceutically acceptable salt thereof.

[0032] In certain embodiments, the cardiovascular disease is selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipidemia, atherosclerosis, hepatic steatosis, metabolic syndrome, and coronary artery disease. In certain embodiments, the cardiovascular disease is familial hypercholesterolemia. In certain embodiments, the cardiovascular disease is autosomal dominant hypercholesterolemia. In certain embodiments, the circulating serum cholesterol level is reduced in the subject. In certain embodiments, the circulating serum LDL-cholesterol level is reduced in the subject. In certain embodiments, the circulating serum VLDL-cholesterol level is reduced in the subject. In certain embodiments, the circulating serum triglyceride level is reduced in the subject. In certain embodiments, the circulating serum lipoprotein A level is reduced in the subject. In certain embodiments, the subject has atherosclerosis. In certain embodiments, atherosclerotic plaque formation is reduced in the subject. In certain embodiments, the subject has a gain-of-function mutation in the PCSK9 gene.

[0033] In certain embodiments, the method comprises administering one additional therapeutic agent, in certain embodiments, the method comprises administering two additional therapeutic agents, in certain embodiments, the method comprises administering three additional therapeutic agents, in certain embodiments, the method comprises administering four additional therapeutic agents.

[0034] In certain embodiments, the additional therapeutic agent is an HMG-CoA reductase inhibitor, an HMG-CoA synthase inhibitor, an HMG-CoA reductase gene expression inhibitor, an HMG-CoA synthase gene expression inhibitor, an MTP / ApoB secretion inhibitor, a CETP inhibitor, a bile acid absorption inhibitor, a cholesterol absorption inhibitor, a cholesterol synthesis inhibitor, a squalene synthetase inhibitor, a squalene epoxidase inhibitor, a squalene cyclase inhibitor, a combined squalene epoxidase / squalene cyclase inhibitor, a fibrate, a niacin ... The additional therapeutic agent is selected from niacin, a combination of niacin and lovastatin, an ion exchange resin, an antioxidant, an ACAT inhibitor, a bile acid sequestrant, a PCSK9 translation inhibitor, an ATP citrate lyase inhibitor, an apoC3 inhibitor, an ANGPTL3 inhibitor, an omega-3 fatty acid, an Lp(a) inhibitor, an apoB inhibitor, an apoA1 inhibitor, a low-density lipoprotein receptor inhibitor, a low-density lipoprotein inhibitor, a diacylglycerol acyltransferase inhibitor, a lysosomal acid lipase inhibitor, and a lecithin cholesterol acyltransferase inhibitor. In certain embodiments, the additional therapeutic agent is an ATP citrate lyase inhibitor. In certain embodiments, the additional therapeutic agent is an apoC3 inhibitor. In certain embodiments, the additional therapeutic agent is an ANGPTL3 inhibitor. In certain embodiments, the additional therapeutic agent is an omega-3 fatty acid. In certain embodiments, the additional therapeutic agent is an Lp(a) inhibitor. In certain embodiments, the additional therapeutic agent is an apoB inhibitor. In certain embodiments, the additional therapeutic agent is an apoA1 inhibitor. In certain embodiments, the additional therapeutic agent is a low-density lipoprotein receptor inhibitor. In certain embodiments, the additional therapeutic agent is a low-density lipoprotein inhibitor. In certain embodiments, the additional therapeutic agent is a diacylglycerol acyltransferase inhibitor. In certain embodiments, the additional therapeutic agent is a lysosomal acid lipase inhibitor. In certain embodiments, the additional therapeutic agent is a lecithin cholesterol acyltransferase inhibitor.

[0035] In certain embodiments, the additional therapeutic agent is voranesolsen, evinacumab, IONIS-ANGPTL3-LRX, icosapent ethyl, eicosapentaenoic acid, eicosapentaenoic acid ester, eicosapentaenoic acid methyl ester, eicosapentaenoic acid ethyl ester, docosahexaenoic acid, docosahexaenoic acid ester, docosahexaenoic acid methyl ester, docosahexaenoic acid ethyl ester, bepedoic acid, ezetimibe, IONIS-APO(a)RX, mipomersen, CSL-112, lomitapide, AAV8.TBG.hLDLR (RGX-501), alipogene tiparvovec (alipogene tiparvovec), prazigastat, sebelipase, ACP-501 / MEDI6012, OM3FA-EE, OM3FA-CA, OM3FA-IPE, alirocumab, evolocumab, bococizumab, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimibe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin. In certain embodiments, the additional therapeutic agent is voranesorsen. In certain embodiments, the additional therapeutic agent is evinacumab. In certain embodiments, the additional therapeutic agent is IONIS-ANGPTL3-LRX. In certain embodiments, the additional therapeutic agent is icosapent ethyl. In certain embodiments, the additional therapeutic agent is eicosapentaenoic acid ethyl ester. In certain embodiments, the additional therapeutic agent is docosahexaenoic acid ethyl ester. In certain embodiments, the additional therapeutic agent is bepedoic acid. In certain embodiments, the additional therapeutic agent is ezetimibe. In certain embodiments, the additional therapeutic agent is IONIS-APO(a)RX. In certain embodiments, the additional therapeutic agent is mipomersen. In certain embodiments, the additional therapeutic agent is CSL-112. In certain embodiments, the additional therapeutic agent is lomitapide. In certain embodiments, the additional therapeutic agent is AAV8.TBG.hLDLR(RGX-501).In certain embodiments, the additional therapeutic agent is alipogene tiparvovec. In certain embodiments, the additional therapeutic agent is prazigastat. In certain embodiments, the additional therapeutic agent is sebelipase. In certain embodiments, the additional therapeutic agent is ACP-501 / MEDI6012. In certain embodiments, the additional therapeutic agent is OM3FA-EE. In certain embodiments, the additional therapeutic agent is OM3FA-CA. In certain embodiments, the additional therapeutic agent is OM3FA-IPE.

[0036] In certain embodiments, PCSK9 is overexpressed in the subject.In certain embodiments, the subject is male.In certain embodiments, the subject is overweight or obese.In certain embodiments, the subject is African American.In certain embodiments, the subject is Caucasian.In certain embodiments, the subject is Hispanic.

[0037] In certain embodiments, the PCSK9 inhibitor and the one or more additional therapeutic agents are administered simultaneously. In other embodiments, the one or more additional therapeutic agents are administered within about 5 minutes to about 168 hours before or after administration of the PCSK9 inhibitor.

[0038] In yet another aspect, the present disclosure provides a method of treating sepsis or septic shock in a subject, the method comprising administering to the subject a compound of the present disclosure (e.g., a compound of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc)) or a pharmaceutically acceptable salt thereof.

[0039] In certain embodiments, the method further comprises co-administering one or more additional therapeutic agents. In certain embodiments, the method comprises administering one additional therapeutic agent. In certain embodiments, the method comprises administering two additional therapeutic agents. In certain embodiments, the method comprises administering three additional therapeutic agents. In certain embodiments, the method comprises administering four additional therapeutic agents.

[0040] In certain embodiments, the additional therapeutic agent is kanamycin, amikacin, tobramycin, dibekacin, gentamicin, sisomicin, netilmicin, streptomycin, neomycin B, C, and E, imipenem, meropenem, ertapenem, doripenem, panipenum, biapenem, razupenem, or the like. penum), tebipenem, lenapenem, tomopenem, thiepenem, ciprofloxacin, garenoxacin, gatifloxacin, gemifloxacin, levofloxacin, cinoxacin, nalidixic acid, moxifloxacin, oxolinic acid, piromidic acid, pipemidic acid, rosoxacin , Enoxacin, Fleroxacin, Lomefloxacin, Nadifloxacin, Norfloxacin, Ofloxacin, Pefloxacin, Rufloxacin, Balofloxacin, Grepafloxacin, Levofloxacin, Pazufloxacin, Sparfloxacin, Temafloxacin, Tosufloxacin, Clinafloxacin, Sitafloxacin, Trovafloxacin, Prulifloxacin, Delafloxacin, JNJ-Q 2, nemonoxacin, zavofloxacin, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, methacycline, minocycline, rolitetracycline, tigecycline, ampicillin, amoxicillin, augmentin, piperacillin, tazobactam, chloramphenicol, and ticarcillin.

[0041] Pharmaceutical Composition The compositions and methods of the present disclosure can be used to treat individuals in need thereof. In certain embodiments, the individual is a mammal, such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or compound is preferably administered as a pharmaceutical composition, for example, comprising the 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 glycols, glycerol, oils (e.g., olive oil), or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are intended for human administration, particularly for invasive administration routes (i.e., routes such as injection or implantation that avoid transport or diffusion through epithelial barriers), the aqueous solution is pyrogen-free or substantially pyrogen-free. The excipient can be selected, for example, to provide delayed release of the drug or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition may be in the form of a dosage unit such as a tablet, capsule (including a sprinkle capsule and a gelatin capsule), granule, lyophilized product for reconstitution, powder, solution, syrup, suppository, injection, 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 a lotion, cream, or ointment.

[0042] Pharmaceutically acceptable carriers may contain physiologically acceptable agents that act, for example, to stabilize, increase the solubility, or increase the absorption of compounds such as the compounds of the present invention. 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 selection of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition may be a self-emulsifying or self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may be a liposome or other polymer matrix, into which, for example, the compounds of the present invention can be incorporated. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively easy to prepare and administer.

[0043] The phrase "pharmaceutically acceptable" is used herein to refer to 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 human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0044] The phrase "pharmaceutically acceptable carrier" as used herein 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 harmful to the patient. Some examples of materials that can function 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 wax; and (9) peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn starch, and the like. (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate, 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) ethyl alcohol; (20) phosphate buffer; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0045] Pharmaceutical compositions (preparations) can be administered to a subject by any of several routes of administration, including, for example, orally (e.g., as drenches, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue, as aqueous or non-aqueous solutions or suspensions); absorption through the oral mucosa (e.g., sublingually); 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). The compounds can also be formulated for inhalation. In certain embodiments, the compounds can be simply dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. 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 the patents cited therein.

[0046] The formulations may be conveniently provided in unit dosage form and may be prepared by any method known in the art of pharmacy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be the amount of compound that produces a therapeutic effect. Generally, out of 100 percent, this amount will range from about 1 percent to about 99 percent of the active ingredient, preferably from about 5 percent to about 70 percent, and most preferably from about 10 percent to about 30 percent.

