Quinolines that modulate serca and their use in the treatment of disease

Abstract

Provided herein are compounds of Formula I, pharmaceutical compositions thereof, and methods of using the same to treat, prevent, or ameliorate one or more symptoms of a neurological disease, a neurodegenerative disorder, or diabetes.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Patent Application Serial No. 16 / 932,832 (Pending), filed July 20, 2020, which is incorporated herein by reference in its entirety. Technical Field

[0003] This article provides information on quinolines, pharmaceutical compositions thereof, and methods for using them to treat, prevent, or improve one or more symptoms of neurological or neurodegenerative diseases or diabetes. This article also provides information on their use in regulating sarcoplasmic reticulum / endoplasmic reticulum Ca2+. 2+ Methods for ATPase (SERCA) activity. Background Technology

[0004] The endoplasmic reticulum (ER) is an organelle that plays a crucial role in several cellular processes essential for cell survival and normal cellular function. These critical processes include intracellular calcium homeostasis, protein secretion, and lipid biosynthesis. (Anelli et al., EMBO J. 2008, 27, 315-327; Pizzo et al., Trends Cell Biol. 2007, 17, 511-517; Ma et al., J. Chem. Neuroanat. 2004, 28, 51-65.)

[0005] Disturbances to ER homeostasis lead to the accumulation of unfolded proteins in the ER, triggering an evolutionarily conserved response known as the unfolded protein response (UPR). (Ron et al., Nat. Rev. Mol. Cell Biol. 2007, 8, 519-529; Malhotra et al., Semin. Cell Dev. Biol. 2007, 18, 716-731.) Disorders leading to ER stress include, for example, dysregulation of cellular redox regulation, glucose deprivation, abnormal calcium regulation in the ER, viral infection, high-fat diets, protein inclusion body diseases (e.g., chronic neurodegenerative diseases), and inclusion body myositis. Kim et al., Nat. Rev. Drug Dis. 2008, 7, 1013-1030; Ma et al., J. Chem. Neuroanat. 2004, 28, 51-65; Ozcan et al., Science 2004, 306, 457-461; Frand et al., Trends Cell Biol. 2000, 10, 203-310. ER stress is associated with a variety of diseases, including neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, polyglutamine disease, and prions), stroke, bipolar disorder, heart disease, atherosclerosis, cancer, diabetes (type 1 and type 2), muscle degeneration, inflammatory diseases, and autoimmune diseases. Kim et al., Nat. Rev. Drug Dis. 2008, 7, 1013-1030; Oyadomari et al., Cell DeathDiffer. 2004, 11, 381-389.

[0006] Sarcoplasmic reticulum / endoplasmic reticulum Ca 2+ ATPase (SERCA) is a major regulator of ER stress and glucose homeostasis in obesity. (Park et al., Proc. Natl. Acad. Sci. U. SA. 2010, 107, 19320-19325.) Obesity disrupts intracellular calcium... 2+ Homeostasis and induction of ER stress. Fu et al., Nature 2011, 473, 528-531. Chronic activation of ER stress is associated with insulin resistance and the development of obesity-related diabetes. Hotamisligil, Cell 2010, 140, 900-917; Kim et al., Nat. Rev. Drug Discov. 2008, 7, 1013-1030. ER Ca2+ was found in small cell lung cancer and non-small cell lung cancer cell lines. 2+ Steady state changes. Bergner et al., J. Exp. Clin. Cancer Res. 2009, 28, 25. Ca2+ is restored via SERCA activation. 2+Homeostasis has been shown to alleviate motor impairment in Parkinson's disease models. (Dahl, Bioorg. Med. Chem. 2017, 25, 53-57). SERCA activation has also been shown to improve memory and coordination in transgenic mouse models of Alzheimer's disease. (Krajnak & Dahl, Bioorg. Med. Chem. Lett. 2018, 28, 1591-1594). Therefore, there is a need for therapeutic agents that can reduce ER stress or restore ER homeostasis for the treatment of diseases caused by ER stress. Summary of the Invention

[0007] This article discloses SERCA modifiers. Certain SERCA modifiers, such as compounds C18, C19, and C20, are compared with other SERCA modifiers (e.g., compounds of formula I, where R...). 2 Compared to amino-substituted phenyl groups (such as C18-C20), they exhibit significantly improved pharmacokinetic properties, such as through C... MAX The AUC and F (%) were measured (see Table 6).

[0008] This article provides a method for treating, preventing, or improving one or more symptoms of endoplasmic reticulum stress-induced disease in a subject, the method comprising administering a compound of formula I to the subject:

[0009]

[0010] Or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs; wherein:

[0011] R 1 and R 2 yes:

[0012] iR 1 It is (a) hydrogen; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (c)–C(O)R 1a –C(O)OR 1a –C(O)NR 1b R 1c –C(NR) 1a )NR 1b R 1c –OR 1a –OC(O)R 1a –OC(O)OR1a –OC(O)NR 1b R 1c –OC(=NR) 1a )NR 1b R 1c –OS(O)R 1a –OS(O)2R 1a –OS(O)NR 1b R 1c –OS(O)2NR 1b R 1c –NR 1b R 1c –NR 1a C(O)R 1d –NR 1a C(O)OR 1d –NR 1a C(O)NR 1b R 1c –NR 1a C(=NR 1d )NR 1b R 1c –NR 1a S(O)R 1d –NR 1a S(O)2R 1d –NR 1a S(O)NR 1b R 1c –NR 1a S(O)2NR 1b R 1c –S(O)R 1a –S(O)2R 1a –S(O)NR 1b R 1c Or –S(O)2NR 1b R 1c And R 2 It is hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic groups; or

[0013] ii.R 1 and R 2 Together with the C and N atoms to which they are directly attached, they form heteroaryl or heterocyclic groups;

[0014] R 3 R 4 R5 R 6 R 7 and R 8 Each is independently (a) hydrogen, cyano, halogroup, or nitro group; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (c)–C(O)R 1a –C(O)OR 1a –C(O)NR 1b R 1c –C(NR) 1a )NR 1b R 1c –OR 1a –OC(O)R 1a –OC(O)OR 1a –OC(O)NR 1b R 1c –OC(=NR) 1a )NR 1b R 1c –OS(O)R 1a –OS(O)2R 1a –OS(O)NR 1b R 1c –OS(O)2NR 1b R 1c –NR 1b R 1c –NR 1a C(O)R 1d –NR 1a C(O)OR 1d –NR 1a C(O)NR 1b R 1c –NR 1a C(=NR 1d )NR 1b R 1c –NR 1a S(O)R 1d –NR 1a S(O)2R 1d –NR 1a S(O)NR 1b R 1c –NR 1a S(O)2NR 1b R 1c –SR 1a –S(O)R1a –S(O)2R 1a –S(O)NR 1b R 1c Or –S(O)2NR 1b R 1c ;

[0015] X is a key, -O-, -NR 1a -、C 1-6 Alkylene, C 2-6 imidene group, C 2-6 Ethyne group, C 3-10 Cycloalkylene, C 6-14 arylene, heteroarylene or heterocyclic; and

[0016] Each R 1a R 1b R 1c and R 1d Independently, it is (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl alkyl, heteroaryl or heterocyclic; (iii) R 1b and R 1c Together with the N atoms to which they are attached, they form heterocyclic groups;

[0017] Each alkyl, alkylene, alkenyl, alkenylene, ynyl, ynylene, cycloalkyl, cycloalkylene, aryl, arylene, aralkyl, heteroaryl, heteroarylene, heterocyclic, and heterocyclic is optionally substituted by one or more substituents Q, in one embodiment by one, two, three, or four substituents Q, wherein each substituent Q is independently selected from (a) oxo, cyano, halo, and nitro; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, and heterocyclic groups, each of which may be further optionally substituented by one or more substituents Q. a In one embodiment, substitution is performed by one, two, three, or four substituents Qa; and (c)–C(O)R a –C(O)OR a –C(O)NR b R c –C(NR) a )NR b R c–OR a –OC(O)R a –OC(O)OR a –OC(O)NR b R c –OC(=NR) a )NR b R c –OS(O)R a –OS(O)2R a –OS(O)NR b R c –OS(O)2NR b R c –NR b R c –NR a C(O)R d –NR a C(O)OR d –NR a C(O)NR b R c –NR a C(=NR d )NR b R c –NR a S(O)R d –NR a S(O)2R d –NR a S(O)NR b R c –NR a S(O)2NR b R c -SR a -S(O)R a -S(O)2R a -S(O)NR b R c and -S(O)2NR b R c , where each R a R b R c and R d Independently (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic groups, each of which may be further optionally substituted by one or more substituents.a In one embodiment, it is substituted by one, two, three, or four substituents Qa; or (iii) R b and R c Together with the N atoms to which they are attached, they form a heterocyclic group, which is further optionally substituented by one or more Q substituents. a In one embodiment, the substitution is performed by one, two, three, or four substituents Qa;

[0018] Each Q a Choose independently from the following groups: (a) oxo, cyano, halogen, and nitro; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl alkyl groups, heteroaryl groups, and heterocyclic groups; and (c)–C(O)R e –C(O)OR e –C(O)NR f R g –C(NR) e )NR f R g –OR e –OC(O)R e –OC(O)OR e –OC(O)NR f R g –OC(=NR) e )NR f R g –OS(O)R e –OS(O)2R e –OS(O)NR f R g –OS(O)2NR f R g –NR f R g –NR e C(O)R h –NR e C(O)OR h –NR e C(O)NR f R g –NR e C(=NR h )NR f R g –NR e S(O)R h –NRe S(O)2R h –NR e S(O)NR f R g –NR e S(O)2NR f R g –SR e –S(O)R e –S(O)2R e –S(O)NR f R g and –S(O)2NR f R g ; where each R e R f R g and R h Independently (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (iii)R f and R g Together with the N atoms to which they are attached, they form heterocyclic groups.

[0019] This article also provides a method for treating, preventing, or improving one or more symptoms of endoplasmic reticulum stress-induced disease in a subject, the method comprising administering to the subject a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof; wherein: R1 is (a) hydrogen or (b) C 1-3 Alkyl; R2 is phenyl, 2-thienyl, 2-furanyl, 2-benzothienyl, or 2-benzofuranyl, wherein R2 is optionally substituted by one or two substituents independently selected from: haloyl, cyano, -O- (C1-C4 alkyl or haloalkyl), C1-C4 alkyl or haloalkyl, –N(CH3)2 and –NH– (C1-C4 alkyl), except for F and NO2; R3 is CH3 or H; R4, R5, R6, R7 and R8 are each independently (a) hydrogen, cyano or haloyl; (b) C 1-4Alkyl, -O-(C1-C4 alkyl), or -N(CH3)2. In another aspect, R2 is phenyl, 2-thienyl, 2-furanyl, 2-benzothienyl, or 2-benzofuranyl, wherein R2 is optionally substituted with -N(CH3)2 or -NH-(C1-C4 alkyl), and the remaining variables are as described in this paragraph. In another aspect, R2 is a phenyl group optionally substituted with -N(CH3)2 or -NH-(C1-C4 alkyl); and the remaining variables are as described in this paragraph.

[0020] This article also provides a method for treating, preventing, or improving one or more symptoms of a disease caused by endoplasmic reticulum stress in a subject, the method comprising administering a compound of formula V to the subject:

[0021]

[0022] Or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs; wherein:

[0023] R 1 (a) Hydrogen; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (c)–C(O)R 1a –C(O)OR 1a –C(O)NR 1b R 1c –C(NR) 1a )NR 1b R 1c –OR 1a –OC(O)R 1a –OC(O)OR 1a –OC(O)NR 1b R 1c –OC(=NR) 1a )NR 1b R 1c –OS(O)R 1a –OS(O)2R 1a –OS(O)NR 1b R 1c –OS(O)2NR 1b R 1c –NR 1b R 1c –NR 1a C(O)R1d –NR 1a C(O)OR 1d –NR 1a C(O)NR 1b R 1c –NR 1a C(=NR 1d )NR 1b R 1c –NR 1a S(O)R 1d –NR 1a S(O)2R 1d –NR 1a S(O)NR 1b R 1c –NR 1a S(O)2NR 1b R 1c –S(O)R 1a –S(O)2R 1a –S(O)NR 1b R 1c Or –S(O)2NR 1b R 1c ;

[0024] R 3 R 4 R 5 R 6 R 7 R 8 and R 9 Each is independently (a) hydrogen, cyano, halogroup, or nitro group; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (c)–C(O)R 1a –C(O)OR 1a –C(O)NR 1b R 1c –C(NR) 1a )NR 1b R 1c –OR 1a –OC(O)R 1a –OC(O)OR 1a –OC(O)NR 1b R 1c –OC(=NR) 1a )NR 1b R 1c –OS(O)R1a –OS(O)2R 1a –OS(O)NR 1b R 1c –OS(O)2NR 1b R 1c –NR 1b R 1c –NR 1a C(O)R 1d –NR 1a C(O)OR 1d –NR 1a C(O)NR 1b R 1c –NR 1a C(=NR 1d )NR 1b R 1c –NR 1a S(O)R 1d –NR 1a S(O)2R 1d –NR 1a S(O)NR 1b R 1c –NR 1a S(O)2NR 1b R 1c –SR 1a –S(O)R 1a –S(O)2R 1a –S(O)NR 1b R 1c Or –S(O)2NR 1b R 1c ;

[0025] Each R 1a R 1b R 1c and R 1d Independently, it is (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl alkyl, heteroaryl or heterocyclic; (iii) R 1b and R 1c Together with the N atoms to which they are attached, they form heterocyclic groups; and

[0026] n is an integer 0, 1, 2, 3, 4, or 5;

[0027] Each alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, and heterocyclic group is optionally substituted by one or more substituents Q, in one embodiment being substituted by one, two, three, or four substituents Q, wherein each substituent Q is independently selected from (a) oxo, cyano, halo, and nitro; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, and heterocyclic groups, each of which may be further optionally substituented by one or more substituents Q. a In one embodiment, substitution is performed by one, two, three, or four substituents Qa; and (c)–C(O)R a –C(O)OR a –C(O)NR b R c –C(NR) a )NR b R c –OR a –OC(O)R a –OC(O)OR a –OC(O)NR b R c –OC(=NR) a )NR b R c –OS(O)R a –OS(O)2R a –OS(O)NR b R c –OS(O)2NR b R c –NR b R c –NR a C(O)R d –NR a C(O)OR d –NR a C(O)NR b R c –NR a C(=NR d )NR b R c –NR a S(O)R d –NR a S(O)2R d –NR a S(O)NR bR c –NR a S(O)2NR b R c -SR a -S(O)R a -S(O)2R a -S(O)NR b R c and -S(O)2NR b R c , where each R a R b R c and R d Independently (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic groups, each of which may be further optionally substituted by one or more substituents. a In one embodiment, it is substituted by one, two, three, or four substituents Qa; or (iii) R b and R c Together with the N atoms to which they are attached, they form a heterocyclic group, which is further optionally substituented by one or more Q substituents. a In one embodiment, the substitution is performed by one, two, three, or four substituents Qa;

[0028] Each Q a Choose independently from the following groups: (a) oxo, cyano, halogen, and nitro; (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl alkyl groups, heteroaryl groups, and heterocyclic groups; and (c)–C(O)R e –C(O)OR e –C(O)NR f R g –C(NR) e )NR f R g –OR e –OC(O)R e –OC(O)OR e –OC(O)NR f R g –OC(=NR) e)NR f R g –OS(O)R e –OS(O)2R e –OS(O)NR f R g –OS(O)2NR f R g –NR f R g –NR e C(O)R h –NR e C(O)OR h –NR e C(O)NR f R g –NR e C(=NR h )NR f R g –NR e S(O)R h –NR e S(O)2R h –NR e S(O)NR f R g –NR e S(O)2NR f R g –SR e –S(O)R e –S(O)2R e –S(O)NR f R g and –S(O)2NR f R g ; where each R e R f R g and R h Independently (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (iii)R f and R g Together with the N atoms to which they are attached, they form heterocyclic groups.

