PCSK9 inhibitors and methods of use thereof

Abstract

Disclosed herein are inhibitors of PCSK9. Also disclosed herein are methods of treating certain diseases and disorders using PCSK9 inhibitors. # imgabs0 #

Description

Background Art

[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 the levels of circulating LDL cholesterol by enhancing the degradation of the LDL receptor independently of its catalytic activity. Secreted PCSK9 binds to the epidermal growth factor domain A (EGFA) of the LDL receptor (LDLR) at the cell surface, and the PCSK9 / LDL receptor complex is internalized into the endosomal / lysosomal compartment. The enhancement of PCSK9 binding affinity to the LDL receptor at the acidic pH of the late endosome / lysosome 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 low 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 onset of septic shock. Septic shock is a usually fatal complication of severe microbial infection (sepsis) that triggers an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis originates from microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharide (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors [Toll-like receptors (TLRs)] and therefore features prominently in the septic inflammatory response (septic shock or sepsis).

[0003] PCSK9 reduces LPS absorption through the liver's LDL receptors, allowing free LPS to overstimulate the body's immune response to pathogens that cause sepsis. Therefore, inhibiting PCSK9 is beneficial in maintaining liver LDL receptors to achieve systemic pathogen clearance and detoxification in response to sepsis. However, there is currently no effective treatment for sepsis or septic shock other than antibiotic therapy.

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

[0005] Two monoclonal antibodies (mAbs) that selectively bind to extracellular PCSK9 and prevent it from interacting with the LDL receptor, alirocumab and evolocumab, have recently been approved by the FDA for use in reducing LDL-C levels. In clinical trials, alirocumab showed a reduction of about 50% in LDL levels compared to placebo. Elbitar et al., Expert Opin Therapeutic Patents 2016 26:1377-1392. Patients taking evolocumab showed a reduction of about 60%-75% in LDL levels. The efficacy of these drugs demonstrates the potential of PCSK9 inhibitors as an effective treatment for those patients with hypercholesterolemia and other cardiovascular diseases. However, both antibody drugs require intravenous administration and may cause allergic reactions or other harmful immune responses in vivo.

[0006] Unlike infections, which may occur occasionally, cardiovascular disease often needs to be managed throughout a person's life. Therefore, ease of dosing and administration becomes a key factor in patient compliance to maintain medication therapy. There is a need for PCSK9 inhibitors that have increased efficacy and are easier to administer that can be achieved with small molecule PCSK9 inhibitors. Summary of the invention

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

[0008]

[0009] in:

[0010] A is selected from H, halo, hydroxy, alkyl, alkylthio, alkenyl, alkoxy, acyloxy

[0011] alkyl, cyano, cycloalkyl, -C(O)OR 6 and -C(O)NR 6 R 7 ;

[0012] B is selected from H, alkyl and halo; or A and B together with the carbon atom to which they are attached are

[0013] Together they form a 5- or 6-membered heteroaryl group;

[0014] X is NR 5 or O;

[0015] Z is cycloalkyl, cycloalkenyl, aryl, heteroaryl or heterocyclyl;

[0016] Each R 6 and R 7are independently H or alkyl;

[0017] Y is selected from aryl, heteroaryl and heterocyclyl; and

[0018] n is 0 or 1.

[0019] In another aspect, the disclosure relates to methods of treating certain diseases and conditions (eg, cardiovascular disease or septic shock) with compounds of Formula (I). DETAILED DESCRIPTION

[0020] The present disclosure relates to PCSK9 inhibitors 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:

[0021]

[0022] in:

[0023] A is selected from H, halo, hydroxy, alkyl, alkylthio, alkenyl, alkoxy, acyloxy

[0024] alkyl, cyano, cycloalkyl, -C(O)OR 6 and -C(O)NR 6 R 7 ;

[0025] B is selected from H, alkyl and halo; or A and B together with the carbon atom to which they are attached are

[0026] Together they form a 5- or 6-membered heteroaryl group;

[0027] X is NR 5 or O;

[0028] Z is cycloalkyl, cycloalkenyl, aryl, heteroaryl or heterocyclyl;

[0029] Each R 6 and R 7 are independently H or alkyl;

[0030] Y is selected from aryl, heteroaryl and heterocyclyl; and

[0031] n is 0 or 1.

[0032] In certain preferred embodiments of Formula I, Z is cycloalkyl.

[0033] In certain embodiments of Formula (I), the structure of the compound is represented by Formula (Ia), (Ib), (Ic) or a pharmaceutically acceptable salt thereof:

[0034]

[0035] in

[0036] R 1 and R 1` R is each independently selected from alkyl, halo, hydroxy, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamide, aryl, heteroaryl, heterocyclyl and aralkyl; or R 1 and R 1` Combine to form an alkylalkene, a cycloalkyl, a cycloalkene, or a heterocyclic group;

[0037] Each R 2 are independently selected from alkyl, halo, hydroxy, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamide, aryl, heteroaryl, heterocyclyl, and aralkyl; and

[0038] n is 1, 2, 3 or 4.

[0039] 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:

[0040]

[0041] 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:

[0042]

[0043] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc), A is selected from 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 other embodiments, A is alkylthio. In other embodiments, A is alkoxy. In other embodiments, A is cycloalkyl. In certain preferred embodiments, A is selected from -SCH3, -SCHF2, and -OCHF2. In 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 alkyl groups.

[0044] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb), or (IIIc), B is H.

[0045] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc), X is NR 5 .

[0046] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc), Y is a heteroaryl or heterocyclic group, preferably a monocyclic heteroaryl group, such as azophenyl, pyridyl, pyrazinyl, pyrimidinyl or thiazolyl. In other embodiments, Y is selected from triazene, Azolyl, iso Monocyclic heteroaryl radicals include oxazolyl, imidazolyl, pyrazolyl and triazolyl.

[0047] In some such embodiments, the monocyclic heteroaryl is unsubstituted or substituted by one or more substituents selected from the group consisting of alkyl, alkylthio, alkoxy, alkoxycarbonyl, amido, carboxyl, cyano, halo, aryl, heteroaryl, heterocyclic, nitro, sulfonamido and alkylthio. In some embodiments, the monocyclic heteroaryl is substituted by 6-membered aryl, heteroaryl or heterocyclic selected from phenyl, pyridyl, 2-hydroxypyridyl, piperidone, 2-hydroxy-1-methylpyridyl, triazolyl, imidazolidinone, pyrimidone, 2-hydroxyisoquinolyl, 3-hydroxypyridazinyl, pyrrolidinone, pyrazolyl and morpholinone. In some embodiments, the monocyclic heteroaryl is substituted by heteroaryl or heterocyclic, which is substituted by one or more substituents selected from the group consisting of halo, CN, alkyl, alkoxy, hydroxy, carboxyl, -CO2 alkyl and tetrazolyl. In certain preferred embodiments, the substituent is disposed in the para position of Y relative to X.

[0048] In other embodiments, Y is a bicyclic heteroaryl such as benzothiazolyl, benzo In some preferred embodiments, the bicyclic heteroaryl is unsubstituted or substituted by one or more substituents selected from the group consisting of alkyl, haloalkyl, hydroxyalkyl, alkylthio, alkoxy, alkoxycarbonyl, acylamino, carboxyl, cyano, halo, heteroaryl, nitro, and sulfonamido. In some preferred embodiments, the bicyclic heteroaryl is unsubstituted or substituted by one or more substituents selected from the group consisting of alkylthio, alkoxycarbonyl, acylamino, carboxyl, halo, and heteroaryl. In some preferred embodiments, Y is -C(O)NR 8 R 9 or -NR 9 C(O)R 10 is substituted with an acylamino substituent, wherein R 8 and R 9 are each independently selected from H, alkyl, heterocyclyl and heteroaryl; or

[0049] R 8 and R 9 Together with the nitrogen atoms to which they are attached, they form a 4-, 5-, 6-, or 7-membered

[0050] membered heterocyclic or heteroaryl ring; and

[0051] R 10 It is an alkyl group.

[0052] In other preferred embodiments, Y is represented by the formula -S(O)2NR 8 R 9 or -NR 9 S(O)2R 10 substituted with a sulfonylamino substituent; wherein

[0053] R 8 and R 9 are each independently selected from H, alkyl and heteroaryl; or

[0054] 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; and

[0055] R 10 It is an alkyl group.

