4',5'-Dihydrospiro[piperidine-4,7'-thieno[2,3-C]pyran] Derivatives as Inhibitors of APOL1 and Methods of Using Same

Abstract

The present disclosure provides at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, compositions comprising same, and methods of using same, including use in the treatment of APOL1-mediated diseases, including pancreatic cancer, focal segmental glomerulosclerosis (FSGS), and / or non-diabetic kidney disease (NDKD).

Description

[Technical field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 307,876, filed February 8, 2022, the contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] The present disclosure provides compounds that can inhibit apolipoprotein L1 (APOL1) and methods of using those compounds to treat APOL1-mediated diseases, such as pancreatic cancer, focal segmental glomerulosclerosis (FSGS) and / or non-diabetic kidney disease (NDKD). In some embodiments, FSGS and / or NDKD are associated with at least one of two common APOL1 genetic variants (G1:S342G:I384M and G2:N388del:Y389del). In some embodiments, pancreatic cancer is associated with elevated APOL1 levels (e.g., elevated APOL1 levels in pancreatic cancer tissue).

[0003] FSGS is a rare kidney disease with an estimated global incidence of 0.2-1.1 / 100,000 / year. FSGS is a disease of the podocyte (the glomerular visceral epithelial cells) that causes proteinuria and progressive decline in kidney function. NDKD is a kidney disease with damage to the podocyte or glomerular vascular bed not caused by diabetes. NDKD is a disease characterized by hypertension and progressive decline in kidney function. Human genetic analysis supports a causal role of G1 and G2 APOL1 variants in inducing kidney disease. Individuals with two APOL1 alleles are at increased risk of developing end-stage kidney disease (ESKD), including primary (idiopathic) FSGS, human immunodeficiency virus (HIV)-associated FSGS, NDKD, arterionephrosclerosis, lupus nephritis, microalbuminuria, and chronic kidney disease. See P. Dummer et al., Semin Nephrol. 35(3):222-236 (2015).

[0004] FSGS and NDKD can be divided into different subgroups based on the underlying etiology. One homogeneous subgroup of FSGS is characterized by the presence of independent common sequence variants in the apolipoprotein L1 (APOL1) gene, called G1 and G2, referred to as "APOL1 risk alleles". G1 codes for a correlated pair of nonsynonymous amino acid changes (S342G and I384M), G2 codes for a two amino acid deletion (N388del:Y389del) near the C-terminus of the protein, and G0 is the ancestral (low risk) allele. A distinct phenotype of NDKD is also found in patients with APOL1 genetic risk variants. In both APOL1-mediated FSGS and NDKD, high levels of proteinuria and rapid loss of kidney function occur in patients with two risk alleles compared to patients with the same disease who have none or only one APOL1 genetic risk variant. Separately, in AMKD, patients with even one risk allele can develop high levels of proteinuria and rapid loss of kidney function. See G. Vajgel et al., J. Rheumatol., November 2019, jrheum.190684.

[0005] APOL1 is a 44 kDa protein that is expressed only in humans, gorillas, and baboons. In humans, the APOL1 gene is expressed in multiple organs, including the liver and kidney. APOL1 is produced primarily by the liver and contains a signal peptide that allows secretion into the bloodstream, where it circulates bound to a subset of high-density lipoproteins. APOL1 also contributes to defense against the invasive parasite Trypanosoma Brucei Brucei (TbBrucei). APOL1 is endocytosed by Tbbrucei and transported to lysosomes, where it is inserted into the lysosomal membrane, forming a pore that results in the parasite's swelling and death.

[0006] The ability to lyse Tb brucei is common to all three APOL1 variants (G0, G1, and G2), but the G1 and G2 APOL1 variants confer additional protection against parasite species that have evolved serum resistance-associated proteins (SRA) that inhibit APOL1 G0. The G1 and G2 APOL1 variants also confer additional protection against Trypanosoma species that cause sleeping sickness. The G1 and G2 variants avoid inhibition by SRA, with G1 conferring additional protection against Tb gambiense (which causes West African sleeping sickness) and G2 conferring additional protection against T brhodesiense (which causes East African sleeping sickness).

[0007] In the kidney, APOL1 is expressed in podocytes, endothelial cells (including glomerular endothelial cells), and some tubular cells. In transgenic mice, podocyte-specific expression of APOL1 G1 or G2 (but not G0) induces structural and functional changes, including albuminuria, renal function loss, podocyte abnormalities, and glomerular sclerosis. Consistent with these data, APOL1 G1 and G2 variants are responsible for inducing and accelerating the progression of FSGS in humans. Individuals carrying APOL1 risk alleles (i.e., homozygous or compound heterozygous for the APOL1 G1 allele or the APOL1 G2 allele) are at increased risk of developing FSGS, and if they do develop FSGS, they are also at risk for rapid decline in renal function. Thus, inhibition of APOL1 may have a beneficial effect in individuals carrying APOL1 risk alleles.

[0008] Although normal plasma concentrations of APOL1 are relatively high and may vary at least 20-fold in humans, circulating APOL1 is not causally related to renal disease. However, renal APOL1 is thought to be responsible for the development of renal diseases, including FSGS and NDKD. Under certain circumstances, the synthesis of APOL1 protein may be increased by approximately 200-fold by proinflammatory cytokines, such as interferon or tumor necrosis factor-α. In addition, several studies have shown that APOL1 protein forms pH-gated Na+ / K+ pores in cell membranes, resulting in a net extrusion of intracellular K+, ultimately activating local and systemic inflammatory responses, cell swelling, and death.

[0009] The risk of ESKD is substantially higher in people of recent sub-Saharan African descent compared with people of European descent. In the United States, ESKD accounts for nearly as many years of life lost in women as breast cancer and more years of life lost in men than colorectal cancer.

[0010] FSGS and NDKD are caused by damage to podocytes, which are part of the glomerular filtration barrier, resulting in proteinuria. Patients with proteinuria are at high risk of developing end-stage kidney disease (ESKD) and proteinuria-related complications such as infection or thromboembolic events. There are no standardized treatment regimens or approved drugs for FSGS or NDKD. Currently, FSGS and NDKD are managed with symptomatic treatments (including blood pressure control using blockers of the renin-angiotensin system), and patients with FSGS and severe proteinuria may be prescribed high-dose steroids. Current treatment options for NDKD are fixed on blood pressure control and blockade of the renin-angiotensin system.

[0011] Corticosteroids, alone or in combination with other immunosuppressants, induce remission in a small number of patients (e.g., remission of proteinuria in a small number of patients), but are also associated with numerous side effects. However, even in patients who initially respond to corticosteroid and / or immunosuppressant treatment, remission is often short-lived. As a result, patients, especially those of modern sub-Saharan African descent who carry two APOL1 risk alleles, rapidly progress to end-stage renal disease (ESRD). Thus, there is an unmet medical need for the treatment of FSGS and NDKD. Specifically, given the evidence that APOL1 plays a causative role in the induction and accelerated progression of renal disease, inhibition of APOL1 should have a positive effect on patients with APOL1-mediated renal disease, especially those who carry two APOL1 risk alleles (i.e., homozygous or compound heterozygous for the G1 or G2 allele). Furthermore, APOL1 is a gene that is aberrantly expressed in multiple cancers (Lin et al., Cell Death and Disease (2021), 12:760). Recently, APOL1 has been found to be abnormally elevated in human pancreatic cancer tissues compared to adjacent tissues and has been associated with poor prognosis in pancreatic cancer patients. In vivo and in vitro experiments have shown that knockdown of APOL1 inhibits cancer cell proliferation and promotes apoptosis of pancreatic cancer cells. [Prior art documents] [Non-patent literature]

[0012] [Non-Patent Document 1] P.Dummer et al.,Semin Nephrol.35(3):222-236(2015) [Non-Patent Document 2] G.Vajgel et al.,J.Rheumatol.,November 2019,jrheum.190684 [Non-Patent Document 3] Lin et al.,Cell Death and Disease(2021),12:760 Summary of the Invention [Means for solving the problem]

[0013] One aspect of the present disclosure provides at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, tautomers of formula I, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, which may be used in the treatment of diseases mediated by APOL1, such as FSGS and NDKD. For example, in some embodiments, the at least one compound is a compound represented by formula I: [ka] A tautomer, a deuterated derivative of the compound or tautomer, or a pharma- ceutically acceptable salt of any of the foregoing, wherein: X1 is selected from S and -CR2a, and X2 is selected from S and -CR2b, One of X1 and X2 is S; When X1 is S, X2 is -CR2b; When X2 is S, X1 is -CR2a; R1 is selected from hydrogen, halogen, cyano, -OH, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 5-8 membered heterocyclyl, and phenyl; The C1-C6 alkyl of R1 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, 5-8 membered heterocyclyl (optionally substituted with 1 to 3 halogen groups), -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, and C1-C4 alkoxy (optionally substituted with 1 to 3 halogen groups); The C1-C6 alkoxy of R1 is optionally substituted with 1 to 3 independently selected from halogen; The C3-C6 cycloalkyl of R1 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), and -C(=O)N(C1-C4 alkyl)2 groups; The phenyl of R1 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), and -C(=O)N(C1-C4 alkyl); R2a is selected from hydrogen, halogen, cyano, -OH, oxo, and C1-C6 alkyl, The C1-C6 alkyl of R2a is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C4 alkoxy; R2b is selected from hydrogen, halogen, cyano, -OH, oxo, and C1-C6 alkyl; each R3a is independently selected from halogen, cyano, -OH, C1-C6 alkoxy, and C1-C6 alkyl (optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH); Two R3a together form an oxo group, each R3b is independently selected from C1-C2 alkyl (optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH); two R3b together form an oxo group; One of R4 and R5 is hydrogen, and the other is selected from C1-C6 alkyl, -C(=O)NH2, -C(=O)O(C1-C4 alkyl), C2-C6 alkynyl, and [ka] is selected from the group consisting of The C1-C6 alkyl of R4 or R5 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), -C(=O)N(C1-C4 alkyl)2, C3-C6 cycloalkyl, 5-10 membered heterocyclyl, phenyl, and 5-10 membered heteroaryl groups; Ring A is selected from C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6 and C10 aryl, and 5- to 10-membered heteroaryl, and Ring A is optionally substituted with 1, 2, 3, 4, or 5 Ra groups; Ra is, for each occurrence, halogen, cyano, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkenyl, C1-C6 haloalkoxy, -C(=O)NRhRi, -NRhRi, -NRhC(=O)Rk, -NRhC(=O)ORk, -NRhC(=O)NRiRj, -NRhS(=O)pRk , -ORk, -OC(=O)Rk, -OC(=O)ORk, -OC(=O)NRhRi, -[O(CH2)q]rO(C1-C6 alkyl), -S(=O)pRk, -S(=O)pNRhRi, -C(=O)ORk, C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6 and C10 aryl, and 5- to 10-membered heteroaryl; The C1-C6 alkyl, C1-C6 alkoxy, and C2-C6 alkenyl of Ra are each independently selected from C6-C10 aryl (optionally substituted with 1 to 3 Rm groups), 5- to 10-membered heterocyclyl (optionally substituted with 1 to 3 Rm groups), 5- to 10-membered heteroaryl (optionally substituted with 1 to 3 Rm groups), cyano, -C(=O)Rk, -C(=O)ORk, -C(=O)NRhRi, -NR optionally substituted with 1 to 3 groups independently selected from -NRhC(=O)Rk, -NRhC(=O)ORk, -NRhC(=O)NRiRj, -NRhS(=O)pRk, -ORk, -OC(=O)Rk, -OC(=O)ORk, -OC(=O)NRhRi, -S(=O)pRk, -S(=O)pNRhRi, and C3-C6 cycloalkyl (optionally substituted with 1 to 3 Rm groups); Each of the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6 and C10 aryl, and 5- to 10-membered heteroaryl of Ra is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, C1-C4 alkyl, C(=O)NRhRi, -NRhRi, -ORk, and oxo; Rh, Ri, and Rj are each independently selected for each occurrence from hydrogen, C1-C4 alkyl, C6-C10 aryl, and C3-C6 carbocycle; Any one of Rh, Ri, and Rj, C1-C4 alkyl, is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH; Rk, for each occurrence, is independently selected from hydrogen, C1-C4 alkyl, 5-10 membered heterocyclyl, and C3-C6 carbocycle; Any one of Rk's C1-C4 alkyl is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH groups; Rm is independently selected for each occurrence from halogen, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, -S(=O)pRk, and -ORk; The C1-C6 alkyl of Rm is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH; k is an integer selected from 0, 1, and 2, where k is 1 when R3a is oxo; m is an integer selected from 0, 1, and 2, where m is 1 when R3b is oxo; p, for each occurrence, is an integer selected from 1 and 2; q and r are integers independently selected from 1, 2, 3, and 4 for each occurrence.

[0014] In some embodiments, at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt of the present disclosure is a compound represented by the following structural formulas IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId: [ka] [ka] Rings A, Ra, R1, and R3a are as defined for formula I above.

[0015] In one aspect of the disclosure, the compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId are selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing.

[0016] In some embodiments, the disclosure provides pharmaceutical compositions comprising at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. In some embodiments, the pharmaceutical compositions may comprise a compound selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. These compositions may further comprise at least one additional pharma- ceutically active ingredient and / or at least one carrier.

[0017] Another aspect of the present disclosure provides a method for treating an APOL1 mediated disease, comprising administering to a subject in need thereof at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, or a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt. In some embodiments, the method comprises administering at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing.

[0018] Another aspect of the present disclosure provides a method of treating an APOL1-mediated cancer (e.g., pancreatic cancer), comprising administering to a subject in need thereof at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, or a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt. In some embodiments, the method comprises administering at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing.

[0019] Another aspect of the present disclosure provides a method of treating APOL1-mediated kidney disease (e.g., ESKD, FSGS, and / or NDKD) comprising administering to a subject in need thereof at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, or a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt. In some embodiments, the method comprises administering at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing.

[0020] In some embodiments, the method of treatment comprises administering to a subject in need thereof at least one additional active agent in the same pharmaceutical composition or as a separate composition with at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. In some embodiments, the method comprises administering at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, together with at least one additional active agent, either in the same pharmaceutical composition or as a separate composition.

[0021] Also provided are methods of inhibiting APOL1, comprising administering to a subject in need thereof at least one compound, tautomer, deuterated derivative, or compound of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of these compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, or a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt. In some embodiments, the method of inhibiting APOL1 comprises administering to a subject in need thereof at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt, or a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, tautomers thereof, deuterated derivatives of these compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] definition The term "APOL1" as used herein means apolipoprotein L1 protein, and the term "APOL1" means apolipoprotein L1 gene.

[0023] The term "APOL1-mediated disease" refers to a disease or condition associated with abnormal APOL1 (e.g., a particular APOL1 genetic variant, elevated APOL1 levels). In some embodiments, the APOL1-mediated disease is an APOL1-mediated renal disease. In some embodiments, the APOL1-mediated disease is associated with patients with two APOL1 risk alleles, e.g., homozygous or compound heterozygous for the G1 allele or the G2 allele. In some embodiments, the APOL1-mediated disease is associated with patients with one APOL1 risk allele.

[0024] The term "APOL1-mediated renal disease" refers to a disease or condition that impairs renal function and can be attributed to APOL1. In some embodiments, APOL1-mediated renal disease is associated with a patient who has two APOL1 risk alleles, for example, homozygous or compound heterozygous for the G1 allele or the G2 allele. In some embodiments, the APOL1-mediated renal disease is selected from ESKD, NDKD, FSGS, HIV-associated nephropathy, arteriosclerosis, lupus nephritis, microalbuminuria, and chronic renal disease. In some embodiments, the APOL1-mediated renal disease is chronic renal disease or proteinuria.

[0025] As used herein, the term "FSGS" means focal segmental glomerulosclerosis, a disease of podocytes (glomerular visceral epithelial cells) that causes proteinuria and progressive decline in kidney function and is associated with two common APOL1 genetic variants (G1:S342G:I384M and G2:N388del:Y389del).

[0026] The term "NDKD" as used herein means non-diabetic kidney disease characterized by severe hypertension and progressive decline in renal function and associated with two common APOL1 genetic variants (G1:S342G:I384M and G2:N388del:Y389del).

[0027] The terms "ESKD" and "ESRD" are used interchangeably herein and refer to end stage renal disease or end stage renal disease. ESKD / ESRD refers to end stage renal disease, i.e., kidney failure, where the kidneys do not function well enough that the patient cannot survive without dialysis or a kidney transplant. In some embodiments, ESKD / ESRD is associated with two APOL1 risk alleles.

[0028] The term "compound", when referring to a compound of the present disclosure, refers to a collection of molecules having the same chemical structure, unless otherwise indicated as a collection of stereoisomers (e.g., a collection of racemates, a collection of cis / trans stereoisomers, or a collection of (E) and (Z) stereoisomers), except that isotopic variations may exist between the constituent atoms of the molecule. Thus, it will be apparent to one of skill in the art that a compound represented by a particular chemical structure containing a deuterium atom as shown also contains a lesser amount of isotopic substitutions having a hydrogen atom at one or more of the designated deuterium positions in the structure. The relative amount of such isotopic substitutions in the compounds of the present disclosure will depend on several factors, including the isotopic purity of the reagents used to make the compound, and the efficiency of isotope incorporation in the various synthetic steps used to prepare the compound. However, as described above, the relative amount of such isotopic substitutions overall will be less than 49.9% of the compound. In other embodiments, the relative amount of such isotopic substitutions overall will be less than 47.5%, less than 40%, less than 32.5%, less than 25%, less than 17.5%, less than 10%, less than 5%, less than 3%, less than 1%, or less than 0.5% of the compounds.

[0029] As used herein, "optionally substituted" is interchangeable with the phrase "substituted or unsubstituted." In general, the term "substituted," whether preceded by the term "optionally," refers to the replacement of a hydrogen radical in a given structure with the radical of a specified substituent. Unless otherwise indicated, an "optionally substituted" group may have a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be either the same or different at all positions. Combinations of substituents envisioned by the present disclosure are those that result in the formation of stable or chemically feasible compounds.

[0030] The term "isotopically modified" refers to a species whose chemical structure differs from a reference compound only in its isotopic composition. In addition, unless otherwise specified, structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structure except for the replacement of hydrogen with deuterium or tritium, or the replacement of carbon with 13C or 14C are within the scope of this disclosure.

[0031] Unless otherwise indicated, structures depicted herein are also intended to include all isomeric forms of the structure, such as racemic mixtures, cis / trans isomers, (Z) and (E) double bond isomers, and geometric (or conformational) isomers, such as (Z) and (E) conformational isomers. Thus, geometric and conformational mixtures of the compounds are within the scope of the disclosure. Unless otherwise specified, all tautomeric forms of the compounds of the disclosure are within the scope of the disclosure.

[0032] The term "tautomer" as used herein refers to one of two or more isomers of a compound that exist together in equilibrium and are readily interchanged by migration of atoms, e.g., hydrogen atoms or groups, within the molecule.

[0033] As used herein, "stereoisomers" refers to enantiomers and diastereomers.

[0034] As used herein, a "deuterated derivative" refers to a compound having the same chemical structure as a reference compound, but with one or more hydrogen atoms replaced by a deuterium atom ("D" or "2H"). It will be recognized that some variation in natural isotopic abundance will occur in the synthesized compound depending on the source of the chemicals used in the synthesis. The concentration of naturally occurring stable hydrogen isotopes is small and insignificant compared to the degree of stable isotopic substitution of the deuterated derivatives described herein, despite this variation. Thus, unless otherwise specified, when a "deuterated derivative" of a compound of the present disclosure is referred to, at least one hydrogen is replaced with deuterium well above its natural isotopic abundance, which is typically about 0.015%. In some embodiments, deuterated derivatives of the disclosure have an isotopic enrichment factor for each deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), or at least 6600 (99% deuterium incorporation).

[0035] The term "isotopic enrichment factor" as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope.

[0036] The term "alkyl" or "aliphatic" as used herein means a straight-chain (i.e., linear or unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated. Unless otherwise specified, an alkyl group contains 1-20 alkyl carbon atoms. In some embodiments, an alkyl group contains 1-10 aliphatic carbon atoms. In some embodiments, an alkyl group contains 1-8 aliphatic carbon atoms. In some embodiments, an alkyl group contains 1-6 alkyl carbon atoms. In some embodiments, an alkyl group contains 1-4 alkyl carbon atoms, in other embodiments, an alkyl group contains 1-3 alkyl carbon atoms, and in still other embodiments, an alkyl group contains 1 or 2 alkyl carbon atoms. In some embodiments, an alkyl group is linear or straight-chained or unbranched. In some embodiments, an alkyl group is branched.

[0037] As used herein, the terms "cycloalkyl" and "cyclic alkyl" refer to a fully saturated monocyclic C3-8 hydrocarbon, or a spirocyclic, fused, or bridged bicyclic or tricyclic C8-14 hydrocarbon, where any individual ring within the bicyclic ring system has 3 to 7 members. In some embodiments, a cycloalkyl is a C3-C12 cycloalkyl. In some embodiments, a cycloalkyl is a C3-C8 cycloalkyl. In some embodiments, a cycloalkyl is a C3-C6 cycloalkyl. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentanyl, and cyclohexyl.