[0047] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the present invention, with a carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present disclosure with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0048] Formulations of the present invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilisates, powders, granules, or solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions, or as elixirs or syrups, or as pastilles (using an inert base such as gelatin and glycerin, or sucrose and acacia), and / or mouthwashes, each containing a predetermined amount of a compound of the present disclosure as an active ingredient. The composition or compound may also be administered as a bolus, electuary, or paste.

[0049] To prepare solid dosage forms for oral administration (such as capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, and the like), 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 starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; and (5) dissolution retardants, such as paraffin. (6) absorption enhancers, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (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. For capsules (including sprinkle capsules and gelatin capsules), tablets, and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0050] Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surfactants, or dispersants. Molded tablets may be made by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0051] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills, and granules, may be optionally divided 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, for example, using hydroxypropylmethylcellulose, other polymer matrices, liposomes, and / or microspheres in various proportions to provide the desired release profile. They may be sterilized, for example, by filtration 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 compositions that release the active ingredient only, or preferentially in a certain portion of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that can 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.

[0052] The liquid dosage form useful for oral administration includes pharmaceutically acceptable emulsion, lyophilized product for reconstitution, microemulsion, solution, suspension, syrup and elixir.In addition to active ingredient, liquid dosage form can contain the inert diluent commonly used in the art, such as water or other solvent, cyclodextrin and its derivatives, solubilizer and emulsifier, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid ester of sorbitan, and their mixtures.

[0053] 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.

[0054] Suspensions may contain, in addition to the active compound, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0055] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and any preservatives, buffers, or propellants that may be required.

[0056] The ointments, pastes, creams and gels may contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

[0057] Powders and sprays can contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, for example, butane and propane.

[0058] Transdermal patch has the additional advantage of providing controlled delivery of the compound of the present disclosure to the body.Such dosage forms can be prepared by dissolving or dispersing active compound in suitable medium.Absorption enhancers can also be used to increase the flux of compound across the skin.The rate of such flux can be controlled by providing a rate-controlling membrane or dispersing compound in a polymer matrix or gel.

[0059] The phrases "parenteral administration" and "administered parenterally," as used herein, refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and epidural and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration include one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into a sterile injectable solution or dispersion immediately 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.

[0060] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (e.g., 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.

[0061] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial action can be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include isotonic agents, such as sugars and sodium chloride, in the compositions. Furthermore, prolonged absorption of injectable pharmaceutical forms can be brought about by the inclusion of agents that delay absorption, such as aluminum monostearate and gelatin.

[0062] In some cases, it is desirable to delay the absorption of drugs from subcutaneous or intramuscular injection in order to prolong the effect of drugs.This can be achieved by using a liquid suspension of crystalline or amorphous material with low water solubility.The absorption rate of the drug then depends on its dissolution rate, which in turn depends on the crystal size and crystalline form.Alternatively, delayed absorption of parenterally administered drug forms can be achieved by dissolving or suspending the drug in an oil vehicle.

[0063] Injectable depot forms are prepared by forming microencapsulated matrices of the target compound in biodegradable polymers such as polylactide-polyglycolide. The drug release rate can be controlled depending on the drug-to-polymer ratio and the properties of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0064] For use in the compounds of the present invention, the active compound may be administered per se or in combination with a pharmaceutically acceptable carrier, as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of the active ingredient.

[0065] The method of introduction can also be provided by a rechargeable or biodegradable device. In recent years, various sustained-release polymeric devices have been developed and tested in vivo for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. Various biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form implants for sustained release of compounds at specific target sites.

[0066] The actual dosage level of the active ingredient in the pharmaceutical composition may be varied to obtain an amount of the active ingredient that is not toxic to the patient and is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration.

[0067] The selected dosage level will depend on a variety of factors, including the activity of the particular compound or combination of compounds, or esters, salts, or amides thereof, used, the route of administration, the time of administration, the rate of excretion of the particular compound used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the particular compound used, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and similar factors well known in the medical arts.

[0068] A physician or veterinarian with ordinary skill in the art can easily determine and prescribe the required therapeutically effective amount of the pharmaceutical composition. For example, a physician or veterinarian can start the dosage of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. A "therapeutically effective amount" refers to the concentration of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of a compound varies depending on the subject's weight, sex, age, and medical history. Other factors that affect the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, other types of therapeutic agents administered in conjunction with the method of the present invention. Multiple administrations of the drug can deliver a larger total dose. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882, incorporated herein by reference).

[0069] Generally, a suitable daily dose of an active compound used in the compositions and methods of the present invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0070] If desired, the effective daily dose of active compound can be administered as 1, 2, 3, 4, 5, 6 or more divided doses that are administered separately at appropriate intervals throughout the day, optionally in unit dosage form.In certain embodiments of the present disclosure, active compound can be administered twice or three times a day.In a preferred embodiment, active compound is administered once a day.

[0071] Patients for this treatment may be any animal in need, including primates, particularly humans; as well as other mammals such as horses, cows, pigs, sheep, cats, and dogs; poultry; and pets in general.

[0072] In certain embodiments, the compounds of the invention may be used alone or co-administered with another type of therapeutic agent.

[0073] The present disclosure includes the use of pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present disclosure.In certain embodiments, the contemplated salts of the present invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts.In certain embodiments, the contemplated salts of the present invention include, but are not limited to, L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts.In certain embodiments, the contemplated salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.In certain embodiments, contemplated salts of the present invention include 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, l-ascorbic acid, l-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-gluconic acid, methylparaben ... These include, but are not limited to, heptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, l-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, l-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, l-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenate.

[0074] Pharmaceutically acceptable acid addition salts may also exist as various solvates, such as, for example, water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may be from the solvent of crystallization, may be inherent in the solvent of preparation or crystallization, or may be associated with such solvent.

[0075] Wetting agents, emulsifying agents, and lubricating agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives, and antioxidants can also be present in the composition.

[0076] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0077] definition Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. Generally, the nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0078] Unless otherwise indicated, the compounds and techniques of the present disclosure are generally carried out according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout this specification.See, for example, "Principles of Neural Science", McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intuitive Biostatistics", Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th ed.", WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th ed.", WH Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th ed.", Sinauer Associates, Inc., Sunderland, MA (2000).

[0079] Chemical terms used herein, unless otherwise defined herein, are used according to conventional usage in the art as exemplified in "The McGraw-Hill Dictionary of Chemical Terms," Parker S., Ed., McGraw-Hill, San Francisco, CA (1985).

[0080] All of the above, and any other publications, patents, and published patent applications mentioned in this application are specifically incorporated herein by reference. In case of conflict, the present specification, including its specific definitions, will control.

[0081] The term "agent" is used herein to refer to a chemical compound (e.g., an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (e.g., a nucleic acid, an antibody (including portions thereof, as well as humanized, chimeric, and human antibodies and monoclonal antibodies), a protein or portion thereof, such as a peptide, lipid, carbohydrate, etc.), or an extract made from biological material such as a bacterial, plant, fungal, or animal (particularly mammalian) cell or tissue. Agents include, for example, agents with known structures and agents with unknown structures. The ability of such agents to inhibit AR or promote AR degradation may make them suitable as "therapeutic agents" in the compounds and compositions of the present disclosure.

[0082] The terms "patient," "subject," or "individual" are used interchangeably and refer to either a human or a non-human animal. These terms include mammals such as humans, primates, livestock animals (including cows, pigs, etc.), companion animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0083] "Treating" a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable, reduction in the extent of the disease, stabilization (i.e., not worsening) of the disease state, prevention of disease spread, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or total). "Treatment" can also mean prolonging survival compared to expected survival in the absence of treatment.

[0084] The term "preventing" is art-recognized and, when used in connection with a condition such as local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure, or any other medical condition, is well understood in the art and includes administration of a composition that reduces the frequency of or delays the onset of symptoms of a medical condition in a subject compared to subjects not receiving the composition. Thus, preventing cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving prophylactic treatment compared to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population relative to an untreated control population, for example, by a statistically and / or clinically significant amount.

[0085] "Administering" or "administration of" a substance, compound, or drug to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, the compound or drug can be administered intravenously, intraarterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, intraocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through the skin tract). The compound or drug can also be suitably introduced by a rechargeable or biodegradable polymeric or other device, e.g., a patch and pump, or formulation, which provides sustained, sustained, or controlled release of the compound or drug. Administration can also be carried out, for example, once, multiple times, and / or over one or more extended periods of time.

[0086] The appropriate method of administering a substance, compound, or agent to a subject also depends, for example, on the age and / or physical condition of the subject, and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability, and toxicity). In some embodiments, the compound or agent is administered to the subject orally, for example, by ingestion. In some embodiments, the orally administered compound or agent is in an extended-release or delayed-release formulation or is administered using a device for such sustained-release or delayed-release.

[0087] As used herein, the phrase "co-administration" refers to any form of administration of two or more different therapeutic agents, in which a second agent is administered while a previously administered therapeutic agent is still effective in the body (e.g., two agents are effective in a patient simultaneously, which may involve a synergistic effect of the two agents). For example, different therapeutic compounds can be administered simultaneously or sequentially, either in the same formulation or in separate formulations. Thus, individuals receiving such treatment can benefit from the combined effects of the different therapeutic agents.

[0088] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or agent is the amount of the drug or agent that has the intended therapeutic effect when administered to a subject. The complete therapeutic effect does not necessarily occur by administering one dose, but may occur only after administering a series of doses. A therapeutically effective amount may be administered in one or more doses. The exact effective amount required for a subject depends, for example, on the subject's size, health, and age, as well as the nature and extent of the condition being treated, such as cancer or MDS. Those skilled in the art can easily determine the effective amount for a given situation through routine experimentation.

[0089] As used herein, the term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances when the event or circumstance occurs and instances when it does not occur. For example, "optionally substituted alkyl" refers to both the alkyl being substituted and the alkyl not being substituted.

[0090] It is understood that the substituents and substitution patterns on the compounds of the present disclosure can be selected by those skilled in the art to provide chemically stable compounds that can be easily synthesized from readily available starting materials using techniques known in the art and the methods described below. When a substituent itself is substituted with two or more groups, it is understood that these multiple groups can be on the same carbon or on different carbons, as long as a stable structure is obtained.