[0056] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc), R 8 and R 9Each is independently selected from H, methyl, ethyl, triazolyl and pyrazolyl. In certain embodiments, R 8 and R 9 One or both of are alkyl, and each alkyl is independently unsubstituted or substituted by one or more substituents selected from the group consisting of methyl, methoxy, carboxyl, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, In other embodiments, R 8 and R 9 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from the group consisting of aziradine, isothiazolidine-1,1-dioxide, azetidine, thiazol-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, 2-oxa-6-azaspiro[3.3]heptane. In certain embodiments, R 8 and R 9 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from the group consisting of 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]dec-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9-diazaspiro[5.5]undecan-3-one, tetrahydropyrrolo[3,4-d]imidazole, pyrimidine, 8-oxa-2-azaspiro[4.5]decane, hexahydro-3H- oxazolo[3,4-a]pyrazin-3-one, 1-oxa-7-azaspiro[3.5]nonane, octahydrocyclopenta[c]pyrrole, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine, 2,7-diazaspiro[4.4]nonane, 2,6-diazaspiro[3.4]octane, 7-oxa-2-azaspiro[3.5]nonane, 1-oxa-8λ 2 -azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, In some embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxyl, and heterocyclic. In some embodiments, the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from the group consisting of methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxyl, and oxetanyl.

[0057] In certain embodiments of Formula (I), (Ia), (Ib), (Ic), (IIa), (IIb), (IIc), (IIIa), (IIIb) or (IIIc), R 1 and R 1` In other embodiments, R 1 and R 1` Combine to form an alkyl olefin (eg, aminoallyl).

[0058] In certain embodiments, the compound is selected from

[0059] or a pharmaceutically acceptable salt thereof.

[0060] 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.

[0061] In another aspect, the present disclosure provides a method of treating a cardiovascular disease or condition 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.

[0062] In certain embodiments, the cardiovascular disease is selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, 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 in the subject is reduced. In certain embodiments, the circulating serum LDL-cholesterol level in the subject is reduced. In certain embodiments, the circulating serum VLDL-cholesterol level in the subject is reduced. In certain embodiments, the circulating serum triglyceride level in the subject is reduced. In certain embodiments, the circulating serum lipoprotein A level in the subject is reduced. In certain embodiments, the subject suffers from atherosclerosis. In certain embodiments, atherosclerotic plaque formation in the subject is reduced. In certain embodiments, the subject has a gain-of-function mutation in the PCSK9 gene.

[0063] 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.

[0064] In certain embodiments, the additional therapeutic agent is selected from HMG-CoA reductase inhibitors, HMG-CoA synthase inhibitors, HMG-CoA reductase gene expression inhibitors, HMG-CoA synthase gene expression inhibitors, MTP / Apo B secretion inhibitors, CETP inhibitors, bile acid absorption inhibitors, cholesterol absorption inhibitors, cholesterol synthesis inhibitors, squalene synthase inhibitors, squalene epoxidase inhibitors, squalene cyclase inhibitors, combined squalene epoxidase / squalene cyclase inhibitors, fibrates, niacin, a combination of niacin and lovastatin, ion exchange resins, antioxidants, ACAT inhibitors, bile acid sequestrants, PCSK9 translation inhibitors, ATP citrate lyase inhibitors, apoC3 inhibitors, ANGPTL3 inhibitors, omega-3 fatty acids, Lp(a) inhibitors, apoB inhibitors, apoA1 inhibitors, low-density lipoprotein receptor inhibitors, low-density lipoprotein inhibitors, diacylglycerol acyltransferase inhibitors, lysosomal acid lipase inhibitors, and lecithin cholesterol acyltransferase inhibitors. 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.

[0065] In certain embodiments, the additional therapeutic agent is selected from volanesorsen, 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, bempedoic acid, ezetimibe, IONIS-APO(a)RX, mipomersen, CSL-112, lomitapide, AAV8.TBG.hLDLR (RGX-501), alipogen tiparvovec), pradigastat, sebelipase, ACP-501 / MEDI6012, OM3FA-EE, OM3FA-CA, OM3FA-IPE, alirocumab, eloumab, bococizumab, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimbe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin. In certain embodiments, the additional therapeutic agent is voranexan. In certain embodiments, the additional therapeutic agent is ivisoumab. 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 ethylester. In certain embodiments, the additional therapeutic agent is docosahexaenoic acid ethylester. In certain embodiments, the additional therapeutic agent is bempedoic 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 aliporin. In certain embodiments, the additional therapeutic agent is purulentastat.In certain embodiments, the additional therapeutic agent is sebacetase. 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.

[0066] 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.

[0067] 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 the administration of the PCSK9 inhibitor.

[0068] In 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.

[0069] 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.

[0070] In certain embodiments, the additional therapeutic agent is selected from the group consisting of kanamycin, amikacin, tobramycin, dibekacin, gentamicin, sisomicin, netilmicin, streptomycin, neomycin B, neomycin C, and neomycin E, imipenum, meropenum, ertapenem, doripenum, panipenum, biapenem, razupenum, tebipenum, lenapenum, tomopenum, thienpenum, ciprofloxacin, garenoxacin, gatifloxacin, gemifloxacin, floxaicin), levofloxacin, cinoxacin, nalidixic acid, moxifloxacin, oxolinic acid, piromidic acid, pipemidic acid, rosofloxacin, enoxacin, fleroxacin, lomefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, rufloxacin, balofloxacin, grefloxacin, levofloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, clinafloxacin, sitafloxacin, trovafloxacin, Prulifloxacin, delafloxacin, JNJ-Q2, nemofloxacin, zabofloxacin, doxycycline, tetracycline, chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, metacycline, minocycline, rolicycline, tigecycline, ampicillin, amoxicillin, augmentin, piperacillin, tazobactam, chloramphenicol, and ticarcillin.

[0071] Pharmaceutical composition

[0072] The compositions and methods disclosed herein can be used to treat individuals in need. In certain embodiments, the individual is a mammal, such as a human or non-human mammal. When applied to an animal (such as a human), the composition or compound is preferably administered as a pharmaceutical composition, which comprises, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions, such as water or physiologically buffered saline or other solvents or vehicles, such as ethylene glycol, glycerol, oil (such as olive oil) or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive administration routes (i.e., routes of transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. For example, an excipient can be selected to achieve delayed release of a medicament or selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form, such as tablets, capsules (including dispersed capsules (sprinkle capsules) and gelatin capsules), granules, lyophiles for reconstruction, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system, such as a skin patch. The composition may also be present in a solution suitable for topical administration, such as a lotion, cream or ointment.

[0073] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent, which, for example, acts to stabilize a compound (such as a compound of the present invention), increase its solubility, or increase its absorption. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier (including 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 drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix, into which, for example, a compound of the present invention may be incorporated. For example, liposomes comprising phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0074] The phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0075] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0076] The pharmaceutical composition (preparation) can be administered to a subject by any of a variety of routes of administration, including, for example, oral administration (e.g., as a drench in the form of an aqueous or non-aqueous solution or suspension, tablets, capsules (including dispersible capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); 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 compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Patents 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.

[0077] The preparation can be conveniently present in unit dosage form and can be prepared by any method known in the pharmaceutical field. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary according to the host being treated, the specific mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be the amount of the compound that produces the therapeutic effect. Usually, the range of this amount (in percentage) will be about 1% to about 99% of the active ingredient, preferably about 5% to about 70%, most preferably about 10% to about 30%.

[0078] The method for preparing these preparations or compositions comprises the step of combining an active compound (such as a compound of the present invention) with a carrier and optionally one or more auxiliary ingredients. In general, the preparation is prepared by uniformly and intimately combining a compound of the present invention with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product.

[0079] Formulations of the invention suitable for oral administration may be in the form of capsules (including dispersible capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilizates, powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a lozenge (using an inert base such as gelatin and glycerin, or sucrose and acacia) and / or as a mouthwash, etc., each containing a predetermined amount of the disclosed compound as the active ingredient. The composition or compound may also be administered in the form of a bolus, electuary or paste.

[0080] To prepare solid dosage forms for oral administration (capsules (including dispersible capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, for example, carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic; (3) wetting agents, Such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retardants, such as paraffin; (6) absorption promoters, such as quaternary ammonium compounds; (7) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (8) absorbents, such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including dispersible capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Solid compositions of a similar type can also be used as fillers in soft-filled and hard-filled gelatin capsules, using excipients such as lactose (lactose / milk sugar) and high molecular weight polyethylene glycols and the like.

[0081] Tablets can be made by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets can be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surfactant or a dispersant. Molded tablets can be prepared by molding a mixture of a powdered compound moistened with an inert liquid diluent in a suitable machine.

[0082] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including dispersible capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the art of pharmaceutical formulation. They may also be formulated to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethylcellulose, other polymer matrices, liposomes and / or microspheres in different proportions that provide the desired release characteristics. They may be sterilized, for example, by filtering through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain an opacifying agent and may be such a composition that they release the active ingredient only or preferentially in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes. If appropriate, the active ingredient may also be in microencapsulated form with one or more of the above-mentioned excipients.

[0083] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may also contain inert diluents commonly used in the art (e.g., water or other solvents), cyclodextrins and derivatives thereof, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuran alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.

[0084] 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.

[0085] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0086] 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 with any preservatives, buffers or propellants that may be required.