[0038] As used herein, the term "cycloalkyl" or "alicyclic" encompasses the term "cycloalkyl" or "cyclic alkyl" and refers to a monocyclic C3-8 hydrocarbon, or a spirocyclic, fused, or bridged bicyclic or tricyclic C8-14 hydrocarbon, which is fully saturated or partially saturated to contain one or more saturated units, but is not aromatic, and any individual ring of the bicyclic ring system has from 3 to 7 members. A bicyclic cycloalkyl includes a combination of a monocyclic carbocyclic ring fused to a phenyl. In some embodiments, a cycloalkyl is a C3-C12 cycloalkyl. In some embodiments, a cycloalkyl is a C3-C10 cycloalkyl. In some embodiments, a cycloalkyl is a C3-C8 cycloalkyl.

[0039] As used herein, the term "heteroalkyl" or "heteroaliphatic" means an alkyl or aliphatic group, as defined above, in which one or two carbon atoms are independently replaced by one or more oxygen, sulfur, nitrogen, phosphorus, or silicon.

[0040] As used herein, the term "alkenyl" refers to a straight-chain (i.e., linear or unbranched) or branched hydrocarbon chain containing one or more double bonds. In some embodiments, an alkenyl group is straight-chain. In some embodiments, an alkenyl group is branched-chain.

[0041] The terms "heterocycle", "heterocyclyl", and "heterocyclic" are used interchangeably herein to refer to non-aromatic (i.e., fully saturated or partially saturated, but not aromatic, as it contains one or more units of unsaturation), monocyclic, or spirocyclic, fused, or bridged bicyclic or tricyclic ring systems in which one or more ring members are independently selected heteroatoms. Bicyclic heterocyclyls include the following combinations of monocyclic rings: monocyclic heteroaryl fused to a monocyclic heterocyclyl; monocyclic heterocyclyl fused to another monocyclic heterocyclyl; monocyclic heterocyclyl fused to a phenyl; monocyclic heterocyclyl fused to a monocyclic cycloalkyl / cycloalkyl; and monocyclic heteroaryl fused to a monocyclic carbocyclyl / cycloalkyl.

[0042] In some embodiments, the heterocycle includes one or more ring atoms substituted with an oxo group (eg, a C=O group, an S=O group, or an SO2 group, etc.).

[0043] In some embodiments, a "heterocycle", "heterocyclyl", "heteroalicyclic", or "heterocyclic" group has 3-14 ring members, where one or more ring members are heteroatoms independently selected from oxygen, sulfur, nitrogen, and phosphorus. In some embodiments, each ring in a bicyclic or tricyclic ring system contains 3-7 ring members. In some embodiments, a heterocycle has at least one unsaturated carbon-carbon bond. In some embodiments, a heterocycle has at least one unsaturated carbon-nitrogen bond. In some embodiments, a heterocycle has one heteroatom independently selected from oxygen, sulfur, nitrogen, silicon, and phosphorus, the quaternized form of any basic nitrogen, or a substitutable nitrogen of a heterocycle, such as N (in the case of 3,4-dihydro-2H-pyrrolyl), NH (in the case of pyrrolidinyl), or NR+ (in the case of N-substituted pyrrolidinyl). In some embodiments, a heterocycle has one heteroatom that is a nitrogen atom. In some embodiments, a heterocycle has one heteroatom that is an oxygen atom. In some embodiments, the heterocycle has two heteroatoms, each independently selected from nitrogen and oxygen. In some embodiments, the heterocycle has three heteroatoms, each independently selected from nitrogen and oxygen. In some embodiments, the heterocyclyl is a 3- to 12-membered heterocyclyl. In some embodiments, the heterocyclyl is a 3- to 10-membered heterocyclyl. In some embodiments, the heterocyclyl is a 3- to 8-membered heterocyclyl. In some embodiments, the heterocyclyl is a 5- to 10-membered heterocyclyl. In some embodiments, the heterocyclyl is a 5- to 8-membered heterocyclyl. In some embodiments, the heterocyclyl is a 5- or 6-membered heterocyclyl. Non-limiting examples of monocyclic heterocyclyls include piperidinyl, piperazinyl, tetrahydropyranyl, azetidinyl, tetrahydrothiophenyl 1,1-dioxide, and the like.

[0044] The term "unsaturated" as used herein means that a moiety has one or more units or degrees of unsaturation. Unsaturation is a situation in which not all of the available valence bonds in a compound are satisfied by substituents, thus causing the compound to contain double or triple bonds.

[0045] The term "alkoxy" or "thioalkyl" as used herein refers to an alkyl group, as previously defined, in which one carbon of the alkyl group is replaced by an oxygen ("alkoxy") or sulfur ("thioalkyl") atom, respectively, provided that the oxygen and sulfur atoms are linked between two carbon atoms. "Cyclic alkoxy" refers to a monocyclic, spirocyclic, bicyclic, bridged bicyclic, tricyclic, or bridged tricyclic hydrocarbon that contains at least one alkoxy group, but is not aromatic. Non-limiting examples of cyclic alkoxy groups include tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, 8-oxabicyclo[3.2.1]octanyl, and oxepanyl.

[0046] As used herein, the terms "haloalkyl", "haloalkenyl" and "haloalkoxy" refer to straight or branched alkyl, alkenyl or alkoxy, respectively, substituted with one or more halogen atoms. Non-limiting examples of haloalkyl groups include -CHF2, -CH2F, -CF3, -CF2-, and perhaloalkyl, such as -CF2CF3. Non-limiting examples of haloalkoxy groups include -OCHF2, -OCH2F, -OCF3, and -OCF2.

[0047] The term "halogen" includes F, Cl, Br, and I, i.e., fluoro, chloro, bromo, and iodo, respectively.

[0048] The term "aminoalkyl" means an alkyl group that is substituted with or contains an amino group.

[0049] As used herein, "amino" refers to a group that is a primary, secondary, or tertiary amine.

[0050] As used herein, a "carbonyl" group refers to C=O.

[0051] As used herein, a "cyano" or "nitrile" group refers to --C.ident.N.

[0052] As used herein, a "hydroxy" group refers to an --OH group.

[0053] As used herein, a "thiol" group refers to -SH.

[0054] As used herein, "tert" and "t-" each refer to tertiary.

[0055] As used herein, an "aromatic group" or "aromatic ring" refers to a chemical group that contains a conjugated planar ring system having delocalized pi orbitals consisting of [4n+2]p orbital electrons, where n is an integer ranging from 0 to 6. Non-limiting examples of aromatic groups include aryl and heteroaryl groups.

[0056] The term "aryl" used alone or as part of a larger moiety, such as "arylalkyl", "arylalkoxy", or "aryloxyalkyl", refers to a monocyclic, or spirocyclic, fused or bridged bicyclic, or tricyclic ring system having a total of 5 to 14 ring members, in which all rings in the system are aromatic rings containing only carbon atoms, and in which each ring of the bicyclic or tricyclic ring system contains 3 to 7 ring members. Non-limiting examples of aryl groups include phenyl (C6) and naphthyl (C10) rings.

[0057] The term "heteroaryl" used alone or as part of a larger moiety, such as "heteroarylalkyl" or "heteroarylalkoxy", refers to a monocyclic, or spirocyclic, fused or bridged, bicyclic, or tricyclic ring system having a total of 5 to 14 ring members, where at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and each ring in the bicyclic and tricyclic ring systems contains 3 to 7 ring members. Bicyclic heteroaryls include the following combinations of monocyclic rings: a monocyclic heteroaryl fused to another monocyclic heteroaryl; and a monocyclic heteroaryl fused to a phenyl. In some embodiments, a heteroaryl group has one or more heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a heteroaryl group has one heteroatom. In some embodiments, a heteroaryl group has two heteroatoms. In some embodiments, a heteroaryl group is a monocyclic ring system having five ring members. In some embodiments, a heteroaryl group is a monocyclic ring system having six ring members. In some embodiments, the heteroaryl is a 3-12 membered heteroaryl. In some embodiments, the heteroaryl is a 3-10 membered heteroaryl. In some embodiments, the heteroaryl is a 3-8 membered heteroaryl. In some embodiments, the heteroaryl is a 5-10 membered heteroaryl. In some embodiments, the heteroaryl is a 5-8 membered heteroaryl. In some embodiments, the heteroaryl is a 5 or 6 membered heteroaryl. Non-limiting examples of monocyclic heteroaryls include pyridinyl, pyrimidinyl, thiophenyl, thiazolyl, isoxazolyl, and the like.

[0058] Non-limiting examples of useful protecting groups for nitrogen-containing groups such as amine groups include, for example, t-butyl carbamate (Boc), benzyl (Bn), tetrahydropyranyl (THP), 9-fluorenylmethyl carbamate (Fmoc), benzyl carbamate (Cbz), acetamide, trifluoroacetamide, triphenylmethylamine, benzylideneamine, and p-toluenesulfonamide.Methods for adding (a process commonly referred to as "protecting") and removing (a process commonly referred to as "deprotecting") such amine protecting groups are well known in the art and are available, for example, in PJ Kocienski, Protecting Groups, Thieme, 1994, and Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Edition (John Wiley & Sons, New York, 1999) and 4th Edition (John Wiley & Sons, New Jersey, 2014), which are incorporated herein by reference in their entirety.

[0059] Non-limiting examples of suitable solvents that may be used in the present disclosure include, but are not limited to, water, methanol (MeOH), ethanol (EtOH), dichloromethane or "methylene chloride" (CH2Cl2), toluene, acetonitrile (MeCN), dimethylformamide (DMF), dimethylsulfoxide (DMSO), methyl acetate (MeOAc), ethyl acetate (EtOAc), heptane, isopropyl acetate (IPAc), tert-butyl acetate (t-BuOAc), isopropyl alcohol (IPA), tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-Me THF), methyl ethyl ketone (MEK), tert-butanol, diethyl ether (Et2O), methyl-tert-butyl ether (MTBE), 1,4-dioxane, and N-methylpyrrolidone (NMP).

[0060] Non-limiting examples of suitable bases that may be used in the present disclosure include, but are not limited to, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), potassium tert-butoxide (KOtBu), potassium carbonate (K2CO3), N-methylmorpholine (NMM), triethylamine (Et3N; TEA), diisopropyl-ethylamine (i-Pr2EtN; DIPEA), pyridine, potassium hydroxide (KOH), sodium hydroxide (NaOH), lithium hydroxide (LiOH), and sodium methoxide (NaOMe; NaOCH3).

[0061] The present disclosure includes pharma- ceutically acceptable salts of the disclosed compounds. A salt of a compound is formed between an acid and a basic group of the compound, such as an amino functional group, or between a base and an acidic group of the compound, such as a carboxyl functional group.

[0062] As used herein, the term "pharmaceutically acceptable" refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic reaction, and the like, commensurate with a reasonable benefit / risk ratio. "Pharmaceutically acceptable salt" refers to any non-toxic salt that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of the present disclosure. Suitable pharmaceutically acceptable salts are, for example, those disclosed in S. M. Berge, et al. J. Pharmaceutical Sciences, 1977, 66, 1-19.

[0063] Acids commonly employed to form pharma- ceutically acceptable salts include inorganic acids such as hydrogen disulfide, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid, and organic acids such as p-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, and acetic acid, and related inorganic and organic acids. Accordingly, such pharma- ceutically acceptable salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, and the like. benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and other salts. In some embodiments, pharma- ceutically acceptable acid addition salts include those formed with mineral acids, such as hydrochloric acid and hydrobromic acid, as well as those formed with organic acids, such as maleic acid.

[0064] Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N+(C1-4 alkyl)4 salts. The present disclosure also contemplates the quaternization of any basic nitrogen-containing group of the compounds disclosed herein. Suitable, non-limiting examples of alkali and alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. Further non-limiting examples of pharmaceutically acceptable salts include ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates. Other suitable, non-limiting examples of pharmaceutically acceptable salts include besylate and glucosamine salts.

[0065] The terms "patient" and "subject" are used interchangeably herein and refer to animals, including humans.

[0066] The terms "effective dose" and "effective amount" are used interchangeably herein and refer to the amount of the compound that produces the desired effect for which it is administered (e.g., amelioration of symptoms of FSGS and / or NDKD, reduction in the severity of FSGS and / or NDKD, or alleviation of symptoms of FSGS and / or NDKD, and / or slowing the progression of FSGS and / or NDKD, or slowing the progression of symptoms of FSGS and / or NDKD). The exact amount of the effective dose will depend on the purpose of treatment and will be ascertainable by the skilled artisan using known techniques (see, e.g., Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

[0067] As used herein, the term "treatment" and its cognates refer to slowing or stopping disease progression. As used herein, "treatment" and its cognates include, but are not limited to, complete or partial remission, lower risk of renal failure (e.g., ESRD), and disease-related complications (e.g., edema, susceptibility to infection, or thromboembolic events). Improvement or reduction in severity of any of these symptoms can be easily assessed according to methods and techniques known in the art or subsequently developed.

[0068] The terms "about" and "approximately," when used in connection with a dose, amount, or weight percentage of a component of a composition or dosage form, include a particular dose, amount, or weight percentage value, or a range of doses, amounts, or weight percentages, that would be recognized by one of skill in the art as providing an equivalent pharmacological effect as that obtained from the particular dose, amount, or weight percentage.

[0069] At least one compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt selected from the compounds of formula I and II, their tautomers, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, may be administered once daily, twice daily, or three times daily, for example, for the treatment of AMKD, including FSGS and / or NDKD. In some embodiments, at least one compound selected from compounds 1-78, their tautomers, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, may be administered once daily, twice daily, or three times daily, for example, for the treatment of AMKD, including FSGS and / or NDKD. In some embodiments, at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is administered once a day. In some embodiments, a compound selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is administered once a day. In some embodiments, at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is administered twice a day. In some embodiments, a compound selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing are administered twice daily.In some embodiments, at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is administered three times per day. In some embodiments, a compound selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is administered three times per day.

[0070] In some embodiments, 2 mg to 1500 mg or 5 mg to 1000 mg of at least one compound selected from Formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, are administered once daily, twice daily, or three times daily. In some embodiments, 2 mg to 1500 mg or 5 mg to 1000 mg of at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from Compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, or pharma- ceutically acceptable salts of any of the foregoing, are administered once daily, twice daily, or three times daily.

[0071] Those skilled in the art will recognize that when the amount of a compound is disclosed, the relative amount of the pharma- ceutically acceptable salt form of the compound is an amount equivalent to the concentration of the free base of the compound. The amounts of compounds, pharma- ceutically acceptable salts, solvates, and deuterated derivatives disclosed herein are based on the free base form of the reference compound. For example, "1000 mg of at least one compound or pharma- ceutically acceptable salt selected from the compound of formula I and its pharma- ceutically acceptable salts" includes 1000 mg of the compound of formula I and a pharma- ceutically acceptable salt of the compound of formula I in a concentration equivalent to 1000 mg of the compound of formula I.

[0072] As used herein, the term "ambient conditions" refers to room temperature, outside air conditions, and uncontrolled humidity conditions.

[0073] Compounds and Compositions In some embodiments, at least one compound selected from formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, its tautomers, deuterated derivatives of the compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, may be used to treat AMKD, including FSGS and NDKD. In some embodiments, the compound of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId may be selected from compounds 1-78, their tautomers, deuterated derivatives of the compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, a pharmaceutical composition comprising at least one compound selected from formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, its tautomers, deuterated derivatives of the compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, may be used to treat AMKD, including FSGS and NDKD. In some embodiments, the pharmaceutical compositions can include a compound selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing.

[0074] In some embodiments of Formula I, [ka] wherein the variable X1 is selected from S and -CR2a, and X2 is selected from S and -CR2b, and wherein one of the variables X1 and X2 is S. In some embodiments of Formula I, the variable X1 is S and the variable X2 is -CR2b. In some embodiments of Formula I, the variable X2 is S and the variable X1 is -CR2a.

[0075] In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the variable R1 is selected from hydrogen, halogen, cyano, -OH, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, 5-8 membered heterocyclyl, and phenyl.

[0076] In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the variable R1 is selected from halogen. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the variable R1 is Cl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the variable R1 is Br. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the variable R1 is I.

[0077] In some embodiments of formula I (including those discussed above that define the variables X1 and X2), the variable R1 is selected from C1-C6 alkyl. In some embodiments of formula I (including those discussed above that define the variables X1 and X2), the variable R1 is C1 alkyl. In some embodiments of formula I (including those discussed above that define the variables X1 and X2), the variable R1 is C2 alkyl.

[0078] In some embodiments of formula I (including those discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is optionally substituted with 1-3 groups independently selected from halogen, cyano, 5-8 membered heterocyclyl (optionally substituted with 1-3 halogen groups), -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, and C1-C4 alkoxy (optionally substituted with 1-3 halogen groups). In some embodiments of formula I (including those discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is optionally substituted with 1-3 groups independently selected from halogen. In some embodiments of formula I (including those discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is optionally substituted with 1-3 groups independently selected from halogen. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is substituted with two halogens. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is substituted with three halogens. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is substituted with one F. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is substituted with two F. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), the C1-C6 alkyl of R1 is substituted with three F. In some embodiments of formula I (including the embodiments discussed above that define the variables X1 and X2), R1 is -CF3. In some embodiments of Formula I (including those discussed above that define the variables X1 and X2), R1 is -CH2CHF2. In some embodiments of Formula I (including those discussed above that define the variables X1 and X2), R1 is -CH2CF3.

[0079] In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the variable R1 is selected from C1-C6 alkoxy. In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the C1-C6 alkoxy of R1 is optionally substituted with 1 to 3 groups independently selected from halogen.

[0080] In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the variable R1 is selected from C3-C6 cycloalkyl. In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the variable R1 is C3 cycloalkyl. In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the C3-C6 cycloalkyl of R1 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), and -C(=O)N(C1-C4 alkyl)2. In some embodiments of Formula I (including the embodiments discussed above that define the variables X1 and X2), the C3-C6 cycloalkyl of R1 is optionally substituted with 1 to 3 groups independently selected from halogen. In some embodiments of Formula I (including the embodiments discussed above that define the variables X1 and X2), the C3-C6 cycloalkyl of R1 is substituted with 1 halogen. In some embodiments of Formula I (including the embodiments discussed above that define the variables X1 and X2), the C3-C6 cycloalkyl of R1 is substituted with 2 halogens. In some embodiments of Formula I (including the embodiments discussed above that define the variables X1 and X2), the C3-C6 cycloalkyl of R1 is substituted with 3 halogens. In some embodiments of Formula I (including the embodiments discussed above that define the variables X1 and X2), R1 is a C4 cycloalkyl substituted with 2 F. In some embodiments of Formula I (including the embodiments discussed above that define the variables X1 and X2), R1 is [ka] It is.

[0081] In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), R1 is selected from phenyl. In some embodiments of Formula I (including those discussed above defining the variables X1 and X2), the phenyl of R1 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), and -C(=O)N(C1-C4 alkyl)2.

[0082] In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the variable R2a is selected from hydrogen, halogen, cyano, -OH, oxo, and C1-C6 alkyl, and the C1-C6 alkyl of R2a is optionally substituted with 1-3 groups independently selected from halogen, cyano, -OH, and C1-C4 alkoxy. In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the variable R2a is hydrogen. In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the variable R2a is selected from C1-C6 alkyl. In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the C1-C6 alkyl of R2a is optionally substituted with 1-3 groups independently selected from halogen, cyano, -OH, and C1-C4 alkoxy. In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the C1-C6 alkyl of R2a is substituted with 1-3 -OH. In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the variable R2a is -CH2OH. In some embodiments of formula I (including those discussed above for defining the variables X1, X2, and R1), the variable R2a is -CHOHCH3.

[0083] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, and R2a), the variable R2b is selected from hydrogen, halogen, cyano, -OH, oxo, and C1-C6 alkyl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, and R2a), the variable R2b is hydrogen.

[0084] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, and R2b), each variable R3a is independently selected from halogen, cyano, -OH, C1-C6 alkyl (optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH), C1-C6 alkoxy, and oxo.

[0085] In some embodiments of formula I (including those embodiments discussed above that define the variables X1, X2, R1, R2a, and R2b), the variable R3a is -OH.

[0086] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, and R2b), when two variables R3a form an oxo, R3b is not oxo. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, and R2b), when two variables R3b form an oxo, R3a is not oxo.

[0087] In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, and R2b), each variable R3a is independently selected from C1-C6 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, and R2b), the variable R3a is C1 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, and R2b), the variable R3a is -CH3. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, and R2b), the C1-C6 alkyl of R3a is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH. In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, and R2b), the C1-C6 alkyl of R3a is optionally substituted with 1 to 3 groups independently selected from halogen. In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, and R2b), the variable R3a is -CHCF2.

[0088] In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, and R2b), each variable R3a is independently selected from C1-C6 alkoxy. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, and R2b), the variable R3a is -OCH3.

[0089] In some embodiments of formula I (including those embodiments discussed above defining the variables X1, X2, R1, R2a, and R2b), the variable R3a is oxo.

[0090] In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, and R3a), the variable R3b is selected from C1-C2 alkyl and oxo. In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, and R3a), the C1-C2 alkyl of R3b is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH.

[0091] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, and R3b), one of R4 and R5 is hydrogen and the other is C1-C6 alkyl, -C(=O)NH2, -C(=O)O(C1-C4 alkyl), C2-C6 alkynyl, and [ka] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, and R3b), the C1-C6 alkyl of R4 or R5 is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl), C1-C4 alkoxy, -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), -C(=O)N(C1-C4 alkyl), C3-C6 cycloalkyl, 5-10 membered heterocyclyl, phenyl, and 5-10 membered heteroaryl.

[0092] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, and R3b), the variable R4 is hydrogen and the variable R5 is [ka] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, and R3b), the variable R4 is selected from: [ka] and the variable R5 is hydrogen.