[0091] As used herein, the term "optionally substituted" refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent, including, but not limited to, hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH-O-alkyl, -OP(O)(O-alkyl) or -CH-OP(O)(O-alkyl). Preferably, "optionally substituted" refers to one to four hydrogen radicals in a given structure being replaced with a substituent as described above. More preferably, one to three hydrogen radicals are replaced with a substituent as described above. It is understood that the substituents may be further substituted.

[0092] As used herein, the term "alkyl" refers to a C1-C 10 Straight chain alkyl group or C1-C 10"Alkyl" refers to saturated aliphatic groups, including, but not limited to, branched alkyl groups. Preferably, "alkyl" refers to a C1-C6 straight chain alkyl group or a C1-C6 branched chain alkyl group. Most preferably, "alkyl" refers to a C1-C4 straight chain alkyl group or a C1-C4 branched chain alkyl group. Examples of "alkyl" include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neo-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl. "Alkyl" groups may be optionally substituted.

[0093] Furthermore, the term "alkyl," as used throughout the specification, examples, and claims, is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, and includes haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0094] As used herein, the term "alkylene" refers to a C1-C 50 Straight chain alkylene group or C1-C 50 It refers to a divalent saturated aliphatic group, including, but not limited to, branched alkylene groups. Furthermore, the term "alkylene," as used throughout the specification, examples, and claims, is intended to include both unsubstituted and substituted alkylene groups, the latter of which refers to alkylene moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups, such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0095] As used herein, the term "heteroalkylene" refers to a divalent aliphatic group in which one or more of the carbon atoms has been replaced with a heteroatom such as N, O, or S, or a moiety such as (C=O). The term includes, but is not limited to, a straight-chain heteroalkylene group of 2 to 50 atoms or a branched-chain heteroalkylene group of 2 to 50 atoms. When a heteroalkylene contains an N atom or another heteroatom having three or more open valence positions, the heteroatom can be unsubstituted (e.g., -N(H)-) or substituted with a substituent known to one of ordinary skill in the art (e.g., alkyl, aralkyl, aryl, etc., as further described herein), or all of the valences of the heteroatom can be occupied by its bond to the heteroalkylene chain (e.g., -N= or =N-). Furthermore, the term "heteroalkylene," as used throughout the specification, examples, and claims, is intended to include both unsubstituted and substituted heteroalkylene groups, the latter of which refers to heteroalkylene moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, and includes haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0096] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0097] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0098] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0099] The term "alkoxy" refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0100] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0101] "C x~y " or "C x ~C y The term "alkyl" when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups containing x to y carbons in the chain. CO alkyl indicates a hydrogen when the group is in a terminal position and a bond when it is internal. 1~6 An alkyl group contains, for example, 1 to 6 carbon atoms in the chain.

[0102] The term "alkylamino," as used herein, refers to an amino group substituted with at least one alkyl group.

[0103] The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0104] The term "amide" as used herein refers to a group.

[0105] [ka] In the formula, R 9 and R 10 each independently represents hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure.

[0106] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, such as a moiety that can be represented by the following formula:

[0107] [ka] In the formula, R 9 , R 10 and R 10 each independently represents a hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached complete a heterocycle with 4 to 8 atoms in the ring structure.

[0108] The term "aminoalkyl," as used herein, refers to an alkyl group substituted with an amino group.

[0109] The term "aralkyl," as used herein, refers to an alkyl group substituted with an aryl group.

[0110] The term "aryl," as used herein, includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is 5- to 7-membered, more preferably 6-membered. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of the rings being aromatic, e.g., the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0111] The term "carbamate" is art-recognized and refers to a group.

[0112] [ka] In the formula, R 9 and R 10 independently represent hydrogen or a hydrocarbyl group.

[0113] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocyclic group.

[0114] The term "carbocycle" includes 5- to 7-membered monocyclic rings and 8- to 12-membered bicyclic rings. Each ring in 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 in a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, can be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings, valence permitting, 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. A "carbocycle" may be substituted at any one or more positions capable of bearing a hydrogen atom.

[0115] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocyclic group.

[0116] The term "carbonate" is art-recognized and refers to the group -OCO2-.

[0117] The term "carboxy," as used herein, refers to a group represented by the formula -CO2H.

[0118] The term "cycloalkyl" includes substituted or unsubstituted non-aromatic monocyclic ring structures, preferably 4-8 membered rings, more preferably 4-6 membered rings. The term "cycloalkyl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of the rings is cycloalkyl and contains a substituent (e.g., R 100 ) is attached to the cycloalkyl ring, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, denzodioxane, tetrahydroquinoline, and the like.

[0119] The term "ester" as used herein refers to the group -C(O)OR 9 In the formula, R 9 represents a hydrocarbyl group.

[0120] The term "ether," as used herein, refers to a hydrocarbyl group linked to another hydrocarbyl group via an oxygen atom. Thus, the ether substituent of a hydrocarbyl group can be hydrocarbyl-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.

[0121] The terms "halo" and "halogen," as used herein, mean halogen and include chloro, fluoro, bromo, and iodo.

[0122] The terms "hetaralkyl" and "heteroaralkyl," as used herein, refer to an alkyl group substituted with a hetaryl group.

[0123] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, wherein the ring structure contains at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of the rings is heteroaromatic; for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, denzodioxane, tetrahydroquinoline, and the like.

[0124] The term "heteroatom," as used herein, means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0125] The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocycle group.

[0126] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to a substituted or unsubstituted non-aromatic ring structure, preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring, which ring structure contains at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of the rings is heterocyclic; for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0127] The term "hydrocarbyl," as used herein, refers to a group bonded through a carbon atom that has no =0 or =S substituents, typically has at least one carbon-hydrogen bond, and a primarily carbon backbone, but may optionally contain heteroatoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbyls for purposes of this application, while substituents such as acetyl (which has an =0 substituent on the linking carbon) and ethoxy (which is linked through an oxygen rather than a carbon) are not. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0128] The term "hydroxyalkyl," as used herein, refers to an alkyl group substituted with a hydroxy group.

[0129] The term "lower," when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups in which there are 10 or fewer atoms, preferably 6 or fewer atoms, in the substituent. For example, "lower alkyl" refers to an alkyl group containing 10 or fewer carbon atoms, preferably 6 or fewer carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent defined herein, whether appearing alone or in combination with other substituents such as hydroxyalkyl and aralkyl, is lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively (in which case, for example, atoms in aryl groups are not counted when counting carbon atoms in an alkyl substituent).

[0130] The terms "polycyclyl," "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 common to two adjacent rings, e.g., the rings are "fused rings." Each ring of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains 3 to 10 atoms, preferably 5 to 7 atoms, in the ring.

[0131] The term "sulfate" is art-recognized and refers to the group -OSO3H, or a pharmaceutically acceptable salt thereof.

[0132] The term "sulfonamido" is art-recognized and refers to a group that can be represented by the general formula:

[0133] [ka] In the formula, R 9 and R 10 independently represent hydrogen or hydrocarbyl.

[0134] The term "sulfoxide" is art-recognized and refers to the group --S(O)--.

[0135] The term "sulfonate" is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof.

[0136] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0137] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more backbone carbons. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is in accordance with the permissible valencies of the substituted atom and substituent, and that the substitution results in a stable compound that does not spontaneously undergo transformation, e.g., by rearrangement, cyclization, elimination, and the like. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. Substituents can include any of the substituents described herein, such as halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, 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. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

[0138] The term "alkylthio," as used herein, refers to an alkyl group substituted with a thiol group.

[0139] The term "thioester" as used herein refers to the group -C(O)SR 9 or -SC(O)R 9 In the formula, R 9 represents a hydrocarbyl.

[0140] The term "thioether," as used herein, is equivalent to an ether where the oxygen has been replaced with a sulfur.

[0141] The term "urea" is art-recognized and has the general formula

[0142] [ka] may be represented by In the formula, R 9 and R 10 independently represent hydrogen or hydrocarbyl.

[0143] The term "modulate," as used herein, includes inhibiting or suppressing a function or activity (eg, cell proliferation), as well as enhancing a function or activity.

[0144] The phrase "pharmaceutically acceptable" is art-recognized. In certain embodiments, this term includes compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0145] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to an acid addition salt or a base addition salt that is suitable or compatible for the treatment of patients.

[0146] The term "pharmaceutically acceptable acid addition salt" as used herein refers to any non-toxic organic or inorganic salt of any base compound represented by Formula I. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, and salicylic acid, and sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Either mono- or di-acid salts can be formed, and such salts can exist in either hydrated, solvated, or substantially anhydrous form. In general, acid addition salts of compounds of Formula I are more soluble in water and various hydrophilic organic solvents and generally exhibit higher melting points compared to their free base forms. The selection of an appropriate salt is known to those skilled in the art. Other pharmaceutically unacceptable salts (e.g., oxalates) may be used, for example, in the isolation of compounds of Formula I for laboratory use or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0147] The term "pharmaceutically acceptable base addition salt," as used herein, refers to a non-toxic organic or inorganic base addition salt of any acid compound represented by Formula I or any of its intermediates. Exemplary inorganic bases that form suitable salts include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines, such as methylamine, trimethylamine, and picoline or ammonia. The selection of an appropriate salt is within the skill of the art.

[0148] Many of the compounds useful in the compounds and compositions of the present disclosure have at least one stereocenter in their structure.This stereocenter may exist in R or S configuration, and the R and S designations are used according to the rules set forth in Pure Appl.Chem.(1976),45,11-30.The present disclosure contemplates all stereoisomeric forms, such as enantiomeric and diastereomeric forms of compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers).See, for example, International Publication No. 01 / 062726.

[0149] Furthermore, certain compounds containing alkenyl groups can exist as Z (zusammen) or E (entgegen) isomers, and in each case the present disclosure includes both mixtures and separate individual isomers.

[0150] Some of the compounds may also exist in tautomeric forms, and such forms, although not explicitly shown in the formulae set forth herein, are intended to be included within the scope of the present disclosure.

[0151] A "prodrug" or "pharmaceutically acceptable prodrug" refers to a compound (e.g., a compound of Formula I) that is metabolized, e.g., hydrolyzed or oxidized, in the host after administration to form a compound of the present disclosure. Typical examples of prodrugs include compounds having a biologically labile or cleavable (protecting) group on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to generate the active compound. Examples of prodrugs that use esters or phosphoramidates as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patent Nos. 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of the present disclosure are metabolized to generate a compound of Formula I. The present disclosure includes within its scope prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in "Design of Prodrugs," Ed. H. Bundgaard, Elsevier, 1985.