[0087] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0088] In addition to the active compounds, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0089] Transdermal patches have the added advantage of providing controlled delivery of the disclosed compound to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0090] The phrase "parenteral administration" as used herein means modes of administration other than enteral and topical administration (usually by injection), and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise a combination of one or more active compounds with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions or sterile powders, which can be reconstituted into sterile injectable solutions or dispersions immediately prior to 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.

[0091] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Appropriate fluidity can be maintained, for example, by the use of coating materials (such as lecithin), by maintaining the desired particle size in the case of dispersions, and by the use of surfactants.

[0092] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. It is possible to ensure that the effects of microorganisms are prevented by including various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol sorbic acid, etc.). It may also be desirable to include isotonic agents, such as sugar, sodium chloride, etc. in the compositions. In addition, the absorption extension of injectable drug forms may be achieved by including agents that delay absorption (such as aluminum monostearate and gelatin).

[0093] In some cases, in order to prolong the effect of the drug, it is desirable to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn can depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is achieved by dissolving or suspending the drug in an oily vehicle.

[0094] Injectable depot forms are prepared by forming a microencapsulated matrix of the subject compound in a biodegradable polymer such as polylactide-polyglycolide. The drug release rate can be controlled according to the ratio of the drug to the polymer and the properties of the specific polymer employed. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by embedding the drug in liposomes or microemulsions compatible with body tissues.

[0095] For use in the compounds of the present invention, the active compound may be administered per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably, 0.5% to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0096] The introduction method can also be provided by a rechargeable or biodegradable device. In recent years, various slow-release polymer devices have been developed and tested in vivo for controlled delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers, including biodegradable and non-degradable polymers, including hydrogels, can be used to form implants to continuously release compounds at specific target sites.

[0097] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0098] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the esters, salts or amides thereof, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0099] A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can start the dosage of the pharmaceutical composition or compound at a level lower than that required to obtain the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means the concentration of the compound sufficient to cause the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's condition, the condition being treated, the stability of the compound, and (if necessary) another type of therapeutic agent administered together with the method of the present invention. A larger total dose can be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine, 13th edition, 1814-1882, incorporated herein by reference).

[0100] In general, a suitable daily dose of the active compound used in the compositions and methods of the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0101] If desired, the effective daily dose of the active compound can be administered at appropriate intervals throughout the day as one, two, three, four, five, six or more sub-doses administered separately, optionally in unit dosage form. In certain embodiments of the present disclosure, the active compound can be administered twice or three times a day. In a preferred embodiment, the active compound will be administered once a day.

[0102] Generally speaking, the patient receiving such treatment is any animal in need thereof, including primates, particularly humans; and other mammals, such as horses, cattle, pigs, sheep, cats, and dogs; poultry; and pets.

[0103] In certain embodiments, the compounds of the invention may be used alone or in combination with another type of therapeutic agent.

[0104] The disclosure includes the pharmaceutically acceptable salt of the compound of the present invention for use in the compositions and methods of the present disclosure. In certain embodiments, the salt of the present invention considered includes but is not limited to alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, the salt of the present invention considered includes but is not limited to L-arginine, phenethylbenzylamine, benzathine, betaine, calcium hydroxide, choline, danol, diethanolamine, diethylamine, 2- (diethylamino) ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, haibamin, 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 salt of the present invention considered includes but is not limited to Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, 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), acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic 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 undecylenic acid salts.

[0105] Pharmaceutically acceptable acid addition salts may also exist in the form of various solvates, such as solvates with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates may also be prepared. The source of such solvates may be derived from the crystallization solvent, inherent in the preparation or crystallization solvent, or exogenous to such solvent.

[0106] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0107] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0108] definition

[0109] 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 and techniques used in connection with the 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.

[0110] Unless otherwise indicated, the compounds and techniques of the present disclosure are generally performed according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout the 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 Edition", WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th Edition", WH Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th Edition", Sinauer Associates, Inc., Sunderland, MA (2000).

[0111] Unless otherwise defined herein, chemical terms used herein are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., ed., McGraw-Hill, San Francisco, CA (1985).

[0112] All of the above and any other publications, patents, and published patent applications mentioned in this application are specifically incorporated by reference. In case of conflict, the present specification, including its specific definitions, will control.

[0113] The term "agent" is used herein to refer to a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biomacromolecule (such as a nucleic acid, an antibody, including portions thereof and humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portions thereof, such as peptides, lipids, carbohydrates), or an extract prepared from a biological material (such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues). Agents include, for example, agents of known structure and agents of unknown structure. 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.

[0114] "Patient", "subject" or "individual" are used interchangeably and refer to humans or non-human animals. These terms include mammals, such as humans, primates, livestock animals (including cattle, pigs, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

[0115] "Treating" a condition or patient means taking steps to obtain a beneficial or desired result, including a clinical result. Beneficial or desired clinical results may include, but are not limited to, alleviation or amelioration of one or more symptoms or symptoms, alleviation of the extent of the disease, stabilization of the disease state (i.e., not worsening), prevention of disease spread, delay or slowing of disease progression, improvement or palliation of the disease state, and detectable or undetectable remission (whether partial or complete). "Treatment" may also refer to prolonging survival compared to the expected survival if not receiving treatment.

[0116] The term "prevention" is art-recognized and well understood in the art when used with respect to a condition such as a local recurrence (e.g., pain), a disease such as cancer, a complex syndrome such as heart failure, or any other medical condition, and includes administering a composition that reduces the frequency of symptoms of, or delays the onset of, a medical condition in a subject relative to a subject not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancer growths in a patient population receiving prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancer growths in a treated population relative to an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0117] "Administering" a substance, compound or medicament to a subject can be performed using one of a variety of methods known to those skilled in the art. For example, the compound or medicament can be administered intravenously, intraarterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinal, intracerebrally, and transdermally (by absorption, such as through a skin catheter). The compound or medicament can also be suitably introduced by a rechargeable or biodegradable polymer device or other device (e.g., patch and pump) or formulation that provides extended, slow or controlled release of the compound or medicament. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0118] The appropriate method of administering a substance, compound, or medicament to a subject will also depend on, for example, the age and / or physical condition of the subject and the chemical and biological properties (e.g., solubility, digestibility, bioavailability, stability, and toxicity) of the compound or medicament. In some embodiments, the compound or medicament is administered orally, e.g., by ingestion to the subject. In some embodiments, the orally administered compound or medicament is an extended release or slow release formulation, or is administered using a device for such slow or extended release.

[0119] As used herein, the phrase "combined administration" refers to any form of administration of two or more different therapeutic agents such that a second agent is administered while the previously administered therapeutic agent is still effective in vivo (e.g., both agents are effective in the patient at the same time, which may include a synergistic effect of the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially in the same formulation or in separate formulations. Thus, an individual receiving such treatment may benefit from the combined effects of the different therapeutic agents.

[0120] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or medicament is an amount of the drug or medicament that will have the desired therapeutic effect when administered to a subject. The full therapeutic effect does not necessarily occur by administering one dose, and may occur only after administering a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount required for a subject will depend, for example, on the size, health, and age of the subject, and the nature and extent of the condition being treated (such as cancer or MDS). A skilled artisan can readily determine the effective amount in a given situation by routine experimentation.

[0121] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where the event or circumstance does not occur. For example, "optionally substituted alkyl" means that the alkyl group may be substituted as well as the alkyl group is not substituted.

[0122] It should be understood that one skilled in the art can select substituents and substitution patterns on the compounds of the present disclosure to produce chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art and those described below. If a substituent is itself substituted with more than one group, it should be understood that these multiple groups can be on the same carbon or on different carbons, as long as a stable structure is obtained.

[0123] As used herein, the term "optionally substituted" refers to one to six hydrogen groups in a given structure being replaced by the radical of a specific substituent, which includes, but is not limited to, hydroxy, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2, or -CH2-OP(O)(O-alkyl)2. Preferably, "optionally substituted" refers to one to four hydrogen groups in a given structure being replaced by the substituents mentioned above. More preferably, one to three hydrogen groups are replaced by the substituents mentioned above. It should be understood that the substituents may be further substituted.

[0124] As used herein, the term "alkyl" refers to a saturated aliphatic group, including but not limited to C1-C 10 Straight chain alkyl group or C1-C 10Branched alkyl groups. Preferably, an "alkyl" group refers to a C1-C6 straight chain alkyl group or a C1-C6 branched chain alkyl group. Most preferably, an "alkyl" group 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, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl or 4-octyl, etc. An "alkyl" group may be optionally substituted.

[0125] Furthermore, the term "alkyl" as used throughout the specification, examples and claims is intended to include both unsubstituted alkyl groups and substituted alkyl groups, the latter referring to alkyl moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone, including halogenated alkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl and the like.

[0126] As used herein, the term "alkylene" refers to a divalent saturated aliphatic group, including but not limited to C1-C 50 Straight chain alkylene group or C1-C 50 Branched chain alkylene groups. In addition, the term "alkylene" as used throughout the specification, examples and claims is intended to include both unsubstituted alkylene groups and substituted alkylene groups, the latter referring to alkylene moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, and the like.