[0093] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from C3-C12 cycloalkyl, 3-12 membered heterocyclyl, C6 and C10 aryl, and 5-10 membered heteroaryl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is (optionally substituted with 1, 2, 3, 4, or 5 Ra groups).

[0094] In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from C3-C12 cycloalkyl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from C3 cycloalkyl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from C4 cycloalkyl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable ring A is [ka] is selected from.

[0095] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from C6 aryl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from C6 aryl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). [ka] is selected from

[0096] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from 5-10 membered heteroaryl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from 5 membered heteroaryl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), the variable ring A is selected from 6 membered heteroaryl (optionally substituted with 1, 2, 3, 4, or 5 Ra groups). In some embodiments of formula I (including those embodiments discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, and R5), variable ring A is [ka] is selected from.

[0097] In some embodiments of Formula I (including the embodiments described above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is independently for each occurrence selected from the group consisting of halogen, cyano, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkenyl, C1-C6 haloalkoxy, -C(=O)NRhRi, -NRhRi, -NRhC(=O)Rk, -NRhC(=O)ORk, -NRhC(=O)NRiRj, -NRhS(=O)pRk, -ORk, -OC(=O)Rk, -OC(=O)ORk, -OC(=O)NRhRi, -[O(CH2)q]rO(C1-C6 alkyl), -S(=O)pRk, -S(=O)pNRhRi, -C(=O)ORk, C3-C12 cycloalkyl, 3-12 membered heterocyclyl, C6 and C10 aryl, and 5-10 membered heteroaryl.

[0098] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is selected from halogen. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is F.

[0099] In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable Ra is selected from C1-C6 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable Ra is C1 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable Ra is -CH3. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable Ra is C2 alkyl.

[0100] In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable Ra is selected from -C(=O)NRhRi. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variables Rh and Ri are each independently selected for each occurrence from hydrogen and C1-C4 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variables Rh and Ri are each hydrogen. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variables Rh and Ri are independently selected from C1-C4 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), one of the variables Rh and Ri is hydrogen and the other is C1-C4 alkyl. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), one of the variables Rh and Ri is hydrogen and the other is -CH3. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variables Rh and Ri are each -CH3.

[0101] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is selected from -ORk. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is independently selected for each occurrence from hydrogen, C1-C4 alkyl, 5-10 membered heterocyclyl, and C3-C6 carbocycle, and the C1-C4 alkyl of any one of Rk is optionally substituted with 1-3 groups independently selected from halogen, cyano, and -OH. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is hydrogen. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is -CH3.

[0102] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is selected from 3-12 membered heterocyclyl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is selected from: [ka] It is.

[0103] In some embodiments of formula I (including the embodiments discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variable Ra is selected from C6 aryl.

[0104] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is selected from 5-10 membered heteroaryl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Ra is [ka] is selected from.

[0105] In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C2-C6 alkenyl of Ra are each optionally substituted with 1-3 groups independently selected from C6-C10 aryl (optionally substituted with 1-3 Rm groups), 5-10 membered heterocyclyl (optionally substituted with 1-3 Rm groups), 5-10 membered heteroaryl (optionally substituted with 1-3 Rm groups), cyano, -C(=O)Rk, -C(=O)ORk, -C(=O)NRhRi, -NRhRi, -NRhC(=O)Rk, -NRhC(=O)ORk, -NRh Optionally substituted with 1 to 3 groups selected from C(=O)NRiRj, -NRhS(=O)pRk, -ORk, -OC(=O)Rk, -OC(=O)ORk, -OC(=O)NRhRi, -S(=O)pRk, -S(=O)pNRhRi, and C3-C6 cycloalkyl (optionally substituted with 1 to 3 Rm groups).

[0106] In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), each of the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C2-C6 alkenyl of Ra is optionally substituted with 1 to 3 groups independently selected from -ORk. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is hydrogen. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is -CH3.

[0107] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), each of the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C2-C6 alkenyl of Ra is optionally substituted with 1-3 groups independently selected from -S(=O)pRk. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable p is 2 and the variable Rk is -CH3.

[0108] In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6, and C10 aryl, and the 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1-3 groups independently selected from halogen, cyano, C1-C4 alkyl, -C(=O)NRhRi, -NRhRi, -ORk, and oxo.

[0109] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6, and C10 aryl, and 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1-3 groups independently selected from halogen. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6, and C10 aryl, and 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1-3 F.

[0110] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6, and C10 aryl, and 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1 to 3 groups independently selected from C1-C4 alkyl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6, and C10 aryl, and 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1 to 3 -CH3 groups.

[0111] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6, and C10 aryl, and the 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1 to 3 groups independently selected from -C(=O)NRhRi. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variables Rh and Ri are each independently selected for each occurrence from hydrogen and C1-C4 alkyl. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variables Rh and Ri are each hydrogen. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), the variables Rh and Ri are independently selected from C1-C4 alkyl. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and ring A), one of the variables Rh and Ri is hydrogen and the other is C1-C4 alkyl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), one of the variables Rh and Ri is hydrogen and the other is -CH3. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), each of the variables Rh and Ri is -CH3.

[0112] In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3-12 membered heterocyclyl, C6, and C10 aryl, and 5-10 membered heteroaryl of Ra are each optionally substituted with 1-3 groups independently selected from -ORk. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is hydrogen. In some embodiments of formula I (including the embodiments discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variable Rk is -CH3.

[0113] In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C3-C12 cycloalkyl, 3- to 12-membered heterocyclyl, C6 and C10 aryl, and 5- to 10-membered heteroaryl of Ra are each optionally substituted with 1-3 oxo.

[0114] In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variables Rh, Ri, and Rj, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, C6-C10 aryl, and C3-C6 cycloalkyl. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the variables Rh, Ri, and Rj, for each occurrence, are each independently selected from hydrogen and C1-C4 alkyl. In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, and Ring A), the C1-C4 alkyl of any one of Rh, Ri, and Rj is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH.

[0115] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, and Rj), the variable Rk, for each occurrence, is independently selected from hydrogen, C1-C4 alkyl, 5-10 membered heterocyclyl, and C3-C6 cycloalkyl. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, and Rj), the variable Rk, for each occurrence, is hydrogen. In some embodiments of Formula I (including those described above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, and Rj), the variable Rk is, for each occurrence, independently selected from C1-C4 alkyl. In some embodiments of Formula I (including those described above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, and Rj), the C1-C4 alkyl of any one of Rk is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH.

[0116] In some embodiments of Formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, Rj, and Rk), the variable Rm, for each occurrence, is independently selected from halogen, cyano, oxo, C1-C6 alkyl, C1-C6 alkoxy, -S(=O)pRk, and ORk. In some embodiments, the C1-C6 alkyl of Rm is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH.

[0117] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, Rj, Rk, and Rm), the variable k is an integer selected from 0, 1, and 2. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, Rj, Rk, and Rm), when R3a is oxo, k is 1.

[0118] In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, ring A, Rh, Ri, Rj, Rk, Rm, and k), the variable m is an integer selected from 0, 1, and 2. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, ring A, Rh, Ri, Rj, Rk, Rm, and k), the variable m is 0. In some embodiments of formula I (including those discussed above that define the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, ring A, Rh, Ri, Rj, Rk, Rm, and k), when R3b is oxo, m is 1.

[0119] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, Rj, Rk, Rm, and m), the variable p, for each occurrence, is an integer selected from 1 and 2. In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, Ring A, Rh, Ri, Rj, Rk, Rm, and m), the variable p is 2.

[0120] In some embodiments of formula I (including those discussed above defining the variables X1, X2, R1, R2a, R2b, R3a, R3b, R4, R5, ring A, Rh, Ri, Rj, Rk, Rm, m, and p), the variables q and r are integers independently selected from 1, 2, 3, and 4 for each occurrence.

[0121] In some embodiments, at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt of the present disclosure is selected from compounds 1-78 shown in Table 1, tautomers thereof, deuterated derivatives of those compounds or tautomers, or pharma- ceutically acceptable salts of any of the foregoing. [ka] ) depicts a bond between two atoms and indicates the location of mixed stereochemistry for a collection of molecules such as a racemic mixture, cis / trans isomers, or (E) / (Z) isomers. An asterisk (e.g., [ka] ) indicates a chiral position in a molecule.

[0122] In some embodiments of the invention, the compound of formula I is selected from the compounds shown in Table 1 below, tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

Table 1-6

Table 1-7

Table 1-8

Table 1-9

[0123] Some embodiments of the present disclosure include derivatives of compounds 1-78, or compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, or pharmaceutically acceptable salts of any of the foregoing. In some embodiments, the derivative is a silicon derivative in which at least one carbon atom in the compounds, tautomers, deuterated derivatives, or pharmaceutically acceptable salts selected from compounds 1-78, or compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharmaceutically acceptable salts of any of the foregoing is replaced by silicon. In some embodiments, the derivative is a boron derivative in which at least one carbon atom in compounds 1-78 or a compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of such compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is replaced by boron. In other embodiments, the derivative is a phosphorus derivative in which at least one carbon atom in compounds 1-78 or a compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of such compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is replaced by phosphorus.

[0124] In some embodiments, the derivative is a silicon derivative in which one carbon atom in the compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, or compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is replaced by silicon or a silicon derivative (e.g., -Si(CH3)2- or -Si(OH)2-). The carbon replaced by silicon may be a non-aromatic carbon. In other embodiments, fluorine is replaced by a silicon derivative (e.g., -Si(CH3)3). In some embodiments, the silicon derivatives of the present invention may include one or more hydrogen atoms replaced by deuterium. In some embodiments, the silicon derivatives of compounds 1-78, or compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of the compounds or tautomers, or pharma- ceutically acceptable salts of any of the foregoing, may have silicon incorporated into a heterocycle.

[0125] In some embodiments, the derivative is a boron derivative in which one carbon atom in a compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, or compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is replaced by a boron derivative or a boron derivative.

[0126] In some embodiments, the derivative is a phosphorus derivative in which one carbon atom in a compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, or compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing, is replaced by phosphorus or a phosphorus derivative.

[0127] Another aspect of the disclosure provides a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of formulas selected from Formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, and compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt selected from Formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered to a patient in need thereof.

[0128] The pharmaceutical composition may further comprise at least one pharma- ceutically acceptable carrier. In some embodiments, the at least one pharma- ceutically acceptable carrier is selected from a pharma- ceutically acceptable vehicle and a pharma- ceutically acceptable adjuvant. In some embodiments, the at least one pharma- ceutically acceptable is selected from a pharma- ceutically acceptable filler, a disintegrant, a surfactant, a binder, and a lubricant.

[0129] It will also be understood that the pharmaceutical compositions of the present disclosure may be employed in combination therapy, i.e., the pharmaceutical compositions described herein may further comprise at least one additional active therapeutic agent. Alternatively, a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt selected from formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, a tautomer, a deuterated derivative of the compound or tautomer, and a pharmaceutically acceptable salt of any of the foregoing, may be administered as a separate composition simultaneously with, prior to, or after a composition comprising at least one other active therapeutic agent. In some embodiments, a pharmaceutical composition comprising at least one compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt selected from compounds 1-78, a tautomer, a deuterated derivative of the compound or tautomer, and a pharmaceutically acceptable salt of any of the foregoing, may be administered as a separate composition simultaneously with, prior to, or after a composition comprising at least one other active therapeutic agent.

[0130] As mentioned above, the pharmaceutical composition disclosed herein may optionally further comprise at least one pharma- ceutically acceptable carrier. The at least one pharma- ceutically acceptable carrier may be selected from adjuvants and vehicles. As used herein, at least one pharma- ceutically acceptable carrier includes any solvent, diluent, other liquid vehicle, dispersing aid, suspending aid, surfactant, isotonicity agent, thickener, emulsifier, preservative, solid binder, and lubricant suitable for the specific dosage form desired. Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. DB Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JC Boylan, 1988 to 1999, Marcel Dekker, New York disclose various carriers used in the formulation of pharmaceutical compositions and known techniques for their preparation. Except to the extent that any conventional carrier is incompatible with the compounds of the present disclosure, such as by producing any undesirable biological effects or otherwise interacting in a deleterious manner with any other components of the pharmaceutical composition, its use is contemplated within the scope of the present disclosure.Non-limiting examples of suitable pharma- ceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates, glycine, sorbic acid, and potassium sorbate), saturated vegetable fatty acids, partial glyceride mixtures of water, salts, and electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars (such as lactose, glucose, and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as carboxymethylcellulose sodium, cellulose acetate, cellulose acetate esters ... Examples of suitable excipients include, but are not limited to, cellulose acetate, ethylcellulose, ethylcellulose and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository wax), oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil), glycols (such as propylene glycol and polyethylene glycol), esters (such as ethyl oleate and ethyl laurate), agar, buffers (such as magnesium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffer, non-toxic compatible lubricants (such as sodium lauryl sulfate and magnesium stearate), colorants, release agents, coating agents, sweeteners, flavors, fragrances, preservatives, and antioxidants.

[0131] Uses of the Compounds and Compositions In some embodiments of the present disclosure, the compounds and pharmaceutical compositions described herein are used to treat FSGS and / or NDKD. In some embodiments, FSGS is mediated by APOL1. In some embodiments, NDKD is mediated by APOL1.

[0132] In some embodiments of the present disclosure, the compounds and pharmaceutical compositions described herein are used to treat cancer. In some embodiments, the cancer is mediated by APOL1.

[0133] In some embodiments of the present disclosure, the compounds and pharmaceutical compositions described herein are used to treat pancreatic cancer, which in some embodiments is mediated by APOL1.

[0134] In some embodiments, the disclosed method comprises administering to a patient in need thereof at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing. In some embodiments, the compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt is selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. In some embodiments, the patient in need thereof carries APOL1 gene variants, i.e., G1;S342G:I384M solution, and G2:N388del:Y389del.

[0135] Another aspect of the present disclosure provides a method of inhibiting APOL1 activity comprising contacting APOL1 with at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. In some embodiments, the method of inhibiting APOL1 activity comprises contacting the APOL1 with at least one compound, tautomer, deuterated derivative, or pharma- ceutically acceptable salt selected from compounds 1-78, tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharma- ceutically acceptable salts of any of the foregoing. EXAMPLES

[0136] In order that the disclosure set forth herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only, and should not be construed as limiting the disclosure in any manner.

[0137] The compounds of the invention can be made according to standard chemical practices or as described herein. The following abbreviations are used throughout the synthetic schemes below and in the descriptions for preparing the compounds of formula I, IIa, IIb, IIc, IId, IIIa, IIIb, IIIc, and IIId, compounds 1-78, tautomers thereof, deuterated derivatives of the compounds and tautomers, and pharma- ceutically acceptable salts of any of the foregoing:

[0138] Abbreviation AcOH = acetic acid ARP = Assay Ready Plates Boc2O = di-tert-butyl dicarbonate CBS = Corey Bakshi Shibata CDMT = 2-chloro-4,6-dimethoxy-1,3,5-triazine Co(OAc)2 = Cobalt(II) acetate DCM = dichloromethane DIBAL-H = diisobutylaluminum hydride DIPEA = N,N-diisopropylethylamine or N-ethyl-N-isopropyl-propan-2-amine DMAP = dimethylaminopyridine DME = dimethoxyethane DMEM = Dulbecco's modified Eagle's medium DMF = Dimethylformamide DMPU = N,N'-Dimethylpropyleneurea DMSO = dimethyl sulfoxide ESI-MS = electrospray ionization mass spectrometry EtOAc = ethyl acetate EtOH = ethanol FBS = fetal bovine serum GCMS = Gas Chromatography Mass Spectrometry HPLC = High Performance Liquid Chromatography IPA = Isopropyl alcohol [Ir{dF(CF3)ppy}2(dtbbpy)]PF6 = [4,4'-bis(1,1-dimethylethyl)-2,2'-bipyridine-N1,N1']bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]iridium(III) hexafluorophosphate LED = Light Emitting Diode LiTMP = lithium tetramethylpiperidide MeCN or ACN = acetonitrile MeI = methyl iodide MeMgBr = methylmagnesium bromide MeMgCl = methylmagnesium chloride MeOAc = methyl acetate MeOH = methanol MsOH = methanesulfonic acid MTBE = methyl tert-butyl ether n-BuLi = n-butyl lithium NBS = n-bromosuccinimide NHPI = N-hydroxyphthalimide NIS=N-iodosuccinimide NMR=nuclear magnetic resonance Pd(dppf)2Cl2 = [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) PP = Polypropylene PPh3 = Triphenylphosphine PTSA = p-toluenesulfonic acid monohydrate SFC = Supercritical Fluid Chromatography TBAF = Tetra-n-butylammonium fluoride TBS = tert-butyldimethylsilyl TEA = triethylamine Tet = tetracycline TFA or TFAA = trifluoroacetic acid TfOH = triflic acid THF = tetrahydrofuran 2-MeTHF = 2-methyltetrahydrofuran TLC = Thin Layer Chromatography TMS = tetramethylsilane TMSCF2Br = (bromodifluoromethyl)trimethylsilane TMSCl = trimethylsilyl chloride EXAMPLES

[0139] Compound synthesis All specific and generic compounds, as well as intermediates disclosed for making those compounds, are considered to be part of the disclosure disclosed herein.

[0140] Preparation S1 2-(3-Thienyl)ethanol (S1) [ka]

[0141] 2-(3-Thienyl)ethanol (S1) was obtained commercially.

[0142] Preparation S2 2-(5-chloro-3-thienyl)ethanol (S2) [ka]

[0143] Step 1. Synthesis of tert-butyl-dimethyl-[2-(3-thienyl)ethoxy]silane (C1) To a solution of 2-(3-thienyl)ethanol (18 g, 140.4 mmol) in DMF (100 mL) were added sequentially imidazole (12 g, 176.3 mmol) and tert-butyl-chloro-dimethyl-silane (24 g, 159.2 mmol). An exotherm was observed. The reaction mixture was stirred at room temperature for 3 h. The reaction was stopped at 90% conversion. The reaction was diluted with MTBE (500 mL) and washed with water (200 mL), 0.5 M HCl (200 mL), water (200 mL), and brine (200 mL). The organic layer was dried, filtered, and concentrated under vacuum. The organic layer was dissolved in heptane and passed through a silica gel plug, which was washed with 1-5% MTBE / heptane. The solvent was removed to give tert-butyl-dimethyl-[2-(3-thienyl)ethoxy]silane C1 (34 g, 99%). 1H NMR (400 MHz, chloroform-d) δ 7.28-7.13 (m, 1H), 7.04-6.91 (m, 2H), 3.80 (t, J = 6.9 Hz, 2H), 2.90-2.75 (m, 2H), 0.88 (s, 9H), -0.00 (s, 6H).

[0144] Step 2. Synthesis of tert-butyl-[2-(5-chloro-3-thienyl)ethoxy]-dimethyl-silane (C2) To a solution of 2,2,6,6-tetramethylpiperidine (36 mL, 213.3 mmol) in tetrahydrofuran (200 mL) cooled to 0° C. was added a solution of hexyllithium (92 mL of 2.3 M, 211.6 mmol). The reaction was stirred at −78° C. for 30 minutes. A solution of tert-butyl-dimethyl-[2-(3-thienyl)ethoxy]silane (34 g, 138.8 mmol) in THF (150 mL) was added to the reaction over 20 minutes. The reaction was stirred at −30° C. for 45 minutes. The reaction was cooled to −78° C. and 1,1,1,2,2,2-hexachloroethane (54 g, 228.1 mmol) was added in small portions. The reaction was allowed to warm to room temperature and stirred overnight. The reaction was quenched with saturated ammonium chloride (125 mL), diluted with water (100 mL), extracted with EtOAc (500 mL), and back-extracted with EtOAc (100 mL). The combined organic layers were washed with 0.5 M HCl (200 mL), water (300 mL), and brine (200 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give the crude product (C2).

[0145] Step 3. Synthesis of 2-(5-chloro-3-thienyl)ethanol (S2) To a solution of tert-butyl-[2-(5-chloro-3-thienyl)ethoxy]-dimethyl-silane C2 (12.5 g, 42.89 mmol) in 2-Me-THF (120 mL) was added TBAF (63 mL of a 1 M solution, 63.00 mmol) in THF. The reaction was stirred at room temperature overnight. The reaction was partitioned between EtOAc (400 mL) and water (400 mL). The layers were separated and the organic layer was extracted with EtOAc (200 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-50% EtOAc in heptane) gave the product 2-(5-chloro-3-thienyl)ethanol S2 (4.5 g, 58%). 1H NMR (300 MHz, chloroform-d) δ 6.82 (d, J=0.9 Hz, 2H), 3.89-3.71 (m, 2H), 2.79 (t, J=6.4 Hz, 2H), 2.05 (s, 1H). LCMS m / z 162.91 [M+H]+.