[0152] The phrase "pharmaceutically acceptable carrier," as used herein, means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, that is useful in formulating a drug for pharmaceutical or therapeutic use.

[0153] The terms "logarithm of solubility," "LogS," or "logS," as used herein, are used in the art to quantify the aqueous solubility of a compound. The aqueous solubility of a compound significantly affects its absorption and distribution properties. Low solubility often results in poor absorption. The LogS value is the unit-removed logarithm (base 10) of the solubility measured in moles / liter.

[0154] As used herein, the term "interact" in the context of an inhibitor relating to one or more residues of PCSK9 refers to a direct or indirect association of the inhibitor with either the backbone of the enzyme or the side chain of any given residue. For example, interactions include covalent bonds, hydrogen bonds, hydrophobic attraction, cation-pi interactions, anion-pi interactions, and others known in the art. [Example]

[0155] Having now generally described the invention, it will be more readily understood by reference to the following examples, which are included solely for the purpose of illustrating certain aspects and embodiments of the disclosure and are not intended to limit the invention.

[0156] Example 1: Preparation of exemplary PCSK9 inhibitors 1 H NMR spectra were performed on a Varian MR-400 spectrometer operating at 400 MHz (proton frequency) and equipped with a 5 mm H / nX broadband probehead with a self-shielded z-gradient coil for inverse detection, a deuterium digital lock channel unit, and a quadrature digital detection unit with transmitter offset frequency shift. Chemical shifts are reported as δ values (ppm) relative to trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in hertz (Hz), and multiplicities are reported using the following abbreviations: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad, and nd = undetermined.

[0157] Example B

[0158] [ka]

[0159] Step A: 1-tert-butyl 3-methyl 3-allylazetidine-1,3-dicarboxylate (2) To a stirred solution of 1-(tert-butyl) 3-methylazetidine-1,3-dicarboxylate (19.3 g, 89.7 mmol) in dry THF (180 mL) at −78° C., LiHMDS (1 M in THF, 179.5 mL, 179.5 mmol) was added dropwise under a nitrogen atmosphere. The reaction mixture was stirred at −78° C. for 1 h, and then allyl bromide (21.7 g, 179.5 mmol) was added to the above reaction mixture at −78° C. The reaction mixture was slowly warmed to room temperature and stirred overnight. Upon completion, the reaction mixture was cooled to 0° C., quenched with saturated aqueous NH4Cl (200 mL), and extracted with EA (150 mL × 3). The combined organic layers were washed with brine (250 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give 1-(tert-butyl) 3-methyl 3-allylazetidine-1,3-dicarboxylate (crude, 22 g, yield: 90%) as a yellow oil, which was used directly in the next step without further purification.

[0160] Step B: tert-butyl 3-allyl-3-(hydroxymethyl)azetidine-1-carboxylate (3) To a stirred solution of 1-(tert-butyl) 3-methyl 3-allylazetidine-1,3-dicarboxylate (22 g, 86 mmol) in dry DCM (300 mL) at −78° C., DIBAl-H (1 M in toluene, 129 mL, 129 mmol) was slowly added and stirred at room temperature for 2 h. After completion, the reaction mixture was cooled to 0° C., quenched with NaOH (1 M in HO, 100 mL), and extracted with DCM (200 mL × 3). The combined organic layers were washed with brine (250 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give tert-butyl 3-allyl-3-(hydroxymethyl)azetidine-1-carboxylate (crude, 15 g, yield: 77%) as a yellow oil, which was used directly in the next step without further purification.

[0161] Step C: tert-Butyl 3-allyl-3-formylazetidine-1-carboxylate (4) To a stirred solution of tert-butyl 3-allyl-3-(hydroxymethyl)azetidine-1-carboxylate (15 g, 66 mmol) in dry DCM (500 mL) at 0 °C, Dess-Martin (56 g, 132 mmol) was added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with NaSO (10%, 200 mL) and aqueous NaHCO (10%, 200 mL) and extracted with DCM (200 mL × 3). The combined organic layers were washed with brine (300 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MTBE / PE = 25%) to give tert-butyl 3-allyl-3-formylazetidine-1-carboxylate (9.2 g, yield: 46%, 3 steps) as a yellow oil.

[0162] Step D: tert-butyl 3-allyl-3-(1-hydroxyallyl)azetidine-1-carboxylate (5) To a stirred solution of tert-butyl 3-allyl-3-formylazetidine-1-carboxylate (9.2 g, 40.8 mmol) in dry THF (200 mL) at −78° C., vinylmagnesium bromide (1 M in THF, 49 mL, 49 mmol) was added, and the reaction mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was cooled to 0° C., quenched with saturated NH4Cl (200 mL), and extracted with EA (200 mL × 3). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give tert-butyl 3-allyl-3-(1-hydroxyallyl)azetidine-1-carboxylate (crude, 10 g, yield: 97%) as a yellow oil, which was used directly in the next step without further purification.

[0163] Step E: tert-Butyl 3-acryloyl-3-allylazetidine-1-carboxylate (6) To a stirred solution of tert-butyl 3-allyl-3-(1-hydroxyallyl)azetidine-1-carboxylate (10 g, 39.5 mmol) in dry DCM (200 mL) at 0 °C, Dess-Martin (25 g, 59 mmol) was added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with NaSO (10%, 100 mL) and aqueous NaHCO (10%, 100 mL) and extracted with DCM (100 mL × 3). The combined organic layers were washed with brine (150 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE = 18%) to give tert-butyl 3-acryloyl-3-allylazetidine-1-carboxylate (4.2 g, yield: 42%, 2 steps) as a yellow oil.

[0164] Step F: tert-Butyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (7) To a stirred solution of tert-butyl 3-acryloyl-3-allylazetidine-1-carboxylate (4.2 g, 16.7 mmol) in toluene (200 mL) was added Grubbs II catalyst (709 mg, 0.8 mmol) under a nitrogen atmosphere. The mixture was stirred at 85 °C for 1 h. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MTBE / PE = 35%) to give tert-butyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (3.1 g, yield: 83%) as a brown solid.

[0165] Step G: 2-Azaspiro[3.4]oct-6-en-5-one (8) To a stirred solution of tert-butyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (2 g, 9.0 mmol) in DCM (15 mL) was added TFA (5 mL) at 0 °C. The mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure to give 2-azaspiro[3.4]oct-6-en-5-one (2 g crude, quantitative), which was used directly in the next step without further purification.

[0166] Step H: Benzyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (9) To a stirred solution of 2-azaspiro[3.4]oct-6-en-5-one (1.1 g, 8.9 mmol) in THF (15 mL) was added TEA (2.7 g, 26.7 mmol) and Cbz-OSU (2.67 g, 10.7 mmol) at 0° C. The mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE=35%) to give benzyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (2 g, yield: 86%, 2 steps) as a yellow oil. LCMS [M+1] + =258.0

[0167] Step I: (R)-Benzyl 7-hydroxy-5-oxo-2-azaspiro[3.4]octane-2-carboxylate (10) A solution of CuCl (46 mg, 0.47 mmol), (S)-TolBINAP (318 mg, 0.47 mmol), and sodium tert-butoxide (45 mg, 0.47 mmol) in THF (10 mL) was stirred at room temperature for 30 min. Bis(pinacolato)diborane (2.96 g, 11.7 mmol) in THF (20 mL) was then added, and the resulting mixture was stirred at room temperature for 10 min. Benzyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (2.0 g, 7.8 mmol) in THF (20 mL) was added to the above solution, followed by MeOH (500 mg, 15.6 mmol). The resulting mixture was stirred at room temperature for 1 h, then water (10 mL) was added, followed by sodium perborate (6.0 g, 39 mmol), and the resulting mixture was vigorously stirred at room temperature for 1 h. The resulting green suspension was filtered through a pad of Celite, poured into a separatory funnel containing saturated aqueous NaHCO3 / Na2SO3 (1 / 1, 50 mL), and extracted with EA (100 mL x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give crude benzyl (R)-7-hydroxy-5-oxo-2-azaspiro[3.4]octane-2-carboxylate (5.4 g, quantitative), which was used directly in the next step without further purification. LCMS [M+1] + =276.0

[0168] Step J: (R)-Benzyl 7-((tert-butyldimethylsilyl)oxy)-5-oxo-2-azaspiro[3.4]octane-2-carboxylate (11) To a stirred solution of benzyl (R)-7-hydroxy-5-oxo-2-azaspiro[3.4]octane-2-carboxylate (2.1 g, 7.8 mmol) in DCM (30 mL) at 0 °C, imidazole (4.3 g, 31.2 mmol) and TBSCl (2.3 g, 15.6 mmol) were added. The resulting mixture was stirred at room temperature for 3 h. After completion, the mixture was diluted with water (50 mL) and extracted with DCM (30 mL × 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE=23%) to give benzyl (R)-7-((tert-butyldimethylsilyl)oxy)-5-oxo-2-azaspiro[3.4]octane-2-carboxylate (1.8 g, yield: 59%, 2 steps) as a yellow oil. LCMS [M+1] + =390.0

[0169] Step K: (5R,7R)-Benzyl 7-((tert-butyldimethylsilyl)oxy)-5-((R)-1,1-dimethylethylsulfinamido)-2-azaspiro[3.4]octane-2-carboxylate (12) To a stirred solution of benzyl (R)-7-((tert-butyldimethylsilyl)oxy)-5-oxo-2-azaspiro[3.4]octane-2-carboxylate (1.7 g, 4.4 mmol) in dry THF (20 mL) at 0 °C, (R)-2-methylpropane-2-sulfinamide (1.06 g, 8.7 mmol) and titanium tetraisopropanolate (5 g, 17.6 mmol) were added. The resulting mixture was stirred at 75 °C overnight. The mixture was cooled to -78 °C, and L-selectride (13.2 mL, 13.2 mmol) was added dropwise. After the addition, the mixture was stirred at -78 °C for 3 h. The mixture was quenched with saturated aqueous NH4Cl, filtered, and extracted with EA (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE=40%) to give benzyl (5R,7R)-7-((tert-butyldimethylsilyl)oxy)-5-(((R)-tert-butylsulfinyl)amino)-2-azaspiro[3.4]octane-2-carboxylate (750 mg, yield: 35%) as a yellow oil. LCMS [M+1] + =495.0