[0127] As used herein, the term "heteroalkylene" refers to a divalent aliphatic group in which one or more carbon atoms have been replaced by 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 2-50 atoms or a branched chain heteroalkylene group of 2-50 atoms. If the heteroalkylene group contains an N atom or another heteroatom with three or more open valence positions, the heteroatom atom may be unsubstituted (e.g., -N(H)-), or it may be substituted with substituents known to those skilled in the art (e.g., alkyl, aralkyl, aryl, etc., as further described herein), or all of the heteroatom valences may be occupied by its bond to the chain of the heteroalkylene group (e.g., -N= or =N-). Furthermore, the term "heteroalkylene" as used throughout the specification, examples and claims is intended to include both unsubstituted heteroalkylene groups and substituted heteroalkylene groups, the latter referring to heteroalkylene moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl and the like.

[0128] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0129] 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 hydrocarbyl C(O)NH-.

[0130] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0131] 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.

[0132] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0133] The term "C x-y " or "C x -C y "" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is intended to include groups containing from x to y carbons in the chain. C0 alkyl represents hydrogen where the group is in a terminal position or a bond if internal. For example, C 1-6 Alkyl groups contain from one to six carbon atoms in the chain.

[0134] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0135] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0136] As used herein, the term "amide" refers to the following group:

[0137]

[0138] Where R 9 and R 10 Each independently represents hydrogen or a hydrocarbon group, or R 9 and R 10 Taken together with the N atom to which they are attached complete the heterocycle having 4 to 8 atoms in the ring structure.

[0139] The terms "amine" and "amino" are art-recognized and refer to unsubstituted and substituted amines and salts thereof, such as the moiety represented by the formula:

[0140]

[0141] Where R 9 , R 10 and R 10 ' each independently represents hydrogen or a hydrocarbon group, or R 9 and R 10 Taken together with the N atom to which they are attached complete the heterocycle having 4 to 8 atoms in the ring structure.

[0142] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0143] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0144] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups, wherein each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes a polycyclic ring system with two or more cyclic rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is aromatic, for example, other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0145] The term "carbamate" is art-recognized and refers to the following group:

[0146]

[0147] Where R 9 and R 10 independently represents hydrogen or a hydrocarbyl group.

[0148] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0149] The term "carbocycle" includes 5-7 membered monocycles and 8-12 membered bicyclic rings. Each ring of the bicyclic carbocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Carbocycles include bicyclic molecules, wherein one, two, or three or more atoms are shared between two rings. The term "fused carbocycle" refers to a bicyclic carbocycle, wherein each ring shares two adjacent atoms with another ring. Each ring of the fused carbocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. 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). Where valence permits, any combination of saturated bicyclic, unsaturated bicyclic, and aromatic bicyclic rings is included in the definition of carbocycle. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo [2.2.1] heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo [4.2.0] oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. "Carbocycle" may be substituted at any position or positions capable of carrying a hydrogen atom.

[0150] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0151] The term "carbonate" is art-recognized and refers to the group -OCO2-.

[0152] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0153] The term "cycloalkyl" includes substituted or unsubstituted non-aromatic monocyclic structures, preferably 4- to 8-membered rings, more preferably 4- to 6-membered rings. The term "cycloalkyl" also includes polycyclic ring systems having two or more cyclic rings, wherein two or more carbon atoms are common to two adjacent rings, wherein at least one ring is a cycloalkyl group and the substituents (e.g., R 100 ) is attached to a cycloalkyl ring, for example, the other cyclic ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, Azoles, thiazoles, pyrazoles, pyridines, pyrazines, pyridazines, pyrimidines, benzodiazoles Denzodioxane, tetrahydroquinoline, etc.

[0154] As used herein, the term "ester" refers to the group -C(O)OR 9 , where R 9 represents a hydrocarbon group.

[0155] As used herein, the term "ether" refers to a hydrocarbyl group connected to another hydrocarbyl group through an oxygen. Thus, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. An ether may 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 may be represented by the general formula alkyl-O-alkyl.

[0156] As used herein, the terms "halo" and "halogen" mean halogen and include chlorine, fluorine, bromine and iodine.

[0157] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a heteroaryl group.

[0158] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings, wherein two or more carbon atoms are shared by two adjacent rings, wherein at least one ring is heteroaromatic, 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, Azoles, thiazoles, pyrazoles, pyridines, pyrazines, pyridazines, pyrimidines, benzodiazoles Alkane, tetrahydroquinoline, etc.

[0159] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

[0160] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0161] The terms "heterocyclyl", "heterocycle" and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3-10 membered rings, more preferably 3-7 membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings, wherein two or more carbon atoms are common to two adjacent rings, wherein at least one ring is heterocyclic, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.

[0162] As used herein, the term "hydrocarbyl" refers to a group bonded by a carbon atom, which does not have a =O or =S substituent, and generally has at least one carbon-hydrogen bond and a main carbon backbone, but may optionally include heteroatoms. Thus, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-nitrophenyl, and even trifluoromethyl are considered hydrocarbyl, but substituents such as acetyl (which has a =O substituent on the connecting carbon) and ethoxy (which is connected by oxygen rather than carbon) are not hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0163] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0164] The term "lower" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy, is intended to include groups in which the substituent has ten or fewer atoms, preferably six or fewer atoms. For example, "lower alkyl" refers to an alkyl group containing ten or fewer carbon atoms, preferably six or fewer carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy substituents defined herein are respectively low acyl, low acyloxy, low alkyl, low alkenyl, low alkynyl or low alkoxy, whether they occur alone or in combination with other substituents, such as in the narration of hydroxyalkyl and aralkyl (in this case, for example, when the carbon atoms in the alkyl substituent are counted, the atoms in the aryl group are not counted).

[0165] The terms "polycyclic group", "polycyclic" and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl) wherein two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings". Each ring of the polycyclic ring may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring contains 3 to 10 atoms, preferably 5 to 7 atoms, in the ring.

[0166] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0167] The term "sulfonamide" is art-recognized and refers to a group represented by the following general formula:

[0168]

[0169] Where R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0170] The term "sulfoxide" is art-recognized and refers to the group -S(O)-.

[0171] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0172] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0173] The term "substituted" refers to the part of the substituent having a hydrogen on one or more carbons of the replacement main chain. It should be understood that "substituted" or "substituted by ... " includes implicit conditions, i.e., such substitutions meet the allowed valence of the substituted atom and the substituent, and the substitution produces a stable compound, for example, the compound will not spontaneously transform such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all allowable substituents of organic compounds. In a broad sense, allowable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For suitable organic compounds, allowable substituents can be one or more and can be the same or different. For purposes of the present invention, heteroatoms (such as nitrogen) can have hydrogen substituents and / or any allowable substituents of organic compounds described herein, and these substituents satisfy the valence of heteroatoms. Substituents may include any of the substituents described herein, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl or aromatic or heteroaromatic moieties. It will be appreciated by those skilled in the art that the moieties substituted on the hydrocarbon chain may themselves be substituted if appropriate.

[0174] As used herein, the term "alkylthio" refers to an alkyl group substituted with a thiol group.

[0175] As used herein, the term "thioester" refers to the group -C(O)SR 9 or -SC(O)R 9 , where R 9 It represents a hydrocarbon group.

[0176] As used herein, the term "thioether" is equivalent to an ether in which the oxygen is replaced by sulfur.

[0177] The term "urea" is art-recognized and can be represented by the general formula Indicates that R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0178] As used herein, the term "modulate" includes inhibiting or suppressing a function or activity (such as cell proliferation) as well as enhancing a function or activity.

[0179] The phrase "pharmaceutically acceptable" is recognized in the art. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0180] As used herein, "pharmaceutically acceptable salt" or "salt" refers to an acid addition salt or a base addition salt that is suitable for or compatible with patient treatment.

[0181] As used herein, the term "pharmaceutically acceptable acid addition salt" means any non-toxic organic or inorganic salt of any base compound represented by Formula I. Exemplary inorganic acids forming suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, and metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids forming suitable salts include monocarboxylic acids, dicarboxylic acids 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. Monoacid or diacid salts can be formed, and such salts can exist in hydrated, solvated or substantially anhydrous forms. Typically, the acid addition salts of the compounds of Formula I are more soluble in water and various hydrophilic organic solvents, and typically exhibit higher melting points compared to their free base forms. The selection of appropriate salts is known to those skilled in the art. Other non-pharmaceutically acceptable salts (eg, oxalates) may be useful, for example, in isolating compounds of Formula I for laboratory use, or for subsequent conversion to pharmaceutically acceptable acid addition salts.

[0182] As used herein, the term "pharmaceutically acceptable base addition salt" means any non-toxic organic or inorganic base addition salt of any acid compound represented by Formula I or any of their 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 appropriate salts is known to those skilled in the art.