[0029] In addition, this article provides a method for treating, preventing, or improving one or more symptoms of a condition, disease, or disorder mediated by sarcoplasmic reticulum / endoplasmic reticulum calcium ATPase (SERCA) in a subject, the method comprising administering to the subject a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0030] This article provides a method for treating, preventing, or improving one or more symptoms of diabetes in a subject, the method comprising administering to the subject a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0031] This article provides a method for increasing glucose tolerance in subjects, comprising administering to the subject a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0032] This article provides a method for treating, preventing, or improving one or more symptoms of Alzheimer's disease in a subject, the method comprising administering to the subject a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0033] This article provides a method for treating, preventing, or improving one or more symptoms of Parkinson's disease in a subject, the method comprising administering to the subject a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0034] This article provides a method for reducing stress in the ER, which involves contacting the ER with a compound of formula I or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs.

[0035] This article provides a method for restoring or maintaining steady state in an ER, which includes contacting the ER with a compound of formula I or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs.

[0036] This article provides Ca for increasing ER. 2+ Methods for determining concentration, comprising contacting ER with a compound of formula I or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs.

[0037] This article provides a method for modulating the activity of SERCA, which involves contacting SERCA with a compound of formula I or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs. Attached Figure Description

[0038] Figure 1 The effects of compound A12 on serum glucose levels in ob / ob mice treated with 50 mg / kg and 75 mg / kg of compound A12 are shown in (A) day 1 and (B) day 21.

[0039] Figure 2 The effects of compounds A12 and B1 on serum glucose levels in ob / ob mice treated daily with 50 mg / kg of compound A12 or B1 for 4 weeks are shown (○ - mediator). -Compound A12; ▲-Compound B1).

[0040] Figure 3 The protocol used to evaluate the effects of the compounds presented herein on diabetic mice is shown.

[0041] Figure 4 (A) shows a pseudo-color two-photon image depicting the Ca2+ of CA1 pyramidal neurons in saline-treated PS1 / APP mice, PS1 / APP mice treated with compound A12 (RD163), and mice not treated with Tg saline. 2+ Response to caffeine (10 mM, 60 seconds); and (B) normalized RyR-Ca morphology of CA1 pyramidal neurons in saline-treated PS1 / APP mice, PS1 / APP mice treated with compound A12, and mice not treated with Tg saline. 2+ reaction.

[0042] Figure 5 The effect of compound A12 on amyloid plaque formation in APP-PS1 mice is shown.

[0043] Figure 6The effects of compound A12 treatment on memory and coordination in an APP-PS1 mouse model of Alzheimer's disease were demonstrated using the Morris water maze (A) and Rotard (B) tests, respectively.

[0044] Figure 7 The effects of A12 treatment on motor dysfunction in rats with 6-OHDA injury are shown. Data are presented in the initiation time test (A), stepping test (B), and cylinder test (C). Detailed Implementation

[0045] To facilitate understanding of the disclosures listed herein, several terms are defined below.

[0046] Generally, the nomenclature used herein and the laboratory procedures of organic chemistry, medicinal chemistry, and pharmacology described herein are those well known and commonly used in the art. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0047] The term "subject" refers to an animal, including but not limited to primates (e.g., humans), cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, or mice. The terms "subject" and "patient" are used interchangeably herein to refer to, for example, a mammalian subject. In one embodiment, the subject is a human.

[0048] The terms “treat,” “treating,” and “treatment” are intended to include alleviating or eliminating a symptom, disease, or ailment, or one or more symptoms of such a symptom, disease, or ailment; alleviating or eradicating one or more causes of such a symptom, disease, or ailment itself.

[0049] The terms “prevent,” “preventing,” and “prevention” are intended to include methods that delay and / or prevent the occurrence of a condition, disease, or disorder and / or its accompanying symptoms; prevent a subject from developing a condition, disease, or disorder; or reduce the risk of a subject developing a condition, disease, or disorder.

[0050] The term "therapeutic effective amount" is intended to include an amount of compound, when administered, sufficient to prevent or to some extent alleviate the development of one or more symptoms of a condition, disease, or disorder being treated. The term "therapeutic effective amount" also refers to an amount of compound that researchers, veterinarians, physicians, or clinicians are seeking to elicit a biological or medical response in a biomolecule (e.g., a protein, enzyme, RNA, or DNA), cell, tissue, system, animal, or human.

[0051] The terms “pharmaceutically acceptable carrier,” “pharmaceuticalally acceptable excipient,” “physiologically acceptable carrier,” or “physiologically acceptable excipient” refer to pharmaceutically acceptable materials, compositions, or media, such as liquid or solid fillers, diluents, solvents, or encapsulating materials. In one embodiment, each component is “pharmaceutically acceptable” in the sense of compatibility with other components of the pharmaceutical formulation and is suitable for contact with human and animal tissues or organs without excessive toxicity, irritation, allergic reactions, immunogenicity, or other problems or complications, in proportion to a reasonable benefit / risk ratio. See Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 7th ed., Rowe et al., The Pharmaceutical Press and the American Pharmaceutical Association: 2012; Handbook of Pharmaceutical Additives, 3rd ed., Ash and Ash, Gower Publishing Company: 2007; and Pharmaceutical Preformulation and Formulation, 2nd ed., Gibson Ed., CRC Press LLC: Boca Raton, FL, 2009.

[0052] The terms "about" or "approximately" mean an acceptable error for a particular value as determined by a person skilled in the art, which will depend in part on how the value was measured or determined. In some embodiments, the terms "about" or "approximately" mean within 1, 2, 3, or 4 standard deviations. In some embodiments, the terms "about" or "approximately" mean within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

[0053] The terms “active ingredient” and “active substance” refer to a compound, alone or in combination with one or more pharmaceutically acceptable excipients, administered to a subject to treat, prevent, or improve one or more symptoms of a condition, disease, or disorder. As used herein, “active ingredient” and “active substance” can be optically active isomers of the compounds described herein.

[0054] The terms “drug,” “therapeutic agent,” and “chemotherapeutic agent” refer to a compound or pharmaceutical composition thereof administered to a subject to treat, prevent, or improve one or more symptoms of a disease, illness, or ailment.

[0055] The term "endoplasmic reticulum stress" or "ER stress" refers to a perturbation of endoplasmic reticulum homeostasis, such as a perturbation of the functional folding of proteins in the endoplasmic reticulum.

[0056] The terms “ER stress syndrome, disease, or disorder,” “ER stress-induced syndrome, disease, or disorder,” “ER stress-related syndrome, disease, or disorder,” or “ER stress-associated syndrome, disease, or disorder” refer to a syndrome, disease, or disorder caused by ER homeostasis disturbances. Specifically, an ER stress syndrome, disease, or disorder is one in which a reduction in ER stress leads to some effect on the underlying syndrome, disease, or disorder; for example, ER stress modulators lead to some improvement in at least some patients currently receiving treatment.

[0057] When used in conjunction with biological materials such as nucleic acids (e.g., DNA or RNA), peptides, and host cells, the terms "naturally occurring" or "naive" refer to materials found in nature and not manipulated by humans. Similarly, "non-naturally occurring" or "non-naive" refer to materials not found in nature or that have been structurally modified or synthesized by humans.

[0058] The term "SERCA" or "sarcoplasmic reticulum Ca" 2+ "ATPase" refers to the sarcoplasmic reticulum / endoplasmic reticulum Ca2+. 2+ ATPases or variants thereof. The term “SERCA variant” is intended to include proteins that are substantially homologous to native SERCA, i.e., proteins with one or more naturally occurring or non-naturally occurring amino acid deletions, insertions, or substitutions compared to the amino acid sequence of native SERCA (e.g., SERCA derivatives, homologs, and fragments). The amino acid sequence of a SERCA variant has at least about 80%, at least about 90%, or at least about 95% identity with native SERCA. SERCA enzymes are classified into at least three classes: SERCA1, SERCA2, and SERCA3. (Stutzmann et al., Pharmacol. Rev. 2011, 63, 700-727; Andersen et al., Acta Physiol. Scand. Suppl. 1998, 643, 45-54.) Class I includes SERCA1a and SERCA1b. Class II includes SERCA2a and SERCA2b. Class III includes SERCA3a, SERCA3b, and SERCA3c.

[0059] The terms “SERCA-mediated condition, disease, or disorder” and “SERCA-mediated condition, disease, or disorder” refer to conditions, diseases, or disorders in which regulation of SERCA activity results in some effect on the underlying condition, disease, or disorder. For example, SERCA agonists result in some improvement in at least some patients who are receiving treatment.

[0060] The term "alkyl" refers to a linear or branched saturated monovalent hydrocarbon group, wherein the alkyl group is optionally substituted by one or more substituents Q as described herein. Unless otherwise stated, the term "alkyl" also covers linear and branched alkyl groups. In some embodiments, the alkyl group is having 1 to 20 (C) groups. 1-20 ), 1 to 15 (C) 1-15 ), 1 to 10 (C 1-10 ) or 1 to 6 (C 1-6 Linear saturated monovalent hydrocarbon groups of carbon atoms, or 3 to 20 (C) 3-20 ), 3 to 15 (c 3-15 ), 3 to 10 (C) 3-10 ) or 3 to 6 (C 3-6 Branched saturated monovalent hydrocarbon groups of carbon atoms. As used in this paper, linear C 1-6 and branched C 3-6 Alkyl groups are also called "lower alkyl groups". Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (including all isomers), n-propyl, isopropyl, butyl (including all isomers), n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl (including all isomers), and hexyl (including all isomers). For example, C 1-6 Alkyl groups refer to linear saturated monovalent hydrocarbon groups with 1 to 6 carbon atoms or branched saturated monovalent hydrocarbon groups with 3 to 6 carbon atoms.

[0061] The term "alkenyl" refers to a linear or branched monovalent hydrocarbon group containing one or more carbon-carbon double bonds, in one embodiment containing one, two, three, four, or five carbon-carbon double bonds, and in another embodiment containing one carbon-carbon double bond. In some embodiments, the alkenyl group is optionally substituted with one or more substituents Q as described herein. The term "alkenyl" also encompasses groups having "cis" and "trans" configurations, or alternatively, "Z" and "E" configurations, as understood by those skilled in the art. As used herein, unless otherwise stated, the term "alkenyl" covers both linear and branched alkenyl groups. For example, C 2-6 Alkenyl refers to a linear unsaturated monovalent hydrocarbon group with 2 to 6 carbon atoms or a branched unsaturated monovalent hydrocarbon group with 3 to 6 carbon atoms. In some embodiments, the alkenyl group has 2 to 20 carbon atoms. 2-20 ), 2 to 15 (C) 2-15 ), 2 to 10 (C)2-10 ) or 2 to 6 (C 2-6 A linear monovalent hydrocarbon group consisting of 3 to 20 carbon atoms. 3-20 ), 3 to 15 (C) 3-15 ), 3 to 10 (C) 3-10 ) or 3 to 6 (C 3-6 Branched monovalent hydrocarbon groups of carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, allyl, butenyl, and 4-methylbutenyl.

[0062] The term "alkynyl" refers to a linear or branched monovalent hydrocarbon group containing one or more carbon-carbon triple bonds. In one embodiment, it contains one, two, three, four, or five carbon-carbon triple bonds; in another embodiment, it contains one carbon-carbon triple bond. In some embodiments, the alkynyl group is optionally substituted by one or more substituents Q as described herein. Unless otherwise stated, the term "alkynyl" also covers linear and branched alkynyl groups. In some embodiments, the alkynyl group has 2 to 20 carbon atoms (C1 to C2). 2-20 ), 2 to 15 (C) 2-15 ), 2 to 10 (C) 2-10 ) or 2 to 6 (C 2-6 A linear monovalent hydrocarbon group consisting of 3 to 20 carbon atoms. 3-20 ), 3 to 15 (C) 3-15 ), 3 to 10 (C) 3-10 ) or 3 to 6 (C 3-6 A branched monovalent hydrocarbon group of a carbon atom. Examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH) and propargyl (-CH₂C≡CH). For example, C 2-6 Alkyne groups refer to linear unsaturated monovalent hydrocarbon groups with 2 to 6 carbon atoms or branched unsaturated monovalent hydrocarbon groups with 3 to 6 carbon atoms.

[0063] The term "cycloalkyl" refers to a cyclic saturated or non-aromatic unsaturated bridged or unbridged monovalent hydrocarbon group, which is optionally substituted by one or more substituents Q as described herein. In some embodiments, the cycloalkyl is a cyclic saturated bridged or unbridged monovalent hydrocarbon group. In some embodiments, the cycloalkyl has from 3 to 20 (C) atoms. 3-20 ), from 3 to 15 (C 3-15 ), from 3 to 10 (C 3-10 ) or from 3 to 7 (C 3-7 ( ) carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decahydronaphthyl and adamantyl.

[0064] The term "aryl" refers to a monocyclic aromatic group and / or a polycyclic monovalent aromatic group containing at least one aromatic ring. In some embodiments, the aryl group has from 6 to 20 (C) groups. 6-20 ), from 6 to 15 (C 6-15 ) or from 6 to 10 (C 6-10 The aryl group includes, but is not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthraceneyl, phenanthryl, pyrene, biphenyl, and terphenyl. In some embodiments, the term "aryl" refers to a bicyclic or tricyclic carbocyclic ring, wherein one ring of the ring is aromatic, while the other rings of the ring may be saturated, partially unsaturated, or aromatic, such as dihydronaphthyl, indenyl, indenyl, or tetrahydronaphthyl (naphthyl). In some embodiments, the aryl group is optionally substituted by one or more substituents Q as described herein.

[0065] The term "aralkyl" or "arylalkyl" refers to a monovalent alkyl group substituted with one or more aryl groups. In some embodiments, the aralkyl group has from 7 to 30 (C) groups. 7-30 ), from 7 to 20 (C 7-20 ) or from 7 to 16 (C 7-16 The carbon atom is aralkyl group. Examples of aralkyl groups include, but are not limited to, benzyl, 1-phenylethyl, 2-phenylethyl, and 3-phenylpropyl. In some embodiments, the aralkyl group is optionally substituted with one or more substituents Q as described herein.

[0066] The term "heteroaryl" refers to a monovalent monocyclic aromatic group or a monovalent polycyclic aromatic group containing at least one aromatic ring, wherein the at least one aromatic ring contains one or more heteroatoms, each of which is independently selected from O, S, N, and P. The heteroaryl group is bonded to the remainder of the molecule through its aromatic ring. Each ring of the heteroaryl group may contain one or two O atoms, one or two S atoms, one to four N atoms, and / or one or two P atoms, provided that the total number of heteroatoms in each ring is four or fewer, and each ring contains at least one carbon atom. In some embodiments, the heteroaryl group has from 5 to 20, from 5 to 15, or from 5 to 10 ring atoms. Examples of monocyclic heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrroleyl, thiadiazolyl, thiazolyl, thiophene, tetrazolyl, triazinyl, and triazolyl. Examples of bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzotriazolyl, benzooxazolyl, furanpyridyl, imidazopyridyl, imidazothiazolyl, indoleazinyl, indoleyl, inzolyl, isobenzofuranyl, isobenzothiaphenyl, isoindoleyl, isoquinolinyl, isothiazolyl, naphthiaridyl, oxazolopyridyl, phthalazinyl, pteridinyl, purine, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalolinyl, quinazolinyl, thiadiazolopyrimidinyl, and thiophenopyridyl. Examples of tricyclic heteroaryl groups include, but are not limited to, acridinel, benzoindolyl, carbazolel, dibenzofuranyl, perimidinyl, phenanthrolyl, phenanthidyl, phenarsazinyl, phenazinyl, phenthiazinyl, phenoxazinyl, and xanthonyl. In some embodiments, the heteroaryl group is optionally substituted with one or more substituents Q as described herein.