[0146] Preparation S3 2-[5-(trifluoromethyl)-3-thienyl]ethanol (S3) [ka]

[0147] Step 1. Synthesis of 2-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]tetrahydropyran (C5) To a mixture of 4-bromo-2-(trifluoromethyl)thiophene (C3) (9 g, 38.96 mmol), dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane; methanesulfonate; N-methyl-2-phenyl-aniline; palladium(2+) (1.8 g, 2.117 mmol), and potassium trifluoro(2-tetrahydropyran-2-yloxyethyl)boranide C4 (10 g, 42.36 mmol) was added toluene (75 mL) and water (25 mL). The top of the reaction was sparged with nitrogen, and then Cs2CO3 (40 g, 122.8 mmol) was added. A reflux condenser was added and the reaction was heated at 100° C. for 48 hours. The reaction was diluted with EtOAc (150 ml) and water (100 ml). The two layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-20% EtOAc in heptane) afforded the product 2-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]tetrahydropyran C5 (9 g, 82%). 1H NMR (300MHz, chloroform-d)δ 7.37(t,J=1.3Hz,1H),7.22(d,J=1.5Hz,1H),4.62(dd,J=4.2,2.8Hz,1H),3.96(dt,J=9.6,6.7Hz,1H),3.75(ddd,J=11.3,8.0,3.4Hz,1H),3.62(d t,J=9.6,6.5Hz,1H),3.55-3.41(m,1H),2.93(t,J=6.6Hz,2H),1.83(ddd ,J=14.2,6.6,3.4Hz,1H),1.73(td,J=9.0,4.2Hz,1H),1.66-1.50(m,4H).

[0148] Step 2. Synthesis of 2-[5-(trifluoromethyl)-3-thienyl]ethanol (S3) To a stirred solution of 2-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]tetrahydropyran C5 (1.8 g, 6.100 mmol) in MeOH (25 mL) was added 4-methylbenzenesulfonic acid (water (1)) (1.2 g, 6.309 mmol) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with water (100 mL) and extracted with MTBE (2×100 mL). The combined organic layers were washed with dilute NaHCO3 (10 mL NaHCO3 and 10 mL water) and brine (10 mL), dried over sodium sulfate, filtered, and evaporated under vacuum to give the crude compound. Purification by silica gel chromatography (gradient: 0-30% EtOAc in heptane) afforded the product 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (820 mg, 69%). 1H NMR (400MHz, chloroform-d)δ 7.35(p,J=1.3Hz,1H),7.23(dt,J=1.7,0.9Hz,1H),3.85(td,J=7.1,6.5,2.7Hz,2H),2.87(td,J=6.4,0.8Hz,2H),2.06(d,J=4.3Hz,1H).

[0149] Preparation S4 2-(5-Bromo-3-thienyl)ethanol (S4) [ka]

[0150] Step 1. Synthesis of 5-bromothiophene-3-carbaldehyde (C7) To a stirred solution of thiophene-3-carbaldehyde C6 (50 g, 40.717 mL, 0.4458 mol) in DMF (500 mL) was added NBS (119.02 g, 0.6687 mol) at 0 °C. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with ice-cold water (600 mL) and extracted with EtOAc (2 x 600 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated. Purification by silica gel chromatography (gradient: 0-2% EtOAc in petroleum ether) afforded the product 5-bromothiophene-3-carbaldehyde C7 (39.2 g, 44%). 1H NMR (400 MHz, chloroform-d) δ 9.77 (s, 1H), 7.99 (d, J = 1.2 Hz, 1H), 7.505 (d, J = 1.6 Hz, 1H).

[0151] Step 2. Synthesis of 2-bromo-4-[(E)-2-methoxyvinyl]thiophene (C8) To a stirred solution of (methoxymethyl)triphenylphosphonium chloride (115.1 g, 0.3358 mol) in diethyl ether (450.00 mL) at 0° C. was added potassium tert-butoxide (1 M in THF) (381 mL of a 1 M solution, 0.3810 mol) dropwise. The reaction was stirred at 0° C. for 1 h. A solution of 5-bromothiophene-3-carbaldehyde C7 (45 g, 0.2215 mol) in diethyl ether (90 mL) was added and the reaction mixture was then stirred at room temperature for 30 min. The reaction mixture was quenched with NH4Cl solution (900 mL) at 0° C. and extracted with EtOAc (2×700 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated. Purification by silica gel chromatography (eluent: petroleum ether) afforded the product 2-bromo-4-[(E)-2-methoxyvinyl]thiophene C8 (44.1 g, 82%). 1H NMR (400 MHz, chloroform-d) δ 7.25 (d, J = 2 Hz, 1H), 7.18 (d, J = 0.8 Hz, 1H), 7.00 (d, J = 1.8 Hz, 1H), 6.91 (d, J = 12.8 Hz, 1H), 6.97 (d, J = 1.2 Hz, 1H), 6.05 (d, J = 6.8 Hz, 1H), 5.72 (d, J = 12.8 Hz, 1H), 5.22 (d, J = 6.4 Hz, 1H), 3.77 (d, J = 2.8 Hz, 3H), 3.64 (d, J = 5.2 Hz, 3H). NMR indicated a 1:1 mixture of E and Z isomers.

[0152] Step 3. Synthesis of 2-(5-bromo-3-thienyl)acetaldehyde (C9) To a stirred solution of 2-bromo-4-[(E)-2-methoxyvinyl]thiophene C8 (14.1 g, 0.0602 mol) in 1,4-dioxane (141.00 mL) was added HCl (4 M in dioxane) (60.200 mL of a 4 M solution, 0.2408 mol) at 0° C. The reaction mixture was stirred at room temperature for 30 min. The reaction was quenched with saturated NaHCO3 solution at 0° C. and extracted with EtOAc. The organic layer was dried over Na2SO4, filtered and concentrated to give 2-(5-bromo-3-thienyl)acetaldehyde C9 (13.1 g, 89%). 1H NMR (400MHz, chloroform-d) δ 9.72 (t, J = 2.4 Hz, 1H), 7.04 (s, 1H), 6.94 (d, J = 1.2 Hz, 1H), 3.66 (d, J = 1.6 Hz, 2H).

[0153] Step 4. Synthesis of 2-(5-bromo-3-thienyl)ethanol (S4) To a stirred solution of 2-(5-bromo-3-thienyl)acetaldehyde C9 (38.5 g, 0.1524 mol) in MeOH (390 mL) was added NaBH4 (13.3 g, 0.3515 mol) at 0 °C. The reaction was stirred for 1 h. The reaction mixture was quenched with ice water (400 mL) and concentrated in vacuo to remove MeOH. The residue was diluted with water (500 mL) and extracted with EtOAc (3 x 300 mL). The separated organic layer was dried over Na2SO4, filtered and concentrated. Purification by column chromatography on neutral alumina (eluent: 35% EtOAc in petroleum ether) gave the product 2-(5-bromo-3-thienyl)ethanol S4 (30.2 g, 84%) 1H NMR (300 MHz, DMSO-d6) δ 7.20 (t, J = 0.9 Hz, 1H), 7.10 (d, J = 1.2 Hz, 1H), 4.64 (q, J = 5.2 Hz, 1H), 3.59-3.55 (m, 2H), 2.67 (t, J = 6.8 Hz, 2H) as a pale yellow liquid.

[0154] Preparation S5 2-(5-Ethyl-3-thienyl)ethanol (S5) [ka]

[0155] Step 1. Synthesis of 2-[2-(5-bromo-3-thienyl)ethoxy]tetrahydropyran (C10) To a stirred solution of 2-(5-bromo-3-thienyl)ethanol S4 (8 g, 0.0328 mol) in THF (80 mL) was added 3,4-dihydro-2H-pyran (3.7696 g, 3.8 mL, 0.0448 mol) and PTSA (259 mg, 0.0015 mol) at room temperature. The reaction mixture was then stirred at room temperature for 16 h. The reaction mixture was treated with saturated aqueous K2CO3 (300 mL) and extracted with EtOAc (2 x 600 mL). The organic layer was dried over Na2SO4, filtered, and concentrated. Purification by silica gel chromatography (gradient: 0-5% EtOAc in petroleum ether) afforded the product 2-[2-[5-bromo-3-thienyl]ethoxy]tetrahydropyran C10 (10.1 g, 90%). 1H NMR (400 MHz, chloroform-d) δ 6.95 (d, J = 1.6 Hz, 1H), 6.92 (d, J = 0.8, 1H), 4.59 (t, J = 2.8 Hz, 1H), 3.94-3.74 (m, 2H), 3.60-3.46 (m, 2H), 2.85 (q, J = 6.4 Hz, 2H), 1.80-1.61 (m, 6H). LCMS m / z 291.03 [M+H]+.

[0156] Step 6. Synthesis of 2-[2-(5-ethyl-3-thienyl)ethoxy]tetrahydropyran (C11) To a stirred solution of 2-[2-(5-bromo-3-thienyl)ethoxy]tetrahydropyran C10 (25 g, 0.0719 mol) in THF (250.00 mL) was added n-BuLi (2.5 M in hexanes) (46.1 mL of a 2.5 M solution, 0.1153 mol) at -76 °C. The reaction was stirred for 1 h. Ethyl iodide (24.832 g, 12.8 mL, 0.1592 mol) was added at -76 °C. The reaction temperature was then allowed to slowly rise to room temperature and then stirred for 16 h. The reaction mixture was quenched with NH4Cl solution (500 mL) and extracted with EtOAc (2 x 300 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated. Purification by silica gel chromatography (gradient: 0-3% EtOAc in petroleum ether) afforded the product 2-[2-(5-ethyl-3-thienyl)ethoxy]tetrahydropyran C11 (13.2 g, 59%). LCMS m / z 241.21 [M+H]+.

[0157] Step 7. Synthesis of 2-(5-ethyl-3-thienyl)ethanol (S5) To a stirred solution of 2-[2-(5-ethyl-3-thienyl)ethoxy]tetrahydropyran C11 (4.4 g, 0.0142 mol) in MeOH (44 mL) was added PTSA (3.0 g, 0.0174 mol) at room temperature and the reaction was stirred for 2 h. The reaction mixture was quenched with saturated aqueous NaHCO3 (150 mL), extracted with EtOAc (2 x 150 mL), dried over Na2SO4, filtered and concentrated. Purification by column chromatography using neutral alumina (eluent: 10% EtOAc in petroleum ether) gave the product 2-(5-ethyl-3-thienyl)ethanol S5 (1.1 g, 45%). 1H NMR(400MHz,DMSO-d6)δ 6.90(d,J=1.2Hz,1H),6.71(d,J=1.2Hz,1H),4.62-4.58(m,1H),3.59-3.55(m,2H),2.77-2.71(m,2H),2.64(t,J=7.2,2H),1.22-1.85(m,3H).

[0158] Preparation S6 2-(5-Ethyl-2-thienyl)ethanol (S6) [ka]

[0159] Step 1. Synthesis of 2-(5-ethyl-2-thienyl)ethanol (S6) To a solution of 2-ethylthiophene C12 (54 g, 466.9 mmol) in anhydrous THF (1 L) at 0 °C, n-BuLi in hexane (255 mL of a 2.2 M solution, 561.0 mmol) was added over 45 min. A pale yellow / orange solution was obtained. The temperature range during the addition was 0-10 °C. The mixture was stirred at room temperature for 30 min. After cooling to 0 °C, a solution of ethylene oxide (200 mL of a 2.9 M solution, 580.0 mmol) was added over 30 min. The reaction was stirred at 0 °C for 2 h and then warmed to room temperature. The reaction mixture was quenched with water (700 mL) and saturated NH4Cl (200 mL) and the THF was evaporated. The product was extracted with EtOAc (1 x 400 mL, 2 x 150 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The organic layer was passed through a silica gel plug, washing with DCM (1000 mL), 80% EtOAc / heptane (2×200 mL), and DCM (2×250 mL) to give 2-(5-ethyl-2-thienyl)ethanol S6 (71.25 g, 93%). 1H NMR (300 MHz, chloroform-d) δ 6.69 (dt, J=3.4, 0.9 Hz, 1H), 6.64 (dt, J=3.3, 1.0 Hz, 1H), 3.84 (t, J=6.3 Hz, 2H), 3.08-2.97 (m, 2H), 2.82 (qd, J=7.5, 1.0 Hz, 2H), 1.31 (t, J=7.5 Hz, 4H).

[0160] Preparation S7 2-[5-(trifluoromethyl)-2-thienyl]ethanol (S7) [ka]

[0161] Step 1. Synthesis of 2-(5-iodo-2-thienyl)ethanol (C14) To a stirred solution of NIS (104.83 g, 0.4680 mol) in DCM (1000 mL) was added 2-(2-thienyl)ethanol C13 (50 g, 0.3900 mol) at 0° C. The reaction was allowed to warm to room temperature and stirred for 16 h. The reaction mixture was diluted with DCM (500 mL), washed with saturated sodium thiosulfate, brine, dried over Na2SO4, and concentrated in vacuo. Purification by column chromatography (eluent: 20% EtOAc in petroleum ether) afforded the product 2-(5-iodo-2-thienyl)ethanol C14 (62 g, 56%). 1H NMR (400 MHz, chloroform-d) δ 7.08 (d, J=3.6 Hz, 1H), 6.57-6.56 (m, 1H), 3.82 (q, J=6 Hz, 2H), 3.05 (q, J=6.4 Hz, 2H). LCMS m / z 254.89 [M+H]+.

[0162] Step 2. Synthesis of 2-[2-(5-iodo-2-thienyl)ethoxy]tetrahydropyran (C15) To a stirred solution of 2-(5-iodo-2-thienyl)ethanol C14 (15 g, 0.0525 mol) and 3,4-dihydro-2H-pyran (6.6284 g, 0.0788 mol) in THF (60 mL) was added PTSA (1.3604 g, 1.2714 mL, 0.0079 mol) at room temperature. The reaction was stirred under pressure with an argon balloon for 16 hours. The reaction mixture was concentrated under reduced pressure. Purification by silica gel chromatography (eluent: 5% EtOAc in petroleum ether) gave the product 2-[2-(5-iodo-2-thienyl)ethoxy]tetrahydropyran C15 (12.8 g, 68%). 1H NMR(400MHz,DMSO-d6)δ 7.14(d,J=3.6Hz,1H),6.64(d,J=3.6Hz,1H),4.59(t,J=3.6Hz,1H),3.80-3.76(m,1H),3.74-3.67(m,1H),3.54 -3.50(m,1H),3.48-3.41(m,1H),3.03(t,J=6Hz,2H),1.75-1.69(m,1H),1.61-1.59(m,1H),1.51-1.42(m,4H).

[0163] Step 3. Synthesis of 2-[2-[5-(trifluoromethyl)-2-thienyl]ethoxy]tetrahydropyran (C16) To a stirred solution of 2-[2-(5-iodo-2-thienyl)ethoxy]tetrahydropyran C15 (10 g, 0.0219 mol) and 2,2-difluoro-2-fluorosulfonyl-methyl acetate (12.63 g, 0.0657 mol) in DMF (40 mL) was added copper(I) bromide dimethylsulfide complex 99% (2.241 g, 0.0109 mol). The reaction was stirred at 100° C. for 16 h. The reaction was allowed to warm to room temperature, diluted with EtOAc (100 mL), filtered, and washed with EtOAc (50 mL). The filtrate was washed with chilled brine, dried over Na2SO4, and concentrated under reduced pressure. Purification by column chromatography using neutral alumina (eluent: 5% EtOAc in petroleum ether) afforded the product C16 2-[2-[5-(trifluoromethyl)-2-thienyl]ethoxy]tetrahydropyran (2.9 g, 41%). 1H NMR (400MHz, chloroform-d)δ 7.25(s,1H),6.82-6.81(m,1H),4.63(t,J=3.6Hz,1H),4.00-3.95(m,1H),3.78-3.75(m,1H),3.64-3.58(m, 1H),3.51-3.48(m,1H),3.12(d,J=6.4Hz,2H),1.90-1.80(m,1H),1.73-1.64(m,1H),1.65-1.51(m,4H).GCMS m / z 280[M]+.

[0164] Step 4. Synthesis of 2-[5-(trifluoromethyl)-2-thienyl]ethanol (S7) To a stirred solution of 2-[2-[5-(trifluoromethyl)-2-thienyl]ethoxy]tetrahydropyran C16 (5.8 g, 0.0170 mol) in MeOH (100 mL) was added PTSA (2.93 g, 0.0170 mol) at room temperature. The reaction was stirred for 16 h. The reaction mixture was concentrated under reduced pressure. Purification by column chromatography using neutral alumina (eluent: 10% EtOAc in petroleum ether) gave the product 2-[5-(trifluoromethyl)-2-thienyl]ethanol S7 (2.3 g, 61%). 1H NMR(400MHz,DMSO-d6)δ 7.52-7.51(m,1H),6.99-6.98(m,1H),4.92(t,J=4.8Hz,1H),3.65-3.61(m,2H),2.98(t,J=6Hz,2H). 19F NMR(376.22MHz,DMSO-d6)δ -53.53(s,3F).GCMS m / z 196.0[M]+.

[0165] Preparation of S8 and S9 2-[5-(chloro)-2-thienyl]propan-1-ol (S8 [enantiomer 1], S9 [enantiomer 2]) [ka]

[0166] Step 1. Synthesis of ethyl 2-(5-chloro-2-thienyl)propanoate (C19) To a stirred solution of ethyl 2-(2-thienyl)propanoate C17 (1 g, 4.1139 mmol) in acetic acid (10 mL) was added N-chlorosuccinimide C18 (549.34 mg, 4.1139 mmol). The reaction mixture was stirred at 100° C. for 1 h. The mixture was concentrated and the resulting crude was diluted with EtOAc (25 mL) and washed with water (10 mL), saturated sodium bicarbonate solution (10 mL), saturated sodium thiosulfate solution (10 mL), and brine solution (10 mL). The organic phase was dried over Na2SO4, filtered, and concentrated to give the crude product. Purification by silica gel chromatography (eluent: 3% EtOAc in petroleum ether) gave the product ethyl 2-(5-chloro-2-thienyl)propanoate C19 (700 mg, 60%). 1H NMR (chloroform-d, 400MHz): δ=6.75-6.73(m,1H), 6.71-6.69(m,1H), 4.20-4.14(m,2H), 3.88-3.73(q,J=6.4Hz,1H), 1.55-1.53(t,J=2.8Hz,3H), 1.30-1.221(m,3H). GCMS m / z 218.0[M]+

[0167] Step 2. Synthesis of 2-(5-chloro-2-thienyl)propan-1-ol (C20) To a stirred solution of ethyl 2-(5-chloro-2-thienyl)propanoate C19 (25 g, 86.877 mmol) in THF (500 mL) was added DIBAL-H (74.135 mL of 25% w / v, 130.32 mmol) dropwise at 0° C. The reaction mixture was stirred at 0° C. for 2 h. The mixture was slowly quenched with saturated NH4Cl solution (300 mL) at 0° C., the suspension was filtered through Celite®, and the Celite® pad was washed with EtOAc (2×200 mL). The filtrate was separated into two layers. The aqueous layer was extracted with EtOAc (2×200 mL). The combined organic layers were washed with brine (200 mL), dried over sodium sulfate, and concentrated. Purification by silica gel chromatography (eluent: 3% EtOAc in petroleum ether) gave 2-(5-chloro-2-thienyl)propan-1-ol C20 (12 g, 72%). 1H NMR (chloroform-d, 400 MHz) δ 6.76-6.75 (d, J = 3.6 Hz, 1H), 6.66-6.65 (dd, J = 4.4 Hz, 1H), 3.71-3.61 (m, 2H), 3.15-3.10 (m, 1H), 1.57-1.52 (m, 1H), 1.34-1.31 (t, J = 6 Hz, 3H). GCMS m / z 176.0 [M]+.

[0168] Step 3. Synthesis of 2-(5-chloro-2-thienyl)propan-1-ol (S8) and (S9) Racemic 2-(5-chloro-2-thienyl)propan-1-ol C20 (12 g, 62.492 mmol) was separated into its constituent enantiomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 30×250 mm; Mobile phase: 10% methanol / hexane mixture (7:3), 90% carbon dioxide. Flow rate: 90 g / min. 2-(5-Chloro-2-thienyl)propan-1-ol S8 (4 g, 35%). 1H NMR (chloroform-d, 400 MHz) δ 6.76-6.75 (d, J = 3.6 Hz, 1H), 6.66-6.65 (dd, J = 3.6 Hz, 1H), 3.73-3.61 (m, 2H), 3.17-3.10 (m, 1H), 1.52-1.49 (t, J = 5.2 Hz, 1H), 1.32-1.30 (d, J = 6.8 Hz, 3H). GCMS: m / z: 176.0 [M] +.

[0169] 2-(5-Chloro-2-thienyl)propan-1-ol S9 (3.75 g, 34%) was obtained. 1H NMR (chloroform-d, 400 MHz) δ 6.76-6.75 (d, J = 4 Hz, 1H), 6.66-6.65 (dd, J = 3.6 Hz, 1H), 3.73-3.61 (m, 2H), 3.15-3.10 (q, J = 6.8 Hz, 1H), 1.51-1.48 (t, J = 5.6 Hz, 1H), 1.33-1.30 (d, J = 7.2 Hz, 3H). GCMS m / z 176.0 [M]+.

[0170] Preparation of S10 and S11 2-(5-Ethyl-2-thienyl)propan-1-ol (S10 Enantiomer 1) and (S11 Enantiomer 2) [ka]

[0171] Step 1. Synthesis of ethyl 2-(5-acetyl-2-thienyl)propanoate (C21) To a stirred solution of ethyl 2-(2-thienyl)propanoate C17 (80 g, 336.92 mmol) in DCM (1500 mL) was added dropwise acetyl chloride (39.671 g, 35.934 mL, 505.38 mmol) at 0 °C, followed by the addition of AlCl3 (67.388 g, 505.38 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. The mixture was slowly quenched with ice water (1000 mL), the two layers were separated, and the aqueous layer was extracted with DCM (2 × 500 mL). The combined organic layers were washed with brine (500 mL) and dried over sodium sulfate. Purification by silica gel chromatography (gradient: 0-5% EtOAc in petroleum ether) afforded the product ethyl 2-(5-acetyl-2-thienyl)propanoate C21 (60 g, 73%). 1H NMR (chloroform-d, 400 MHz) δ 7.56-7.54 (t, J = 4.0 Hz, 1H), 6.99-6.98 (m, 1H), 4.20-4.14 (m, 2H), 4.01-3.96 (q, J = 7.2 Hz, 1H), 2.52 (s, 3H), 1.60-1.56 (d, J = 7.2 Hz, 3H), 1.28-1.23 (m, 3H). LCMS m / z 227.1 [M+H]+.