[0183] Many compounds that can be used in the compounds and compositions of the present disclosure have at least one stereocenter in their structure. The stereocenter can exist in R or S configuration, and the R and S symbols are used according to the rules described in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms of compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers), such as enantiomeric and diastereomeric forms. See, for example, WO01 / 062726.

[0184] Furthermore, certain compounds containing alkenyl groups may exist as Z (same side) or E (different side) isomers. In each case, the present disclosure includes both the mixture and the individual isomers.

[0185] Some compounds may also exist in tautomeric forms. Although not explicitly indicated in the formulas described herein, such forms are intended to be included within the scope of the present disclosure.

[0186] "Prodrug" or "pharmaceutically acceptable prodrug" refers to a compound that is metabolized (e.g., hydrolyzed or oxidized) in a host after administration to form a method of the present disclosure (e.g., a compound of formula I). ​​Typical examples of prodrugs include compounds having biologically unstable or cleavable (protective) groups on the functional portion of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminized, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated or dephosphorylated to produce an active compound. Examples of prodrugs using esters or phosphoramidates as biologically unstable or cleavable (protective) groups are disclosed in U.S. Patents 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 produce compounds 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.

[0187] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filtrate, diluent, excipient, solvent or encapsulating material, which can be used to formulate a medicament for medical or therapeutic use.

[0188] As used herein, the term "logarithm of solubility", "LogS" or "logS" is used in the art to quantify the water solubility of a compound. The water solubility of a compound significantly affects its absorption and distribution characteristics. Low solubility is usually accompanied by poor absorption. The LogS value is the unit peel logarithm (base 10) of the solubility measured in moles / liter.

[0189] As used herein, the term "interaction" in the context of an inhibitor associated with one or more residues of PCSK9 refers to direct or indirect binding of the inhibitor to the backbone of the enzyme or to the side chain of any given residue. For example, interactions include covalent bonds, hydrogen bonds, hydrophobic attraction, cation-π interactions, anion-π interactions, and other interactions well known in the art.

[0190] Example

[0191] Having now generally described the invention, the invention will be more readily understood by reference to the following examples, which are included merely for the purpose of illustrating certain aspects and embodiments of the disclosure and are not intended to limit the invention.

[0192] Example 1: Preparation of Exemplary PCSK9 Inhibitors

[0193] 1 H NMR spectroscopy was performed on a Varian MR-400 spectrometer operating at 400 MHz (proton frequency) equipped with: self-shielded z-gradient coil 5 mm 1H / nX reverse detection broadband probe, deuterium digital lock channel unit, orthogonal digital detection unit with transmitter offset frequency shift. Chemical shifts are reported as δ values ​​in ppm relative to trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in Hertz (Hz) and the following abbreviations are used to report multiplicities (s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad, nd = not determined).

[0194] Example B

[0195]

[0196] Step A: 1-tert-butyl 3-methyl 3-allylazetidine-1,3-dicarboxylate (2)

[0197] Under nitrogen atmosphere, at -78 ° C, LiHMDS (1M THF solution, 179.5 mL, 179.5 mmol) was added dropwise to a stirred solution of azetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (19.3 g, 89.7 mmol) in anhydrous THF (180 mL). 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. After completion, the reaction mixture was cooled to 0 ° C, quenched with saturated NH4Cl aqueous solution (200 mL), and extracted with EA (150 mL × 3). The combined organic layers were washed with brine (250 mL), dried over Na2SO4, 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.

[0198] Step B: tert-Butyl 3-allyl-3-(hydroxymethyl)azetidine-1-carboxylate (3)

[0199] At -78 ° C, DIBAl-H (1M in toluene, 129 mL, 129 mmol) was slowly added to a stirred solution of 3-allylazetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (22 g, 86 mmol) in anhydrous DCM (300 mL) and stirred at room temperature for 2 h. After completion, the reaction mixture was cooled to 0 ° C and quenched with NaOH (1M in H2O solution, 100 mL) and extracted with DCM (200 mL × 3). The combined organic layer was washed with brine (250 mL), dried over Na2SO4, 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.

[0200] Step C: tert-Butyl 3-allyl-3-formylazetidine-1-carboxylate (4)

[0201] At 0 ° C, Dess-Martin (56g, 132mmol) was added to a stirred solution of 3-allyl-3-(hydroxymethyl)azetidine-1-carboxylic acid tert-butyl ester (15g, 66mmol) in anhydrous DCM (500mL), and the mixture was stirred at room temperature for 2h. The reaction mixture was quenched with Na2SO3 (10%, 200mL) and NaHCO3 aqueous solution (10%, 200mL), extracted with DCM (200mL×3). The combined organic layer was washed with brine (300mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MTBE / PE=25%) to give 3-allyl-3-formylazetidine-1-carboxylic acid tert-butyl ester (9.2g, yield: 46%, three steps) as a yellow oil.

[0202] Step D: tert-Butyl 3-allyl-3-(1-hydroxyallyl)azetidine-1-carboxylate (5)

[0203] At -78 ° C, to a stirred solution of 3-allyl-3-formylazetidine-1-carboxylic acid tert-butyl ester (9.2g, 40.8mmol) in anhydrous THF (200mL), vinylmagnesium bromide (1M THF solution, 49mL, 49mmol) was added, and the reaction mixture was stirred at room temperature for 2h. After completion, the reaction mixture was cooled to 0 ° C, quenched with saturated NH4Cl (200mL), and extracted with EA (200mL×3). The combined organic layer was washed with brine (200mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain 3-allyl-3-(1-hydroxyallyl)azetidine-1-carboxylic acid tert-butyl ester (crude, 10g, yield: 97%), which was a yellow oil that was directly used in the next step without further purification.

[0204] Step E: tert-Butyl 3-acryloyl-3-allylazetidine-1-carboxylate (6)

[0205] At 0 ° C, Dess-Martin (25g, 59mmol) was added to a stirred solution of 3-allyl-3-(1-hydroxyallyl)azetidine-1-carboxylic acid tert-butyl ester (10g, 39.5mmol) in anhydrous DCM (200mL), and the mixture was stirred at room temperature for 2h. The reaction mixture was quenched with Na2SO3 (10%, 100mL) and NaHCO3 aqueous solution (10%, 100mL), extracted with DCM (100mL×3). The combined organic layer was washed with brine (150mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE=18%) to obtain 3-acryloyl-3-allylazetidine-1-carboxylic acid tert-butyl ester (4.2g, yield: 42%, two steps) as a yellow oil.

[0206] Step F: tert-Butyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (7)

[0207] Under nitrogen atmosphere, Grubbs-II catalyst (709mg, 0.8mmol) was added to a stirred solution of 3-acryloyl-3-allylazetidine-1-carboxylic acid tert-butyl ester (4.2g, 16.7mmol) in toluene (200mL). The mixture was stirred at 85 ° C for 1h. 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 obtain 5-oxo-2-azaspiro [3.4] oct-6-ene-2-carboxylic acid tert-butyl ester (3.1g, yield: 83%) as a brown solid.

[0208] Step G: 2-Azaspiro[3.4]oct-6-en-5-one (8)

[0209] 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-ene-5-one (2 g crude, quantitative), which was used directly in the next step without further purification.

[0210] Step H: Benzyl 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylate (9)

[0211] To a stirred solution of 2-azaspiro[3.4]oct-6-ene-5-one (1.1 g, 8.9 mmol) in THF (15 mL) at 0°C was added TEA (2.7 g, 26.7 mmol) and Cbz-OSU (2.67 g, 10.7 mmol). The mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE=35%) to give 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylic acid benzyl ester (2 g, yield: 86%, two steps) as a yellow oil. LCMS[M+1] + =258.0

[0212] Step I: (R)-7-Hydroxy-5-oxo-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (10)

[0213] 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. Then a solution of bis(pinacol)diborane (2.96 g, 11.7 mmol) in THF (20 mL) was added, and the resulting mixture was stirred at room temperature for 10 min. A solution of 5-oxo-2-azaspiro[3.4]oct-6-ene-2-carboxylic acid benzyl ester (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, followed by water (10 mL), followed by sodium perborate (6.0 g, 39 mmol), and the resulting mixture was stirred vigorously at room temperature for 1 h. The resulting green suspension was filtered through a celite pad, poured into a separatory funnel containing a saturated NaHCO3 / Na2SO3 aqueous solution (1 / 1, 50 mL), and extracted with EA (100 mL×3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give crude (R)-7-hydroxy-5-oxo-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (5.4 g, quantitative), which was used directly in the next step without further purification. LCMS [M+1] + =276.0

[0214] Step J: (R)-7-((tert-butyldimethylsilyl)oxy)-5-oxo-2-azaspiro[3.4]octane-2-yl)- Benzyl formate (11)