[0067] The term "heterocyclic group" or "heterocycle" refers to a monovalent monocyclic non-aromatic ring system or a monovalent polycyclic ring system containing at least one non-aromatic ring, wherein one or more of the non-aromatic ring atoms are heteroatoms, each of which is independently selected from O, S, N, and P; and the remaining ring atoms are carbon atoms. In some embodiments, the heterocyclic group or heterocyclic group has 3 to 20, 3 to 15, 3 to 10, 3 to 8, 4 to 7, or 5 to 6 ring atoms. The heterocyclic group is bonded to the rest of the molecule through its non-aromatic ring. In some embodiments, the heterocyclic group is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may be spirocyclic, fused, or bridged, and wherein nitrogen or sulfur atoms may optionally be oxidized, nitrogen atoms may optionally be quaternized, and some rings may be partially or fully saturated or aromatic. The heterocyclic group can be attached to the main structure at any heteroatom or carbon atom that results in a stable compound. Examples of heterocyclic groups include, but are not limited to, nitrogen-containing heterocyclic groups. Benzyl, benzodioxolyl, benzofuranonyl, benzopyranonyl, benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothiophenyl, benzothiopyranyl, benzooxazinyl, β-carbolinyl, chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl, dihydrobenzisothiazinyl, dihydrobenzisothiazinyl, dihydrofuranyl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl, dihydropyridine The following groups are listed: 1,4-dithiaalkyl, dihydropyrimidinyl, dihydropyrroleyl, dioxolaneyl, 1,4-dithiaalkyl, furanoneyl, imidazoalkyl, imidazolinyl, dihydroindolyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothiophenyl, isobenzodihydropyranyl, isocoumarinyl, isodihydroindolyl, isothiazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinoneyl, oxazolidinyl, oxiranyl, piperazinyl, piperidinyl, 4-piperidinoneyl, pyrazolidinyl, pyrazolinyl, pyrroleyl, quininecycloyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothiophenyl, thiamorpholinyl, thiazolinyl, tetrahydroquinolinyl, and 1,3,5-trithiaalkyl. In some embodiments, the heterocyclic group is optionally substituted with one or more substituents Q as described herein.

[0068] The terms “halogen,” “halogen,” or “halogenated group” refer to fluorine, chlorine, bromine, and / or iodine.

[0069] The term "optionally substituted" is intended to mean that a group or substituent such as alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, and heterocyclic groups may be substituted by one or more substituents Q, each of which is independently selected, for example, (a) oxo (=O), cyano (-CN), halogroup, and nitro (-NO2); (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, and heterocyclic groups, each of which is further substituted by one or more substituents Qa, wherein in one embodiment, the substituents Qa are one, two, three, four, or five. a Substitution; and (c)-C(O)R a -C(O)OR a -C(O)NR b R c -C(NR) a )NR b R c -OR a -OC(O)R a -OC(O)OR a -OC(O)NR b R c -OC(=NR) a )NR b R c -OS(O)R a -OS(O)2R a -OS(O)NR b R c -OS(O)2NR b R c -NR b R c -NR a C(O)R d -NR a C(O)OR d -NR a C(O)NR b R c -NR a C(=NR d )NR b R c -NR a S(O)R d-NR a S(O)2R d -NR a S(O)NR b R c -NR a S(O)2NR b R c -P(O)R a R d -P(O)(OR) a )R d -P(O)(OR) a (OR) d ), -SR a -S(O)R a -S(O)2R a -S(O)NR b R c and -S(O)2NR b R c , where each R a R b R c and R d Independently (i) hydrogen; (ii) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic groups, each of which is optionally substituted by one or more substituents Qa, and in one embodiment by one, two, three, or four substituents Q. a Replace; or (iii)R b and R c The N atoms to which they are attached are connected to form a heteroaryl or heterocyclic group, each of which is optionally substituted by one or more substituents Qa, and in one embodiment by one, two, three or four substituents Q. a Substitution. As used herein, all substituted groups described herein are "optionally substituted" unless otherwise specified.

[0070] In one implementation, each substituent Q a Independently select from the following groups: (a) oxo, cyano, halogen, and nitro; and (b) C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15aralkyl, heteroaryl and heterocyclyl; and (c)–C(O)R e , –C(O)OR e , –C(O)NR f R g , –C(NR e )NR f R g , –OR e , –OC(O)R e , –OC(O)OR e , –OC(O)NR f R g , –OC(=NR e )NR f R g , –OS(O)R e , –OS(O)2R e , –OS(O)NR f R g , –OS(O)2NR f R g , –NR f R g , –NR e C(O)R h , –NR e C(O)OR h , –NR e C(O)NR f R g , –NR e C(=NR h )NR f R g , –NR e S(O)R h , –NR e S(O)2R h , –NR e S(O)NR f R g , –NR e S(O)2NR f R g , –P(O)R e R h , –P(O)(OR e )R h , –P(O)(OR e )(OR h ), –SR e , –S(O)R e , –S(O)2R e , –S(O)NR f R gand –S(O)2NR f R g ; where each R e R f R g and R h Independently for (i) hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-14 Aryl, C 7-15 Aryl, heteroaryl, or heterocyclic; or (ii)R f and R g Together with the N atoms to which they are attached, they form heteroaryl or heterocyclic groups.

[0071] In some embodiments, "optical activity" and "enantiomeric activity" refer to an aggregate of molecules in which there is an enantiomeric excess of not less than about 50%, not less than about 70%, not less than about 80%, not less than about 90%, not less than about 91%, not less than about 92%, not less than about 93%, not less than about 94%, not less than about 95%, not less than about 96%, not less than about 97%, not less than about 98%, not less than about 99%, not less than about 99.5%, or not less than about 99.8%. In some embodiments, the compound comprises about 95% or more of the desired enantiomer and about 5% or less of the less preferred enantiomer based on the total weight of the two enantiomers discussed.

[0072] In describing optically active compounds, the prefixes R and S are used to indicate the absolute configuration of the compound with respect to one or more of its chiral centers. (+) and (-) are used to indicate the optical rotation of the compound, i.e., the direction in which the plane of polarization is rotated by the compound. The (-) prefix indicates that the compound is left-handed, meaning it rotates the plane of polarization to the left or counterclockwise. The (+) prefix indicates that the compound is right-handed, meaning it rotates the plane of polarization to the right or clockwise. However, the optical rotation symbols (+) and (-) are independent of the absolute configurations R and S of the compound.

[0073] The term "isotope variant" refers to a compound that contains an isotope in a non-natural proportion on one or more atoms of the atoms constituting the compound. In some embodiments, the "isotope variant" of the compound contains one or more isotopes in a non-natural proportion, including but not limited to hydrogen (…). 1 H), deuterium ( 2 H), tritium ( 3 H), carbon-11 ( 11 C), Carbon-12 ( 12 C), Carbon-13 ( 13C), Carbon-14 ( 14 C) Nitrogen-13 ( 13 N), nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), Oxygen-14 ( 14 O), Oxygen-15 ( 15 O), Oxygen-16 ( 16 O), Oxygen-17 ( 17 O), Oxygen-18 ( 18 O), Fluorine-17 ( 17 F), Fluorine-18 ( 18 F), Phosphorus-31 ( 31 P), Phosphorus-32 ( 32 P), Phosphorus-33 ( 33 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-35 ( 35 S), sulfur-36 ( 36 S), Chlorine-35 ( 35 Cl), Chlorine-36 ( 36 Cl), Chlorine-37 ( 37 Cl), Bromine-79 ( 79 Br), bromine-81 ( 81 Br), Iodine-123 ( 123 I), iodine-125( 125 I), iodine-127( 127 I), iodine-129( 129 I) and Iodine-131 131 I). In some embodiments, the "isotope variant" of the compound is in a stable form, i.e., non-radioactive. In some embodiments, the "isotope variant" of the compound contains one or more isotopes in non-natural proportions, including but not limited to hydrogen (I). 1 H), deuterium ( 2 H), carbon-12 ( 12 C), Carbon-13 ( 13 C) Nitrogen-14 ( 14 N), nitrogen-15 ( 15 N), Oxygen-16 ( 16 O), Oxygen-17 ( 17 O), Oxygen-18 ( 18 O), Fluorine-17 ( 17 F), Phosphorus-31 ( 31 P), sulfur-32 ( 32 S), sulfur-33 ( 33 S), sulfur-34 ( 34 S), sulfur-36 (36 S), Chlorine-35 ( 35 Cl), Chlorine-37 ( 37 Cl), Bromine-79 ( 79 Br), bromine-81 ( 81 Br) and iodine-127 127 I). In some embodiments, the "isotopic variant" of the compound is in an unstable form, i.e., radioactive. In some embodiments, the "isotopic variant" of the compound contains one or more isotopes in non-natural proportions, including but not limited to tritium (I). 3 H), carbon-11 ( 11 C), Carbon-14 ( 14 C) Nitrogen-13 ( 13 N), Oxygen-14 ( 14 O), Oxygen-15 ( 15 O), Fluorine-18 ( 18 F), Phosphorus-32 ( 32 P), Phosphorus-33 ( 33 P), sulfur-35 ( 35 S), Chlorine-36 ( 36 Cl), Iodine-123 ( 123 I), iodine-125( 125 I), iodine-129( 129 I) and Iodine-131 131 I). It should be understood that in compounds as provided herein, any hydrogen can be, for example, where feasible to a person skilled in the art. 2 H, or any carbon can be, for example 13 C, or any nitrogen can be, for example 15 N, or any oxygen, can be, for example... 18 O. In some embodiments, the "isotopic variant" of the compound contains a non-natural proportion of deuterium (D).

[0074] The term "solvate" refers to a complex or aggregate formed of one or more solute molecules (such as the compounds provided herein) and one or more solvent molecules (present in stoichiometric or non-stoichiometric amounts). Suitable solvents include, but are not limited to, water, methanol, ethanol, n-propanol, isopropanol, and acetic acid. In some embodiments, the solvent is pharmaceutically acceptable. In one embodiment, the complex or aggregate is in a crystalline form. In another embodiment, the complex or aggregate is in a non-crystalline form. When the solvent is water, the solvate is a hydrate. Examples of hydrates include, but are not limited to, hemihydrates, monohydrates, dihydrates, trihydrates, tetrahydrates, and pentahydrates.

[0075] The phrase “enantiomer, mixture of enantiomers, mixture of two or more diastereomers, or isotopic variants thereof; or pharmaceutically acceptable salts, solvates, hydrates or prodrugs thereof” has the same meaning as the following phrases: “(i) an enantiomer, mixture of enantiomers, mixture of two or more diastereomers, or isotopic variant of the compound mentioned herein; (ii) a pharmaceutically acceptable salt, solvate, hydrate or prodrug of the compound mentioned herein; (iii) a pharmaceutically acceptable salt, solvate, hydrate or prodrug of an enantiomer, mixture of enantiomers, mixture of two or more diastereomers, or isotopic variant of the compound mentioned herein.”

[0076] In one embodiment, this document provides a compound of formula (I) or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or pharmaceutically acceptable salts, solvates, hydrates, or prodrugs thereof; wherein said variables are as described above. In another embodiment, this document provides a compound of formula (V) or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or pharmaceutically acceptable salts, solvates, hydrates, or prodrugs thereof; wherein said variables are as described above. In yet another embodiment, this document provides compounds selected from the group consisting of:

[0077]

[0078]

[0079]

[0080] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0081] In another embodiment, this document provides compounds selected from the group consisting of:

[0082]

[0083]

[0084] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0085] In yet another embodiment, compounds selected from the group consisting of:

[0086]

[0087]

[0088] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0089] In yet another embodiment, compounds selected from the group consisting of:

[0090]

[0091] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0092] In yet another embodiment, compounds selected from the group consisting of:

[0093]

[0094]

[0095] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0096] In yet another embodiment, compounds selected from the group consisting of:

[0097]

[0098] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0099] In another embodiment, compounds selected from the group consisting of:

[0100]

[0101]

[0102] And their isotopic variants; and their pharmaceutically acceptable salts, solvates, hydrates and prodrugs.

[0103] In some embodiments, the compounds provided herein exhibit activity as SERCA agonists. In some embodiments, the compounds provided herein exhibit activity as allosteric SERCA modulators. In some embodiments, the compounds provided herein exhibit activity as SERCA2b agonists. In some embodiments, the compounds provided herein exhibit activity as allosteric SERCA2b modulators.

[0104] In some embodiments, the compounds provided herein exhibit activity that reduces ER stress. In some embodiments, the compounds provided herein exhibit activity that increases ER calcium. 2 +Concentration of activity.

[0105] The compounds provided herein are intended to cover all possible stereoisomers unless a specific stereochemistry is specified. In the case of compounds containing alkenyl or alkenyl groups, the compounds may exist as one of geometric cis / trans (or Z / E) isomers or a mixture of geometric cis / trans (or Z / E) isomers. Where the structural isomers are interconvertible, the compounds may exist as a single tautomer or a mixture of tautomers. This can manifest as proton tautomerism in compounds containing, for example, imino, ketone, or oxime groups; or as so-called valence tautomerism in compounds containing aromatic moieties. Thus, a single compound may exhibit more than one type of isomerism.

[0106] The compounds provided herein may be enantiomerically pure, such as a single enantiomer or a single diastereomer, or a mixture of stereoisomers, such as a mixture of enantiomers, for example, a racemic mixture of two enantiomers; or a mixture of two or more diastereomers. Therefore, those skilled in the art will recognize that, for compounds subjected to intra-intracellular epimerization, application of the compound in its (R) form is equivalent to application of the compound in its (S) form. Conventional techniques for the preparation / separation of single enantiomers include synthesis from suitable optically pure precursors, asymmetric synthesis from achiral starting materials, or resolution of mixtures of enantiomers, for example, chiral chromatography, recrystallization, resolution, diastereomer salt formation, or derivatization into diastereomer adducts followed by separation.

[0107] When the compounds provided herein contain an acidic or basic moiety, they may also be provided as pharmaceutically acceptable salts (see Berge et al., J. Pharm. Sci. 1977, 66, 1-19; and “Handbook of Pharmaceutical Salts, Properties, and Use,” edited by Stahl and Wermuth; Wiley-VCH and VHCA, Zurich, 2002).

[0108] Suitable acids for preparing pharmaceutically acceptable salts include, but are not limited to, acetic acid, 2,2-dichloroacetic acid, acetylated amino acids, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, boric acid, (+)-camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, and cyclamic acid. Cyclohexanesulfonic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glucoheponic acid, D-gluconic acid, D-glucuronic acid, L-glutamic acid, α-oxoglutarate, glycolic acid, hippuric acid, hydrobromic acid, hydroiodic acid, (+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid, maleic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, palmitic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid, saccharic acid (acid), salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanate, p-toluenesulfonic acid, undecenoic acid and valeric acid.