[0172] Step 2. Synthesis of ethyl 2-(5-ethyl-2-thienyl)propanoate (C22) To a stirred solution of ethyl 2-(5-acetyl-2-thienyl)propanoate C21 (60 g, 245.79 mmol) in TFA (400 mL) was added triethylsilane (42.870 g, 58.9 mL, 368.69 mmol) dropwise at 0 °C. The reaction mixture was stirred at room temperature for 4 h. The reaction was concentrated, quenched with ice-cold water (500 mL), and extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, and concentrated to give the crude product. Purification by silica gel chromatography (gradient: 0-3% EtOAc in petroleum ether) afforded the product, ethyl 2-(5-ethyl-2-thienyl)propanoate C22 (50 g, 82%). 1H NMR (chloroform-d, 400 MHz) δ 6.73-6.72 (dd, J=3.6 Hz, 1H), 6.62-6.60 (m, 1H), 4.18-4.13 (m, 2H), 3.93-3.88 (q, J=7.2 Hz, 1H), 2.82-2.78 (m, 2H), 1.55-1.53 ​​(d, J=7.2 Hz, 3H) 1.30-1.23 (m, 6H). LCMS m / z 213.2 [M+H]+.

[0173] Step 3. Synthesis of 2-(5-ethyl-2-thienyl)propan-1-ol (C23) To a stirred solution of ethyl 2-(5-ethyl-2-thienyl)propanoate C22 (50 g, 200.18 mmol) in THF (1000 mL) was added DIBAL-H (25% in toluene) (227.75 mL of 25% w / v, 400.36 mmol) dropwise at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. The mixture was slowly quenched with saturated NH4Cl solution (500 mL) at 0 °C and extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, and concentrated. Purification by silica gel chromatography (gradient: 0-5% EtOAc in petroleum ether) afforded the product 2-(5-ethyl-2-thienyl)propan-1-ol C23 (31 g, 89%). 1H NMR (chloroform-d, 400 MHz): δ 6.69-6.68 (d, J=3.6 Hz, 1H), 6.64-6.62 (m, 1H), 3.72-3.60 (m, 2H), 3.18-3.13 (q, J=6.8 Hz, 1H), 2.83-2.77 (m, 2H), 1.61-1.5 (m, 1H), 1.35-1.28 (m, 6H). LCMS m / z 171.02 [M+H]+.

[0174] Step 4. Synthesis of 2-(5-ethyl-2-thienyl)propan-1-ol (S10) and (S11) Racemic 2-(5-ethyl-2-thienyl)propan-1-ol C23 (31 g, 178.06 mmol) was separated into its constituent enantiomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 30×250 mm; Mobile phase: 10% methanol / hexane mixture (7:3), 85% carbon dioxide. 2-(5-ethyl-2-thienyl)propan-1-ol S10 (13.45 g, 43%). 1H NMR (chloroform-d, 400 MHz): δ = 6.69-6.68 (d, J = 3.2 Hz, 1H), 6.63-6.62 (d, J = 3.2 Hz, 1H), 3.73-3.61 (m, 2H), 3.19-3.14 (q, J = 6.8 Hz, 1H), 2.83-2.78 (m, 2H), 1.54-1.47 (m, 1H), 1.35-1.27 (m, 6H). LCMS m / z 171.1 [M+H]+.

[0175] 2-(5-Ethyl-2-thienyl)propan-1-ol S11 (11.35 g, 37%) was obtained. 1H NMR (chloroform-d, 400 MHz): δ 6.68-6.67 (d, J = 3.6 Hz, 1H), 6.63 (d, J = 3.6 Hz, 1H), 3.73-3.61 (m, 2H), 3.20-3.12 (m, 1H), 2.83-2.77 (q, J = 7.6 Hz, 2H), 1.54-1.45 (m, 1H), 1.33-1.27 (m, 6H). LCMS m / z 171.1 [M+H]+.

[0176] Preparation S12 2-(5-Chloro-2-thienyl)ethanol (S12) [ka]

[0177] 2-(5-Chloro-2-thienyl)ethanol (S12) was obtained commercially.

[0178] Preparation S13 1-(2-Methylsulfonylethyl)pyrazole-4-carbaldehyde (S13) [ka]

[0179] Preparation of 1-(2-methylsulfonylethyl)pyrazole-4-carbaldehyde (S13) A solution of 1H-pyrazole-4-carbaldehyde C25 (10 g, 104.1 mmol), 11-methylsulfonylethylene C24 (10 mL, 114.2 mmol) and K2CO3 (25 g, 180.9 mmol) in THF (200 mL) was stirred at 60° C. After stirring overnight, the mixture was cooled to room temperature and concentrated to dryness. The product was suspended in diethyl ether (100 mL) to triturate the product and stirred for 2 h. The product was filtered and dried overnight to give 11-(2-methylsulfonylethyl)pyrazole-4-carbaldehyde S13 (20280 mg, 83%). 1H NMR(400MHz,DMSO-d6)δ 9.80(s,1H),8.54(d,J=0.7Hz,1H),8.05(d,J=0.7Hz,1H),4.64(t,J=6.8Hz,2H),3.80-3.67(m,2H),2.96(d,J=0.7Hz,3H).LCMS m / z 203.01[M+H]+.

[0180] Preparation S14 1-[2-[tert-Butyl(dimethyl)silyl]oxyethyl]pyrazole-4-carbaldehyde (S20) [ka]

[0181] Step 1. Synthesis of tert-butyl-(2-iodoethoxy)-dimethyl-silane (C27) To a stirred solution of 2-iodoethanol C26 (2 g, 0.0116 mol) and imidazole (1.58 g, 0.0232 mol) in DCM (40 mL) was added tert-butyl-chloro-dimethyl-silane (1.9 g, 0.0126 mol) at 0 °C. The reaction was allowed to warm to room temperature and stirred for 4 h. The reaction mixture was diluted with DCM (100 mL), washed with saturated NaHCO3 and brine, dried over Na2SO4, and concentrated under reduced pressure to give tert-butyl-(2-iodoethoxy)-dimethyl-silane C27 (2.5 g, 68%). 1H NMR (400 MHz, chloroform-d) δ 3.83 (t, J = 6.8 Hz, 2H), 3.20 (t, J = 6.8 Hz, 2H), 0.90 (s, 9H), 0.08 (s, 6H).

[0182] Step 2. Synthesis of 1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]pyrazole-4-carbaldehyde (S14) To a solution of 1H-pyrazole-4-carbaldehyde C25 (20 g, 208.1 mmol) and K2CO3 (115 g, 832.1 mmol) in MeCN (200 mL) was added tert-butyl-(2-iodoethoxy)-dimethyl-silane C27 (65 g, 227.1 mmol). The reaction was heated to 80° C. The reaction was stirred for 5 h. The reaction was cooled to 50° C. and stirred for 16 h. The reaction mixture was warmed, filtered, and the solids were washed with MeCN (200 mL). The solids were discarded. The filtrate was concentrated. The residue was partitioned between EtOAc (400 mL) and water (400 mL). The organic phase was separated, washed with water (400 mL) and brine (400 mL), dried over MgSO4, filtered, and concentrated. Purification by silica gel chromatography (800 g column, 0-80% EtOAc in hexanes) afforded the product 1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]pyrazole-4-carbaldehyde S14 (46 g, 87%) as a pale yellow oil. 1H NMR (300 MHz, chloroform-d) δ 9.86 (s, 1H), 7.98 (s, 2H), 4.25 (dd, J = 5.5, 4.5 Hz, 2H), 3.96 (dd, J = 5.5, 4.5 Hz, 2H), 0.83 (s, 9H), -0.06 (s, 6H). LCMS m / z 255.14 [M+H]+.

[0183] Preparation S15 1-Methyltriazole-4-carbaldehyde (S15) [ka] 1-Methyltriazole-4-carbaldehyde (S15) was obtained from a commercial source.

[0184] Preparation S16 (S)-2-Ethynyl-4-oxopiperidine-1-carboxylate tert-butyl (S16) [ka]

[0185] Step 1. Synthesis of tert-butyl 2-ethynyl-4-oxo-2,3-dihydropyridine-1-carboxylate (C29) To a solution of 4-methoxypyridine C28 (30.00 g, 274.91 mmol, 27.78 mL, 1.0 equiv) and Boc2O (66.00 g, 302.40 mmol, 69.47 mL, 1.1 equiv) in THF (500 mL) was added ethynylmagnesium bromide (0.5 M solution, 825 mL, 1.5 equiv) dropwise at 0 °C. The reaction was stirred at 25 °C for 3 h. TLC (petroleum ether:ethyl acetate = 5:1) showed that material A was consumed, then the reaction was quenched with aqueous HCl (1.5 L, 1 M) under a 0 °C bath. The mixture was stirred at 25 °C for 0.5 h and extracted with ethyl acetate (500 mL x 3). The organic layer was washed with brine (1 L x 2), dried over Na2SO4, filtered and concentrated to give a residue. Purification by silica gel chromatography (0–10% EtOAc in petroleum ether) afforded the product, tert-butyl 2-ethynyl-4-oxo-2,3-dihydropyridine-1-carboxylate C29 (38.00 g, 171.75 mmol, 62.47% yield) as a yellow solid. 1H NMR (400 MHz, chloroform-d) δ 7.72 (br s, 1H), 5.40 (d, J = 8.4 Hz, 1H), 5.31 (br s, 1H), 2.85 (dd, J = 6.4 Hz, J = 16.4 Hz, 1H), 2.61 (d, J = 16.4 Hz, 1H), 2.28 (d, J = 2.4 Hz, 1H), 1.56 (s, 9H).

[0186] Step 2. Synthesis of tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate (C30) To a solution of tert-butyl 2-ethynyl-4-oxo-2,3-dihydropyridine-1-carboxylate C29 (38.00 g, 171.75 mmol, 1.0 equiv) in AcOH (400 mL) was added Zn powder (82.21 g, 1.26 mol, 7.3 equiv) in several portions within 5 min at 25 °C. The reaction was stirred at 55 °C for 4 h under N2. TLC (petroleum ether:ethyl acetate = 5:1, KMnO4) showed a major new spot. The reaction was filtered and the cake was carefully washed with ethyl acetate (500 mL x 3). All the filtrates were concentrated in vacuum to give a residue. The residue was poured into 600 mL of ice water and extracted with ethyl acetate (500 mL x 3). The organic layers were combined, washed with saturated sodium bicarbonate solution (500 mL×3) and brine (500 mL×2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give the crude product, which was purified by silica gel column chromatography (petroleum ether / ethyl acetate=100:1 to 20:1) to give tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate C30 (30.00 g, 131.68 mmol, 76.67% yield) as a white solid. 1H NMR (400 MHz, chloroform-d) δ 5.44 (br s, 1H), 4.23 (d, J = 11.2 Hz, 1H), 3.57-3.49 (m, 1H), 2.70 (dd, J = 6.8 Hz, J = 14.4 Hz, 1H), 3.54-2.40 (m, 3H), 2.41 (d, J = 2.4 Hz, 1H), 1.50 (s, 9H). LCMS m / z 168.2 [M-55]+.

[0187] Step 3. Synthesis of (S)-tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate (S16) Racemic tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate C30 (50 g, 223.9 mmol) was separated into its constituent enantiomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 20×250 mm; Mobile phase: 40% methanol (containing 5 mM ammonia), 60% carbon dioxide. The second eluting peak was concentrated in vacuo to give (S)-tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate S16 (22.5 g, 89%). 1H NMR (chloroform-d, 300 MHz): δ 5.45 (s, 1H), 4.24 (d, J = 13.3 Hz, 1H), 3.54 (dt, J = 13.3, 8.3 Hz, 1H), 2.93-2.24 (m, 5H), 1.51 (s, 9H). Note: The stereochemistry of S16 was confirmed by synthesizing compound 2 by an alternative method. The data were convergent.

[0188] Preparation S17 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)piperidin-4-one (S17) [ka]

[0189] Step 1. Synthesis of 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)-2,3-dihydropyridin-4-one (C33) 1-Methyltriazole-4-carbaldehyde S15 (3.1 g, 27.90 mmol) in MeOH (60 mL) was treated with (2,4-dimethoxyphenyl)methanamine C31 (4.5 mL, 29.95 mmol) and stirred at room temperature until 1H NMR indicated complete imine formation. [(Z)-3-Methoxy-1-methylene-allyloxy]-trimethyl-silane C32 (10.5 mL, 53.93 mmol) was then added and the reaction was stirred at room temperature for 1 h. The reaction was quenched with 1 M HCl (8 mL) and stirred for 10 min. The mixture was then made basic with saturated sodium bicarbonate and extracted with DCM (4 x 150 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give the crude product. The crude product was purified by silica gel column chromatography (0 to 60% 20% MeOH / DCM in EtOAc) to give 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)-2,3-dihydropyridin-4-one C33 (5.66 g, 58%) as a yellow solid. 1H NMR (400MHz, chloroform-d)δ 7.54(s,1H),7.24(dd,J=7.7,0.8Hz,1H),7.09(dd,J=7.6,0.9Hz,1H),6.50(d,J=7.7Hz,2H),4.96(dd,J=7.5,1.1Hz,1H),4.93- 4.88(m,1H),4.59-4.35(m,2H),4.08(s,3H),3.85(s,6H),2.96(dd,J=16.3,7.4Hz,1H),2.51(ddd,J=16.3,2.5,1.2Hz,1H).LCMS m / z 329.1[M+H]+.

[0190] Step 2. Synthesis of 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)piperidin-4-one (S17) CuBr (360 mg, 2.510 mmol) in THF (35 mL) was cooled to -10 °C and lithium tri-tert-butoxyaluminum hydride (26 mL of a 1 M solution in THF, 26.00 mmol) was added slowly. The mixture was stirred at -10 °C for 45 min, resulting in a dark brown solution. This solution was then added slowly to a solution of 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)-2,3-dihydropyridin-4-one C33 (5.66 g, 16.28 mmol) in THF (30 mL), which was cooled to 0 °C. The reaction was stirred at 0 °C for 1 h, then quenched with citric acid (16 mL 2 M aqueous solution, 32.00 mmol), basified with 2 M NaOH until pH reached 10, and diluted with DCM (150 mL). The organic layer was separated and the aqueous solution was extracted again with DCM (3×100 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. This material was purified by silica gel column chromatography (isocratic EtOAc) to give 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)piperidin-4-one S17 (5.0 g, 87%) as a thick yellow oil. 1H NMR (400MHz, chloroform-d)δ 7.42(s,1H),7.32(d,J=8.3Hz,1H),6.48(dd,J=8.3,2.4Hz,1H),6.44(d,J=2.4Hz,1H),4.35(t,J=5.9Hz,1H),4.09(s,3H),3.80(s,3H),3.7 7(s,3H),3.63-3.53(m,2H),2.95(dt,J=12.2,6.0Hz,1H),2.84(ddt,J=14.5,5.4,0.9Hz,1H),2.74-2.66(m,2H),2.50(t,J=6.1Hz,2H).LCMS m / z 331.13[M+H]+.

[0191] Intermediate S18-S29 Intermediates S18-S29 (see Table 2) were prepared in two steps using the appropriate aldehydes and similar methods described above for intermediate S17. Aldehydes were either prepared by the methods described above or obtained from commercial sources. [Table 2-1] [Table 2-2] [Table 2-3]

[0192] Preparation S30 4-Oxo-2-phenyl-piperidine-1-carboxylate tert-Butyl (S30) [ka]

[0193] tert-Butyl 4-oxo-2-phenyl-piperidine-1-carboxylate (S30) was obtained from a commercial source.

[0194] Preparation S31 2-Cyclopropyl-4-oxo-piperidine-1-carboxylate benzyl (S31) [ka]

[0195] Preparation of 2-cyclopropyl-4-oxo-piperidine-1-carboxylate benzyl (S31) CuI (1.6 g, 8.401 mmol) in THF (11 mL) at −78° C. under nitrogen atmosphere was treated with cyclopropylmagnesium bromide (18 mL of a 0.5 M solution in THF, 9.00 mmol). The mixture was stirred at −78° C. for 30 min, then diethyloxonio(trifluoro)boranuide (801 μL, 6.490 mmol) was added. The mixture was stirred at −78° C. for another 10 min. 4-Oxo-2,3-dihydropyridine-1-carboxylate benzyl C34 (1 g, 4.324 mmol) in THF (4 mL) was then added, and the mixture was continued to stir at −78° C. for 2 h. The reaction was quenched with saturated aqueous NH4Cl (3 mL), the aqueous phase was separated, and extracted with EtOAc (3×20 mL). The organic layer was dried over sodium sulfate and concentrated in vacuo. This material was purified by silica gel column chromatography (0-50% EtOAc in heptane) to afford benzyl 2-cyclopropyl-4-oxo-piperidine-1-carboxylate S31 (463 mg, 36%). 1H NMR (300MHz, chloroform-d)δ 7.36(d,J=3.3Hz,5H),5.16(d,J=2.4Hz,2H),4.42(d,J=13.5Hz,1H),3.94(s,1H),3.46(ddd,J=13.8,11.8,4.0Hz,1H),2.65(dd,J=14.4,6.7 Hz,1H),2.57-2.31(m,3H),0.91(dddd,J=12.8,9.7,7.9,4.8Hz,1H),0.69-0.54(m,1H),0.53-0.32(m,2H),0.28(dt,J=9.4,4.8Hz,1H).LCMS m / z 274.12[M+H]+.

[0196] compound 1 2-Chloro-2'-(2-methylpyrimidin-5-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine](1) [ka]

[0197] Step 1. Synthesis of 2-chloro-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(2-methylpyrimidin-5-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (C35) 1-[(2,4-dimethoxyphenyl)methyl]-2-(2-methylpyrimidin-5-yl)piperidin-4-one S25 (120 mg, 0.3515 mmol) and 2-(5-chloro-3-thienyl)ethanol S2 (70 mg, 0.4046 mmol) were dissolved in dioxane (1 mL), cooled in an ice bath, and triflic acid (90 μL, 1.017 mmol) was added. The reaction mixture was allowed to warm to room temperature. After 3 h, the reaction mixture was diluted with 1M NaOH and EtOAc (30 mL each), the organic layer was separated, dried over sodium sulfate, and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-10% MeOH in DCM) afforded the major product, 2-chloro-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(2-methylpyrimidin-5-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C35 (83 mg, 49%) as an oil. 1H NMR (400 MHz, chloroform-d) δ 8.75(s,2H),7.18(d,J=8.2Hz,1H),6.59(s,1H),6.46(dd,J=8.2,2.4Hz,1H) ,6.42(d,J=2.4Hz,1H),4.06-3.90(m,2H),3.82(s,3H),3.77(s,3H),3.71-3 .61(m,1H),3.57(d,J=13.2Hz,1H),3.01(d,J=13.1Hz,1H),2.97-2.87(m,1H ),2.75(s,3H),2.72-2.46(m,3H),2.15-2.02(m,2H),1.99-1.77(m,2H).LCMS m / z 486.26 [M+H]+. NMR was consistent with a pair of enantiomers. Relative trans stereochemistry is assumed.

[0198] Step 2. Synthesis of 2-chloro-2'-(2-methylpyrimidin-5-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (1) 2-Chloro-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(2-methylpyrimidin-5-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C35 (76 mg, 0.1564 mmol) was dissolved in TFA (1 mL) and HO (200 μL) and heated to 90 °C. After 3 h by LCMS, the reaction mixture was cooled to room temperature and diluted with 1 M NaOH and EtOAc (30 mL each). The organic layer was separated, dried over sodium sulfate, and concentrated to an oil. Purification by silica gel chromatography (gradient: 0-10% MeOH in DCM) afforded the product 2-chloro-2'-(2-methylpyrimidin-5-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 1 (34 mg, 63%) as a white solid. 1H NMR (400MHz, chloroform-d)δ 8.69(s,2H),6.61(s,1H),4.15(dd,J=11.6,2.5Hz,1H),4.05-3.91(m,2H),3.32-3.19(m,1H), 3.14-3.01(m,1H),2.75(s,3H),2.72-2.56(m,2H),2.20-2.04(m,2H),1.91-1.70(m,2H).LCMS m / z 336.03[M+H]+.

[0199] compound 2 (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (2[enantiomer 2]) [ka]

[0200] Step 1. Synthesis of 2-chloro-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (C36) 1-[(2,4-dimethoxyphenyl)methyl]-2-(1-methyltriazol-4-yl)piperidin-4-one S25 (3.5 g, 9.905 mmol) in DCM (50 mL) was cooled to 0 °C and treated with a solution of 2-(5-chloro-3-thienyl)ethanol S2 (1.82 g, 11.19 mmol) in DCM (10 mL). Triflic acid (2.6 mL, 29.38 mmol) was added slowly and the reaction was allowed to warm to room temperature. After 30 min, the reaction was carefully quenched with saturated sodium bicarbonate solution and the organics were separated by filtration through a phase separator. The organics were concentrated in vacuo and the material was purified by silica gel chromatography (gradient: 20-100% EtOAc in heptane). The first eluting peak gave 2-chloro-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C36 (3.1 g, 65%) as an off-white foam. 1H NMR (400 MHz, chloroform-d) δ 7.53(s,1H),7.23(d,J=8.3Hz,1H),6.56(s,1H),6.45(dd,J=8.3,2.4Hz,1H),6. 40(d,J=2.4Hz,1H),4.07(s,3H),4.02(dd,J=11.7,2.9Hz,1H),3.95(dd,J=6.0,4 .9Hz,2H),3.79(s,3H),3.74(s,3H),3.41(dd,J=184.3,13.8Hz,2H),2.90-2.80 (m,1H),2.67-2.50(m,3H),2.31(dt,J=13.9,2.8Hz,1H),1.98-1.85(m,3H).LCMS m / z 475.32[M+H]+.