[0215] At 0 ° C, imidazole (4.3 g, 31.2 mmol) and TBSCl (2.3 g, 15.6 mmol) were added to a stirred solution of (R)-7-hydroxy-5-oxo-2-azaspiro [3.4] octane-2-carboxylic acid benzyl ester (2.1 g, 7.8 mmol) in DCM (30 mL) at 0 ° C. 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 layer was 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 (R)-7-((tert-butyldimethylsilyl)oxy)-5-oxo-2-azaspiro [3.4] octane-2-carboxylic acid benzyl ester (1.8 g, yield: 59%, two steps) as a yellow oil. LCMS [M+1] + =390.0

[0216] Step K: (5R,7R)-7-((tert-butyldimethylsilyl)oxy)-5-((R)-1,1-dimethylethyloxy Benzyl 2-azaspiro[3.4]octane-2-carboxylate (12)

[0217] To a stirred solution of (R)-7-((tert-butyldimethylsilyl)oxy)-5-oxo-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (1.7 g, 4.4 mmol) in anhydrous THF (20 mL) at 0°C, (R)-2-methylpropane-2-sulfinamide (1.06 g, 8.7 mmol) and titanium tetraisopropoxide (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 by saturated NH4Cl aqueous solution, 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

[0218] Step L: (5R,7R)-5-((R)-1,1-dimethylethylsulfonamido)-7-hydroxy-2-azaspiro[3.4]octane Benzyl 2-oxanecarboxylate (13)

[0219] To a stirred solution of (5R,7R)-7-((tert-butyldimethylsilyl)oxy)-5-(((R)-tert-butylsulfinyl)amino)-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (800 mg, 1.62 mmol) in THF (3 mL) was added TBAF (1 M in THF, 2.42 mL, 2.42 mmol) and stirred for 2 h at room temperature. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=3%) to give (5R,7R)-5-(((R)-tert-butylsulfinyl)amino)-7-hydroxy-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (600 mg, yield: 97%) as a yellow oil. LCMS[M+1] + =381.0

[0220] Step M: (5R,7S)-5-((R)-1,1-dimethylethylsulfonylamino)-7-(1,3-dioxoisoindoline- Benzyl 2-azaspiro[3.4]octane-2-carboxylate (14)

[0221] To a stirred solution of (5R,7R)-5-(((R)-tert-butylsulfinyl)amino)-7-hydroxy-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (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 nitrogen atmosphere at 0° C. After 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 benzyl (5R,7S)-5-((R)-1,1-dimethylethylsulfonamido)-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

[0222] Step N: (5R,7S)-5-amino-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-yl Benzyl formate (15)

[0223] To a stirred solution of benzyl (5R,7S)-5-((R)-1,1-dimethylethylsulfonamido)-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (600 mg, 1.2 mmol) in DCM (5 mL) at 0 °C was added dihydrochloric acid HCl. Alkane (4M di The mixture was stirred at room temperature for 30 min. The reaction mixture was concentrated under reduced pressure to give (5R,7S)-5-amino-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (610 mg crude product, quantitative), which was used directly in the next step without further purification. LCMS[M+1] + =406.0

[0224] Step O: (5R,7S)-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-7-(1,3- Benzyl dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylate (16)

[0225] To a stirred solution of (5R,7S)-5-amino-7-(1,3-dioxoisoindolin-2-yl)-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (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 Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=2%) to give benzyl (5R,7S)-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

[0226] Step P: (5R,7S)-7-amino-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)- Benzyl 2-azaspiro[3.4]octane-2-carboxylate (17)

[0227] To a stirred solution of benzyl (5R,7S)-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) and stirred for 1 h at room temperature. 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 Na2SO4, filtered and concentrated under reduced pressure to give benzyl (5R,7S)-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

[0228] 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)

[0229] To a stirred solution of (5R,7S)-7-amino-5-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-2-azaspiro[3.4]octane-2-carboxylic acid benzyl ester (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 Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM=8%) to give benzyl (5R,7S)-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 was carried out using standard conditions to afford the title compound.

[0230] Embodiment 13

[0231]

[0232] Step A: Benzyl cyclopent-3-en-1-ylcarbamate

[0233] DPPA (140 g, 508.8 mmol) was added dropwise to a solution of cyclopent-3-ene-1-carboxylic acid (51.8 g, 462.5 mmol) in toluene (780 mL) and Et3N (56.1 g, 555.6 mmol). 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 layer was washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product of cyclopent-3-ene-1-ylcarbamic acid benzyl ester (126 g, quantitative), which was a yellow oil. The crude product was directly used in the next step without further purification.

[0234] LCMS [M+H] + =218.1.

[0235] Step B: Benzyl ((1R,3r,5S)-6-oxabicyclo[3.1.0]hexan-3-yl)carbamate

[0236] At 0 ° C, m-CPBA (85%, 50 g, 290.3 mmol) was added to a solution of cyclopent-3-ene-1-ylcarbamic acid benzyl ester (35 g, 161.3 mmol) in DCM (875 mL). The mixture was stirred at 0 ° C for 30 min. After completion, the reactant was quenched with Na2SO3 (aqueous solution) and NaHCO3 (aqueous solution) and extracted with DCM (800 mL × 2). The combined organic phase was washed with brine (800 mL × 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA / PE = 30%) to give ((1R, 3r, 5S) -6- oxabicyclo [3.1.0] hex-3-yl) benzyl carbamate (32.5 g, 86.7%) as a white solid. LCMS [M + H] + =234.1

[0237] 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)

[0238] A mixture of benzyl ((1R, 3r, 5S)-6-oxabicyclo[3.1.0]hex-3-yl)carbamate (32.5 g, 139.5 mmol) and (S)-1-phenylethyl-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.

[0239] Step D: tert-Butyl ((1R,3R,4R)-3-hydroxy-4-(((S)-1-phenylethyl)amino)cyclopentyl)carbamate ester

[0240] 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 at room temperature for 3 hours under a hydrogen atmosphere. 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

[0241] Step E: tert-Butyl ((1R,3R,4R)-3-amino-4-hydroxycyclopentyl)carbamate

[0242] 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)2 (6.17 g). The reaction mixture was stirred at 60 °C for 20 hours under a hydrogen atmosphere. 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

[0243] Step F: ((1R,3R,4R)-3-((6-(dimethylcarbamoyl)benzo[d]thiazol-2-yl)amino)-4-hydroxy tert-Butyl (cyclopentyl)carbamate

[0244] 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 for 6 h under a nitrogen atmosphere. After completion, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic layer was 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 light yellow solid. LCMS [M+H] + =421.3

[0245] Step G: 2-(((1R,2R,4R)-4-amino-2-hydroxycyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole- 6-Formamide

[0246] 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 dihydrochloric acid (HCl) at room temperature. alkane (2M, 5mL) solution. After addition, the reaction mixture was stirred at room temperature for 1h. 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 (669mg, quantitative) as the HCl salt, which was used in the next step without further purification. LCMS[M+H] + =321.1

[0247] Step H: 2-(((1R,2R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-hydroxycyclopentyl)amino)-N,N- Dimethylbenzo[d]thiazole-6-carboxamide

[0248] 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 K2CO3 (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 (eluted 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

[0249] Step I: 2-(((1R,4R)-4-((5-chloropyrimidin-2-yl)amino)-2-oxocyclopentyl)amino)-N,N-dimethyl Benz[d]thiazole-6-carboxamide

[0250] 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 portions at 0°C. After 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 solution (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 (eluted 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 Step J: 2-(((1R,4S)-4-((5-chloropyrimidin-2-yl)amino)-2-(cyano methyl)cyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide

[0251] At 0 ° C, NaH (52 mg, 1.3 mmol) was added to a solution of diethyl (cyanomethyl)phosphonate (230.7 mg, 1.3 mmol) in THF (3.2 mL). After addition, the mixture was stirred at 0 ° C for 30 minutes. Then a solution of 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 min. After completion, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (15 mL×2). The combined organic layer was 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 (eluted 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 1HNMR(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).

[0252] Example D

[0253] N,N-Dimethyl-2-[(1R,4S)-(Z)-2-(2-aminoethylidene)-4-(5-chloro-2-pyrimidinylamino)cyclopentane [amino]-1,3-benzothiazole-6-carboxamide

[0254]

[0255] The title compound can be prepared using 2-(((1R,4S)-4-((5-chloropyrimidin-2-yl)amino)-2-(cyanomethylene)cyclopentyl)amino)-N,N-dimethylbenzo[d]thiazole-6-carboxamide using reducing conditions.

[0256] Example E

[0257]

[0258]

[0259] Step A: (S)-2,3-Dihydro-1H-inden-1-ol (2)

[0260] Under nitrogen atmosphere, a solution of R-CBS (1.4 g, 5.15 mmol) and (CH3)2S-BH3 (0.48 mL, 0.95 mmol) was stirred at room temperature for 10 minutes. DCM (33 mL) and (CH3)2S-BH3 (28.4 mL, 56.82 mmol) were then added to the solution at -5 °C. A solution of 2,3-dihydro-1H-indene-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 addition, the reaction mixture was stirred at -5 °C for 2 hours. The mixture was quenched by CH3OH (40 mL) and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel, eluting with EtOAc / petroleum ether = 0%-10% 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. 1H NMR(400MHz,DMSO-d6)δ7.34-7.31(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).