[0109] Suitable bases for preparing pharmaceutically acceptable salts include, but are not limited to, inorganic bases such as magnesium hydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, or sodium hydroxide; and organic bases such as primary, secondary, tertiary, and quaternary amines, aliphatic and aromatic amines, including but not limited to L-arginine, phenethylbenzylamine, benzathine penicillin, choline, tannin, diethanolamine, diethylamine, dimethylamine, dipropylamine, diisopropylamine, 2-(diethylamino)ethanol, and ethanolamine. Ethylamine, ethylenediamine, isopropylamine, N-methylglucosamine, hydrabamine, 1H-imidazolium, L-lysine, morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine, piperazine, propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine, pyridine, quinine ring, quinoline, isoquinoline, secondary amine, triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucosamine, 2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

[0110] The compounds described herein can also be provided as prodrugs, which are functional derivatives of compounds of, for example, formula V, and are readily converted into the parent compound in vivo. Prodrugs are often useful because, in some cases, they may be easier to administer than the parent compound. They can become bioavailable through oral administration, while the parent drug cannot. Prodrugs may also have enhanced solubility in pharmaceutical compositions compared to the parent compound. Prodrugs can be converted into the parent drug through various mechanisms, including enzymatic processes and metabolic hydrolysis. See Harper, Progress in Drug Research 1962, 4, 221-294; Morozowich et al., “Design of Biopharmaceutical Properties through Prodrugs and Analogs,” Roche, ed., APHA Acad. Pharm. Sci. 1977; “Bioreversible Carriers in Drug: Drug Design, Theory and Application,” Roche, ed., APHA Acad. Pharm. Sci. 1987; “Design of Prodrugs,” Bundgaard, Elsevier, 1985; Wang et al., Curr. Pharm. Design 1999, 5, 265-287; Pauletti et al., Adv. Drug Delivery Rev. 1997, 27, 235-256; Mizen et al., Pharm. Biotech. 1998, 11, 345-365; Gaignault et al., Pract. Med. Chem. 1996, 671-696; Asgharnejad in “Transport Processes in Pharmaceutical Systems,” Amidon et al., eds., Marcell Dekker, 185-218, 2000; Balant et al., Eur. J. Drug Metab. Pharmacokinet. 1990, 15, 143-53; Balimane and Sinko, Adv. Drug Delivery Rev. 1999, 39, 183-209; Browne, Clin. Neuropharmacol. 1997, 20, 1-12; Bundgaard, Arch. Pharm. Chem.1979, 86, 1-39; Bundgaard, Controlled Drug Delivery 1987, 17, 179-96; Bundgaard, Adv. Drug Delivery Rev. 1992, 8, 1-38; Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130; Fleisher et al., Methods Enzymol. 1985, 112, 360-381; Farquhar et al., J. Pharm. Sci. 1983, 72, 324-325; Freeman et al., J. Chem. Soc., Chem. Commun. 1991, 875-877; Friis and Bundgaard, Eur.; J. Pharm. Sci. 1996, 4, 49-59; Gangwar et al., Des. Biopharm. Prop. Prodrugs Analogs, 1977, 409-421; Nathwani and Wood, Drugs 1993, 45, 866-94; Sinhababu and Thakker, Adv.Drug Delivery Rev. 1996, 19, 241-273; Stella et al., Drugs 1985, 29, 455-73; Tan et al., Adv.Drug Delivery Rev.1999, 39, 117-151; Taylor, Adv.Drug Delivery Rev.1996, 19, 131-148; Valentino and Borchardt, Drug Discovery Today 1997, 2, 148-155; Wiebe and Knaus, Adv.Drug Delivery Rev. 1999, 39, 63-80; and Waller et al., Br. J. Clin. Pharmac. 1989, 28, 497-507.

[0111] The compounds described herein can be prepared, isolated, or obtained by any method known to those skilled in the art, and the following examples are merely representative and do not exclude other related procedures.

[0112] In one embodiment, for example, the compound of formula I is prepared as shown in Scheme I by a coupling reaction of amine I-1 with compound I-2 having a leaving group L, optionally in the presence of a coupling agent, to form compound I. In some embodiments, L is a hydroxyl or a halogroup. In some embodiments, L is a hydroxyl, fluorine, chlorine, bromine, or iodine.

[0113] Option I

[0114]

[0115] In another embodiment, for example, the compound of formula V is prepared as shown in scheme Ia by a coupling reaction of amine I-1 with compound V-2 having a leaving group L, optionally in the presence of a coupling agent, to form compound V. In some embodiments, L is a hydroxyl or a halogroup. In some embodiments, L is a hydroxyl, fluorine, chlorine, bromine, or iodine.

[0116] Solution Ia

[0117]

[0118] Examples of suitable coupling agents include, but are not limited to, carbodiimides (e.g., N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC hydrochloride), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methyl iodide (EDC methyl iodide), 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide p-toluenesulfonate, N,N'-diisopropylcarbodiimide (DIC), and 1, 3-Dicyclohexylcarbodiimide (DCC), 1,1'-carbonyldiimidazole (CDI), bis(2-oxo-3-oxazolyl)phosphine chloride (BOP-Cl), 2-chloro-1,3-dimethylimine-6-fluorophosphate (CIP), tribromotri(dimethylamino)phosphonium hexafluorophosphate, tribromopyrrolidone-based phosphonium hexafluorophosphate (PyBroP), O-(7-azabenzotriazol-1-yl)-N,N,N',N',N'-tetramethylureonium hexafluorophosphate (HATU), O-(7 (-azabenzotriazol-1-yl)-N,N,N',N',N'-tetramethylureon tetrafluoroborate (TATU), (7-azabenzotriazol-1-yloxy)tripyrrolidone phosphonium hexafluorophosphate (PyAOP), (benzotriazol-1-yloxy)tripyrrolidone phosphonium hexafluorophosphate (PyBOP), (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP reagent), N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)urea Onyx hexafluorophosphate (HBTU), O-(benzotriazol-1-yl)-N,N,N',N',N'-tetramethylureon tetrafluoroborate (TBTU), O-(benzotriazol-1-yl)-N,N,N',N',N'-bis(tetramethylene)ureon hexafluorophosphate (HBPyU), O-(benzotriazol-1-yl)-N,N,N',N'-bis(pentylene)ureon hexafluorophosphate, acetic anhydride, SOCl2, PCl3, POCl3, PCl5, and mixtures thereof.

[0119] In one embodiment, this document provides a pharmaceutical composition comprising a compound provided herein, including a compound of formula (I) or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof; and a pharmaceutically acceptable excipient.

[0120] The disclosed compounds can be administered to patients in various forms depending on the chosen route of administration, as will be understood by those skilled in the art. The disclosed compounds can be administered, for example, via oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration, and pharmaceutical compositions are formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and local administration. Parenteral administration can be achieved through continuous infusion over a selected time period.

[0121] The disclosed compounds can be suitably formulated into pharmaceutical compositions for administration to a subject. The pharmaceutical compositions of this teaching optionally include one or more pharmaceutically acceptable carriers and / or diluents for use therein, such as lactose, starch, cellulose, and dextrose. Other excipients may also be included, such as flavoring agents; sweeteners; and preservatives, such as methylparaben, ethylparaben, propylparaben, and butylparaben. A more complete list of suitable excipients can be found in the Handbook of Pharmaceutical Excipients (5th edition, Pharmaceutical Press (2005)). Those skilled in the art will know how to prepare formulations suitable for various routes of administration. Conventional procedures and ingredients for selecting and preparing suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003–20th edition) and the United States Pharmacopeia: National Formulary (USP 24NF19), published in 1999. Carriers, diluents, and / or excipients are acceptable in the sense that they are compatible with other components of the pharmaceutical composition and harmless to the recipient.

[0122] Typically, for oral therapeutic use, the disclosed compounds may be blended with excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, etc.

[0123] For parenteral administration, solutions of the compounds used in the disclosed methods are typically prepared in water appropriately mixed with a surfactant such as hydroxypropyl cellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof, with or without alcohol, as well as in oils. Under normal storage and use conditions, these preparations contain preservatives to prevent microbial growth.

[0124] Generally, for injection applications, it is appropriate to use sterile aqueous solutions or dispersions and sterile powders of the compounds used in the disclosed methods for the provisional preparation of sterile injectable solutions or dispersions.

[0125] The pharmaceutical compositions described herein can be applied topically to the skin, orifices, or mucous membranes. Topical application, as used herein, includes dermal (inner), conjunctival, intracorneal, intraocular, ocular, ear, transdermal, nasal, vaginal, urethral, ​​respiratory, and rectal applications.

[0126] The pharmaceutical compositions described herein can be formulated in any dosage form suitable for surface application to achieve local or systemic effects, including emulsions, solutions, suspensions, creams, gels, hydrogels, ointments, dusting powders, dressings, elixirs, lotions, suspensions, tinctures, pastes, foams, films, aerosols, irrigates, sprays, suppositories, bandages, and dermal patches. Surface formulations of the pharmaceutical compositions described herein may also include liposomes, micelles, microspheres, nanosystems, and mixtures thereof.

[0127] Pharmaceutically acceptable carriers and excipients suitable for use in the surface formulations provided herein include, but are not limited to, aqueous media, water-miscible media, non-aqueous media, antimicrobial agents or preservatives that inhibit microbial growth, stabilizers, solubility enhancers, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, wetting agents or emulsifiers, complexing agents, masking agents or chelating agents, penetration enhancers, cryoprotectants, lyophilization protectants, thickeners, and inert gases.

[0128] The pharmaceutical compositions described herein can be provided in the form of ointments, creams, and gels. Suitable ointment carriers include oily or hydrocarbon carriers, including lard, benzoic acid lard, olive oil, cottonseed oil and other oils, and white petrolatum; emulsifiable or absorbent carriers, such as hydrophilic petrolatum, hydroxystearin sulfate, and anhydrous lanolin; water-removable carriers, such as hydrophilic ointments; water-soluble ointment carriers, including polyethylene glycol of various molecular weights; and emulsion carriers, whether water-in-oil (W / O) or oil-in-water (O / W) emulsions, including cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid (see Remington: The Science and Practice of Pharmacy, ibid.). These carriers are emollients but often require the addition of antioxidants and preservatives.

[0129] Suitable cream bases can be oil-in-water or water-in-oil. Suitable cream media can be washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, also known as the "internal" phase, typically consists of petrolatum and fatty alcohols (such as cetyl alcohol or stearyl alcohol). The aqueous phase usually, but not necessarily, exceeds the volume of the oil phase and usually contains a humectant. Emulsifiers in cream formulations can be nonionic surfactants, anionic surfactants, cationic surfactants, or amphoteric surfactants.

[0130] Gels are semi-solid suspension systems. Single-phase gels contain organic macromolecules that are substantially uniformly distributed throughout the liquid carrier. Suitable gelling agents include, but are not limited to, cross-linked acrylic polymers such as carbomer, carboxylated polyalkylene compounds, and... Hydrophilic polymers, such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers, and polyvinyl alcohol; cellulose polymers, such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methylcellulose; gums, such as astragalus gum and xanthan gum; sodium alginate; and gelatin. To prepare a homogeneous gel, dispersants such as alcohols or glycerols may be added, or the gelling agent may be dispersed by grinding, mechanical mixing, and / or stirring.

[0131] In one embodiment, this document provides a method for treating, preventing, or improving one or more symptoms of a SERCA-mediated condition, disease, or disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers, a mixture of two or more diastereomers, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0132] In some implementations, the condition, disease, or ailment mediated by SERCA is mediated by SERCA2a. In some implementations, the condition, disease, or ailment mediated by SERCA is mediated by SERCA2b.

[0133] In some implementations, SERCA-mediated conditions, diseases, and disorders are cardiovascular diseases, cancer, diabetes, inflammatory diseases, metabolic diseases, or neurological diseases. In other implementations, SERCA-mediated conditions, diseases, and disorders are heart disease, stroke, stenosis, restenosis, diseases associated with vascular smooth muscle cell proliferation, diseases associated with neointimal formation, diseases associated with calmodulin-PP2B, diseases associated with NFAT, arteriovenous fistula failure, heart disease, heart-related diseases, urinary incontinence, cancer, asthma, pulmonary hypertension, chronic obstructive pulmonary disease, diabetes, neurodegenerative diseases, bipolar disorder, atherosclerosis, muscle degeneration, or autoimmune diseases.

[0134] In yet another embodiment, this document provides a method for treating, preventing, or improving one or more symptoms of diabetes in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, such as a compound of formula I or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or pharmaceutically acceptable salts, solvates, hydrates, or prodrugs thereof. In one embodiment, the diabetes is type 1. In one embodiment, the diabetes is type 2.

[0135] In yet another embodiment, this document provides a method for improving glucose tolerance in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers thereof, a mixture of two or more diastereomers thereof, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0136] In yet another embodiment, this document provides a method for treating, preventing, or improving one or more symptoms of hepatic steatosis in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers thereof, a mixture of two or more diastereomers thereof, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0137] In yet another embodiment, this document provides a method for treating, preventing, or improving one or more symptoms of obesity in a subject, the method comprising administering to the subject a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers thereof, a mixture of two or more diastereomers thereof, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof.

[0138] In another embodiment, this document provides a method for promoting thermogenesis in a subject, the method comprising administering to the subject a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers thereof, a mixture of two or more diastereomers thereof, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.

[0139] In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a primate other than a human, livestock (such as a cow), a sporting animal, or a pet (such as a horse, dog, or cat).

[0140] Conditions, diseases, or disorders that can be treated with the compounds provided herein include, but are not limited to: (1) inflammatory or allergic diseases, including systemic allergic reactions and hypersensitivity reactions, atopic dermatitis, urticaria, drug allergies, insect bite allergies, food allergies (including celiac disease, etc.) and mastocytosis; (2) inflammatory bowel diseases, including Crohn's disease, ulcerative colitis, ileitis, and enteritis; (3) vasculitis and Behcet's syndrome; (4) psoriasis and inflammatory skin diseases, including dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria, viral skin lesions (including viral skin lesions derived from human papillomavirus, HIV, or RLV infection), bacterial, flugal, and other parasitic skin lesions, and lupus erythema; (5) asthma and respiratory allergic diseases, including allergic asthma, exercise-induced asthma, allergic rhinitis, otitis media, allergic conjunctivitis, and allergic rhinitis. (6) Lung diseases and chronic obstructive pulmonary disease; (7) Autoimmune diseases, including arthritis (including rheumatoid arthritis and psoriatic arthritis), systemic lupus erythematosus, type I diabetes, myasthenia gravis, multiple sclerosis, Graves' disease and glomerulonephritis; (8) Transplant rejection (including allogeneic transplant rejection and graft-versus-host disease), for example, skin transplant rejection, solid organ transplant rejection, bone marrow transplant rejection; (9) Fever; (10) Cardiovascular diseases, including acute heart failure, hypotension, Hypertension, angina pectoris, myocardial infarction, cardiomyopathy, congestive heart failure, atherosclerosis, coronary artery disease, restenosis and vascular stenosis; (10) cerebrovascular diseases, including traumatic brain injury, stroke, ischemia-reperfusion injury and aneurysm; (11) breast cancer, skin cancer, prostate cancer, cervical cancer, uterine cancer, ovarian cancer, testicular cancer, bladder cancer, lung cancer, liver cancer, laryngeal cancer, oral cancer, colon cancer and gastrointestinal cancer (e.g., esophageal cancer, stomach cancer, pancreatic cancer), brain cancer, thyroid cancer, hematologic cancer and lymphatic system cancer; (1 2) Fibrosis, connective tissue diseases and sarcoidosis; (13) Genital and reproductive disorders, including erectile dysfunction; (14) Gastrointestinal disorders, including gastritis, ulcers, nausea, pancreatitis and vomiting; (15) Nervous system disorders, including Alzheimer's disease; (16) Sleep disorders, including insomnia, narcolepsy, sleep apnea syndrome and Pickwick syndrome; (17) Pain; (18) Kidney disorders; (19) Eye disorders, including glaucoma; and (20) Infectious diseases, including HIV.