[0201] Step 2. Synthesis of 2-chloro-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (C37) In a large microwave vial, 2-chloro-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]C36 (3.1 g, 6.416 mmol) in water (4 mL, 222.0 mmol) and TFA (14 mL, 181.7 mmol) was heated to 95 °C. After 10 h, the reaction was bright pink and LCMS indicated consumption of starting material. The reaction was cooled to room temperature and the mixture was concentrated using a rotary evaporator to remove volatiles. The remaining solution was diluted with DCM and quenched slowly with saturated sodium bicarbonate solution until the pink color disappeared and the pH reached 9. The organic layer was separated, filtered through a pad of celite, and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0 to 12% MeOH in DCM) afforded 2-chloro-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C37 (1.7 g, 80%) as a tan foam. 1H NMR (400MHz, chloroform-d)δ 7.47(s,1H),6.58(s,1H),4.39(dd,J=11.8,2.6Hz,1H),4.06(s,3H),3.95(t,J=5.5Hz,2H),3.27(td,J=12.5,2.8Hz,1H),3.05(ddd,J= 12.2,4.7,2.2Hz,1H),2.61(td,J=5.4,3.0Hz,2H),2.36(dt,J=13.8,2.7Hz,1H),2.06(dq,J=14.0,2.6Hz,1H),1.95-1.78(m,2H).LCMS m / z 325.1[M+H]+.

[0202] Step 3. Synthesis of (2R,4R)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (C38 [enantiomer 1]) and (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (2 [enantiomer 2]) 2-Chloro-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]C37 (1.09 g, 2.89 mmol) was separated into its constituent enantiomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 20×250 mm; Mobile phase: 15% methanol (containing 5 mM ammonia), 85% carbon dioxide. Peak A was concentrated via rotary evaporator to give (2R,4R)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] C38 [enantiomer 1] (435 mg, 42%) as an off-white foam. 1H NMR (400MHz, chloroform-d)δ 7.46(s,1H),6.59(s,1H),4.39(dd,J=11.8,2.7Hz,1H),4.06(s,3H),3.95(t,J=5.5Hz,2H),3.27(td,J=12.5,2.8Hz,1H),3.05(ddd,J= 12.2,4.7,2.1Hz,1H),2.61(td,J=5.4,3.2Hz,2H),2.36(dt,J=13.7,2.7Hz,1H),2.06(dq,J=13.9,2.6Hz,1H),1.95-1.78(m,2H).LCMS m / z 325.14[M+H]+.

[0203] Peak B was concentrated in vacuo to give (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 2 [enantiomer 2] (455 mg, 45%) as a white solid. The structure and stereochemistry were confirmed via X-ray crystallography. 1H NMR (400MHz, chloroform-d)δ 7.42(s,1H),6.58(s,1H),4.35(dd,J=11.7,2.6Hz,1H),4.06(s,3H),3.99-3.93(m,2H),3.24(td,J=12.4,2.7Hz,1H),3.02(ddd,J=1 2.2,4.8,2.2Hz,1H),2.61(td,J=5.4,3.8Hz,2H),2.36(dt,J=13.6,2.7Hz,1H),2.05(dq,J=13.8,2.5Hz,1H),1.88-1.74(m,2H).LCMS m / z 325.14[M+H]+.

[0204] compound 3 2'-Cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine](3) [ka]

[0205] Step 1. Synthesis of 2'-cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-1'-benzyl carboxylate (C39) 2-Cyclopropyl-4-oxo-piperidine-1-carboxylate benzyl S31 (57 mg, 0.1989 mmol) and 2-(5-ethyl-2-thienyl)ethanol S6 (40 mg, 0.2432 mmol) were dissolved in DCM (1 mL). The mixture was cooled to -20°C and to it was added trifluoromethanesulfonic acid (53 μL, 0.5989 mmol). The mixture was stirred at -20°C for 30 minutes and then quenched with a saturated aqueous solution of sodium bicarbonate. The mixture was extracted with DCM (2 x 3 mL). The combined organic layers were dried and purified by silica gel chromatography (gradient: 0-5% 7M ammonia in DCM) to give benzyl 2'-cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-1'-carboxylate C39 (69 mg, 77%) as a pair of trans enantiomers. LCMS m / z 412.21 [M+H]+.

[0206] Step 2. Synthesis of 2'-cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] (3) In a microwave vial, 2'-cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-1'-benzyl carboxylate C39 (69 mg, 0.153 mmol) was dissolved in MeOH (2 mL). Ammonium formate (89 mg, 1.411 mmol) was added followed by Pd / C (22 mg of 10 wt%, 0.02067 mmol). The mixture was heated to 140 °C in a microwave for 10 min. The reaction mixture was filtered through a pad of Celite® and the filtrate was concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-10 of 7 M ammonia in MeOH in DCM) afforded 2'-cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]3 (24 mg, 38%). 1H NMR (300MHz, chloroform-d)δ 6.50(d,J=1.1Hz,1H),4.04-3.78(m,2H),3.12-2.86(m,2H),2.86-2.63(m,4H),2.18-1.92(m,2H),1.91-1.79(m,2H) ,1.65(dd,J=13.6,11.4Hz,1H),1.28(t,J=7.5Hz,3H),0.81-0.62(m,1H),0.57-0.29(m,2H),0.24-0.06(m,2H).LCMS m / z 278.17[M+H]+.

[0207] Compounds 4~28 Compounds 4-28 (see Table 3) were prepared by methods similar to compounds 1, 2, or 3, with modifications apparent to one of skill in the art. Thiopheneethanols and piperidones were prepared by methods described above or obtained from commercial sources. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8]

[0208] compound 29 2'-Cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] (29 [enantiomer 1]) [ka]

[0209] 2'-Cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] 3 (24 mg, 0.07600 mmol) was separated into its constituent enantiomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 20×250 mm; Mobile phase: 40% methanol (containing 5 mM ammonia), 60% carbon dioxide. Peak A was concentrated via rotary evaporator to give 2'-cyclopropyl-2-ethyl-spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] 29 (4.6 mg, 38%) as a single enantiomer. 1H NMR (400MHz, chloroform-d)δ 6.54(d,J=1.2Hz,1H),3.98-3.75(m,2H),3.19(d,J=10.8Hz,2H),2.88-2.64(m,4H),2.46(td,J=10.3,3.8Hz,1H),2.1 9-2.00(m,3H),1.99-1.87(m,1H),1.29(t,J=7.5Hz,3H),1.07-0.78(m,2H),0.65-0.35(m,3H),0.34-0.18(m,1H).LCMS m / z 278.17[M+H]+.

[0210] Preparation S32 (2S,4S)-2'-bromo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl ester (S32) [ka]

[0211] Step 1. Synthesis of (2S)-2'-bromo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl ester (C40) A mixture of (S)-tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate S16 (400 mg, 1.792 mmol), azidomethyl(trimethyl)silane (248 mg, 1.919 mmol), copper(II) sulfate (86 mg, 0.5388 mmol), and sodium ascorbate (316 mg, 1.794 mmol) in DMF (16 mL) was heated to 50° C. The solution was stirred overnight and then cooled to room temperature where the mixture was diluted with ethyl acetate and water. The organic layer was washed with water (5×), brine, dried over sodium sulfate, and concentrated in vacuo to give crude (S)-tert-butyl 4-oxo-2-[1-(trimethylsilylmethyl)triazol-4-yl]piperidine-1-carboxylate (631 mg). LCMS m / z 353.27 [M+H]+. This material was dissolved in dioxane (10.8 mL) and 2-(5-bromo-3-thienyl)ethanol S4 (510 mg, 2.463 mmol) was added. The reaction was placed in an ice bath and triflic acid (952 μL, 10.74 mmol) was added. The reaction was allowed to warm to room temperature overnight. The reaction mixture was diluted with DCM and washed with saturated sodium bicarbonate and brine. The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude product, (2S)-2'-bromo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (791 mg, 100%); LCMS m / z 441.12 [M+H]+, was redissolved in DCM (10.8 mL) and treated with BocO (782 mg, 3.583 mmol) and DIPEA (937 μL, 5.377 mmol). After 2 h, the solvent was removed in vacuo. Purification by silica gel chromatography (gradient: 0-30% EtOAc in heptane) afforded the product (2S)-2'-bromo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylic acid C40 (660 mg, 68%). LCMS m / z 541.21 [M+H]+.

[0212] Step 2. Synthesis of (2S)-2'-bromo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl ester (S32) To a solution of (2S)-2'-bromo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate C40 (660 mg, 1.219 mmol) in THF (9.9 mL) was added TBAF (1.4 mL of a 1 M solution, 1.400 mmol) at 0° C. After 30 min, the reaction was quenched with water, diluted with DCM and the organic layer was collected through a phase separator and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-50% EtOAc in heptane) afforded the product tert-butyl (2S)-2'-bromo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylic acid S32 (390 mg, 57%) as a 3:1 mixture of diastereomers. LCMS m / z 469.16 [M+H]+.

[0213] Compounds 30 and 31 (2S)-2'-Cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (30) and (2S,4S)-2'-Cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (31) [ka]

[0214] Preparation of (2S)-2'-cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (30) and (2S,4S)-2'-cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (31) (2S)-tert-butyl 2'-bromo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate S32 (117 mg, 0.2069 mmol), cyclopropylboronic acid (22 mg, 0.2561 mmol), Pd(dppf)Cl2 (17 mg, 0.02082 mmol), and K2CO3 (57 mg, 0.4124 mmol) were taken up in DME (1.7 mL) and the mixture was purged with nitrogen for 5 minutes before being heated to reflux overnight. The reaction was diluted with DCM and water and the organic layer was separated through a phase separator and dried in vacuum. Purification by reverse phase chromatography (gradient: 0-100% MeCN in water with 0.1% TFA) afforded tert-butyl (2S)-2'-cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate (35 mg, 30%). LCMS m / z 431.35 [M+H]+ This intermediate was then dissolved in HCl (720 μL of a 4 M solution in dioxane, 2.880 mmol). After stirring for 30 min, the mixture was concentrated via rotary evaporator. Purification by reverse-phase HPLC (Method: purification by C18 Waters Sunfire column (30 × 150 mm, 5 micron). Gradient: MeCN in HO containing 0.1% trifluoroacetic acid) afforded one tube's worth of a 60:40 diastereomeric mixture of (2S)-2'-cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 30 (10.50 mg, 11%) as trifluoroacetate salts.1H NMR (400MHz, chloroform-d)δ 7.90(d,J=5.3Hz,1H),6.49(dd,J=14.1,0.8Hz,1H),5.12-4.84(m,1H),4.12(d,J=8.9Hz,3 H),4.03-3.73(m,2H),3.54(dd,J=45.9,12.9Hz,2H),3.01-2.15(m,6H),2.11-1.95(m,1H), 1.14-0.91 (m, 2H), 0.80-0.59 (m, 2H). The major product, another tubeful of single diastereomer (2S,4S)-2'-cyclopropyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 31 (5 mg, 10%) was obtained as the trifluoroacetate salt. 1H NMR (400MHz, chloroform-d)δ 9.69(d,J=116.3Hz,2H),7.89(s,1H),6.47(d,J=0.8Hz,1H),4.93(d,J=12 .3Hz,1H),4.10(s,3H),3.98-3.91(m,2H),3.66-3.55(m,1H),3.48(d,J=1 2.4Hz,1H),2.68-2.62(m,3H),2.43-2.28(m,2H),2.21(d,J=14.7Hz,1H), 2.03(ttd,J=8.3,5.0,0.8Hz,1H),1.06-0.95(m,2H),0.74-0.65(m,2H).

[0215] compound 32 (2S)-2'-Methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (32) [ka]

[0216] Preparation of (2S,4S)-2'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (32) This compound was made under the conditions of compound 31, but using methylboronic acid, to give (2S)-2'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 32 (3.5 mg, 10%) as the trifluoroacetate salt. 1H NMR (400MHz, chloroform-d)δ 9.74(d,J=118.8Hz,2H),7.88(s,1H),6.47(d,J=1.2Hz,1H),4.93(d,J=12.3Hz,1H),4.10(s,3H),3.96(t,J=5.5Hz,2H),3.61( t,J=12.6Hz,1H),3.48(d,J=12.2Hz,1H),2.70-2.58(m,3H),2.49-2.37(m,4H),2.36-2.28(m,1H),2.22(d,J=14.6Hz,1H).LCMS m / z 305.16[M+H]+.

[0217] compound 33 (2S,4S)-2'-Iodo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (33) [ka]

[0218] Step 1. Synthesis of (2S)-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl ester (C41) A mixture of (S)-tert-butyl 2-ethynyl-4-oxopiperidine-1-carboxylate S16 (500 mg, 2.239 mmol), azidomethyl(trimethyl)silane (356 μL, 2.397 mmol), copper(II) sulfate (107 mg, 0.6704 mmol), sodium ascorbate (400 mg, 2.271 mmol) in DMF (20 mL) was heated to 50° C. After stirring overnight, the mixture was cooled to room temperature, concentrated, and redissolved in DCM / water. The organic layer was collected through a phase separator and concentrated to give crude (S)-4-oxo-2-[1-(trimethylsilylmethyl)triazol-4-yl]piperidine-1-carboxylic acid (789 mg, 100%). LCMS 353.23 [M+H]+. This intermediate was dissolved in dioxane (10 mL). 2-(3-Thienyl)ethanol S1 (574 mg, 4.478 mmol) was added and the reaction was cooled to 0° C. Triflic acid (792 μL, 8.94 mmol) was added and the reaction was allowed to warm to room temperature. After 1 h, the mixture was diluted with DCM and washed with saturated sodium bicarbonate and brine. The organic layer was dried over sodium sulfate and concentrated in vacuo to give crude (2S)-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran], LCMS m / z 363.22 [M+H]+, which was immediately redissolved in DCM (10 mL) and treated with DIPEA (1.17 mL, 6.716 mmol) and Boc2O (977 mg, 4.477 mmol). After 30 min, the solvent was removed in vacuo. Purification by silica gel chromatography (gradient: 0-30% EtOAc in heptane) afforded (2S)-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl C41 (510 mg, 49%) as a mixture of diastereomers. LCMS m / z 463.31[M+H]+.

[0219] Step 2. Synthesis of tert-butyl (2S,4S)-2'-iodo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate (C42) A solution of (2S)-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl C41 (475 mg, 1.027 mmol) and NIS (305 mg, 1.356 mmol) in chloroform (3.4 mL) and acetic acid (1.1 mL) was stirred at room temperature overnight. The reaction was quenched with saturated sodium bicarbonate solution and diluted with DCM. The organic phase was collected by filtration through a phase separator and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient: 0 to 50% EtOAc in heptane) afforded the desired diastereomer as peak B, (2S,4S)-2'-iodo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate tert-butyl C42 (370 mg, 61%). 1H NMR (400MHz, chloroform-d)δ 7.30(s,1H),6.88(s,1H),5.25(dd,J=9.9,6.5Hz,1H),3.96-3.89(m,1H),3.89-3.85(m,4H),3.43(ddd,J=13.9,9.6,5.7Hz,1H),2.65-2. 54(m,3H),2.46(ddd,J=14.5,6.5,1.7Hz,1H),2.26(dt,J=15.5,7.8Hz,1H),1.92(ddd,J=14.5,5.7,3.9Hz,1H),1.42(s,9H),0.14(s,9H).

[0220] Step 3. Synthesis of (2S,4S)-2'-iodo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (33) To a solution of (2S,4S)-tert-butyl 2'-iodo-2-(1-((trimethylsilyl)methyl)-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate C42 (240 mg, 0.4078 mmol) in THF (3.6 mL) was added TBAF (110 μL, 0.3732 mmol). The reaction was stirred at room temperature for 5 h and then quenched with saturated sodium bicarbonate solution and DCM. The organic phase was separated, dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-50% EtOAc in heptane) gave the product tert-butyl (2S,4S)-2'-iodo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-carboxylate (160 mg, 57%). LCMS m / z 517.26 [M+H]+. 50 mg of this intermediate was dissolved in dioxane (1.2 mL) and treated with HCl (1 mL of a 4 M solution in dioxane, 4.000 mmol). After 30 min, the solvent was removed in vacuo. Purification by reverse phase HPLC. Method: C18 Waters Sunfire column (30x150 mm, 5 microns). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid. (2S,4S)-2'-iodo-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 33 was obtained as the trifluoroacetate salt (12 mg, 55%). 1H NMR (400MHz, chloroform-d)δ 9.69(d,J=115.5Hz,2H),7.87(s,1H),6.96(s,1H),4.93(d,J=12.3Hz,1H),4.09(s,3H),3 .92(td,J=5.6,2.6Hz,2H),3.68-3.37(m,2H),2.82-2.50(m,3H),2.50-2.03(m,3H).LCMS m / z 416.85[M+H]+.

[0221] compound 34 (2S,4S)-2'-Chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (34) [ka]

[0222] Step 1. Synthesis of 1-((2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one (C43) To a solution of (2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 25 (150 mg, 0.4525 mmol) in DCM (2 mL) was added TEA (130 μL, 0.9327 mmol) and TFAA (90 μL, 0.6475 mmol). The reaction mixture was stirred for 2 h, diluted with EtOAc and saturated NaHCO3, the organic layer was dried and concentrated to give an oil. Purification by silica gel chromatography (gradient: 0-60% EtOAc in heptane) afforded the product 1-((2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoro-ethanone C43 (160 mg, 84%) as a white foam. LCMS m / z 421.09[M+H]+.

[0223] Step 2. Synthesis of (2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'(5'H)-one (C44) A solution of 1-((2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoro-ethanone C43 (160 mg, 0.3802 mmol), cobalt(II) acetate tetrahydrate (10 mg, 0.04015 mmol), and N-hydroxyphthalimide (25 mg, 0.1533 mmol) in ACN (3 mL) was vacuum purged three times with an oxygen balloon. The flask was heated to 60° C. under an oxygen atmosphere. After 4 h, the reaction mixture was diluted with water and EtOAc, and the organic layer was dried and concentrated to give an oil. Purification by silica gel chromatography (gradient: 0 to 80% EtOAc in heptane) afforded (2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'(5'H)-one C44 (30 mg, 18%) as a white solid. 1H NMR (400 MHz, chloroform-d) δ 7.23(s, 1H), 6.13(s, 1H), 5.67-5.53(m, 1H), 4.44-4.26(m, 2H), 4.18-4.01(m, 1H), 3.95-3.73(m, 1H), 2.85-2.70(m, 1H), 2.70-2.56(m, 2H), 2.33(s, 3H), 2.14-1.99(m, 1H).LCMS m / z 435.04[M+H]+.

[0224] Step 3. Synthesis of (2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (34) To a solution of (R)-(+)-2-methyl-CBS-oxazaborolidine (15 μL of a 1M solution in toluene, 0.0150 mmol) in MTBE (0.4 mL) at 0° C., borane tetrahydrofuran (140 μL of a 1M solution in THF, 0.1400 mmol) was added. After 2 min, a solution of (2S,4S)-2′-chloro-2-(3-methylisoxazol-5-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7′-thieno[2,3-c]pyran]-4′(5′H)-one C44 (30 mg, 0.06899 mmol) in MTBE (1 mL) was added and the reaction mixture was stirred at 0° C. for 1 h. The reaction was quenched with 1M HCl (0.3 mL) and stirred for 30 min. 6M NaOH (0.5 mL) was added and the mixture was stirred overnight. The reaction mixture was diluted with EtOAc and water and the organic layer was concentrated to give an oil. Purification by reverse phase HPLC (Method: C18 Waters Sunfire column (30×150 mm, 5 micron). Gradient: MeCN in H2O containing 10 mM ammonium hydroxide) gave (2S,4S)-2'-chloro-2-(3-methylisoxazol-5-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 34 (20 mg, 82%) as an oil. 1H NMR (400MHz, chloroform-d)δ 6.86(s,1H),6.01(s,1H),4.48(t,J=2.9Hz,1H),4.41(dd,J=11.6,2.6Hz,1H),3.99(qd,J=12.4,2.8Hz,2H),3.20(td,J=12. 4,2.6Hz,1H),3.06(ddd,J=12.2,4.8,2.2Hz,1H),2.30(s,3H),2.20-2.09(m,1H),1.98-1.85(m,2H),1.76-1.64(m,2H).LCMS m / z 341.03[M+H]+.