[0261] Step B: (R)-2-(2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (4)

[0262] To a solution of (S)-2,3-dihydro-1H-indene-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 DIAD (11.1 g, 54.8 mmol) dropwise at 0°C under nitrogen atmosphere. After the addition, the reaction mixture was stirred at room temperature overnight. The reaction solution was then concentrated under reduced pressure. The residue was purified by silica gel flash chromatography using t The product was purified by eluting with BuOMe / petroleum ether = 0% - 11% to obtain (R)-2-(2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (4) (3.3 g, 34%) as a yellow oil. 1 HNMR (400MHz, DMSO-d6) δ7.84 (s, 4H), 7.29 (d, J = 7.6Hz, 1H), 7.24-7.20 (m, 1H), 7.13-7.1 1(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).

[0263] Step C: 2-((1R)-3-bromo-2,3-dihydro-1H-inden-1-yl)isoindoline-1,3-dione (5)

[0264] 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) under nitrogen atmosphere at room temperature. The reaction mixture was stirred at 80°C for 2 hours. 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 EtOAc / petroleum ether = 0%-9% 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.

[0265] Step D: 2-((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)isoindole Phyline-1,3-dione (6)

[0266] 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 DMBNH2 (3.13 g, 18.77 mmol) in DCM (40 mL) was added Et3N (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 Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel, eluting with 50% NH3-CH3OH / DCM=0%-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.

[0267] Step E: (3R)-N1-(2,4-dimethoxybenzyl)-2,3-dihydro-1H-indene-1,3-diamine (7)

[0268] 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 CH3OH (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 solution (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 flash chromatography on silica gel eluting with 50% NH3-CH3OH / DCM=0%-6% to give (3R)-N 1 -(2,4-dimethoxybenzyl)-2,3-dihydro-1H-indene-1,3-diamine (7) (700 mg, 78%), as a yellow oil. LCMS [M+H] + =299.5.

[0269] Step F: 6'-(((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)amino)- 2H-[1,3'-Bipyridyl]-2-one (9)

[0270] Under nitrogen atmosphere, at room temperature, (3R)-N 1A solution of -(2,4-dimethoxybenzyl)-2,3-dihydro-1H-indene-1,3-diamine (7) (400 mg, 1.34 mmol), 6'-chloro-2H-[1,3'-bipyridyl]-2-one (8) (277 mg, 1.34 mmol) and t-BuONa (258 mg, 2.68 mmol) in DMA (5 mL) was added. t BuxPhosPdG3 (119 mg, 0.134 mmol). 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 layer was 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%-7% to give 6'-(((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridine]-2-one (9) (440 mg, 70%) as a brown solid. LCMS[M+H] + =469.6. 1 H 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).

[0271] Step G: 6'-(((1R)-3-amino-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridine]-2- Ketone (10)

[0272] A solution of 6'-(((1R)-3-((2,4-dimethoxybenzyl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridine]-2-one (9) (440 mg, 0.94 mmol) in CF3COOH (6 mL) was stirred at 80°C for 48 hours. 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 crude product (290 mg) was used in the next step without further purification. LCMS[M+H] + =319.5.

[0273] Step H: 6'-(((1R)-3-((5-chloropyrimidin-2-yl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H- [1,3'-Bipyridyl]-2-one (12)

[0274] To a solution of 6'-(((1R)-3-amino-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridyl]-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 hours. 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 flash chromatography on silica gel, eluting with CH3OH / DCM=0%-3% to give 6'-(((1R)-3-((5-chloropyrimidin-2-yl)amino)-2,3-dihydro-1H-inden-1-yl)amino)-2H-[1,3'-bipyridinyl]-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.

[0275] Example 2: Biological Activity of Exemplary PCSK9 Inhibitors

[0276] Human recombinant PCSK9 was expressed as follows :

[0277] Protein sequence :

[0278]

[0279] The following assays are used to identify and evaluate PCKS9 inhibitors that effectively inhibit PCSK9 function.

[0280] Example: PCSK9 SPR Assay

[0281] At 25 °C in Biacore TM Surface plasmon resonance data were collected on a T200 or 3000 system (GE Healthcare). Streptavidin was immobilized on a CM5 (GE Healthcare) or CMD500d sensor chip (XanTec Bioanalytics) using standard amine coupling chemistry at 25°C with HBS-N (10 mM HEPES, 0.15 M NaCl, pH 7.4) as the running buffer. Briefly, the carboxymethyl dextran surface was activated by injecting 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. To capture streptavidin, 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. The residual activated groups were blocked by 7min injection of 1M ethanolamine (pH 8.5). By injecting 150 μL of protein diluted to 16 μg / mL in HBS-N, 0.05% tween-20, 0.1mM CaCl2, the PCSK9 protein of Avi tagging was captured on the streptavidin surface. The typical surface density obtained was 8000RU-10000RU. The SPR binding data were obtained with a flow rate of 30 μL / min using the appropriate dilution series of each compound, with a capture time of 100s and a dissociation time of 300s. The running buffer used for compound binding studies was HBS-N, 0.05% tween-20, 0.1mM CaCl2, 4% DMSO. The data not including the volume effect were corrected for DMSO. Using standard processing procedures, all data were double-quoted for blank injection and reference surface, and data processing and kinetic fitting were performed using Scrubber software version 2.0c (biological software (BioLogic Software)). A simple 1:1 binding model was fitted to the data to determine k a , k d and K D value.

[0282] The ability of PCSK9 inhibitors, and pharmaceutically acceptable salts thereof disclosed herein, to bind and inhibit PCSK9 was established using representative PCSK9 inhibitors of the present disclosure listed in the following table:

[0283] PCSK9 affinity (nM) Classification <![CDATA[K D =2,000-20,000nM]]> + <![CDATA[K D =200-2,000nM]]> ++ <![CDATA[K D <200nM]]> +++

[0284]

[0285]

[0286]

[0287] Incorporated by Reference

[0288] All publications and patents mentioned herein are hereby incorporated by reference in their entirety to the same extent 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.

[0289] Equivalent solutions

[0290] Although specific embodiments of the present invention have been discussed, the above description is illustrative rather than restrictive. Many variations of the present invention will become apparent to those skilled in the art by reading this specification and the claims below. The full scope of the present invention should be determined by reference to the claims together with the full scope of their equivalents and the specification together with such variations.

Claims

1. A compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof: in: A is selected from H, halo, hydroxy, alkyl, alkylthio, alkenyl, alkoxy, acyloxy, cyano, cycloalkyl, -C(O)OR 6 and -C(O)NR 6 R 7 ; 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-membered or 6-membered heteroaryl; X is NR 5 or O; Z is cycloalkyl, cycloalkenyl, aryl, heteroaryl or heterocyclyl; Each R 6 and R 7 are independently H or alkyl; Y is selected from aryl, heteroaryl and heterocyclyl; and n is 0 or 1.

2. The compound according to claim 1, wherein Z is cycloalkyl.

3. The compound according to claim 1 or 2, wherein the structure of the compound is represented by formula (Ia), (Ib), (Ic) or a pharmaceutically acceptable salt thereof: in R 1 and R 1` R is each independently selected from alkyl, halo, hydroxy, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamide, aryl, heteroaryl, heterocyclyl and aralkyl; or R 1 and R 1` Combine to form an alkylalkene, a cycloalkyl, a cycloalkene, or a heterocyclic group; Each R 2 are independently selected from alkyl, halo, hydroxy, carboxyl, acyl, ester, thioester, alkoxy, phosphoryl, amino, amide, cyano, nitro, azido, alkylthio, alkenyl, alkynyl, cycloalkyl, heterocyclylalkyl, heteroaralkyl, sulfonamide, aryl, heteroaryl, heterocyclyl, and aralkyl; and n is 1, 2, 3 or 4.

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

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

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

7. The compound according to claim 6, wherein A is H.

8. The compound according to claim 6, wherein A is an alkyl group.

9. The compound according to claim 6, wherein A is alkylthio.

10. The compound according to claim 6, wherein A is alkoxy. The compound according to claim 6 , wherein A is cycloalkyl.

12. The compound according to claim 6, wherein A is selected from -SCH3, -SCHF2 and -OCHF2.

13. The compound of claim 6, wherein A is a halo group (eg, a chloro group).

14. 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, which is unsubstituted or substituted with one or more alkyl groups.

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

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

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

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

19. The compound according to claim 18, wherein Y is selected from azophenyl, pyridonyl, pyridinyl, pyrazinyl, pyrimidinyl and thiazolyl.

20. The compound according to claim 18, wherein Y is selected from triazene, Azolyl, iso oxazolyl, imidazolyl, pyrazolyl and triazolyl.