[0141] Depending on the condition, disease, or ailment to be treated and the condition of the subject, the compounds or pharmaceutical compositions provided herein may be administered via oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisional injection or infusion, subcutaneous injection or implantation), inhalation, nasal, vaginal, rectal, sublingual, or topical (e.g., percutaneous or local) routes of administration, and may be formulated alone or in appropriate dose units with pharmaceutically acceptable excipients, carriers, adjuvants, and media suitable for each route of administration. Administration of the compounds or pharmaceutical compositions provided herein in the form of reservoir formulations in which the active ingredient is released over a predetermined time period is also provided.

[0142] When treating, preventing, or improving one or more symptoms of the conditions, diseases, or disorders described herein, appropriate dose levels are generally from about 0.001 to 100 mg per kilogram of subject body weight per day (mg / kg daily), from about 0.01 to about 75 mg / kg daily, from about 0.1 to about 50 mg / kg daily, from about 0.5 to about 25 mg / kg daily, or from about 1 to about 20 mg / kg daily, and these dose levels may be administered in a single dose or multiple doses. Within this range, the dose may be from about 0.005 to about 0.05, from about 0.05 to about 0.5 mg / kg daily, from about 0.5 to about 5.0 mg / kg daily, from about 1 to about 15 mg / kg daily, from about 1 to about 20 mg / kg daily, or from about 1 to about 50 mg / kg daily.

[0143] For oral administration, the pharmaceutical compositions provided herein can be formulated in tablet form containing from about 1.0 to about 1,000 mg of active ingredient. In one embodiment, they are formulated in tablet form containing about 1, about 5, about 10, about 15, about 20, about 25, about 50, about 75, about 100, about 150, about 200, about 250, about 300, about 400, about 500, about 600, about 750, about 800, about 900, and about 1,000 mg of active ingredient for symptomatic dose adjustment in the patients being treated. The pharmaceutical compositions can be administered at a frequency of 1 to 4 times daily (including once, twice, three times, and four times daily).

[0144] However, it should be understood that the specific dose level and frequency of administration for any particular patient can vary and will depend on a variety of factors, including the activity of the specific compound used, the metabolic stability and duration of action of the compound, age, weight, general health status, sex, diet, administration pattern and time, excretion rate, drug combination, severity of the specific disease, and the host receiving the therapy.

[0145] In one embodiment, this document provides a method for reducing stress in the ER, the method comprising contacting the ER with an effective amount of a compound provided herein, such as a compound of formula I or its enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or pharmaceutically acceptable salts, solvates, hydrates, or prodrugs thereof. In one embodiment, the ER stress is caused by ER stress. 2+ Caused by steady-state disturbances.

[0146] In yet another embodiment, this document provides a method for restoring or maintaining steady state in an ER, the method comprising contacting the ER with an effective amount of a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers thereof, a mixture of two or more diastereomers thereof, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.

[0147] In yet another implementation, this paper provides Ca for improving ER. 2+ Methods for determining concentration, comprising contacting the ER with an effective amount of the compounds provided herein, such as compounds of formula I or their enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or pharmaceutically acceptable salts, solvates, hydrates, or prodrugs thereof.

[0148] In yet another embodiment, this document provides a method for modulating the activity of SERCA, the method comprising contacting SERCA with an effective amount of a compound provided herein, such as a compound of formula I or an enantiomer thereof, a mixture of enantiomers thereof, a mixture of two or more diastereomers thereof, or an isotopic variant thereof; or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.

[0149] In some embodiments, the SERCA is SERCA1. In some embodiments, the SERCA is SERCA2. In some embodiments, the SERCA is SERCA3.

[0150] In some embodiments, the SERCA is SERCA1a. In some embodiments, the SERCA is SERCA1b. In some embodiments, the SERCA is SERCA2a. In some embodiments, the SERCA is SERCA2b. In some embodiments, the SERCA is SERCA3a. In some embodiments, the SERCA is SERCA3b. In some embodiments, the SERCA is SERCA3c.

[0151] The compounds provided herein, such as compounds of formula I or their enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs, may also be used in combination with other agents or therapies applicable to the treatment, prevention, or improvement of one or more symptoms of a disease, ailment, or disorder to which the compounds provided herein are applicable.

[0152] Other suitable therapeutic agents may include, but are not limited to: (1) alpha-adrenergic agents; (2) antiarrhythmic agents; (3) antiatherosclerotic agents, such as ACAT inhibitors; (4) antibiotics, such as anthracyclines, bleomycin, mitomycin, actinomycin D, and priloin; (5) anticancer agents and cytotoxic agents, such as alkylating agents, such as nitrogen mustard, alkyl sulfonates, nitrosoureas, ethyleneimine, and triazine; (6) anticoagulants, such as acenitroprusside, argatroban, bivalirudin, lepirudine, fondaparin, heparin, phenindone, warfarin, and (7) Antidiabetic agents, such as biguanides (e.g., metformin), glucosidase inhibitors (e.g., acarbose), insulin, megglitinide (e.g., repaglinide), sulfonylureas (e.g., glimepiride, glibenclamide, and glipizide), thiozolidinediones (e.g., troglitazone, rosiglitazone, and pioglitazone), and PPAR-γ agonists; (8) Antifungal agents, such as amorolfine, amphotericin B, anisoflavone, bifonazole, butenafine, butonazole, caspofungin, cyclophosphamide, etc. Piroctone ketone, clotrimazole, econazole, fenticonazole, ferulic acid, fluconazole, isoconazole, itraconazole, ketoconazole, micafungin, miconazole, naftifine, natamycin, nystatin, oxconazole, rivanazole, posaconazole, schizomycin, sertaconazole, thioconazole, terbinafine, terconazole, tiaconazole, and voriconazole; (9) anti-inflammatory drugs, such as nonsteroidal anti-inflammatory agents, such as aceclofenac, acimetidine, amoxicillin, aspirin, azapromide, benorilate, bromofenac, carbofenac, celecoxib, choline magnesium salicylate, diclofenac. Acid, diflunisal, etordosulfuric acid, etoricoxib, faislamine, fenbufen, fenprofen, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketoroxyprofen, lornoxicam, loxoprofen, luminicoxib, meclofenamic acid, meloxicam, aminopyrine, methyl salicylate, magnesium salicylate, nabumetone, naproxen, nimesulide, oxyphenbutazone, parecoxib, phenylbutazone, piroxicam, salicylates (10) Antimetabolites, such as folic acid antagonists, purine analogs and pyrimidine analogs; (11) Antiplatelet agents, such as GPIIb / IIIa blockers (e.g., abciximab, eptifibatide and tirofiban), P2Y(AC) antagonists (e.g., clopidogrel, ticlopidine and CS-747), cilostazol, dipyridamole and aspirin; (12) Antiproliferative agents, such as methotrexate, FK506 (tacrolimus) and mycophenolate mofetil; (13) Anti-TNF antibodies or soluble TNF receptors, such as etanercept, rapamycin and leflunimide.(14) aP2 inhibitors; (15) β-adrenergic agents, such as carvedilol and metoprolol; (16) bile acid sequestrants, such as cholestyramine; (17) calcium channel blockers, such as amlodipine besylate; (18) chemotherapeutic agents; (19) cyclooxygenase 2 (COX-2) inhibitors, such as celecoxib and rofecoxib; (20) cyclosporine; (21) cytotoxic drugs, such as azathioprine and cyclophosphamide; (22) diuretics, such as chlorothiazide, hydrochlorothiazide, flumethiazide, bendroflumethiazide, and methylchlorothiazide. Trichloromethiazide, polythiazide, benzylthiazide, ethacrynic acid, ticrynafen, chlorthalidone, furosemide, mozolomide, bumetanide, triamterene, amiloride, and spironolactone; (23) endothelial peptide converting enzyme (ECE) inhibitors, such as amiphenone phosphate; (24) enzymes, such as L-asparaginase; (25) factor VIIa inhibitors and factor Xa inhibitors; (26) farnesyltransferase inhibitors; (27) fibrates; (28) growth factor inhibitors, such as PDGF activity regulators; (29) growth hormone secretagogues; (30) HMG CoA reductase inhibitors, such as pravastatin, lovastatin, atorvastatin, simvastatin, NK-104 (also known as pitavastatin, nivastatin, or nibevastatin) and ZD-4522 (also known as rosuvastatin, atorvastatin, or visastatin); neutral peptide chain endopeptidase (NEP) inhibitors; (31) hormonal agents, such as glucocorticoids (e.g., cortisone), estrogen / antiestrogenic drugs, androgen / antiandrogen drugs, progestins and luteinizing hormone-releasing hormone antagonists, and octreotide acetate; (32) immunosuppressants; (33) mineralocorticoid receptor antagonists, such as spironolactone and eplerenone; (34) microtubule-disruptors. (35) Agents, such as sea sucrose; (36) Microtubule stabilizers, such as paclitaxel, docetaxel and acetylcholine AF; (37) MTP inhibitors; (38) Nicotinic acid; (39) Phosphodiesterase inhibitors, such as PDE III inhibitors (e.g., cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil and vardenafil); (40) Plant-derived products, such as periwinkle alkaloids, epipodophyllotoxin and taxanes; (41) Platelet-activating factor (PAF) antagonists; (42) Platinum coordination complexes, such as cisplatin, saline and carboplatin; (43) Potassium channel openers; (44) Isoprenyl protease inhibitors; (5) Protein tyrosine kinase inhibitors;(45) Renin inhibitors; (46) Squalene synthase inhibitors; (47) Steroids, such as aldosterone, beclomethasone, betamethasone, deoxycorticosterone acetate, fludrocortisone, hydrocortisone (cortisol), prednisolone, prednisone, methylprednisolone, dexamethasone, and triamcinolone; (48) TNF-α inhibitors, such as tenidap; (49) Thrombin inhibitors, such as hirudin; (50) Thrombolytic agents, such as aniplasm, ralteplase, teneplastase, tissue plasminogen activator (tPA), and rehydration solution. Group tPA, streptokinase, urokinase, prourokinase and anisole plasminogen activator-streptokinase complex (APSAC); (51) thromboxane receptor antagonists, such as ifetroban; (52) topoisomerase inhibitors; (53) angiopeptidase inhibitors (NEP-ACE dual inhibitors), such as omapatirat and gemmotirat; and (54) various other miscellaneous agents, such as hydroxyurea, procarbazine, mitotane, hexamethylpyrimidine and gold compounds.

[0153] In some embodiments, other therapies that may be used in combination with the compounds provided herein include, but are not limited to, surgery, endocrine therapy, biological response modifiers (e.g., interferon, interleukin, and tumor necrosis factor (TNF)), hyperthermia and cryotherapy, and agents to reduce any adverse reactions (e.g., antiemetics).

[0154] Other agents or drugs of this kind may be administered concurrently or sequentially with the compounds provided herein, such as compounds of Formula I or their enantiomers, mixtures of enantiomers, mixtures of two or more diastereomers, or isotopic variants; or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs, in the same manner and amounts commonly used therein. When the compounds provided herein are used concurrently with one or more other drugs, pharmaceutical compositions containing other drugs in addition to the compounds provided herein may be used, but this is not required. Therefore, pharmaceutical compositions provided herein include those containing one or more other active ingredients or therapeutic agents in addition to the compounds provided herein.

[0155] Example

[0156] This disclosure will be further understood through the following non-limiting embodiments.

[0157] As used herein, the symbols and conventions used in these processes, methods, and embodiments, whether or not specific abbreviations are specifically defined, are consistent with those used in contemporary scientific literature such as the Journal of the American Chemical Society or the Journal of Biological Chemistry. Specifically, but not limited to, the following abbreviations may be used in the embodiments and throughout the patent specification: g (gram); mg / ml (milligram); mL (milliliters); μL (microliters); M (moles); mM (millimoles); μM (micromoles); mol (moles); mmol (millimoles); hr or hrs (hours or hours); and min (minutes).

[0158] For all the following embodiments, standard procedures and methods known to those skilled in the art can be used. Unless otherwise stated, all temperatures are expressed in °C (degrees Celsius). Unless otherwise stated, all procedures are performed at room temperature.

[0159] Bioassay

[0160] ER stress cell survival assay

[0161] CSM14.1 cells were maintained at 32°C in complete medium containing Dulbecco modified Eagle's medium (DMEM) with 10% fetal bovine serum (FSB), 1% L-glutamine, 100 IU / mL penicillin, and 100 μg / mL streptomycin. Cells were recovered from the culture by trypsin digestion and then seeded at a concentration of 1000 cells / well in 20 μL of DMEM assay medium containing 2% FSB, 100 IU / mL penicillin, and 100 μg / mL streptomycin in 384-well plates (Greiner #781098). Seeding was performed using a MultiDrop Combi reagent seeder. The plates were incubated overnight at 32°C.

[0162] use The Beckman Coulter 2000 liquid processor was used to prepare the test compounds by sequentially diluting them twofold in 100% DMSO. Dosage-response profiles were obtained for 10 concentrations of the test compounds. Using an FX liquid processor (Beckman Coulter), 2.5 μL of the test compound was transferred from a 100% DMSO serial dilution plate to an intermediate plate containing 47.5 μL of DMEM assay medium containing 2% FBS, 100 IU / mL penicillin, and 100 μg / mL streptomycin, and mixed. To reduce or eliminate interference from compound precipitation, 6 μL of the diluted compound was immediately transferred to the assay plate to achieve a high compound concentration of 100 μM in 99% DMEM assay medium and 1% DMSO. After incubating the assay plate for 2 hours, 4 μL of 112.5 μM carotenoid (TG) (diluted to DMSO stock solution in assay TC medium) was dispensed into each well using a MultiDrop Combi reagent planter to obtain a final concentration of approximately 15 μM TG. 4 μL of tissue medium containing only the medium was manually transferred to each control well using a 16-channel electronic pipette. After incubating the plate overnight (approximately 16 to 24 hours), (Promega) (16 μL) was added to all wells and luminescence was measured. High luminescence indicates cell viability.

[0163] Alternatively, the test compound is tested at a single compound concentration (e.g., 2 μM) to determine its effect on cell survival compared to the medium.

[0164] Biological results are summarized in Table 1, where each compound was tested at 2 μM, and where A represents values ​​greater than 50%, B represents values ​​between 10% and 50%, C represents values ​​between 1% and 10%, and D represents values ​​not greater than 1%.

[0165] Table 1

[0166]

[0167] Cell rescue assay

[0168] Protection against TG-induced cell death. Human embryonic kidney (HEK293) cells were grown in Dulbecco modified Eagle medium (DMEM) containing 10% FBS and 1% antibiotic antifungal solution (ABAM). Mouse neuroblastoma (N2a) cells were grown in 1:1 DMEM containing 5% FBS and 1% ABAM. Cells were grown in a humidified environment at 10% CO2 in an incubator. Cells grown in 96-well plates were exposed to the test compound (20 μM) for 2 hours, followed by the addition of beta-carotene (15 μM for HEK293 cells and 1 μM for N2a cells) to induce ER stress. After incubation in a cell culture incubator for 24 hours, the beta-carotene was added to the wells. Reagent (10% v / v). Add Fluorescence readings were acquired 2 hours after the reagent was applied. Cell viability was calculated as a percentage of relative fluorescence units (RFU) compared to the control. Control cells treated with the medium showed similar viability to untreated cells.

[0169] Protection against hydrogen peroxide-induced cell death. N2a cells grown in 96-well plates were exposed to the test compound (40 μM) for 2 h, followed by the addition of hydrogen peroxide (200 μM). After incubation with hydrogen peroxide for 40 min, the test compound was added to the wells. Reagent (10% v / v). Add Fluorescence readings were acquired 2 hours after the reagent was applied. Cell viability was calculated as a percentage of relative fluorescence units (RFU) compared to the control. Control cells treated with the medium showed similar viability to untreated cells.