[0225] Preparation S33 (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'(5'H)-one (S33) [ka]

[0226] Step 1. Synthesis of 1-((2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one (C45) A solution of (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]2 (2.2 g, 6.613 mmol) in DCM (30 mL) was treated with TEA (3.6 mL, 25.83 mmol) followed by TFAA (1.1 mL, 7.914 mmol). After 20 min, the reaction mixture was diluted with saturated sodium bicarbonate and DCM. The organic layer was separated via a phase separator and concentrated in vacuo to give 1-((2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one C45 (2.8 g, 84%) as a yellow semi-solid. 1H NMR (400 MHz, chloroform-d) δ 7.60 (s, 1H), 6.58 (s, 1H), 5.51 (t, J = 9.2 Hz, 1H), 4.08 (s, 3H), 3.97-3.80 (m, 4H), 2.94 (t, J = 13.1 Hz, 1H), 2.59 (t, J = 5.5 Hz, 2H), 2.44 (ddd, J = 34.2, 17.1, 8.0 Hz, 2H), 2.07 (d, J = 14.7 Hz, 1H).19F NMR (376 MHz, chloroform-d) δ -69.99. LCMS m / z 421.14 [M+H]+.

[0227] Step 2. Synthesis of (2S,4S)-2'-chloro-2-methyl-6-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (S33) To a solution of 1-((2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one C45 (660 mg, 1.556 mmol) in ACN (18 mL) was added cobalt(II) acetate tetrahydrate (70 mg, 0.2810 mmol) and N-hydroxyphthalimide (135 mg, 0.8276 mmol). The reaction was purged and evacuated with oxygen (3x) and heated to 45°C under an oxygen balloon. After 2.5 h, the reaction was diluted with water and partitioned with DCM. The organics were collected through a phase separator and concentrated via rotary evaporator. Purification by silica gel chromatography (gradient: heptane, 0 to 65% EtOAc) afforded (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (366 mg, 49%) as a white foam. 1H NMR (300MHz, chloroform-d)δ 7.63(s,1H),7.19(s,1H),5.55(t,J=8.8Hz,1H),4.44-4.24(m,2H),4.09(s,3H),3.91(d, LCMS m / z 435.04[M+H]+.

[0228] compound 35 (2S,4S)-2'-Chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (35) [ka]

[0229] Synthesis of (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (35) A solution of (S)-(-)-2-methyl-CBS-oxazaborolidine (145 μL of a 1 M solution in THF, 0.1450 mmol) in MTBE (4 mL) was cooled to 0° C. and treated with borane tetrahydrofuran (1000 μL of a 1 M solution in THF, 1.00 mmol). A solution of (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (200 mg, 0.4172 mmol) in MTBE (1.5 mL) was then added and the reaction was stirred at 0° C. After 15 min, the reaction was quenched with 2M HCl, the ice bath was removed, and the partitioned mixture was stirred vigorously overnight. The reaction was diluted with DCM and the organics were collected via a phase separator and concentrated via rotary evaporator. Purification by silica gel chromatography: 0-70% EtOAc in Heptane) afforded 1-((2S,4S)-2'-chloro-4'-hydroxy-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one (140 mg, 74%) as a white foam. 1H NMR (300 MHz, chloroform-d) δ 7.62 (s, 1H), 6.86 (s, 1H), 5.53-5.41 (m, 1H), 4.47 (dt, J = 9.4, 3.3 Hz, 1H), 4.10 (s, 3H), 3.94 (qd, J = 12.3, 3.3 Hz, 4H), 2.91 (t, J = 13.1 Hz, 1H), 2.65-2.38 (m, 2H), 2.19 (d, J = 14.4 Hz, 1H), 2.07 (d, J = 3.8 Hz, 1H).19F NMR (282 MHz, chloroform-d) δ -69.96. LCMS m / z 437.11 [M+H]+. This material was then dissolved in MeOH (3 mL) and treated with NaOH (2 mL of a 2 M solution, 4.000 mmol) at room temperature. After 5 min, the reaction was diluted with water and DCM and filtered through a phase separator to elute the organics, which were concentrated in vacuo. The residue was taken up in DCM (3 mL) and HCl (95 μL of a 4 M solution, 0.3800 mmol) was added dropwise.The material was concentrated, redissolved in MeOH, and concentrated again (2 times). The pale yellow film was then dissolved in water, transferred to a vial, frozen in a dry ice bath at -78°C, and lyophilized over the weekend to give (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol 35 (97.1 mg, 59%), hydrochloride salt as a yellow solid. 1H NMR (300MHz, methanol-d4)δ 8.07(s,1H),6.94(s,1H),4.83(dd,J=12.5,3.2Hz,1H),4.51(t,J=3.8Hz,1 H),4.14-4.04(m,4H),3.87(dd,J=12.2,4.1Hz,1H),3.59(td,J=13.1,3.3H z,1H),3.49-3.38(m,1H),2.62(dt,J=14.8,2.9Hz,1H),2.40(dd,J=14.7,2 .8Hz,1H),2.31(dd,J=14.9,12.6Hz,1H),2.15(td,J=14.0,4.8Hz,1H).LCMS m / z 340.98[M+H]+.

[0230] compound 36 (2S,4S)-2'-Chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (36) [ka]

[0231] Synthesis of (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (36) This compound was made according to the conditions for compound 35, but using (R)-(+)-2-methyl-CBS-oxazaborolidine solution (1M in toluene), to give (2S,4S)-2'-chloro-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol hydrochloride salt 36 (134 mg, 78%) as a white solid. 1H NMR (300MHz, methanol-d4)δ 8.07(s,1H),6.95(s,1H),4.91(dd,J=12.4,3.2Hz,1H),4.51(t,J=3.5Hz,1H),4.13(s,3H),3.98(ddd,J=51.0,12.3,3. 5Hz,2H),3.60-3.37(m,2H),2.61(dt,J=14.5,2.9Hz,1H),2.47-2.34(m,2H),2.04(ddd,J=15.0,13.2,4.9Hz,1H).LCMS m / z 341.07[M+H]+.

[0232] Compounds 37 and 38 (2S,4S)-2'-chloro-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (37) [DIAST-1] and (38) [DIAST-2] [ka]

[0233] Step 1. Synthesis of 1-((2S,4S)-2'-chloro-4'-hydroxy-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one (C46) [DIAST-1] and (C47) [DIAST-2] A solution of (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (30 mg, 0.06565 mmol) in 2-MeTHF (1 mL) was cooled to 0 °C and treated dropwise with MeMgBr (26 μL of a 3.4 M solution in 2-MeTHF, 0.08840 mmol). After 15 min at 0 °C, the reaction was quenched with saturated ammonium chloride, diluted with water, and extracted with DCM through a phase separator. The organics were concentrated via rotary evaporator. Purification by silica gel chromatography (gradient: 0-80% EtOAc in heptane) afforded 1-((2S,4S)-2'-chloro-4'-hydroxy-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one C46[DIAST-1] (8 mg, 26%) as a white solid. LCMS m / z 451.24[M+H]+. The second eluting peak gave 1-((2S,4S)-2'-chloro-4'-hydroxy-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one C47 [DIAST-2] (9 mg, 29%) as a clear film. 1H NMR (300MHz, chloroform-d)δ 7.61(s,1H),6.86(s,1H),5.59(t,J=9.0Hz,1H),4.08(s,3H),3.89(s,2H),3.82-3.67(m,2H),3.03(t ,J=12.8Hz,1H),2.48(dt,J=18.5,10.1Hz,2H),2.27(s,1H),1.96(d,J=14.9Hz,1H),1.44(s,3H).LCMS m / z 433.2(M+H-18)+.

[0234] Step 2. Synthesis of (2S,4S)-2'-chloro-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (37) [DIAST-1] and (38) [DIAST-2] 1-((2S,4S)-2'-chloro-4'-hydroxy-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl)-2,2,2-trifluoroethan-1-one (C46) [DIAST-1] (8 mg) and (C47) [DIAST-2] (9 mg) were lifted separately in MeOH (1 mL) and treated with NaOH (0.4 mL of a 2 M solution, 0.8000 mmol). After 10 min at room temperature, the reactions were diluted with water, passed through a phase separator and extracted with DCM (2x). The organics were concentrated in vacuo to give the respective diastereomers. (2S,4S)-2'-chloro-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (37) [DIAST-1] (6.6 mg, 28%) was obtained as a white foam. 1H NMR (300MHz, chloroform-d)δ 7.44(s,1H),6.86(s,1H),4.33(dd,J=11.7,2.6Hz,1H),4.07(s,3H),3.80(q,J=11.9Hz,2H),3.31(td,J=12.3,2.9Hz,1H),3.09-3. 00(m,1H),2.52-2.42(m,1H),2.00(m,J=2.6Hz,1H),1.87(td,J=13.0,4.6Hz,1H),1.72(dd,J=13.9,11.7Hz,1H),1.43(s,3H).LCMS m / z 355.07[M+H]+. (2S,4S)-2'-chloro-4'-methyl-2-(1-methyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (38) [DIAST-2] (6.9 mg, 29%) was obtained as a white foam.1H NMR (300MHz, chloroform-d)δ 7.42(s,1H),6.86(s,1H),4.41(dd,J=11.6,2.6Hz,1H),4.07(s,3H),3.78(s,2H),3.21(td,J=12.4,2.6Hz,1H),3.11-2.97(m ,1H),2.38-2.23(m,2H),2.14(dd,J=14.1,2.6Hz,1H),1.92(dd,J=13.4,11.6Hz,1H),1.77-1.63(m,1H),1.45(s,3H).ESI-MS m / z 355.07[M+H]+.

[0235] Compounds 39, 40, and 41

[0236] (2'S,7S)-2-chloro-4-(difluoromethyl)-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidin]-4-ol (39) [DIAST-1] and (40) [DIAST-2] and (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (41) [ka]

[0237] Preparation of (2'S,7S)-2-chloro-4-(difluoromethyl)-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidin]-4-ol (39) [DIAST-1] and (40) [DIAST-2] and (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (41) In a small microwave vial, a solution of (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (30 mg, 0.06565 mmol) and PPh3 (21 mg, 0.08007 mmol) in MeCN (500 μL) was added at room temperature to DMPU (18 μL, 0.1494 mmol) followed by [bromo(difluoro)methyl]-trimethyl-silane (19 μL). The resulting solution was heated at 60° C. overnight. The reaction was cooled to room temperature and treated with KOH (300 μL of a 1 M solution, 0.3000 mmol) to form the deprotected product after 10 min. The reaction was diluted with DCM and water, and the organics were collected via phase separator and concentrated via rotary evaporator. Purification by normal phase chromatography (gradient: 0-15% MeOH in DCM) afforded the diastereomers as two separate peaks. Peak A afforded (2'S,7S)-2-chloro-4-(difluoromethyl)-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidin]-4-ol 39 [DIAST-1] (5.3 mg, 19%) as a pale orange foam. 1H NMR (400MHz, methanol-d4)δ 7.82(s,1H),6.98(d,J=1.0Hz,1H),5.95(t,J=55.3Hz,1H),4.27(dd,J=11.8,2.7Hz,1H),4.08(s,4H),3.80(dd,J=12.2,2.9Hz,1H),3. 24-3.14(m,1H),3.01(ddd,J=12.7,4.7,2.0Hz,1H),2.52(dt,J=13.9,2.6Hz,1H),2.02(dd,J=13.9,2.6Hz,1H),1.91-1.76(m,2H).LCMS m / z 391.07[M+H]+.

[0238] Peak B gave (2'S,7S)-2-chloro-4-(difluoromethyl)-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidin]-4-ol 40[DIAST-2] (5.2 mg, 20%) as a semi-solid. 1H NMR (400 MHz, methanol-d4) δ 7.81(s,1H),6.97(d,J=1.1Hz,1H),5.95(t,J=55.2Hz,1H),4.29(dd,J=11. 8,2.5Hz,1H),4.10(d,J=12.3Hz,1H),4.08(s,3H),3.77(ddd,J=12.2,3.3, 1.8Hz,1H),3.17(td,J=12.6,2.6Hz,1H),3.05-2.98(m,1H),2.32-2.20(m, 2H),1.91(dd,J=13.5,11.8Hz,1H),1.76(ddd,J=14.0,12.6,4.7Hz,1H).19F NMR(376MHz, methanol-d4)δ -131.30(d,J=281.2Hz),-136.94(d,J=281.2Hz).LCMS m / z 391.07[M+H]+.

[0239] The reaction did not go to completion, so some impure deprotected starting material was isolated. Purification by reverse phase chromatography (gradient: 0-100% MeCN in water with 0.1% TFA) afforded (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one 41 as the trifluoroacetate salt. LCMS m / z 339.21 [M+H]+.

[0240] compound 42 (2'S,7S)-2-Chloro-4-methoxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylic acid (42) [ka]

[0241] Step 1. Synthesis of (2'S,7S)-2-chloro-4-hydroxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate tert-butyl ester (C48) A solution of (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (80 mg, 0.1751 mmol) in MeOH (1 mL) was treated with NaOH (500 μL of a 2 M solution, 1.000 mmol). After 20 min at room temperature, LCMS indicated complete consumption of the product and formation of intermediate 2. The reaction was diluted with DCM (1.5 mL), treated with Boc2O (75 mg, 0.3436 mmol) and stirred at room temperature overnight. The reaction was diluted with water and passed through a phase separator to extract with DCM (2×). The organics were concentrated in vacuo then dissolved in MeOH (1 mL) and treated with NaBH4 (20 mg, 0.5286 mmol). After 10 min, the reaction was quenched with water, diluted with DCM and extracted through a phase separator (3x). The organics were concentrated. Purification by normal phase chromatography (gradient: 0-100% EtOAc in heptane) afforded (2'S,7S)-2-chloro-4-hydroxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate tert-butyl C48 (43 mg, 52%). LCMS m / z 441.12[M+H]+.

[0242] Step 2. Synthesis of (2'S,7S)-2-chloro-4-methoxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylic acid (C49) (2'S,7S)-2-chloro-4-hydroxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate tert-butyl C48 (43 mg, 0.09041 mmol) in 2-MeTHF (1.5 mL) was treated with NaH (12 mg of 60% w / w in mineral oil, 0.3000 mmol) and stirred for 30 min. MeI (20 μL, 0.3213 mmol) was then added. After 2.5 h, the reaction was quenched with water, diluted with DCM, and extracted (2 times) through a phase separator. The organic layer was concentrated via rotary evaporator, dissolved in DCM (1 mL), and treated with HCl (300 μL of a 4 M solution in dioxane, 1.200 mmol). After 1.5 h, the reaction was basified with 2M NaOH and extracted with DCM (2x) through a phase separator. The organic layer was concentrated in vacuo and purified by normal phase chromatography (gradient: 0-15% MeOH in DCM) to give (2'S,7S)-2-chloro-4-methoxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C49 (25 mg, 78%) as a mixture of diastereomers. LCMS m / z 355.03 [M+H]+.

[0243] Step 3. Synthesis of (2'S,7S)-2-chloro-4-methoxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylic acid (42) (2'S,7S)-2-chloro-4-methoxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C49 (25 mg) was separated into its constituent enantiomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 10 x 250 mm; Mobile phase: 40% ethanol (5 mM ammonia), 60% carbon dioxide. Peak B gave (2'S,7S)-2-chloro-4-methoxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 42 (9.9 mg, 59%) as a pale yellow film. 1H NMR (300MHz, chloroform-d)δ 7.38(s,1H),6.79(s,1H),4.40(dd,J=11.6,2.7Hz,1H),4.16-3.91(m,6H),3.43(s,3H),3.19(td,J=12.5,2.7Hz,1H),3.08-2.97(m,1H) ),2.33(dt,J=13.4,2.7Hz,1H),2.10(dd,J=14.1,2.6Hz,1H),1.93(dd,J=13.4,11.7Hz,1H),1.69(ddd,J=13.9,12.5,4.7Hz,1H).LCMS m / z 355.03[M+H]+.

[0244] compound 43 (2'S,7S)-2-Chloro-4,4-dideuterio-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine] (43) [ka]

[0245] Step 1. Synthesis of (2'S,7S)-2-chloro-4-deuterio-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidin]-4-ol) (C50) To an oven-dried vial under argon was added LiAlD4 (16.9 mg, 0.4026 mmol) and diethyl ether (1.5 mL). The solution was cooled to 0 °C and (2S,4S)-2'-chloro-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (32 mg, 0.06476 mmol) was added as a solution in diethyl ether (1.5 mL) and THF (1 mL). The reaction was allowed to warm to room temperature and stirred for 30 min before being carefully quenched with H2O (10 mL) and the pH adjusted with 2M NaOH. The mixture was extracted with DCM (3 x 10 mL), passed through a phase separator and concentrated to give the crude C50 product.

[0246] Step 2. Synthesis of (2'S,7S)-2-chloro-4,4-dideuterio-2'-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine] (43) The crude C50 product in CDCl3 (1 mL) was treated with deuterated (triethyl)silane (70 μL, 0.4399 mmol) and TFA (330 μL, 4.283 mmol) and stirred for 45 min. TfOH (30 μL, 0.3390 mmol) was added and the reaction was stirred at room temperature for 60 min, at which point it was quenched into water and DCM. The pH of the aqueous layer was adjusted to >10 by addition of 2M NaOH and extracted with DCM (3 times). The organic layer was dried over sodium sulfate and concentrated. Purification by reverse phase HPLC (Method: purification by C18 Waters Sunfire column (30 × 150 mm, 5 micron). Gradient: MeCN in HO containing 5 mM HCl) gave (2′S,7S)-2-chloro-4,4-dideuterio-2′-(1-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4′-piperidine] 43 (19.7 mg, 82%) as the hydrochloride salt. 1H NMR (400MHz, methanol-d4)δ 8.13(s,1H),6.76(s,1H),4.87(dd,J=12.4,3.2Hz,1H),4.13(s,3H),4.02(d,J=2.4Hz,2H),3.56(td,J=13.1,3.2Hz,1H),3.42(ddd,J=12.9 ,4.7,2.1Hz,1H),2.55(dt,J=14.6,2.9Hz,1H),2.41(dd,J=14.7,12.5Hz,1H),2.37-2.29(m,1H),2.15(ddd,J=14.9,13.4,4.7Hz,1H).LCMS m / z 327.24[M+H]+.

[0247] Preparation S34 1-[2-Bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoroethanone (S34) [ka]

[0248] Step 1. Synthesis of 2-bromo-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (C51) This compound was made following similar conditions as step 1 for compound 2, but using thiopheneethanol S4. The major product was 2-bromo-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (1.27 g, 77%) as a pair of enantiomers assuming trans stereochemistry. LCMS m / z 519.2[M+H]+.

[0249] Step 2. Synthesis of 2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (C52) This compound was prepared under similar conditions as in step 2 of compound 2, using 2-bromo-1'-[(2,4-dimethoxyphenyl)methyl]-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]C51 (500 mg, 0.8449 mmol) as starting material. 2-Bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]C52 (312 mg) was obtained. LCMS m / z 369.07[M+H]+.

[0250] Step 3. 1-[2-Bromo-2'-(1-methylthiazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoro-ethanone (S34) To a solution of 2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]C52 (312 mg) in DCM (10 mL) was added TEA (383 mg, 3.785 mmol) and the solution was cooled to 0°C. TFAA (238 mg, 1.133 mmol) was added and the reaction was allowed to warm to room temperature. After 1 h, the reaction mixture was diluted with saturated sodium bicarbonate solution and DCM. The organics were separated via phase separator and concentrated via rotary evaporator. Purification by normal phase chromatography (gradient: 0 to 60% EtOAc in heptane) afforded 1-[2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoro-ethanone S34 (240 mg, 58%). 1H NMR (400MHz, chloroform-d)δ 7.64(d,J=17.1Hz,1H),6.75(s,1H),5.55(d,J=9.8Hz,1H),4.10(s,3H),3.99-3. 74(m,4H),2.98(t,J=13.1Hz,1H),2.70-2.36(m,4H),2.10(d,J=19.5Hz,1H).LCMS m / z 465.15[M+H]+.

[0251] Preparation S35 1-[2-Bromo-4-hydroxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoroethanone (S35) [ka]

[0252] Step 1. Synthesis of 2'-bromo-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (C53) A solution of 1-[2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoro-ethanone S34 (240 mg, 0.5040 mmol) in MeCN (9.7 mL) was treated with cobalt(II) acetate tetrahydrate (13 mg, 0.05219 mmol) and N-hydroxyphthalimide (38 mg, 0.2329 mmol). The reaction was purged and evacuated with oxygen (3x) and heated to 45°C under an oxygen balloon. After 8 h, the reaction was diluted with water and partitioned with DCM. The organics were collected through a phase separator and concentrated via rotary evaporator. Purification by silica gel chromatography (gradient: 0 to 60% EtOAc in heptane) afforded the product 2'-bromo-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C53 (130 mg, 52%). 1H NMR (400 MHz, chloroform-d) δ 7.67 (s, 1H), 7.37 (s, 1H), 5.57 (t, J = 9.1 Hz, 1H), 4.18-3.86 (m, 6H), 3.17 (dd, J = 14.6, 11.0 Hz, 1H), 2.75-2.57 (m, 2H), 2.15 (d, J = 14.2 Hz, 1H).19F NMR (376 MHz, chloroform-d) δ -69.96. LCMS m / z 479.06 [M+H]+.