[0170] Step L: (5R,7R)-benzyl 5-((R)-1,1-dimethylethylsulfinamido)-7-hydroxy-2-azaspiro[3.4]octane-2-carboxylate (13) To a stirred solution of benzyl (5R,7R)-7-((tert-butyldimethylsilyl)oxy)-5-(((R)-tert-butylsulfinyl)amino)-2-azaspiro[3.4]octane-2-carboxylate (800 mg, 1.62 mmol) in THF (3 mL) was added TBAF (1 M in THF, 2.42 mL, 2.42 mmol) at room temperature and stirred for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=3%) to give benzyl (5R,7R)-5-(((R)-tert-butylsulfinyl)amino)-7-hydroxy-2-azaspiro[3.4]octane-2-carboxylate (600 mg, yield: 97%) as a yellow oil. LCMS [M+1] + =381.0

[0171] Step M: (5R,7S)-benzyl 5-((R)-1,1-dimethylethylsulfinamido)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (14) To a stirred solution of benzyl (5R,7R)-5-(((R)-tert-butylsulfinyl)amino)-7-hydroxy-2-azaspiro[3.4]octane-2-carboxylate (600 mg, 1.58 mmol) in THF (15 mL) was added PPh3 (828 mg, 3.16 mmol), isoindoline-1,3-dione (348 mg, 2.37 mmol), and DIAD (638 mg, 3.16 mmol) under a nitrogen atmosphere at 0 °C. After the addition, the mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=2%) to give (5R,7S)-benzyl 5-((R)-1,1-dimethylethylsulfinamido)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (750 mg, yield: 93%) as a yellow oil. LCMS [M+1] + =510.0

[0172] Step N: (5R,7S)-Benzyl 5-amino-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (15) To a stirred solution of (5R,7S)-benzyl 5-((R)-1,1-dimethylethylsulfinamido)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (600 mg, 1.2 mmol) in DCM (5 mL) was added HCl in dioxane (4 M in dioxane, 5 mL) at 0° C. The mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure to give (5R,7S)-benzyl 5-amino-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (610 mg crude, quantitative), which was used directly in the next step without further purification. LCMS [M+1] + =406.0

[0173] Step O: (5R,7S)-Benzyl 5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (16) To a stirred solution of (5R,7S)-benzyl 5-amino-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (350 mg, 0.86 mmol) in DMSO (3.5 mL) was added 2-chloro-N,N-dimethylbenzo[d]thiazole-6-carboxamide (312 mg, 1.3 mmol) and DIEA (333 mg, 2.58 mmol) at room temperature. The resulting mixture was stirred at 90 °C overnight. The mixture was diluted with water (30 mL) and extracted with EA (30 mL × 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=2%) to give (5R,7S)-benzyl 5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (300 mg, yield: 57%) as a yellow oil. LCMS [M+1] + =610.3

[0174] Step P: (5R,7S)-Benzyl 7-amino-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-2-azaspiro[3.4]octane-2-carboxylate (17) To a stirred solution of (5R,7S)-benzyl 5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (300 mg, 0.49 mmol) in EtOH (3 mL) was added hydrazine hydrate (246 mg, 4.9 mmol) at room temperature and stirred for 1 h. After completion, the mixture was filtered, and the filtrate was diluted with water (50 mL) and extracted with EA (30 mL × 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give (5R,7S)-benzyl 7-amino-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-2-azaspiro[3.4]octane-2-carboxylate (240 mg crude, quantitative), which was used directly in the next step without further purification. LCMS [M+1] + =480.3

[0175] Steps Q and R: N,N-dimethyl-2-{(5R,7S)-7-(5-chloro-2-pyrimidinylamino)-2-aza-5-spiro[3.4]octylamino}-1,3-benzothiazole-6-carboxamide (Example B) To a stirred solution of (5R,7S)-benzyl 7-amino-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-2-azaspiro[3.4]octane-2-carboxylate (240 mg, 0.5 mmol) in DMSO (3 mL) was added 2,5-dichloropyrimidine (112 mg, 0.75 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 2 h. After completion, the mixture was diluted with water (30 mL) and extracted with EA (50 mL × 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=8%) to give (5R,7S)-benzyl 7-((5-chloropyrimidin-2-yl)amino)-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-2-azaspiro[3.4]octane-2-carboxylate (260 mg, yield: 88%) as a yellow solid. LCMS [M+1] + =592.0. Deprotection of the benzyl carbamate using standard conditions gives the title compound.

[0176] Example 13

[0177] [ka]

[0178] Step A: Benzyl cyclopent-3-en-1-ylcarbamate To a solution of cyclopent-3-ene-1-carboxylic acid (51.8 g, 462.5 mmol) and EtN (56.1 g, 555.6 mmol) in toluene (780 mL) was added DPPA (140 g, 508.8 mmol) dropwise. The reaction mixture was stirred at room temperature for 2 hours, and then BnOH (74.9 g, 693.8 mmol) was added dropwise to the mixture. The reaction mixture was heated to 90 °C and stirred overnight under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was poured into water (3 L) and extracted with ethyl acetate (500 mL × 3). The combined organic layers were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude benzyl cyclopent-3-en-1-ylcarbamate (126 g, quantitative) as a yellow oil. The crude product was used directly in the next step without further purification. LCMS[M+H] + =218.1.

[0179] Step B: Benzyl ((1R,3r,5S)-6-oxabicyclo[3.1.0]hexan-3-yl)carbamate To a solution of benzyl cyclopent-3-en-1-ylcarbamate (35 g, 161.3 mmol) in DCM (875 mL) was added m-CPBA (85%, 50 g, 290.3 mmol) at 0 °C. The mixture was stirred at 0 °C for 30 min. After completion, the reaction was quenched with aqueous NaSO and NaHCO and extracted with DCM (800 mL × 2). The combined organic phase was washed with brine (800 mL × 2), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE = 30%) to give benzyl ((1R,3r,5S)-6-oxabicyclo[3.1.0]hexan-3-yl)carbamate (32.5 g, 86.7%) as a white solid. LCMS [M+H] + =234.1

[0180] Step C: Benzyl ((1S,3S,4S)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate (5a) and Benzyl ((1R,3R,4R)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate (5b) A mixture of benzyl ((1R,3r,5S)-6-oxabicyclo[3.1.0]hexan-3-yl)carbamate (32.5 g, 139.5 mmol) and (S)-1-phenylethan-1-amine (33.8 g, 279 mmol) was stirred at 120 °C overnight, and then MeOH (1.5 L) and (Boc)2O (30.4 g) were added. After stirring at room temperature for 30 min, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA (3% NH3.MeOH) / PE = 78%) to give a mixture of 5a and 5b (18.2 g, 36.9%) as a yellow solid. LCMS [M+H] + =355.3.

[0181] Step D: tert-butyl ((1R,3R,4R)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate To a solution of benzyl ((1R,3R,4R)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate (18.2 g, 51.4 mmol) in MeOH (360 mL) was added (Boc)2O (11.8 g, 54 mmol) and Pd / C (4.6 g). The reaction mixture was stirred under a hydrogen atmosphere at room temperature for 3 hours. After completion, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE=78%) to give tert-butyl ((1R,3R,4R)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate (8.5 g, 50%) as a colorless oil. LCMS [M+H] + =321.2

[0182] Step E: tert-butyl ((1R,3R,4R)-3-amino-4-hydroxycyclopentyl)carbamate To a solution of tert-butyl ((1R,3R,4R)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate (18.5 g, 57.8 mmol) in MeOH (360 mL) was added Pd(OH) (6.17 g). The reaction mixture was stirred under a hydrogen atmosphere at 60° C. for 20 hours. Upon completion, the reaction mixture was filtered and the filtrate was concentrated to give tert-butyl ((1R,3R,4R)-3-amino-4-hydroxycyclopentyl)carbamate (11 g, 88%) as a white solid. LCMS [M+H] + =217.1

[0183] Step F: tert-butyl ((1R,3R,4R)-3-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-4-hydroxycyclopentyl)carbamate To a solution of tert-butyl ((1R,3R,4R)-3-amino-4-hydroxycyclopentyl)carbamate (700 mg, 3.22 mmol) in DMSO (15 mL) was added DIPEA (1.25 g, 9.66 mmol) and 2-chloro-N,N-dimethylbenzo[d]thiazole-6-carboxamide (931 mg, 3.85 mmol). The mixture was heated to 80° C. and stirred under a nitrogen atmosphere for 6 hours. After completion, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layers were washed with brine (30 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 50 / 1 to 20 / 1) to give tert-butyl ((1R,3R,4R)-3-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-4-hydroxycyclopentyl)carbamate (880 mg, 65%) as a pale yellow solid. LCMS [M+H] + =421.3

[0184] Step G: 2-(((1R,2R,4R)-4-amino-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide To a solution of tert-butyl ((1R,3R,4R)-3-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-4-hydroxycyclopentyl)carbamate (880 mg, 2.09 mmol) in DCM (25 mL) was added HCl in dioxane (2 M, 5 mL) at room temperature. After the addition, the reaction mixture was stirred at room temperature for 1 hour. After completion, the mixture was concentrated under reduced pressure to give 2-(((1R,2R,4R)-4-amino-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (669 mg, quantitative) as the HCl salt, which was used in the next step without further purification. LCMS [M+H] + =321.1

[0185] Step H: 2-(((1R,2R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide To a solution of 2-(((1R,2R,4R)-4-amino-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (669 mg, 2.09 mmol) in DMSO (10 mL) was added KCO (865 mg, 6.27 mmol) and 2,5-dichloropyrimidine (309 mg, 2.09 mmol). The mixture was heated to 80 °C and stirred overnight under a nitrogen atmosphere. After cooling to room temperature, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (20 mL × 2). The combined organic layers were washed with brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting with DCM / MeOH = 50 / 1 to 20 / 1) to give 2-(((1R,2R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (600 mg, 66.4%) as a white solid. LCMS [M+H] + =433.3