[0170] The results are summarized in Table 2, where A represents values ​​greater than 50% cell rescue, B represents values ​​between 10% and 50% cell rescue, C represents values ​​between 1% and 10% cell rescue, and D represents values ​​no greater than 1% cell rescue.

[0171] Ca 2+ -ATPase assay

[0172] Compared to the control, Ca2+ was performed using microsomal preparations derived from HEK293 cells at a range of calcium concentrations corresponding to physiological ranges. 2+ -ATPase assay. An NADH-linked enzyme-coupled ATPase assay was used for 96-well microplates to measure the rate of ATP hydrolysis over a range of calcium concentrations in the presence of the test compound, where V... max The calcium dependence of ATPase was determined by fitting it to a Hill function. Each well contained 2 μg or 7 μg of SR vesicles (optimized for skeletal or cardiac SR, respectively), 50 mM MOPS (pH 7.0), 100 mM KCl, 5 mM MgCl2, 1 mM EGTA, 0.2 mM NADH, 1 mM phosphoenolpyruvate, 5 IU pyruvate kinase, 5 IU lactate dehydrogenase, and 3.5 μg / mL A23187 (calcium ion carrier). CaCl2 was added to [Ca 2+ Release to a specific value. Measurements were initiated after adding 5 mM ATP and read in a SpectraMax Plus microplate spectrophotometer. Compounds A12, A13, and C19 increased baseline ATPase activity by 10–15%; and compounds C18 and C20 increased ATPase activity by more than 15%.

[0173] Table 2

[0174]

[0175]

[0176] SERCA agonists against [Ca 2+ ] ER Determining the impact

[0177] The effect of the test compound (compound A12) on [Ca] was evaluated in HeLa cells overexpressing BI-1. 2+ ] ER The effects of BI-1 on the diabetic state and its reduction [Ca] 2+ ] ER For direct measurement of ER Ca 2+ Content, using genetically encoded Ca 2+ The indicator was ER cameleon. HeLa cells were transfected with an ER-cameleon-encoding plasmid for 2 days prior to analysis. Cells were treated with the test compound for 24 hours and then subjected to Ca2+-free treatment after the addition of carotenoids to deplete ER reserves. 2+ Imaging was performed in HBSS. Emission ratio imaging of the ERcameleon was performed using a 430 / 24 excitation filter, a 450nm dichroic mirror, and two emission filters (475 / 40 for CFP and 535 / 25 for YFP). The fluorescence ratio of YFP / CFP is relative to the ERcameleon. 2+ A measure of level. Compound A12 significantly restored [Ca]. 2+ ]ER.

[0178] Determining the effect of SERCA agonists on serum glucose levels

[0179] Ob / ob mice (10 weeks old, n=3) were intraperitoneally (ip) injected with 100 μL of a solution containing 0 mg / kg (the medium), 10 mg / kg, or 50 mg / kg of the test compound selected from compounds A12, C18, C19, or C20, once daily for a total of 5 days. The protocol used to evaluate the effect of the test compound is shown in [details omitted]. Figure 3 Fasting glucose was measured at baseline and 10 hours after administration of the test compounds. Glucose levels in blood samples drawn from the tail vein were measured using a OneTouch Ultra 2Meter (LifeScan, Inc.). Compounds A12, C18, C19, and C20 significantly reduced blood glucose as early as day 2; and ob / ob mice maintained the same low glucose levels as lean mice for up to one week after the last injection of the test compounds.

[0180] Determine the effect of SERCA agonists on improved glucose and insulin tolerance.

[0181] Both glucose tolerance tests (GTT) and insulin tolerance tests (ITT) were performed 10 hours after fasting and 2 hours after A12 injection, with baseline serum glucose measured before the start of the test. For the GTT, D-glucose dissolved in 0.9% NaCl was administered intraperitoneally at a dose of 1 g / kg. Serum glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes after glucose administration. For the ITT, insulin was administered intraperitoneally at a dose of 1 IU / kg. Serum glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes after insulin administration. Glucose levels in blood samples drawn from the tail vein were measured using a OneTouch Ultra 2Meter (LifeScan, Inc.). In ob / ob mice, a glucose tolerance test (GTT) on day 7 post-injection showed that the hyperglycemic response to intraperitoneal glucose stimulation was significantly reduced in ob / ob mice treated with compound A12 compared with ob / ob mice treated with the mediated substance; and an insulin tolerance test (ITT) on day 10 post-injection showed that insulin-stimulated glucose deposition was strongly enhanced in ob / ob mice treated with compound A12 compared with ob / ob mice treated with the mediated substance.

[0182] Determining the effects of SERCA agonists on glucose and lipid metabolism

[0183] The expression of key genes involved in gluconeogenesis and adipogenesis was measured. RNA was isolated from liver samples of ob / ob mice, and mRNA expression of indicated genes was quantified using mouse-specific primers in a 7500 real-time PCR system (Applied Biosystems) with iTaq Fast SYBR Green Supermix (Bio-Rad) with ROX. Gene expression was normalized to 18S. RNA was isolated using Trizol (Invitrogen). cDNA was generated using a high-capacity cDNA reverse transcription kit (Applied Biosystems). Real-time PCR was performed in a 7500 real-time PCR system (Applied Biosystems) with mouse-specific primers using iTaq Fast SYBR Green Supermix (Bio-Rad) with ROX. Gene expression was normalized to 18S. The isolated liver tissue was homogenized in RIPA buffer containing protease inhibitors and phosphatase inhibitors (Roche). Protein samples were matched to their protein concentrations, and 30 μg of each sample was applied to SDS-PASE and transferred to a nitrocellulose membrane. The membrane was then incubated with the appropriate phosphor or a total primary antibody specific to the desired protein, followed by incubation with a suitable secondary antibody conjugated to horseradish peroxidase (Pierce), and the signal intensity was visualized by chemiluminescence (Pierce). Films from at least four independent experiments were scanned, and the density of the immunoreaction bands was evaluated using NIH Image software. GAPDH (Santa Cruz Biotechnology) was used as a loading control.

[0184] Compound A12 significantly reduced the mRNA expression of glucose-6-phosphatase (G6PAse) and phosphoenolpyruvate carboxykinase (PEPCK), both known candidates involved in glucose homeostasis. Compound A12 also significantly reduced the mRNA expression of numerous adipogenesis genes, such as stearoyl-CoA desaturase-1 (SCD1), diacylglycerol acyltransferase 2 (DGAT2), fatty acid synthase (FASn), and acetyl-CoA and sterol regulatory element-binding protein 1c (SREBP1c). Compound A12 increased the expression of the transcription factor peroxisome proliferator-activated receptor α (PPARα) and its target peroxisome proliferator-activated receptor γ coactivator-1α (PGC1α) (known for its involvement in lipid oxidation and mitochondrial biogenesis). These data suggest that compound A12 affects glucose and lipid homeostasis, thus indicating that this compound mediates energy homeostasis in the liver of obese mice.

[0185] Protection of human pancreatic islets under type 1 diabetes-related stress

[0186] Human islet microtissue treated with cytokines was used to simulate type 1 diabetes in vitro. This study tested the effects of compounds on β-cell viability in human islets under cytokine stress. The model consisted of human islet microtissue from HLA-A2 positive donors, grown and isolated in 96-well plates (one microtissue per well). The microtissue was treated for 7 days with a mixture of cytokines (IL1-β 5 ng / mL, IFN-γ 25 ng / mL, TNF-α 25 ng / mL) and either a compound (5 μM) or a mediator. Six microtissues were treated for each compound or mediator. On day 7, total ATP content was measured using CellTiter-Glo (Promega). Compound A17 increased viability by 10–20% compared to baseline; compounds A12, A13, A19, C19, and C20 increased viability by 20–50%; and compound C18 increased viability by more than 50%.

[0187] Determining the effect of SERCA agonists on reducing ER stress

[0188] Protein samples were prepared from the livers of ob / ob mice treated with the mediator (ob) or ob / ob mice treated with 50 mg / kg of the test compound (compound A12). Proteins were analyzed and quantified by Western blotting. Bands were normalized to GAPDH and expressed as 100% of ob / ob + mediator (ob). Liver tissue was homogenized in ice-cold tissue lysis buffer using a benchtop homogenizer. The homogenized sample was centrifuged at 8,000 × g for 20 min at 4 °C. The lipid layer was removed, and the supernatant was transferred to Eppendorf tubes. After centrifugation at 16,000 × g for 60 min at 4 °C, the supernatant was normalized to the same concentration and boiled in 1 × Laemmli buffer at 100 °C for 5 min. The lysates were cooled to room temperature and then loaded for Western blotting analysis. The protein lysates were resolved on an SDS-polyacrylamide gel and transferred to a PVDF membrane at 100 V at 4 °C for 2 h. The membrane was blocked in 10% blocking reagent and incubated overnight at 4°C with primary antibody in Tris-buffered saline / Tween (TBST) / 10% blocking reagent. After incubation, the membrane was washed three times in TBST for 20 minutes each time and incubated for 1 hour at room temperature with secondary antibody in TBST / 10% blocking reagent. The membrane was washed three times for 20 minutes each time and developed using a chemiluminescence assay system. To peel the membrane for use with another primary antibody, the membrane was stirred at 50°C for 20 minutes in a box containing peeling buffer (TBS containing 2% SDS and 100 mM 2-mercaptoethanol, pH 7.5). The membrane was washed three times for 20 minutes each time, then blocked and incubated with primary antibody.

[0189] Administration of the test compounds, acting as SERCA agonists, resulted in improved ER function, as evidenced by expressed protein biomarkers of ER stress. Compounds A12 and C18 significantly reduced phosphorylation of PKR-like ER kinase (PERK) and e1F2α. Dephosphorylation of PERK and e1F2α indicated a mitigation of the ER stress response. The test compounds also significantly reduced the expression of the pro-apoptotic transcription factor C / EBP homolog (CHOP), suggesting that the test compounds may also be involved in attenuating ER stress-induced apoptosis.

[0190] SERCA activation

[0191] The NADH-linked enzyme-coupled ATPase assay characterized the test compounds at a range of concentrations. Each well contained 2 mg or 7 mg of SR vesicles (optimized for bone or cardiac SR, respectively), 50 mM MOPS (pH 7.0), 100 mM M KCl, 5 mM MgCl2, 1 mM EGTA, 0.2 mM NADH, 1 mM phosphoenolpyruvate, 5 IU pyruvate kinase, 5 IU lactate dehydrogenase, and 3.5 mg / mL A23187 (calcium ion carrier), with CaCl2 added to release free [Ca2+] to a specific value. Assays were initiated after adding 5 mM ATP and read out using a SpectraMax Plus microplate spectrophotometer, with ATPase activity fitted using the Hill function. EC 50 The value was determined to be the maximum V. max The concentration of the test compound required for 50% activation.

[0192] Biological results are summarized in Table 3, where for EC50 values, A represents values ​​less than 20 μM, B represents values ​​between 20 μM and 100 μM, and C represents values ​​greater than 100 μM; and for 10 μM, V max The increase is represented by A', which is greater than 50%, B', which is between 20% and 50%, and C', which is no greater than 20%.

[0193] Selective determination

[0194] Compound A12 was tested in duplicate at 10 μM against a set of 164 biological targets frequently tested in the pharmaceutical and biotechnology industries. The methods used for each target were adapted from scientific literature to maximize reliability and reproducibility. Reference standards were run as an essential part of each assay to ensure the validity of the results. Of the 164 targets, compound A12 is only reactive to human adenosine A. 2A Human serotonin (5-hydroxytryptamine) 5-HT 2BRabbit monoamine transporter and human adrenaline (NET) transporter showed significant responses.

[0195] Table 3

[0196]

[0197] 1. CSR: Heart SR.

[0198] 2. SSR: Skeletal SR.

[0199] Treatment of Alzheimer's disease (AD) and Parkinson's disease (PD)

[0200] Mouse model: PS1 / APP mice (PS1M146V and APPSWE) were used (Howlett et al., Brain Res. 2004, 1017, 130-136). Age-matched NTg controls were located on the same background strain (C57b16 / J9).

[0201] Drug administration: Compound A12 was administered intraperitoneally (IP, 10 mg / kg in sterile water) to AD-Tg and non-Tg mice twice daily for 4 weeks. For TASTPM, administration began at 5 months (consistent with the onset of moderate plaque formation and cognitive deficits). Control mice were administered 0.9% saline daily.

[0202] Brain slice preparation: Mice were deeply anesthetized with halothane and rapidly decapitated. The brain was quickly extracted, and 300 μm or 400 μm thick transverse hippocampal slices were cut into ice-cold oxygenated artificial cerebrospinal fluid (aCSF) with the following composition (in mM): 125 NaCl, 2.5 KCl, 1.25 KH2PO4, 1.2 MgSO4, 2 CaCl2, 10 dextrose, and 25 NaHCO3.

[0203] Behavioral testing in an Alzheimer's disease model: Compound A12 was administered intraperitoneally (IP, 10 mg / kg) to APPSWE / PSEN1dE9 double transgenic mice, daily for 4 weeks (5 days a week) starting at 4 months of age. Mice were tested after the last administration of A12 or the carrier solution, according to the dosing regimen. In short, a visible platform was presented to the mice, and then removed. The distance the mice swam to find the platform was measured; shorter distances indicated increased memory. Figure 6 As shown in (A), in the hidden platform test, mice treated with A12 showed a reduction in total distance compared to mice treated with the medium. Motor coordination, strength, and balance were assessed using the Rotarod test. Figure 6(B) Mice were first trained until they could remain on a rotating bar at 16 rpm for three consecutive 120-second trials. The next day, the mice were placed back on the bar for a single trial at 18 rpm (maximum duration 120 seconds). The duration a mouse could remain on the bar was recorded, and the mouse was then returned to its cage. The bar speed was then increased to 21 rpm, and another test was performed on all mice. This process was repeated for bar speeds of 24 rpm, 27 rpm, 30 rpm, 33 rpm, and 36 rpm. All speeds were repeated twice for mice in different groups, with 30-minute intervals between each test. The average values ​​from the two tests were analyzed. The longer a mouse remained on the rotating bar, the better its motor coordination, strength, and balance.

[0204] Ca 2+ Imaging: Intraneural Ca2+ imaging of single neurons in brain slice preparations was performed using a custom video-rate multiphoton imaging system based on an upright Olympus BX51 microscope frame. 2+ Imaging. Ca2+ was injected into individual neurons via a patch pipette. 2 + Indicator: Bi-fura-2 (50 μM). Laser excitation was provided by a 100 fs pulse at 780 nm (80 MHz) from a Ti:sapphire laser (MaiTai Broadband, Spectra-Physics). The laser beam was scanned by a general scanning galvanometer (General Scanning Lumonics) allowing for rapid (7.9 kHz) bidirectional scanning on the x-axis and by a conventional linear galvanometer on the y-axis to provide a full-frame scan rate of 30 frames / second. The laser beam was focused onto the tissue using an Olympus 40x water immersion objective (numerical aperture 0.8). The emitted fluorescence was detected by a wide-field photomultiplier tube (electron tube) to derive a video signal, which was captured and analyzed using Video Savant 5.0 software (IO Industries). Background-corrected images were further analyzed using MetaMorph software. For clarity, the results are presented inversely to make [Ca 2+ The increase in [] corresponds to an increasing rate. The percentage change is calculated as [(F / ΔF)⁻¹] × 100, where F is the mean resting fluorescence at baseline, and ΔF is the decrease in fluorescence reflecting Ca release. Differences between the drug and saline treatment groups were assessed for significance using two-way ANOVA and Scheffe post-hoc analysis (p < 0.05). For the measurement of Ca... 2+ The dataset of the reaction excludes the cell nucleus.