21. according to the compound described in any one of claim 18 to 20, wherein said monocyclic heteroaryl is unsubstituted or is substituted by one or more substituents selected from the group consisting of alkyl, alkylthio, alkoxy, alkoxycarbonyl, acylamino, carboxyl, cyano, halo, aryl, heteroaryl, heterocyclic radical, nitro, sulfonamido and alkylthio.

22. The compound of claim 21, wherein the monocyclic heteroaryl is substituted with a 6-membered aryl, heteroaryl or heterocyclic group selected from phenyl, pyridyl, 2-hydroxypyridyl, piperidonyl, 2-hydroxy-1-methylpyridyl, triazolyl, imidazolidinonyl, pyrimidonyl, 2-hydroxyisoquinolyl, 3-hydroxypyridazinyl, pyrrolidinonyl, pyrazolyl and morpholinonyl.

23. The compound of claim 21, wherein the monocyclic heteroaryl is substituted with a heteroaryl or heterocyclic radical, wherein the heteroaryl or heterocyclic radical is substituted with one or more substituents selected from the group consisting of halo, CN, alkyl, alkoxy, hydroxy, carboxyl, -CO2 alkyl, and tetrazolyl.

24. A compound according to any one of claims 21 to 23, wherein the substituent is disposed in the para position of Y relative to X.

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

26. The compound according to claim 25, wherein Y is selected from benzothiazolyl, benzo oxazolyl, benzimidazolyl, triazolopyridinyl, thiazolopyridinyl, quinolyl and quinoxalinyl.

27. according to the compound described in claim 25 or 26, wherein said bicyclic heteroaryl is unsubstituted or is substituted by one or more substituents selected from following: alkyl, haloalkyl, hydroxyalkyl, alkylthio, alkoxyl, alkoxycarbonyl, acylamino, carboxyl, cyano, halo, heteroaryl, nitro and sulfonamido.

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

29. A compound according to any one of claims 1 to 28, wherein Y is represented by the formula -C(O)NR 8 R 9 or -NR 9 C(O)R 10 is substituted with an acylamino substituent, wherein 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; and R 10 It is an alkyl group.

30. A compound according to any one of claims 1 to 28, wherein Y is represented by the formula -S(O)2NR 8 R 9 or -NR 9 S(O)2R 10 substituted with a sulfonylamino substituent; wherein 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; and R 10 It is an alkyl group.

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

32. A compound according to any one of claims 29 to 31, wherein R 8 and R 9 One or both of are alkyl, and each alkyl is independently unsubstituted or substituted by one or more substituents selected from the group consisting of methyl, methoxy, carboxyl, cyano, hydroxy, dimethylamino, ethoxycarbonyl, phenyl, methoxyphenyl, Oxazolyl, tetrazolyl, 2-methyl-tetrazolyl, triazolyl, 1-methyltriazolyl, 4-methyltriazolyl and 2,4-dihydro-3H-1,2,4-triazol-3-onyl.

33. The compound according to claim 29 or 30, wherein R 8 and R 9 Together with the nitrogen atom to which they are attached they form a heterocyclic ring selected from the group consisting of aziridine, isothiazolidine-1,1-dioxide, azetidine, thiazol-4(5Hn)-one, morpholine, piperidine, piperazine, pyrrolidine, thiomorpholine-1,1-dioxide, 2-oxa-6-azaspiro[3.3]heptane.

34. The compound according to claim 29 or 30, wherein R 8 and R 9 Together with the nitrogen atom to which they are attached, they form a heterocyclic ring selected from the group consisting of 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]dec-3-one, tetrahydro-1,7-naphthyridine, 1-oxa-4,9-diazaspiro[5.5]undecan-3-one, tetrahydropyrrolo[3,4-d]imidazole, pyrimidine, 8-oxa-2-azaspiro[4.5]decane, hexahydro-3H- oxazolo[3,4-a]pyrazin-3-one, 1-oxa-7-azaspiro[3.5]nonane, octahydrocyclopenta[c]pyrrole, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine, 2,7-diazaspiro[4.4]nonane, 2,6-diazaspiro[3.4]octane, 7-oxa-2-azaspiro[3.5]nonane, 1-oxa-8λ 2 -azaspiro[4.5]decane, 2-oxa-6-azaspiro[3.3]heptane, tetrahydrofuran, Oxadiazole, triazole, pyridone, tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3(2H)-one, piperidone, 3,6-diazabicyclo[3.1.1]heptane, 5-oxa-2,7-diazaspiro[3.5]nonane, pyrazole and pyridazin-3(2H)-one.

35. A compound according to claim 33 or 34, wherein the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from the group consisting of alkyl, alkoxycarbonyl, halo, hydroxy, cyano, carboxyl and heterocyclyl.

36. A compound according to claim 33 or 34, wherein the heterocyclic ring is unsubstituted or substituted with one or more substituents selected from the group consisting of methyl, ethoxycarbonyl, halo, hydroxy, cyano, carboxyl and oxetanyl.

37. A compound according to any one of claims 2 to 36, wherein R 1 and R 1` Combine to form a heterocyclic group (eg, azetidinyl).

38. A compound according to any one of claims 2 to 36, wherein R 1 and R 1` Combine to form an alkyl olefin (eg, aminoallyl).

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

40. The method of claim 1, wherein the compound is selected from or a pharmaceutically acceptable salt thereof.

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

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

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

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

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

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

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

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

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

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

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

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

55. The method of claim 54, wherein the method comprises administering an additional therapeutic agent.

56. The method of claim 54, wherein the method comprises administering two additional therapeutic agents.

57. The method of claim 54, wherein the method comprises administering three additional therapeutic agents.

58. The method of any one of claims 54 to 57, wherein the additional therapeutic agent is selected from the group consisting of 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 / Apo B secretion inhibitors, CETP inhibitors, bile acid absorption inhibitors, cholesterol absorption inhibitors, cholesterol synthesis inhibitors, squalene synthase inhibitors, squalene epoxidase inhibitors, squalene cyclase inhibitors, combined squalene epoxidase / squalene cyclase inhibitors, fibrates, niacin, a combination of niacin and lovastatin, ion exchange resins, antioxidants, ACAT inhibitors, bile acid sequestrants, PCSK9 translation inhibitors, ATP citrate lyase inhibitors, apoC3 inhibitors, ANGPTL3 inhibitors, omega-3 fatty acids, Lp(a) inhibitors, apoB inhibitors, apoA1 inhibitors, low-density lipoprotein receptor inhibitors, low-density lipoprotein inhibitors, diacylglycerol acyltransferase inhibitors, lysosomal acid lipase inhibitors, and lecithin cholesterol acyltransferase inhibitors.

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

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

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

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

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

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

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

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

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

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

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

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

71. The method of any one of claims 54 to 70, wherein the additional therapeutic agent is selected from the group consisting of volanesan, ivizumab, IONIS-ANGPTL3-LRX, icosapentaenoic acid ethyl, eicosapentaenoic acid, eicosapentaenoic acid ester, icosapentaenoic acid methyl, icosapentaenoic acid ethyl, docosahexaenoic acid, docosahexaenoic acid ester, docosahexaenoic acid methyl, docosahexaenoic acid ethyl, bempedoic acid, ezetimibe, IONIS-APO(a)RX, mipomersen, CSL-112, lomitapide, AAV8.TBG. hLDLR (RGX-501), alipegin, prilocastat, sebacetin, ACP-501 / MEDI6012, OM3FA-EE, OM3FA-CA, OM3FA-IPE, alirocumab, evolocumab, bercocilizumab, RG7652, LY3015014, mAb316P, berberine, quercetin, ezetimibe, policosanol, BMS-962476, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

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

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

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

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

76. The method of claim 71, wherein the additional therapeutic agent is eicosapent ethyl.

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

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

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

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

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

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

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

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

85. The method of claim 71, wherein the additional therapeutic agent is alipegin.

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

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

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

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

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

91. 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 condition is associated with PCSK9.

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

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

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

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

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

98. The method of any one of claims 42 to 95, wherein the subject is Spanish.

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

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

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

103. The method of claim 102, wherein the method comprises administering an additional therapeutic agent.

104. The method of claim 102, wherein the method comprises administering two additional therapeutic agents.

105. The method of claim 102, wherein the method comprises administering three additional therapeutic agents.

106. The method of any one of claims 102 to 105, wherein the additional therapeutic agent is selected from the group consisting of kanamycin, amikacin, tobramycin, dibekacin, gentamicin, sisomicin, netilmicin, streptomycin, neomycin B, neomycin C and neomycin E, imipenem, meropenem, ertapenem, doripenem, panipenem, biapenem, azupenem, tebipenem, lenapenem, tomopenem, ceempenem, ciprofloxacin, gareoxacin, gatifloxacin, gemifloxacin, levofloxacin, cinoxacin, nalidixic acid, moxifloxacin, oxolinic acid, piromidic acid, pipemidic acid, rosofloxacin, enoxacin, enoxacin, natrium ... or a combination thereof.

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

108. The method of any one of claims 102 to 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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