[0186] Step I: 2-(((1R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-oxocyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide To a solution of 2-(((1R,2R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (600 mg, 1.39 mmol) in DCM (40 mL) was added Dess-Martin periodinane (1.18 g, 2.78 mmol) in small portions at 0 °C. After the addition, the mixture was stirred at 0 °C for 1 h. After completion, 10% Na2SO3 solution was added to the mixture and extracted with ethyl acetate (40 mL × 2). The combined organic layers were washed with saturated aqueous sodium bicarbonate (20 mL × 2) and brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting with DCM / MeOH = 50 / 1 to 20 / 1) to give 2-(((1R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-oxocyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (280 mg, 46.8%) as a yellow solid. LCMS [M+H] + =431.0

[0187] Step J: 2-(((1R,4S)-4-((5-chloropyrimidin-2-yl)amino)-2-(cyanomethylene)cyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide To a solution of diethyl (cyanomethyl)phosphonate (230.7 mg, 1.3 mmol) in THF (3.2 mL) was added NaH (52 mg, 1.3 mmol) at 0° C. After the addition, the mixture was stirred at 0° C. for 30 minutes. Then, 2-(((1R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-oxocyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (280 mg, 0.65 mmol) in THF (3.2 mL) was added dropwise, and the mixture was stirred at 0° C. for 30 minutes. After completion, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (15 mL×2). The combined organic layers were washed with brine (15 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluting with DCM / MeOH = 50 / 1 to 20 / 1) to give 2-(((1R,4S)-4-((5-chloropyrimidin-2-yl)amino)-2-(cyanomethylene)cyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide (190 mg, yield: 64.5%) as a yellow solid. LCMS [M+H] + =454.3 1 HNMR(400MHz,DMSO-d6)δ:9.05-9.00(s,1H),8.39-8.29(m,2H),7.88-7.81(m,2H),7.51-7.43(m,1H),7.34-7.31(m,1H),5. 76-5.68(m,1H),5.29-5.09(m,1H),4.41-4.34(m,1H),3.18-3.09(m,1H),2.96(s,6H),2.75-2.58(m,2H),2.33-1.71(m,1H).

[0188] Example D N,N-Dimethyl-2-[(1R,4S)-(Z)-2-(2-aminoethylidene)-4-(5-chloro-2-pyrimidinylamino)cyclopentylamino]-1,3-benzothiazole-6-carboxamide

[0189] [ka]

[0190] The title compound can be prepared by using 2-(((1R,4S)-4-((5-chloropyrimidin-2-yl)amino)-2-(cyanomethylene)cyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide using reduction conditions.

[0191] Example E

[0192] [ka]

[0193] Step A: (S)-2,3-dihydro-1H-inden-1-ol (2) A solution of R-CBS (1.4 g, 5.15 mmol) and (CH)S-BH (0.48 mL, 0.95 mmol) was stirred at room temperature under a nitrogen atmosphere for 10 minutes. Then, DCM (33 mL) and (CH)S-BH (28.4 mL, 56.82 mmol) were added to this solution at −5° C. A solution of 2,3-dihydro-1H-inden-1-one (1) (5 g, 37.88 mmol) in DCM (30 mL) was added dropwise to the above solution while maintaining the reaction temperature below 0° C. After the addition, the reaction mixture was stirred at −5° C. for 2 hours. The mixture was quenched with CHOH (40 mL) and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluting with 0% to 10% EtOAc / petroleum ether to give (S)-2,3-dihydro-1H-inden-1-ol (2) (4.9 g, 96%) as a colorless oil. LCMS [M+H-OH] + =117.3. 1 H NMR(400MHz,DMSO-d6)δ 7.34-7.31(m,1H),7.23-7.16(m,3H),5.20(br.,s.,1H),5.04(t,J=6.8Hz,1H ),2.94-2.87(m,1H),2.74-2.66(m,1H),2.36-2.28(m,1H),1.81-1.72(m,1H).

[0194] Step B: (R)-2-(2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (4) To a solution of (S)-2,3-dihydro-1H-inden-1-ol (2) (4.9 g, 36.57 mmol), isoindoline-1,3-dione (3) (6.99 g, 47.5 mmol), and PPh3 (14.4 g, 54.8 mmol) in THF (102 mL) was added dropwise DIAD (11.1 g, 54.8 mmol) under a nitrogen atmosphere at 0°C. After the addition, the reaction mixture was stirred at room temperature overnight. The reaction solution was then concentrated under reduced pressure. The residue was t Purification by silica gel flash chromatography eluting with BuOMe / petroleum ether = 0% to 11% gave (R)-2-(2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (4) (3.3 g, 34%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δ 7.84(s,4H),7.29(d,J=7.6Hz,1H),7.24-7.20(m,1H),7.13-7.11(m,2H),5. 74(t,J=8.0Hz,1H),3.18-3.14(m,1H),2.96-2.92(m,1H),2.47-2.40(m,2H).

[0195] Step C: 2-((1R)-3-bromo-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (5) To a solution of (R)-2-(2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (4) (3.3 g, 12.5 mmol) and NBS (2.45 g, 13.8 mmol) in CCl4 (32 mL) was added AIBN (0.2 g, 1.25 mmol) at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at 80 °C for 2 h. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluting with 0% to 9% EtOAc / petroleum ether to give 2-((1R)-3-bromo-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (5) (3.2 g, 75%) as a yellow oil.

[0196] Step D: 2-((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (6) To a solution of 2-((1R)-3-bromo-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (5) (3.2 g, 9.38 mmol) and DMBNH (3.13 g, 18.77 mmol) in DCM (40 mL) was added EtN (0.69 mL, 9.38 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM (20 mL) and washed with brine (50 mL). The organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluting with 50% NH3-CH3OH / DCM (0% to 3%) to give 2-((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (6) (1.3 g, 32%) as a yellow solid. LCMS [M+H] + =429.5.

[0197] Step E: (3R)-N1-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-indene-1,3-diamine (7) To a solution of 2-((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (6) (1.3 g, 3 mmol) in THF (20 mL) and CHOH (20 mL) was added N2H4-H2O (1.82 mL, 30 mmol) at 0 °C. The reaction mixture was stirred at room temperature overnight. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (50 mL) and washed with water (30 mL), saturated aqueous NaHCO3 (30 mL), and brine (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluting with 50% NH3-CH3OH / DCM = 0% to 6% to give (3R)-N 1-(2,4-Dimethoxybenzyl)-2,3-dihydro-1H-indene-1,3-diamine (7) (700 mg, 78%) was obtained as a yellow oil. LCMS [M+H] + =299.5.

[0198] Step F: 6'-(((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (9) (3R)-N in DMA (5 mL) 1 To a solution of -(2,4-dimethoxybenzyl)-2,3-dihydro-1H-indene-1,3-diamine (7) (400 mg, 1.34 mmol), 6'-chloro-2H-[1,3'-bipyridin]-2-one (8) (277 mg, 1.34 mmol) and t-BuONa (258 mg, 2.68 mmol), t BuxPhosPdG3 (119 mg, 0.134 mmol) was added at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at 100 °C overnight. After cooling to room temperature, the mixture was diluted with water (30 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluting with CH3OH / DCM = 0% to 7% to give 6'-(((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (9) (440 mg, 70%) as a brown solid. LCMS [M+H] + =469.6. 1H NMR(400MHz,DMSO-d6)δ7.98(d,J=2.8Hz,1H),7.65-7.63(m,1H),7.51-7. 43(m,3H),7.34-7.22(m,5H),6.65(d,J=8.8Hz,1H),6.55(d,J=2.4Hz,1H), 6.52-6.46(m,2H),6.31-6.28(m,1H),5.45-5.39(m,1H),4.16(t,J=7.6Hz, 1H), 3.78 (s, 3H), 3.76-3.72 (m, 5H), 2.89-2.82 (m, 1H), 1.70-1.62 (m, 1H).

[0199] Step G: 6'-(((1R)-3-amino-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (10) A solution of 6'-(((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (9) (440 mg, 0.94 mmol) in CF3COOH (6 mL) was stirred at 80 °C for 48 h. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was diluted with water (10 mL) and extracted with DCM (10 mL × 2). The aqueous layer was adjusted to pH = 8 with saturated NaHCO3 solution and extracted with DCM (10 mL × 5). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product (290 mg) was used in the next step without further purification. LCMS [M+H] + =319.5.

[0200] Step H: 6'-(((1R)-3-((5-chloropyrimidin-2-yl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (12) To a solution of 6'-(((1R)-3-amino-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (10) (crude, 290 mg, 0.91 mmol) and 2,5-dichloropyrimidine (11) (150 mg, 1 mmol) in DMSO (5 mL) was added K2CO3 (377 mg, 2.73 mmol) at room temperature under a nitrogen atmosphere. The reaction mixture was stirred at 80 °C for 3 h. After cooling to room temperature, the mixture was diluted with water (40 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluting with CH3OH / DCM = 0% to 3% to give 6'-(((1R)-3-((5-chloropyrimidin-2-yl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridin]-2-one (12) (120 mg, 31%) as a yellow solid. LCMS [M+H] + =431.5. Example E can be separated from its isomers by chromatography.

[0201] Example 2: Biological Activity of Exemplary PCSK9 Inhibitors Human recombinant PCSK9 was expressed as follows: Protein sequence: QEDEDGDYEELVLALRSEEDGLAEAPEHGTTATFHRCAKDPWRLPGTYVVVLKEETHLSQSERTARRLQAQAARRGYLTKILHVFHGLLPGFLVKMSGDLLELALKLPHVDYIEEDSSVFAQSIPWNLERITPPRYRADEYQPPDGGSLVEVYLLDTSIQSDHREIEGRVMVT DFENVPEEDGTRFHRQASKCDSHGTHLAGVVSGRDAGVAKGASMRSLRVLNCQGKGTVSGTLIGLEFIRKSQLVQPVGPLVVLLPLAGGYSRVLNAACQRLARAGVVLVTAAGNFRDDACLYSPASAPEVITVGATNAQDQPVTLGTLGTGTNFGRCVDLFAPGEDIIGASSDCS TCFVSQSGTSQAAAHVAGIAAMMLSAEPELTLAELRQRLIHFSAKDVINEAWFPEDQRVLTPNLVAALPPSTHGAGWQLFCRTVWSAHSGPTRMATAVARCAPDEELLSCSSFSRSGKRRGERMEAQGGKLVCRAHNAFGGEGVYAIARCCLLPQANCSVHTAPPAEASMGTR VHCHQQGHVLTGCSSHWEVEDLGTHKPPVLRPRGQPNQCVGHREASIHASCCHAPGLECKVKEHGIPAPQEQVTVACEEGWTLTGCSALPGTSHVLGAYAVDNTCVVRSRDVSTTGSTSEGAVTAVAICCRSRHLAQASQELQSGSGGLNDIFEAQKIEWHENLYFQGHHHHHH

[0202] The following assay methods were used to identify and evaluate PCSK9 inhibitors effective in inhibiting PCSK9 function.