[0205] Aβ deposition

[0206] Mice were perfused with ice-cold PBS (3 mL) via the heart, followed by perfusion with 4% paraformaldehyde (5 mL). The brain was extracted and fixed overnight in a 30% sucrose cryoprotectant solution. 40 μm thick coronal hippocampal sections were cut on a cryostat and collected in TBS (0.1 M Tris, 0.9% saline, pH 7.4).

[0207] Thioflavin S staining: Free-floating hippocampal sections were washed with TBS (4 x 3 min). The sections were then immersed in 0.5% thioflavin S (in 50 / 50 ethanol / distilled water, Sigma-Aldrich) for 10 min, followed by washing with 50% ethanol for 2 x 3 min. The sections were washed again with TBS (2 x 3 min), mounted with minimal drying, and covered with a coverslip in the presence of a fading-resistant mounting medium, PVA-DABCO, for microscopic examination.

[0208] Confocal images of immunolabeled tissues were obtained using 4X and 10X objectives on an Olympus Fluoview confocal microscope. The density of amyloid plaques was quantified by averaging the percentage of stained positive areas in the hippocampus and cortex from 3–5 sections from each animal using MetaMorph software (Molecular Devices), with a threshold set higher than background staining, as determined by software parameters and operator confirmation. The background threshold intensity did not differ significantly across animal strains or treatment conditions (p > 0.05). The experimenters were unaware of the animal strains and treatment conditions.

[0209] In vivo treatment with compound A12 restored ER Ca in AD mice. 2+ Signal transduction. A) Pseudo-color two-photon images depict the CA2+ response of saline-treated PS1 / APP (bottom, left) and compound A12-treated (middle) CA1 pyramidal neurons to caffeine (10 mM, 60 s). The right image shows the caffeine response from a non-Tg control neuron. B) Peak Ca2+ from (A). 2+ The bar graph shows the normalized RyR-Ca2+ response in PS1 / APP (red) treated with compound A12 compared to saline-treated PS1 / APP (black) and non-Tg saline-treated neurons.

[0210] The results are shown in Figure 4 and Figure 5 middle. Figure 4 In vivo treatment with compound A12 (RD 163) showed that ER Ca2+ in AD mice was restored. 2+ Signal transduction. Figure 5 The study showed that APP-PS1 mice treated with compound A12 (RD 163) for 4 weeks (10 mg / kg, ip) had fewer amyloid plaques stained with thioflavin S compared to APP-PS1 mice treated with saline. The mice were approximately 6 months old.

[0211] Behavioral testing in a rat model of Parkinson's disease impaired by 6-hydroxydopamine (6-OHDA). Male Wistar rats were trained for stepping and initiation time (IT) tests 6 days prior to treatment with 6-OHDA or NaCl (Day 6). Animals in groups 1 through 3 were given the mediator [NaCl, or a mixture of DMSO (10%) / Tween 80 (10%) / water (80%)] once daily for 16 days prior to 6-OHDA treatment. Animals from group 4 were given A12 (10 mg / kg) once daily. All treatments were administered via the intraperitoneal route once daily except Sundays. Surgery was performed at D0 under ketamine (50 mg / kg) and toluene-1,000 (10 mg / kg). Animals received a unilateral injection of 6 μl of 6-OHDA (sigmaAldrich) in the left substantia nigra pars compacta. Local analgesia was achieved during surgery via subcutaneous lidocaine injection. At the end of the surgery, animals were treated with buprenorphine (0.05 mg / kg, SC). On day 11, animals were given L-DOPA + benserazide or A12, followed by a motor inability test. For the IT test, only one of the two forelimbs was allowed free movement, and the time required to initiate a plane-to-plane movement was recorded using 180 seconds as the break point. Figure 7 As seen in (A), the starting time of the injured animals was increased with the use of 6-OHDA, while A12 reduced the starting time. In the stepping test, the experimenter held the rat and allowed only one of its two forelimbs to move freely above a plane. The other hand held the unmonitored forelimb still, leaving one paw touching the table. The experimenter then moved the animal slowly forward. The number of steps taken by the right paw was counted. Figure 7 (B) shows the results of the stepping test using L-DOPA and A12. The number of adjusted steps in the injured animal decreased sharply, while A12 increased the number of adjusted steps. In the cylinder test, the animal was placed in a Plexiglas cylinder and immediately videotaped for 15 minutes. During this period, the number of times the ipsilateral, contralateral, and both paws simultaneously (double paw contact) contacted the cylinder wall was recorded and then expressed as a percentage of the total number of contacts. Figure 7 (C) Detail the results of the A12 treatment in the cylinder test. 6-OHDA animals showed a reduced number of exposures, while A12 increased the number of exposures.

[0212] Pharmacokinetics in Sprague Dawley rats

[0213] The pharmacokinetics of the test compound (compound A12) were evaluated in Sprague Dawley rats. The compound was prepared at 1 mg / mL in DMSO / Tween 80 / water (10 / 10 / 80, V / V / V / V / ), and administered in triplicate at 1 mg / kg intravenously (iv) or 2 mg / kg orally (PO). Blood was drawn into EDTA-containing tubes at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, and 8 h, and plasma was harvested by centrifugation. Plasma (25 μL) was treated with acetonitrile (125 μL) containing an internal standard. The sample was then centrifuged at 4,000 rpm for 5 min in a benchtop centrifuge, and the filtrate was collected. The filtrate was injected onto a Thermo Betasil C18 HPLC column (5 μL, 50 × 2.1 mm). Mobile phase A was water containing 0.1% formic acid. Mobile phase B was acetonitrile containing 0.1% formic acid. Separation was achieved using a gradient from 90% A / 10% B to 5% A / 95% B over 7 minutes. An API Sciex 4000 equipped with a turbine ion spray source was used for all analytical measurements. A positive ion MRM method was developed. The peak area of ​​the product ion was measured using the peak area of ​​the internal standard. Data were fitted using WinNonLin (Pharsight Corporation, Mountain View, CA). The oral pharmacokinetic characteristics of compound A12 are: T1 / 2: 1.17 hr; C 最 Large: 0.26 μM; AUC last: 0.41 μM·hr; CL obs : 251 mL / min / kg; and F%: 13.22.

[0214] Brain barrier penetration of compound A12 was also determined in mice at a dose of 10 mg / kg IP in a similar manner. Mouse brains were harvested 1 hour after administration. The brain-to-plasma ratio was 2.6.

[0215] The pharmacokinetics of compounds A13, A17, and A19 were evaluated in Sprague Dawley rats. Compound A13 was tested in 0.2% DMA / 0.5% DMA. Prepared in 99.3% brine. Compounds A17 and A18 were prepared in 0.2% DMA / 2% brine. Prepared in 97.8% saline solution. The results are summarized in Table 4.

[0216] The pharmacokinetics of compound A13 were evaluated in dogs. Compound A13 was tested in 0.2% DMA / 0.5% DMA. Prepared in 99.3% saline. Results are summarized in Table 5. For pharmacokinetic evaluation, the test compound was administered intravenously (IV) (1 mg / kg body weight) to three male beagle dogs and orally (PO) (10 mg / kg body weight) to three male beagle dogs. Blood (approximately 1.0 mL) was collected via the femoral vein at 0, 0.017, 0.083, 0.5, 1, 2, 4, 6, 8, and 24 hours post-administration into tubes containing K3EDTA anticoagulant. Plasma samples were analyzed by LC / MS / MS. Analytical results were confirmed using quality control samples to obtain intra-assay variability. >66% of the quality control samples had an accuracy between 80-120% of one or more known values. A standard set of pharmacokinetic parameters was generated from the concentration data, including the area under the curve (AUC). 0-t and AUC 0-无穷大 Elimination half-life, clearance rate, volume of distribution and bioavailability (based on AUC0-t), and maximum plasma concentration (C0; C 最大 ) and the time to reach maximum plasma concentration (T) 最大 ).

[0217]

[0218]

[0219] Pharmacokinetics in CD1 mice

[0220] The pharmacokinetics of the test compound were evaluated in CD1 mice. The compound was prepared at 1 mg / mL in DMSO / Tween 80 / water (10 / 10 / 80, V / V / V / V / ), and administered in triplicate at 2 mg / kg intravenously (iv) or 10 mg / kg orally (PO). Blood was drawn into EDTA-containing tubes at 0.25 h, 0.5 h, 1 hr, 2 hr, 4 hr, 8 hr, and 24 hr, and plasma was harvested by centrifugation. 10 μL of plasma containing 50% acetonitrile in water (5 μL) was added to 200 μL of ACN containing an internal standard. The sample was vortexed for 30 seconds. After centrifugation at 4 °C and 4,000 rpm for 15 min, the supernatant was diluted 3-fold with water. Then, 20 μL of the diluted supernatant was injected into LC / MS / MS for quantitative analysis. The sample was injected onto an Agilent ZORBAX XDB-Phenyl 5μm column (50 × 2.10 mm). Mobile phase A was water containing 0.1% formic acid. Mobile phase B was acetonitrile containing 0.1% formic acid. Separation was achieved using a gradient from 70% A / 30% B to 0% A / 100% B over 2.10 min. A Shimadzu LCMS-8050 equipped with a turbine ion spray source was used for all analytical measurements. A positive ion MRM method was developed. The peak area of ​​the product ion was measured using the peak area of ​​the internal standard. Data were fitted using WinNonLin (Pharsight Corporation, Mountain View, CA). The results are shown in Table 6.

[0221] Table 6

[0222]

[0223]

[0224] Compound Synthesis

[0225] 3-Methyl-N-(2-methylquinoline-8-yl)butyramide A1

[0226] 3-Methylbutyryl chloride (120 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A1. ESI-MS: m / z 243 [M+H] +.

[0227] N-(2-methylquinoline-8-yl)pentylamide A2

[0228] 2,2-Dimethylpropionyl chloride (120 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A2. ESI-MS: m / z 243 [M+H] + .

[0229] 3,3-Dimethyl-N-(2-methylquinoline-8-yl)butyramide A3

[0230] 3,3-Dimethylbutyryl chloride (134 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A3. ESI-MS: m / z 257 [M+H] + .

[0231] 4-(tert-butyl)-N-(2-methylquinoline-8-yl)benzamide A4

[0232] 4-tert-butylbenzoyl chloride (197 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A4. ESI-MS: m / z 319 [M+H] + .

[0233] 4-Butyl-N-(2-methylquinoline-8-yl)benzamide A5

[0234] 4-Butylbenzoyl chloride (197 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A5. ESI-MS: m / z 319 [M+H] + .

[0235] 4-Fluoro-N-(quinoline-8-yl)benzamide A6

[0236] 4-Fluorobenzoyl chloride (158 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 8-aminoquinoline (144 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A6. ESI-MS: m / z 267 [M+H] + .

[0237] 3-Fluoro-N-(quinoline-8-yl)benzamide A7

[0238] 3-Fluorobenzoyl chloride (158 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 8-aminoquinoline (144 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A7. ESI-MS: m / z 267 [M+H] + .

[0239] 4-Methoxy-N-(2-methylquinoline-8-yl)benzamide A8

[0240] 4-Methoxybenzoyl chloride (171 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A8. ESI-MS: m / z 293 [M+H] + .

[0241] 2-Methoxy-N-(2-methylquinoline-8-yl)benzamide A9

[0242] 2-Methoxybenzoyl chloride (171 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A9. ESI-MS: m / z 293 [M+H] + .

[0243] 2-Ethoxy-N-(2-methylquinoline-8-yl)benzamide A10

[0244] 2-Ethoxybenzoyl chloride (185 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A10. ESI-MS: m / z 307 [M+H] + .

[0245] 4-Isopropoxy-N-(quinoline-8-yl)benzamide A11

[0246] 4-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (144 mg, 1.0 mmol) was added to the solution, and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A11. ESI-MS: m / z 307 [M+H] + .

[0247] 4-Isopropoxy-N-(2-methylquinoline-8-yl)benzamide A12

[0248] 4-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution, and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A12. ESI-MS: m / z 321 [M+H] + .

[0249] 3-Isopropoxy-N-(2-methylquinoline-8-yl)benzamide A13

[0250] 3-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution, and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A13. ESI-MS: m / z 321 [M+H] + .

[0251] 2-((5-methoxyquinoline-8-yl)carbamoyl)benzoic acid A14

[0252] Phthalic anhydride (148 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 5-methoxyquinoline-8-amine (174 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 20 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A14. ESI-MS: m / z 323 [M+H] + .

[0253] 2,6-Difluoro-N-(2-methylquinoline-8-yl)benzamide A15

[0254] 2,6-Difluorobenzoyl chloride (177 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A15. ESI-MS: m / z 299 [M+H] + .

[0255] 4-Cyano-2-fluoro-N-(2-methylquinoline-8-yl)benzamide A16

[0256] 4-Cyano-2-fluorobenzoic acid (165 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A16. ESI-MS: m / z 306 [M+H] + .

[0257] 2-Chloro-4-methyl-N-(2-methylquinoline-8-yl)benzamide A17

[0258] 2-Chloro-4-methylbenzoic acid (171 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution, and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A17. ESI-MS: m / z 311 [M+H] + .

[0259] 3-Chloro-4-methoxy-N-(2-methylquinoline-8-yl)benzamide A18

[0260] 3-Chloro-4-methoxybenzoic acid (187 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A18. ESI-MS: m / z 327 [M+H]+ .

[0261] 2-Methoxy-3-methyl-N-(2-methylquinoline-8-yl)benzamide A19

[0262] 2-Methoxy-3-methylbenzoic acid (166 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A19. ESI-MS: m / z 307 [M+H] + .

[0263] 2,3,4-Trifluoro-N-(2-methylquinoline-8-yl)benzamide A20

[0264] 2,3,4-Trifluorobenzoyl chloride (195 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A20. ESI-MS: m / z 317 [M+H] + .

[0265] N-(quinolin-8-yl)-1-naphthylcarboxamide A21

[0266] 1-Naphthoyl chloride (191 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 8-aminoquinoline (144 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A21. ESI-MS: m / z 299 [M+H] + .

[0267] 2-(4-Chlorophenyl)-N-(2-methylquinoline-8-yl)acetamide A22

[0268] 4-Chlorophenylacetyl chloride (189 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A22. ESI-MS: m / z 311 [M+H] + .

[0269] 3-(4-Methoxyphenyl)-N-(2-methylquinoline-8-yl)propionamide A23

[0270] 3-(4-methoxyphenyl)propionyl chloride (199 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A23. ESI-MS: m / z 321 [M+H] + .

[0271] 5-(4-Methoxyphenyl)-N-(2-methylquinoline-8-yl)isoxazole-3-carboxamide A24

[0272] 5-(4-methoxyphenyl)isoxazole-3-carboxylic acid (219 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (158 mg, 1.0 mmol) was added to the solution, and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A24. ESI-MS: m / z 360 [M+H] + .

[0273] 6-Chloro-N-(2-methylquinoline-8-yl)nicotinamide A25

[0274] 6-Chloronicotinyl chloride (176 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A25. ESI-MS: m / z 298 [M+H] + .

[0275] 3-Chloro-N-(2-methylquinoline-8-yl)benzo[b]thiophene. 2-Formamide A26

[0276] 3-Chlorobenzothiophene-2-carbonyl chloride (231 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A26. ESI-MS: m / z 353 [M+H] + .

[0277] N-(2-methylquinoline-8-yl)benzofuran-2-carboxamide A27

[0278] Method 1. Benzofuran-2-carbonyl chloride (181 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at a cooled (0 °C) temperature. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A27. ESI-MS: m / z 303 [M+H] + .