[0203] Example: PCSK9 SPR Assay Surface plasmon resonance data were collected on a Biacore™ T200 or 3000 system (GE Healthcare) at 25°C. Streptavidin was immobilized onto a CM5 (GE Healthcare) or CMD500d sensor chip (XanTec Bioanalytics) using standard amine coupling chemistry with HBS-N (10 mM HEPES, 0.15 M NaCl, pH 7.4) as the running buffer at 25°C. Briefly, the carboxymethyldextran surface was activated by injecting a 1:1 ratio of 0.4 M 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) / 0.1 M N-hydroxysuccinimide (NHS) at a flow rate of 10 μL / min for 12 min. For streptavidin capture, the protein was diluted to 0.2 mg / mL in 10 mM sodium acetate (pH 4.5) and captured by injecting 100 μL onto the activated chip surface. Remaining activated groups were blocked by a 7-minute injection of 1 M ethanolamine (pH 8.5). Avi-tagged PCSK9 protein was captured onto the streptavidin surface by injecting 150 μL of protein diluted to 16 μg / mL in HBS-N, 0.05% Tween-20, and 0.1 mM CaCl2. Typical surface densities obtained were 8,000–10,000 RU. SPR binding data were obtained using an appropriate dilution series of each compound at a flow rate of 30 μL / min, with a 100-second capture time and a 300-second dissociation time. The running buffer for compound binding studies was HBS-N, 0.05% Tween-20, 0.1 mM CaCl2, and 4% DMSO. Data were corrected for DMSO excluded volume effects. All data were double-referenced against a blank injection and a reference surface using standard procedures, and data processing and kinetic fitting were performed using Scrubber software, version 2.0c (BioLogic Software). Data were fitted using a simple 1:1 binding model to obtain k a , k d and K. D value was determined.

[0204] The ability of the PCSK9 inhibitors of the present disclosure, and pharmaceutically acceptable salts thereof, to bind to and inhibit PCSK9 was established using representative PCSK9 inhibitors of the present disclosure listed in the table below:

[0205] [Table 1]

[0206] [Table 2-1]

[0207] [Table 2-2]

[0208] Incorporation by Reference All publications and patents mentioned herein are incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0209] equivalent While specific embodiments of the invention have been described, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification and the following claims. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

1. A compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof: 【Chemical 1】 [In the formula, A is H, halo, hydroxy, alkyl, alkylthio, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR ６ , and —C(O)NR ６ R ７ Selected from: B is selected from H, alkyl, and halo; or A and B together with the carbon atom to which they are attached form a 5- or 6-membered heteroaryl; X is NR ５ or O; Z is cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; Each R ６ and R ７ is independently H or alkyl; Y is selected from aryl, heteroaryl, and heterocyclyl; n is 0 or 1.

2. The compound of claim 1 wherein Z is cycloalkyl.

3. 3. The compound of claim 1 or 2, wherein the structure of the compound is represented by formula (Ia), (Ib), (Ic), or a pharmaceutically acceptable salt thereof: 【Chemistry 2】 [In the formula, R １ and R １’ are each independently selected from alkyl, halo, hydroxyl, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amido, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamido, aryl, heteroaryl, heterocyclyl, and aralkyl; or R １ and R １’ combine to form an alkylalkene, cycloalkyl, cycloalkene, or heterocyclyl; Each R ２ is independently selected from alkyl, halo, hydroxyl, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amido, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamido, aryl, heteroaryl, heterocyclyl, and aralkyl; n is 1, 2, 3 or 4.

4. 4. The compound of claim 3, wherein the structure of the compound is represented by formula (IIa), (IIb), (IIc), or a pharmaceutically acceptable salt thereof: 【Chemistry 3】

5. 4. The compound of claim 3, wherein the structure of the compound is represented by formula (IIIa), (IIIb), (IIIc), or a pharmaceutically acceptable salt thereof: 【Chemistry 4】

6. A is H, hydroxy, alkylthio, alkyl, alkoxy, acyloxy, cyano, cycloalkyl, —C(O)OR ６ , and —C(O)NR ６ R ７ The compound according to any one of claims 1 to 5, selected from:

7. 7. The compound of claim 6, wherein A is H.

8. 7. The compound of claim 6, wherein A is alkyl.

9. 7. The compound of claim 6, wherein A is alkylthio.

10. 7. The compound of claim 6, wherein A is alkoxy.

11. 7. The compound of claim 6, wherein A is cycloalkyl.

12. A is -SCH ３ , -SCHF ２ , and -OCHF ２ 7. The compound of claim 6, selected from:

13. 7. The compound of claim 6, wherein A is halo (e.g., chloro).

14. 7. The compound of claim 6, wherein A and B, together with the carbon atoms to which they are attached, form a pyrrolyl or thienyl ring that is unsubstituted or substituted with one or more alkyls.

15. The compound of any one of claims 1 to 14, wherein B is H.

16. X is NR ５ The compound according to any one of claims 1 to 15,

17. The compound of any one of claims 1 to 16, wherein Y is heteroaryl or heterocyclyl.

18. The compound of any one of claims 1 to 17, wherein Y is monocyclic heteroaryl.

19. 19. The compound of claim 18, wherein Y is selected from pyridyl, pyridonyl, pyridinyl, pyrazinyl, pyrimidinyl, and thiazolyl.

20. 19. The compound of claim 18, wherein Y is selected from triazenyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, and triazolyl.

21. 21. The compound of any one of claims 18-20, wherein the monocyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from alkyl, alkylthio, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, aryl, heteroaryl, heterocyclyl, nitro, sulfonamido, and alkylthio.

22. 22. The compound of claim 21, wherein the monocyclic heteroaryl is substituted with a 6-membered aryl, heteroaryl, or heterocyclyl selected from phenyl, pyridinyl, 2-hydroxypyridinyl, piperidinonyl, 2-hydroxy-1-methylpyridinyl, triazolyl, imidazolidinonyl, pyrimidonyl, 2-hydroxyisoquinolinyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl, and morpholinonyl.

23. The monocyclic heteroaryl may be halo, CN, alkyl, alkoxy, hydroxy, carboxy, —CO ２ 22. The compound of claim 21, which is substituted with heteroaryl or heterocyclyl substituted with one or more substituents selected from alkyl, and tetrazolyl.

24. 24. The compound of any one of claims 21 to 23, wherein the substituent is positioned para to Y relative to X.

25. The compound of any one of claims 1 to 17, wherein Y is a bicyclic heteroaryl.

26. 26. The compound of claim 25, wherein Y is selected from benzothiazolyl, benzoxazolyl, benzimidazolyl, triazolopyridinyl, thiazolopyrindinyl, quinolinyl, and quinoxalinyl.

27. 27. The compound of claim 25 or 26, wherein the bicyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from alkyl, haloalkyl, hydroxyalkyl, alkylthio, alkoxy, alkoxycarbonyl, amido, carboxy, cyano, halo, heteroaryl, nitro, and sulfonamido.

28. 27. The compound of claim 25 or 26, wherein the bicyclic heteroaryl is unsubstituted or substituted with one or more substituents selected from alkylthio, alkoxycarbonyl, amido, carboxy, halo, and heteroaryl.

29. Y is a group of the formula -C(O)NR ８ R ９ or -NR ９ C(O)R １０ and is substituted with an amide substituent of the formula: R ８ and R ９ are each independently selected from H, alkyl, heterocyclyl, and heteroaryl; or R ８ and R ９ together with the nitrogen atom to which they are attached form a 4-, 5-, 6-, or 7-membered heterocyclic or heteroaryl ring; R １０ The compound of any one of claims 1 to 28, wherein is alkyl.

30. Y is a group of the formula -S(O) ２ NR ８ R ９ or -NR ９ S (O) ２ R １０ and is substituted with a sulfonamide substituent of the formula: R ８ and R ９ are each independently selected from H, alkyl, and heteroaryl; or R ８ and R ９ together with the nitrogen atom to which they are attached form a 4-, 5-, 6-, or 7-membered heterocyclic ring; R １０ The compound of any one of claims 1 to 28, wherein is alkyl.

31. R ８ and R ９ 31. The compound of claim 29 or 30, wherein each is independently selected from H, methyl, ethyl, triazolyl, and pyrazolyl.

32. R ８ and R ９ is alkyl, and each alkyl is independently unsubstituted or substituted with one or more substituents selected from methyl, methoxy, carboxy, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, oxadiazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl, and 2,4-dihydro-3H-1,2,4-triazol-3-onyl.

33. R ８ and R ９ together with the nitrogen atom to which they are attached form a heterocyclic ring selected from azirazine, isothiazolidine-1,1-dioxide, azetinidine, thiazol-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, 2-oxa-6-azaspiro[3.3]heptane.

34. R ８ and R ９ together with the nitrogen atom to which they are attached, form 2,8-diazaspiro[5,5]undecene, tetrahydroimidazo[1,2-a]pyrazine, octahydropyrazino[2,1-c][1,4]oxazine, tetrahydropyrido[3,4-d]pyrimidine, 2-oxa-8-azaspiro[4.5]decane, tetrahydropyrrolo[3,4-c]pyrazole, thiomorpholine, 2-oxa-7-azaspiro[3.5]nonane, 2,8-diazaspiro[4.5]decan-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9 ... Azaspiro[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 31. The compound of claim 29 or 30, wherein the heterocyclic ring is selected from azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, oxadiazole, triazole, pyridinone, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3(2H)-one, piperidinone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5]nonane, pyrazole, and pyridazin-3(2H)-one.

35. 35. The compound of claim 33 or 34, wherein the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxy, and heterocyclyl.

36. 35. The compound of claim 33 or 34, wherein the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxy, and oxetanyl.

37. R １ and R １’ 37. The compound of any one of claims 2 to 36, wherein: combine to form a heterocyclyl (e.g., azetidinyl).

38. R １ and R １’ 37. The compound of any one of claims 2 to 36, wherein: combine to form an alkylalkene (e.g., aminoallyl).