[0279] Method 2. Benzofuran-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product A27. ESI-MS: m / z 303 [M+H] + .

[0280] 2-(quinolin-8-yl)-2,3-dihydrophthalazine-1,4-dione A28

[0281] Phthalic anhydride (148 mg, 1.0 mmol) was dissolved in acetic acid (5 mL). Quinoline-8-ylhydrazine (159 mg, 1.0 mmol) was added to this solution, and the solution was heated to 80 °C. After heating with stirring for 8 hours, the mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A28. ESI-MS: m / z 290 [M+H] + .

[0282] 2-(quinolin-8-yl)isoindoline-1,3-dione A29

[0283] Phthalic anhydride (148 mg, 1.0 mmol) was dissolved in acetic acid (5 mL). 8-Aminoquinoline (144 mg, 1.0 mmol) was added to this solution, and the solution was heated to 110 °C. After heating with stirring for 20 hours, the mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound A29. ESI-MS: m / z 275 [M+H] + .

[0284] 5-Bromo-N-(2-methylquinoline-8-yl)thiophene-2-carboxamide B1

[0285] Method 1. 5-Bromo-2-thiophene carbamate (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound B1. ESI-MS: m / z 348 [M+H] + .

[0286] Method 2. 5-Bromo-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product B1. ESI-MS: m / z 348 [M+H] + .

[0287] N-(2-methylquinoline-8-yl)benzo[b]thiophene-2-carboxamide B2

[0288] Method 1. Benzo[b]thiophene-2-carbonyl chloride (196 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound B2. ESI-MS: m / z 319 [M+H] + .

[0289] Method 2. 1-Benzothiophene-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product B2. ESI-MS: m / z 319 [M+H] + .

[0290] 3-Cyano-N-(quinoline-8-yl)benzamide B3

[0291] 3-Cyanobenzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 8-aminoquinoline (144 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound B3. ESI-MS: m / z 274 [M+H]+.

[0292] 4-Cyano-N-(2-methylquinoline-8-yl)benzamide B4

[0293] 4-Cyanobenzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound B4. ESI-MS: m / z 288 [M+H] + .

[0294] 4-Bromo-N-(2-methylquinoline-8-yl)benzamide C1

[0295] 4-Bromobenzoyl chloride (219 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C1. ESI-MS: m / z 342 [M+H] + .

[0296] 2-Fluoro-N-(2-methylquinoline-8-yl)benzamide C2

[0297] 2-Fluorobenzoyl chloride (159 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C2. ESI-MS: m / z 281 [M+H] + .

[0298] 2-Nitro-N-(2-methylquinoline-8-yl)benzamide C3

[0299] 2-Nitrobenzoyl chloride (186 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C3. ESI-MS: m / z 308 [M+H] + .

[0300] 3-Trifluoromethoxy-N-(2-methylquinoline-8-yl)benzamide C4

[0301] 3-Trifluoromethoxybenzoyl chloride (225 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C4. ESI-MS: m / z 347 [M+H] + .

[0302] 2-Trifluoromethyl-N-(2-methylquinoline-8-yl)benzamide C5

[0303] 2-Trifluoromethylbenzoyl chloride (209 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C5. ESI-MS: m / z 331 [M+H] + .

[0304] 3-Fluoro-N-(2-methylquinoline-8-yl)benzamide C6

[0305] 3-Fluorobenzoyl chloride (159 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C6. ESI-MS: m / z 281 [M+H] + .

[0306] 3-Nitro-N-(2-methylquinoline-8-yl)benzamide C7

[0307] 3-Nitrobenzoyl chloride (186 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C7. ESI-MS: m / z 308 [M+H] + .

[0308] 4-Nitro-N-(2-methylquinoline-8-yl)benzamide C8

[0309] 4-Nitrobenzoyl chloride (186 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C8. ESI-MS: m / z 308 [M+H] + .

[0310] 2-Chloro-N-(2-methylquinoline-8-yl)benzamide C9

[0311] 2-Chlorobenzoyl chloride (175 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C9. ESI-MS: m / z 297 [M+H] + .

[0312] 4-Trifluoromethyl-N-(2-methylquinoline-8-yl)benzamide C10

[0313] 4-Trifluoromethylbenzoyl chloride (209 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C10. ESI-MS: m / z 331 [M+H] + .

[0314] 4-Trifluoromethoxy-N-(2-methylquinoline-8-yl)benzamide C11

[0315] 4-Trifluoromethoxybenzoyl chloride (225 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C11. ESI-MS: m / z 347 [M+H] + .

[0316] 3-Trifluoromethyl-N-(2-methylquinoline-8-yl)benzamide C12

[0317] 3-Trifluoromethylbenzoyl chloride (209 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C12. ESI-MS: m / z 331 [M+H] + .

[0318] 4-Ethoxy-N-(2-methylquinoline-8-yl)benzamide C13

[0319] 4-Ethoxybenzoyl chloride (185 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C13. ESI-MS: m / z 307 [M+H] + .

[0320] 4-Fluoro-N-(2-methylquinoline-8-yl)benzamide C14

[0321] 4-Fluorobenzoyl chloride (159 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C14. ESI-MS: m / z 281 [M+H] + .

[0322] 3-Chloro-N-(2-methylquinoline-8-yl)benzamide C15

[0323] 3-Chlorobenzoyl chloride (175 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C15. ESI-MS: m / z 297 [M+H] + .

[0324] 3-Bromo-N-(2-methylquinoline-8-yl)benzamide C16

[0325] 3-Bromobenzoyl chloride (219 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C16. ESI-MS: m / z 342 [M+H] + .

[0326] 4-Chloro-N-(2-methylquinoline-8-yl)benzamide C17

[0327] 4-Chlorobenzoyl chloride (175 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C17. ESI-MS: m / z 297 [M+H] + .

[0328] 4-N,N-Dimethylamino-N-(2-methylquinoline-8-yl)benzamide C18

[0329] NN-Dimethylaminobenzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C18. ESI-MS: m / z 306 [M+H] + .

[0330] 4-(Ethylamino)-N-(2-methylquinoline-8-yl)benzamide C19

[0331] 4-(ethylamino)benzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C19. ESI-MS: m / z 306 [M+H] + .

[0332] 4-(isopropylamino)-N-(2-methylquinoline-8-yl)benzamide C20

[0333] 4-(isopropylamino)benzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C20. ESI-MS: m / z 320 [M+H] + .

[0334] 4-(isopropylthio)-N-(2-methylquinoline-8-yl)benzamide C21

[0335] 4-(isopropylthio)benzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C21. ESI-MS: m / z 337 [M+H] + .

[0336] 4-(Ethylthio)-N-(2-methylquinoline-8-yl)benzamide C22

[0337] 4-(ethylthio)benzoyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a solution of 8-aminoquinalidine (158 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL) at 0 °C. After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound C22. ESI-MS: m / z 323 [M+H] + .

[0338] 8-(4-Isopropoxybenzamido)quinoline-2-carboxylic acid E1

[0339] 4-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline-2-carboxylic acid (188 mg, 1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound E1. ESI-MS: m / z 351 [M+H] + .

[0340] 8-(3-Isopropoxybenzamido)quinoline-2-carboxylic acid E2

[0341] 3-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline-2-carboxylic acid (188 mg, 1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound E2. ESI-MS: m / z 351 [M+H] + .

[0342] 8-(5-bromothiophene-2-carboxamido)quinoline-2-carboxylic acid E3

[0343] 5-Bromo-2-thiophene carboxyl chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 8-aminoquinoline-2-carboxylic acid (188 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound E3. ESI-MS: m / z 378 [M+H] + .

[0344] 8-(benzo[b]thiophene-2-carboxamido)quinoline-2-carboxylic acid E4

[0345] Benzo[b]thiophene-2-carbonyl chloride (196 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 8-aminoquinoline-2-carboxylic acid (188 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound E4. ESI-MS: m / z 349 [M+H] + .

[0346] N-(2-(3,5-dimethyl-1H-pyrazol-1-yl)quinolin-8-yl)-3-isopropoxybenzamide F1

[0347] 3-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)- N ′-Ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) was dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 2-(3,5-dimethyl-1H-pyrazol-1-yl)quinoline-8-amine (238 mg, 1.0 mmol) was added to this solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound F1. ESI-MS: m / z 401 [M+H] + .

[0348] N-(2-(3,5-dimethyl-1H-pyrazol-1-yl)quinolin-8-yl)-4-isopropoxybenzamide F2

[0349] 4-Isopropoxybenzoic acid (180 mg, 1.0 mmol) and N-(3-dimethylaminopropyl)- N ′-Ethylcarbodiimide hydrochloride (230 mg, 1.2 mmol) was dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (260 μL, 1.5 mmol) was added and the solution was stirred for 10 min. 2-(3,5-dimethyl-1H-pyrazol-1-yl)quinoline-8-amine (238 mg, 1.0 mmol) was added to this solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound F2. ESI-MS: m / z 401 [M+H] + .

[0350] 5-Bromo-N-(2-(3,5-dimethyl-1H-pyrazol-1-yl)quinoline-8-yl)thiophene-2-carboxylic acid F3

[0351] 5-Bromo-2-thiophene carbamate chloride (1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 2-(3,5-dimethyl-1H-pyrazol-1-yl)quinoline-8-amine (238 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound F3. ESI-MS: m / z 428 [M+H] + .

[0352] N-(2-(3,5-dimethyl-1H-pyrazol-1-yl)quinoline-8-yl)benzo[b]thiophene-2-carboxamide F4

[0353] Benzo[b]thiophene-2-carbonyl chloride (196 mg, 1.0 mmol) was dissolved in dichloromethane (2 mL) and cooled to 0 °C. This solution was added dropwise to a cooled (0 °C) solution of 2-(3,5-dimethyl-1H-pyrazol-1-yl)quinoline-8-amine (238 mg, 1.0 mmol) and N,N-diisopropylethylamine (260 μL, 1.5 mmol) in dichloromethane (5 mL). After the addition was complete, the mixture was warmed to room temperature and stirred for 2.5 hr. The mixture was diluted with water and extracted with 2 volumes of dichloromethane. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give compound F4. ESI-MS: m / z 399 [M+H] + .

[0354] N-(2-methylquinolin-8-yl)thiophene-2-carboxamide G1

[0355] 2-Thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G1. ESI-MS: m / z 269 [M+H] + .

[0356] 5-Methyl-N-(2-methylquinoline-8-yl)thiophene-2-carboxamide G2

[0357] 5-Methyl-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G2. ESI-MS: m / z 283 [M+H] + .

[0358] 3-Methyl-N-(2-methylquinoline-8-yl)thiophene-2-carboxamide G3

[0359] 3-Methyl-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G3. ESI-MS: m / z 283 [M+H] + .

[0360] 5-Chloro-N-(2-methylquinoline-8-yl)thiophene-2-carboxamide G4

[0361] 5-Chloro-2-thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G4. ESI-MS: m / z 303 [M+H] + .

[0362] 5-Acetyl-N(2-methylquinoline-8-yl)thiophene-2-carboxamide G5

[0363] 5-Acetyl-2-thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G5. ESI-MS: m / z 311 [M+H] + .

[0364] 3,5-Dibromo-N-(2-methylquinoline-8-yl)thiophene-2-carboxamide G6

[0365] 3,5-Dibromothiophene-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G6. ESI-MS: m / z 427 [M+H] + .

[0366] 4,5-Dibromo-N-(2-methylquinoline-8-yl)thiophene-2-carboxamide G7

[0367] 4,5-Dibromothiophene-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G7. ESI-MS: m / z 427 [M+H] + .

[0368] 4-Bromo-N-(2-methylquinoline-8-yl)thiafen-2-formamide G8

[0369] 4-Bromo-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product G8. ESI-MS: m / z 348 [M+H] + .

[0370] 5-Bromo-N-(quinolin-8-yl)thiophene-2-carboxamide H1

[0371] 5-Bromo-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 6-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H1. ESI-MS: m / z 334 [M+H] + .

[0372] N-(quinolin-8-yl)thiophene-2-carboxamide H2

[0373] 2-Thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H2. ESI-MS: m / z 255 [M+H] + .

[0374] 5-Methyl-N-(quinolin-8-yl)thiafen-2-formamide H3

[0375] 5-Methyl-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinalidine (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H3. ESI-MS: m / z 269 [M+H] + .

[0376] 3-Methyl-N-(quinolin-8-yl)thiophene-2-carboxamide H4

[0377] 3-Methyl-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H4. ESI-MS: m / z 269 [M+H] + .

[0378] 5-Chloro-N-(quinolin-8-yl)thiophene-2-carboxamide H5

[0379] 5-Chloro-2-thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H5. ESI-MS: m / z 289 [M+H] + .

[0380] 5-Acetyl-N-(quinolin-8-yl)thiophene-2-carboxamide H6

[0381] 5-Acetyl-2-thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H6. ESI-MS: m / z 297 [M+H] + .

[0382] N-(quinolin-8-yl)benzofuran-2-carboxamide H7

[0383] Benzofuran-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H7. ESI-MS: m / z 289 [M+H] + .

[0384] 5-Nitro-N-(quinoline-8-yl)thiafen-2-formamide H8

[0385] 5-Nitro-2-thiophenecarboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 6-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H8. ESI-MS: m / z 300 [M+H] + .

[0386] 4-Bromo-N-(quinolin-8-yl)thiophene-2-carboxamide H9

[0387] 4-Bromo-2-thiophenic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H9. ESI-MS: m / z 334 [M+H] + .

[0388] 3,5-Dibromo-N-(quinolin-8-yl)thiophene-2-carboxamide H10

[0389] 3,5-Dibromothiophene-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected, and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H10. ESI-MS: m / z 413 [M+H] + .

[0390] 4,5-Dibromo-N-(quinolin-8-yl)thiophene-2-carboxamide H11

[0391] 4,5-Dibromothiophene-2-carboxylic acid (1.0 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.2 mmol) were dissolved in dimethylformamide (5 mL). N,N-diisopropylethylamine (1.5 mmol) was added and the solution was stirred for 10 min. 8-Aminoquinoline (1.0 mmol) was added to the solution and the mixture was stirred for 20 h. The mixture was diluted with water and extracted with 2 volumes of ethyl acetate. The organic layer was collected and the solvent was removed by rotary evaporation. The residue was purified by preparative reversed-phase HPLC using a water-acetonitrile gradient to give the desired product H11. ESI-MS: m / z 413 [M+H] + .

[0392] *****

[0393] The embodiments described above are provided to give those skilled in the art a complete disclosure and description of how to manufacture and use the claimed embodiments, and are not intended to limit the scope of the disclosure herein. Modifications that are obvious to those skilled in the art are intended to fall within the scope of the appended claims. All publications, patents, and patent applications referenced in this specification are incorporated herein by reference as if each such publication, patent, or patent application were specifically and individually indicated to be incorporated herein by reference.

Claims

1. A pharmaceutical composition comprising: i) Compounds selected from the following structural formulas: ； Or its pharmaceutically acceptable salt; and ii) Pharmaceutically acceptable carriers, excipients, or diluents.

2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is in solid form.

3. Use of the pharmaceutical composition according to claim 1 in the preparation of a medicament for treating Alzheimer's disease or Parkinson's disease in a subject.

4. Use of a pharmaceutical composition in the preparation of a medicament for treating one or more symptoms of diabetes in a subject of need, said pharmaceutical composition comprising: i) Compounds selected from the following structural formulas: ； Or its pharmaceutically acceptable salt; and ii) Pharmaceutically acceptable carriers, excipients, or diluents.

5. The use as claimed in claim 4, wherein the diabetes is type 1.

6. The use as claimed in claim 4, wherein the diabetes is type 2.

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