39. The compound of any one of claims 2 to 38, wherein n is 0.

40. The compound is 【Chemistry 5】 or a pharmaceutically acceptable salt thereof.

41. 41. A pharmaceutical composition comprising a compound according to any one of claims 1 to 40 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

42. 41. A method of treating a cardiovascular disease or disorder in a subject, comprising administering to the subject a compound of any one of claims 1 to 40, or a pharmaceutically acceptable salt thereof.

43. 43. The method of claim 42, wherein the cardiovascular disease or disorder is selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome, and coronary artery disease.

44. 44. The method of claim 42 or 43, wherein the cardiovascular disease or disorder is familial hypercholesterolemia.

45. 44. The method of claim 42 or 43, wherein the cardiovascular disease or disorder is autosomal dominant hypercholesterolemia.

46. 46. The method of any one of claims 42 to 45, wherein the level of circulating serum cholesterol is reduced in the subject.

47. 47. The method of any one of claims 42 to 46, wherein the level of circulating serum LDL-cholesterol is reduced in the subject.

48. 48. The method of any one of claims 42 to 47, wherein the level of circulating serum VLDL-cholesterol is reduced in the subject.

49. 49. The method of any one of claims 42 to 48, wherein circulating serum triglyceride levels are reduced in the subject.

50. 50. The method of any one of claims 42 to 49, wherein the level of circulating serum lipoprotein A is reduced in the subject.

51. 51. The method of any one of claims 42 to 50, wherein the subject has atherosclerosis.

52. 52. The method of any one of claims 42 to 51, wherein atherosclerotic plaque formation is reduced in the subject.

53. The method of any one of claims 42 to 52, wherein the subject has a gain-of-function mutation in the PCSK9 gene.

54. 54. The method of any one of claims 42 to 53, further comprising co-administering one or more additional therapeutic agents.

55. 55. The method of claim 54, comprising administering one additional therapeutic agent.

56. 55. The method of claim 54, comprising administering two additional therapeutic agents.

57. 55. The method of claim 54, comprising administering three additional therapeutic agents.

58. The additional therapeutic agent is an HMG-CoA reductase inhibitor, an HMG-CoA synthase inhibitor, an HMG-CoA reductase gene expression inhibitor, an HMG-CoA synthase gene expression inhibitor, an MTP / ApoB secretion inhibitor, a CETP inhibitor, a bile acid absorption inhibitor, a cholesterol absorption inhibitor, a cholesterol synthesis inhibitor, a squalene synthetase inhibitor, a squalene epoxidase inhibitor, a squalene cyclase inhibitor, a combined squalene epoxidase / squalene cyclase inhibitor, a fibrate, niacin, niacin and lovastatin, and 58. The method of any one of claims 54-57, wherein the active ingredient is selected from a combination of, an ion exchange resin, an antioxidant, an ACAT inhibitor, a bile acid sequestrant, a PCSK9 translation inhibitor, an ATP citrate lyase inhibitor, an apoC3 inhibitor, an ANGPTL3 inhibitor, an omega-3 fatty acid, an Lp(a) inhibitor, an apoB inhibitor, an apoA1 inhibitor, a low density lipoprotein receptor inhibitor, a low density lipoprotein inhibitor, a diacylglycerol acyltransferase inhibitor, a lysosomal acid lipase inhibitor, and a lecithin cholesterol acyltransferase inhibitor.

59. 59. The method of any one of claims 54 to 58, wherein the additional therapeutic agent is an ATP citrate lyase inhibitor.

60. 59. The method of any one of claims 54-58, wherein the additional therapeutic agent is an apoC3 inhibitor.

61. 59. The method of any one of claims 54 to 58, wherein the additional therapeutic agent is an ANGPTL3 inhibitor.

62. 59. The method of any one of claims 54 to 58, wherein the additional therapeutic agent is an omega-3 fatty acid.

63. 59. The method of any one of claims 54-58, wherein the additional therapeutic agent is an Lp(a) inhibitor.

64. 59. The method of any one of claims 54-58, wherein the additional therapeutic agent is an apoB inhibitor.

65. 59. The method of any one of claims 54 to 58, wherein the additional therapeutic agent is an apoA1 inhibitor.

66. 59. The method of any one of claims 54 to 58, wherein the additional therapeutic agent is a low density lipoprotein receptor inhibitor.

67. 59. The method of any one of claims 54-58, wherein the additional therapeutic agent is a low density lipoprotein inhibitor.

68. 59. The method of any one of claims 54-58, wherein the additional therapeutic agent is a diacylglycerol acyltransferase inhibitor.

69. 59. The method of any one of claims 54 to 58, wherein the additional therapeutic agent is a lysosomal acid lipase inhibitor.

70. 59. The method of any one of claims 54-58, wherein the additional therapeutic agent is a lecithin cholesterol acyltransferase inhibitor.

71. The additional therapeutic agent is voranesolsen, evinacumab, IONIS-ANGPTL3-LRX, icosapent ethyl, eicosapentaenoic acid, eicosapentaenoic acid ester, eicosapentaenoic acid methyl ester, eicosapentaenoic acid ethyl ester, docosahexaenoic acid, docosahexaenoic acid ester, docosahexaenoic acid methyl ester, docosahexaenoic acid ethyl ester, bepedoic acid, ezetimibe, IONIS-APO(a)RX, mipomersen, CSL-112, lomitapide, AAV8.TBG.

71. The method of any one of claims 54 to 70, wherein the agonist is selected from hLDLR (RGX-501), alipogene tiparvovec, prazigastat, sebelipase, ACP-501 / MEDI6012, OM3FA-EE, OM3FA-CA, OM3FA-IPE, alirocumab, evolocumab, bococizumab, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimibe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

72. 72. The method of claim 71, wherein the additional therapeutic agent is volanesorsen.

73. 72. The method of claim 71, wherein the additional therapeutic agent is evinacumab.

74. 72. The method of claim 71, wherein the additional therapeutic agent is IONIS-ANGPTL3-LRX.

75. 72. The method of claim 71, wherein the additional therapeutic agent is icosapent ethyl.

76. 72. The method of claim 71, wherein the additional therapeutic agent is eicosapentaenoic acid ethyl ester.

77. 72. The method of claim 71, wherein the additional therapeutic agent is docosahexaenoic acid ethyl ester.

78. 72. The method of claim 71, wherein the additional therapeutic agent is bepedoic acid.

79. 72. The method of claim 71, wherein the additional therapeutic agent is ezetimibe.

80. 72. The method of claim 71, wherein the additional therapeutic agent is IONIS-APO(a)RX.

81. 72. The method of claim 71, wherein the additional therapeutic agent is mipomersen.

82. 72. The method of claim 71, wherein the additional therapeutic agent is CSL-112.

83. 72. The method of claim 71, wherein the additional therapeutic agent is lomitapide.

84. 72. The method of claim 71, wherein the additional therapeutic agent is AAV8.TBG.hLDLR (RGX-501).

85. 72. The method of claim 71, wherein the additional therapeutic agent is alipogen tiparvovec.

86. 72. The method of claim 71, wherein the additional therapeutic agent is prazigastat.

87. 72. The method of claim 71, wherein the additional therapeutic agent is sebelipase.

88. 72. The method of claim 71, wherein the additional therapeutic agent is ACP-501 / MEDI6012.

89. 72. The method of claim 71, wherein the additional therapeutic agent is OM3FA-EE.

90. 72. The method of claim 71, wherein the additional therapeutic agent is OM3FA-CA.

91. 72. The method of claim 71, wherein the additional therapeutic agent is OM3FA-IPE.

92. The method of any one of claims 42 to 91, wherein the disease or disorder is associated with PCSK9.

93. The method of any one of claims 42 to 92, wherein PCSK9 is overexpressed in the subject.

94. 94. The method of any one of claims 42 to 93, wherein the subject is male.

95. 95. The method of any one of claims 42 to 94, wherein the subject is overweight or obese.

96. 96. The method of any one of claims 42 to 95, wherein the subject is African American.

97. 96. The method of any one of claims 42 to 95, wherein the subject is Caucasian.

98. 96. The method of any one of claims 42-95, wherein the subject is of Hispanic descent.

99. 99. The method of any one of claims 42-98, wherein the PCSK9 inhibitor and the one or more additional therapeutic agents are administered simultaneously.

100. 101. The method of claim 100, wherein the one or more additional therapeutic agents are administered within about 5 minutes to about 168 hours before or after administration of the PCSK9 inhibitor.

101. 41. A method of treating sepsis or septic shock in a subject, comprising administering to the subject a compound of any one of claims 1 to 40.

102. 102. The method of claim 101, further comprising co-administering one or more additional therapeutic agents.

103. 103. The method of claim 102, comprising administering one additional therapeutic agent.

104. 103. The method of claim 102, comprising administering two additional therapeutic agents.

105. 103. The method of claim 102, comprising administering three additional therapeutic agents.

106. The additional therapeutic agent is selected from the group consisting of kanamycin, amikacin, tobramycin, dibekacin, gentamicin, sisomicin, netilmicin, streptomycin, neomycin B, C, and E, imipenem, meropenem, ertapenem, doripenem, panipenem, biapenem, razupenem, tebipenem, lenapenem, tomopenem, thiepenem, ciprofloxacin, gaclopramide ... Lenoxacin, gatifloxacin, gemifloxacin, levofloxacin, cinoxacin, nalidixic acid, moxifloxacin, oxolinic acid, piromidic acid, pipemidic acid, losoxacin, enoxacin, fleroxacin, lomefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, rufloxacin, balofloxacin, grepa 106. The method of any one of claims 102-105, wherein the antiviral agent is selected from floxacin, levofloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, clinafloxacin, sitafloxacin, trovafloxacin, prulifloxacin, delafloxacin, JNJ-Q2, nemonoxacin, zavofloxacin, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, methacycline, minocycline, rolitetracycline, tigecycline, ampicillin, amoxicillin, augmentin, piperacillin, tazobactam, chloramphenicol, and ticarcillin; or a combination thereof.

107. 107. The method of any one of claims 102-106, wherein the PCSK9 inhibitor and the one or more additional therapeutic agents are administered simultaneously.

108. 107. The method of any one of claims 102-106, wherein the one or more additional therapeutic agents are administered within about 5 minutes to about 168 hours before or after administration of the PCSK9 inhibitor.

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