[0253] Step 2. Synthesis of 1-[2-bromo-4-hydroxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoroethanone (S35) To a flask containing MTBE (1.67 mL) at 0° C. under N2 was added (R)-(+)-2-methyl-CBS-oxazaborolidine solution (50 μL of a 1 M solution in toluene, 0.0500 mmol), followed by dropwise addition of borane tetrahydrofuran (506 μL of a 1 M solution in THF, 0.5060 mmol). A solution of 2'-bromo-2-(1-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C53 (125 mg, 0.2530 mmol) in MTBE (850 μL) was then added dropwise. The reaction mixture was stirred for 15 min at 0° C. and then slowly quenched with 1 M HCl. The resulting mixture was allowed to warm to room temperature and stirred overnight. The organic layer was separated, washed with water and brine, dried over sodium sulfate, and concentrated. Purification by silica gel chromatography (gradient: 0-60% EtOAc in heptane) afforded the product 1-[2-bromo-4-hydroxy-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoro-ethanone S35 (84 mg, 69%) as a mixture of stereoisomers. 1H NMR (400MHz, chloroform-d)δ 7.61(s,1H),6.98(d,J=6.0Hz,1H),5.67-5.36(m,1H),4.48(dq,J=9.3,3.1Hz,1H),4.08(d,J=1.9Hz,3H),4.01-3.7 5(m,4H),2.98(dt,J=57.0,13.0Hz,1H),2.66-2.35(m,2H),2.19(dd,J=8.9,5.0Hz,1H),1.96(d,J=14.9Hz,1H).LCMS m / z 481.08[M+H]+.

[0254] Compounds 44 and 45 2-Bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidin]-4-ol (44) and (45) [ka]

[0255] Synthesis of 2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidin]-4-ol (44) and (45) S35 was purified to give two product peaks, each of which was taken up separately in DCM (700 μL) and treated with NaOH (127 μL of a 2 M solution, 0.2540 mmol). After stirring for 30 min, the organics were collected through a phase separator and dried to give 2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidin]-4-ol 44 (3.6 mg, 54%). 1H NMR(400MHz,DMSO-d6)δ 7.89(s,1H),7.07(s,1H),5.42(d,J=6.6Hz,1H),4.43(d,J=5.1Hz,1H),4.14-3.86(m,5H),3.62(dd,J=11 .7,5.6Hz,1H),3.08-2.76(m,2H),2.17(d,J=13.4Hz,1H),2.02(d,J=13.5Hz,1H),1.76-1.54(m,2H)LCMS m / z 385.09 [M+H]+, and 2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidin]-4-ol 45 (1.8 mg, 60%). 1H NMR(400MHz,DMSO-d6)δ 7.90(s,1H),7.07(s,1H),5.40(s,1H),4.41(s,1H),4.18-3.79(m,5H),3.64(dd,J=11.8,5.1Hz ,1H),3.07-2.73(m,2H),2.29(d,J=13.3Hz,1H),1.92(d,J=13.3Hz,1H),1.76-1.46(m,2H).LCMS m / z 385.09[M+H]+.

[0256] compound 46 2-(3,3-Difluorocyclobutyl)-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (46) [ka]

[0257] Synthesis of 2-(3,3-difluorocyclobutyl)-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (46) To a vial was added Ir[df(CF3)ppy]2(dtbbpy)PF6 (2 mg, 0.001981 mmol), NiCl2 glyme (4 mg, 0.01820 mmol), and 4-tert-butyl-2-(4-tert-butyl-2-pyridyl)pyridine (5 mg, 0.01863 mmol) as solids under an inert atmosphere. A solution of 1-[2-bromo-2'-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-yl]-2,2,2-trifluoro-ethanone S34 (80.68 mg, 0.1578 mmol) in DME (1 mL) was added, followed by a solution of bis(trimethylsilyl)silyl-trimethyl-silane (45 mg, 0.1810 mmol), 2,6-dimethylpyridine (38 mg, 0.3546 mmol), and 3-bromo-1,1-difluoro-cyclobutane (207.5 μL, 1.578 mmol) in DME (1 mL). The vial was sealed and irradiated overnight in a Sigma SynLED photoreactor. The reaction vial was uncapped, diluted with water (2 mL) and DCM (2 mL), and stirred for several minutes. The biphasic mixture was passed through parallel hydrophobic filter plates. The organic layer was evaporated to give crude C54. To this was added methanol (1.050 mL) and NaOH (283.2 μL 6 M solution, 1.699 mmol). The resulting mixture was stirred at −55° C. for 20 min. The reaction mixture was evaporated at 40° C. via Genevac. Water (2 mL) and DCM (2 mL) were added and the mixture was passed through a phase separator. The organic layer was concentrated in vacuo. Purification by reverse phase HPLC (Method: C18 Waters Sunfire column (30×150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) gave 2-(3,3-difluorocyclobutyl)-2′-(1-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4′-piperidine] 46 (8.6 mg, 10%) as the trifluoroacetate salt.1H NMR (400MHz, methanol-d4)δ 8.04(s,1H),6.70(d,J=0.9Hz,1H),4.86(dd,J=12.5,3.1Hz,1H),4.12(s,3H),4.09-3.93(m,2H), 3.66-3.49(m,2H),3.40(ddd,J=12.8,4.6,2.1Hz,1H),3.16-2.87(m,2H),2.76-2.01(m,8H).LCMS m / z 381.28[M+H]+.

[0258] Preparation S37 2-Bromo-2'-phenyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate tert-butyl ester (S36) [ka]

[0259] Step 1. Synthesis of tert-butyl 2'-phenylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate (C55) To a solution of tert-butyl 4-oxo-2-phenyl-piperidine-1-carboxylate S30 (3 g, 10.90 mmol) and 2-(3-thienyl)ethanol S1 (2 g, 15.60 mmol) in dioxane (15 mL) was added triflic acid (2.95 mL, 33.32 mmol) dropwise at 0° C. The reaction was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was concentrated via rotary evaporator to give crude 2′-phenylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4′-piperidine] (3 g, 45%). LCMS m / z 285.96 [M+H]+. This material was taken up in ACN (50 mL) and treated with Boc2O (1.5 g, 6.873 mmol) and TEA (1 g, 9.882 mmol) and the reaction was stirred overnight. The reaction mixture was then concentrated, diluted with EtOAc and washed with saturated sodium bicarbonate solution. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-100% EtOAc in heptane) afforded tert-butyl 2'-phenylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate C55 (2.5 g, 72%). LCMS m / z 386.14 [M+H]+.

[0260] Step 2. Synthesis of tert-butyl 2-bromo-2'-phenyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate (S36) 2'-Phenylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate tert-butyl C55 (600 mg, 1.556 mmol) was dissolved in ACN (20 mL) and DMAP (20 mg, 0.1637 mmol) was added followed by NBS (280 mg, 1.573 mmol). The reaction was stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate and diluted with water and ethyl acetate. The organic layer was washed with water, dried over sodium sulfate and concentrated. Purification by silica gel chromatography (gradient: 0-100% EtOAc in heptane) afforded 2-bromo-2'-phenyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-1'-carboxylate tert-butyl S36 (700 mg, 80%) as a mixture of stereoisomers. LCMS 463.94[M+H]+.

[0261] Compound 47-50 Compounds 47-50 (see Table 4) were prepared by a similar method to compound 46, with modifications that would be apparent to one of skill in the art based on the protecting groups present. All compounds were isolated as trifluoroacetate salts. Alkyl halides were obtained from commercial sources. [Table 4]

[0262] Preparation S37 (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (S37) [ka]

[0263] Step 1. Synthesis of (2S)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (C56) (S)-2-ethynyl-4-oxopiperidine-1-carboxylate tert-Butyl S16 (3.4 g, 15.23 mmol) and 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (3.5 g, 17.30 mmol) were dissolved in dioxane (51 mL) and cooled to 0° C. Triflic acid (4 mL, 45.20 mmol) was added dropwise and the reaction was allowed to warm to room temperature. After 6 h, the solution was diluted with DCM and quenched with 2M Na2CO3. The mixture was extracted with DCM (3 eq. v), dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-20% MeOH in DCM) afforded the product (2S)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] C56 (3.8 g, 56%) as a mixture of stereoisomers. LCMS m / z 302.06 [M+H]+.

[0264] Step 2. Synthesis of (2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (C57) To a solution of 2,4-dimethoxybenzaldehyde (2.75 g, 16.55 mmol) in DCE (43 mL) was added (2S)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] C56 (3.7 g, 8.271 mmol) and sodium triacetoxyborohydride (5.2 g, 24.65 mmol). The reaction mixture was stirred for 30 min and quenched with saturated sodium bicarbonate. The solution was diluted with DCM and the organic layer was collected through a phase separator. The solvent was removed in vacuo. Purification by silica gel chromatography (gradient: 0-60% EtOAc in heptane) afforded (2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] C57 (3.4 g, 86%) as a mixture of diastereomers. LCMS m / z 452.14[M+H]+

[0265] Step 3. Synthesis of (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (S37) To a solution of (2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (3.4 g, 7.087 mmol) in THF (48 mL) was added n-BuLi (4.25 mL of a 2.5 M solution in hexanes, 10.62 mmol) under nitrogen at -78°C. The reaction was stirred at -78°C for 30 min, then TMSCl (7.8 mL of a 1 M solution in THF, 7.800 mmol) was added. The reaction was allowed to warm to room temperature and after 30 min was quenched with saturated ammonium chloride solution and diluted with water. The mixture was extracted with DCM (3x) and the organic layer was dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-50% EtOAc in heptane) separated the two diastereomers. Peak B gave the desired (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] S37 (1.1 g, 59%). 1H NMR (300 MHz, chloroform-d) δ 7.38(d,J=8.2Hz,1H),7.09(d,J=1.4Hz,1H),6.54-6.43(m,2H),4.29(d,J=13.3Hz, 1H),3.88(td,J=5.6,1.9Hz,2H),3.81(d,J=5.4Hz,6H),3.65(d,J=13.3Hz,1H),3.40 (dd,J=11.6,2.8Hz,1H),2.77(dt,J=12.0,3.6Hz,1H),2.65(dt,J=6.5,3.3Hz,2H),2 .43-2.28(m,2H),2.08(dd,J=14.0,11.6Hz,1H),1.95-1.85(m,2H),0.17(s,9H).19F NMR(282MHz,chloroform-d)δ -55.26.

[0266] compound 51 ((2S)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol (51) [ka]

[0267] Step 1. Synthesis of ((2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol (C58) (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] To a solution of S37 (350 mg, 0.6590 mmol) in THF (5 mL) was added n-BuLi (430 μL of a 2.3 M solution in hexanes, 0.9890 mmol) at −78° C. under nitrogen. The reaction was stirred for 30 min and then DMF (51 μL, 0.6587 mmol) was added. The reaction was allowed to warm slowly to room temperature and then quenched with NH4Cl and diluted with DCM. The organic layer was collected through a phase separator and dried to give the crude aldehyde (363 mg), which was immediately dissolved in MeOH (700 μL) and DCM (2.7 mL) and treated with sodium borohydride (264 μL, 6.594 mmol). After 15 min, the reaction was quenched with saturated sodium bicarbonate solution, diluted with DCM, and the organics were extracted through a phase separator. The volatiles were removed in vacuo. Purification by silica gel chromatography (gradient: 0-70% EtOAc in heptane) afforded ((2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol C58 (113 mg, 31%). 1H NMR (300MHz, chloroform-d)δ 7.19(s,1H),6.39-6.25(m,2H),4.46(d,J=3.3Hz,2H),4.12(d,J=13.3Hz,1H),3.82-3.72(m,2H),3.64(d,J=5.2Hz,6H),3.48(d,J=13.3Hz, 1H),3.24(dd,J=11.6,2.8Hz,1H),2.66-2.38(m,3H),2.17(td,J=12.4,3.3Hz,2H),2.01-1.84(m,1H),1.83-1.62(m,2H),0.00(s,9H).LCMS m / z 554.13[M+H]+.

[0268] Step 2. Synthesis of ((2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol (C59) ((2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol To a solution of C58 (115 mg, 0.2077 mmol) in THF (1.75 mL) was added TBAF (56 μL, 0.1900 mmol) at 0° C. The reaction was allowed to warm to room temperature and after 1 h was quenched with saturated sodium bicarbonate solution and diluted with DCM. The organics were collected through a phase separator and concentrated by rotary evaporation. Purification by silica gel chromatography (gradient: 0 to 70% EtOAc in heptane) afforded the product ((2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol C59 (95 mg, 92%). 1H NMR (300MHz, chloroform-d)δ 7.39(d,J=8.2Hz,1H),6.67-6.36(m,2H),4.66(d,J=5.0Hz,2H),4.40-4.10(m,1H) ,4.07-3.34(m,10H),2.85-2.52(m,3H),2.52-2.24(m,3H),2.24-1.76(m,3H).LCMS m / z 482.16[M+H]+.

[0269] Step 3. Synthesis of ((2S)-1-(2,4-dimethoxybenzyl)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol (C60) To a solution of ((2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol C59 (95 mg, 0.1973 mmol), copper(II) sulfate (15 mg, 0.09398 mmol), and sodium ascorbate (15 mg, 0.07533 mmol) in DMF (400 μL) was added azidomethyl(trimethyl)silane (124 μL of a 1.5 M solution, 0.1860 mmol). The reaction was heated at 40° C. for 4 h, cooled to room temperature, and continued stirring for an additional 48 h. The reaction was quenched with saturated sodium bicarbonate solution and DCM, and the organic layer was collected through a phase separator. The solvent was removed in vacuo to give the crude TMS protected intermediate LCMS m / z 611.2 [M+H]+. The crude reaction mixture was dissolved in THF (950 μL) and TBAF (70 μL, 0.2375 mmol) was added at 0° C. The reaction was stirred for 3 h, at which point complete conversion was observed. The reaction was quenched with saturated sodium bicarbonate solution, diluted with DCM and passed through a phase separator. The organics were dried via rotary evaporator and purified by silica gel chromatography (gradient: 0-70% EtOAc in heptane) to give the product ((2S)-1-(2,4-dimethoxybenzyl)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol C60 (52 mg, 46%). 1H NMR (300MHz, chloroform-d)δ 7.56(s,1H),7.25(d,J=8.2Hz,1H),6.54-6.30(m,2H),4.65(s,2H),4.05(d,J=20.9Hz,6H),3.78(d,J=15.4Hz,6H),3.67(d,J=13.8Hz,1H),3 .21(d,J=13.8Hz,1H),2.87(s,1H),2.75(dt,J=7.2,3.6Hz,2H),2.58(td,J=11.1,5.1Hz,1H),2.34(d,J=13.9Hz,1H),2.08-1.91(m,4H).LCMS m / z 539.24[M+H]+.

[0270] Step 4. Synthesis of ((2S)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol (51) A mixture of ((2S)-1-(2,4-dimethoxybenzyl)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)methanol C60 (45 mg, 0.08355 mmol) in water (78 μL, 4.330 mmol) and TFA (227 μL, 2.946 mmol) was heated to 90° C. Upon completion, the reaction was cooled to room temperature, diluted with DCM, and then slowly poured into 6M aqueous NaOH. The organic layer was collected through a phase separator and dried in vacuum. The organic layer was separated through a phase separator and the solvent was blown off. Purification by silica gel chromatography (gradient: 0–20% MeOH in DCM) afforded the product ((2S)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2′-(trifluoromethyl)-4′,5′-dihydrospiro[piperidine-4,7′-thieno[2,3-c]pyran]-3′-yl)methanol 51 (15.6 mg, 48%). 1H NMR (300MHz, chloroform-d)δ 7.45(s,1H),4.64(d,J=1.4Hz,2H),4.37(dd,J=11.7,2.6Hz,1H),4.02(d,J=17.2Hz,5H),3.25(td,J=12.4,2.7Hz,1H),3.00(d dd,J=12.3,4.9,2.1Hz,1H),2.75(t,J=5.5Hz,2H),2.36(dt,J=13.6,2.7Hz,1H),2.04(q,J=2.7Hz,1H),1.95-1.68(m,2H).LCMS m / z 389.18[M+H]+.

[0271] Compounds 52 and 53 1-((2S)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)-1l3-ethan-1-ol (52) [DIAST-1] and (53) [DIAST-2] [ka]

[0272] Step 1. Synthesis of (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-carbaldehyde (C61) To a solution of 2-[1'-[(2,4-dimethoxyphenyl)methyl]-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'-yl]ethynyl-trimethyl-silane (1.1 g, 2.078 mmol) in THF (15 mL) was added n-BuLi (1.4 mL of a 2.3 M solution in hexanes, 3.220 mmol) under N2 at -78 °C. The reaction was allowed to stir at -78 °C for 30 min and then DMF (240 μL, 3.100 mmol) was added. The reaction was allowed to warm to room temperature over 30 min, quenched with saturated ammonium chloride solution and diluted with DCM. The organic layer was collected through a phase separator and concentrated to give crude (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-carbaldehyde C61 (460 mg, 40%), which was carried on directly to the next step. LCMS m / z 552.1[M+H]+

[0273] Step 2. Synthesis of 1-((2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)ethan-1-ol (C62) The crude (2S)-1-(2,4-dimethoxybenzyl)-2'-(trifluoromethyl)-2-((trimethylsilyl)ethynyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-carbaldehyde C61 (460 mg) was dissolved in THF (15 mL) and cooled to 0 °C. Methylmagnesium chloride (1.22 mL of a 3.4 M solution in THF, 4.148 mmol) was added. The reaction was stirred for 20 min and complete conversion was observed. To this reaction mixture was added TBAF (4.1 mL of a 1 M solution, 4.100 mmol). After 2 h, the reaction was quenched with saturated sodium bicarbonate solution and diluted with DCM. The organic phase was collected through a phase separator and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient: 0-70% EtOAc in heptane) afforded the product 1-((2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)ethan-1-ol C62 (345 mg, 83%). 1H NMR (300 MHz, chloroform-d) δ 7.39(d,J=8.2Hz,1H),6.66-6.39(m,2H),5.27(p,J=7.7,6.9Hz,1H),4.30(d, J=13.5Hz,1H),3.99-3.73(m,8H),3.63(d,J=13.5Hz,1H),3.50-3.38(m,1H),3 .00(dq,J=16.4,5.5Hz,1H),2.77(dq,J=14.2,5.8Hz,2H),2.51-2.25(m,3H),2 .12(ddd,J=14.1,11.6,8.8Hz,1H),2.01-1.80(m,3H),1.58-1.44(m,3H).LCMS m / z 496.07[M+H]+.

[0274] Step 3. Synthesis of 1-((2S)-1-(2,4-dimethoxybenzyl)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)ethan-1-ol (C63) To a solution of 1-((2S)-1-(2,4-dimethoxybenzyl)-2-ethynyl-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)ethan-1-ol C62 (340 mg, 0.6721 mmol), copper(II) sulfate (54 mg, 0.3383 mmol), and sodium ascorbate (67 mg, 0.3365 mmol) in DMF (1.4 mL) was added azidomethyl(trimethyl)silane (422 μL of a 1.5 M solution, 0.6330 mmol). The reaction was heated at 40° C. for 4 h, cooled to room temperature, and continued stirring for an additional 48 h. The reaction was quenched with saturated sodium bicarbonate solution and DCM, and the organic layer was collected through a phase separator. The solvent was removed in vacuo to give the crude TMS protected intermediate. LCMS m / z 625.11 [M+H]+. The crude reaction mixture was dissolved in THF (3.3 mL) and TBAF (238 μL, 0.8070 mmol) was added at 0° C. The reaction was stirred for 3 h, at which point complete conversion was observed. The reaction was quenched with saturated sodium bicarbonate solution, diluted with DCM and passed through a phase separator. The organics were dried via rotary evaporator and purified by silica gel chromatography (gradient: 0 to 70% EtOAc in heptane) to give the product 1-((2S)-1-(2,4-dimethoxybenzyl)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)ethan-1-ol C63 (290 mg, 74%).1H NMR (300MHz, chloroform-d) δ 7.56(s,1H),7.24(s,1H),6.56-6.36(m,2H),5.32(s,2H),5.25(s,1H),4.04 (d,J=31.0Hz,6H),3.79(d,J=15.8Hz,6H),3.67(d,J=13.8Hz,1H),3.22(d,J= 13.8Hz,1H),3.03(d,J=5.9Hz,0H),2.88-2.71(m,2H),2.57(d,J=11.9Hz,1H) ,2.36(d,J=13.8Hz,1H),2.15-1.85(m,4H),1.51(dd,J=6.7,2.7Hz,3H).LCMS m / z 553.05[M+H]+.

[0275] Step 4. Synthesis of 1-((2S)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)-11-3-ethan-1-ol (52) [DIAST-1] and (53) [DIAST-2] A solution of 1-((2S)-1-(2,4-dimethoxybenzyl)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)ethan-1-ol C63 (278 mg, 0.5031 mmol) in water (470 μL, 26.09 mmol) and TFA (1.36 mL, 17.65 mmol) was heated to 90 °C for 3 h. The reaction mixture was cooled to room temperature, diluted with DCM, and then slowly poured into 6 M aqueous NaOH. The organic layer was separated by passing through a phase separator and concentrated. Purification by silica gel chromatography (gradient: 0-20% MeOH in DCM) afforded a 1:1 diastereomeric mixture. This material was separated into its constituent diastereomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 10×250 mm; Mobile phase: 20% IPA with 5 mM ammonia, 80% carbon dioxide. 1-((2S)-2-(1-methyl-1H-1,2,3-triazol-4-yl)-2′-(trifluoromethyl)-4′,5′-dihydrospiro[piperidine-4,7′-thieno[2,3-c]pyran]-3′-yl)-113-ethan-1-ol 52 [DIAST-1] (20.3 mg, 20%). LCMS m / z 403.09 [M+H]+. 1-((2S)-2-(1-Methyl-1H-1,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-3'-yl)-113-ethan-1-ol 53 [DIAST-2] (5.6 mg, 5%). LCMS m / z 403.09[M+H]+.

[0276] Preparation S38 (2'S,7S)-2-Chloro-2'-ethynylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (S38) [ka]

[0277] Step 1. Synthesis of (2S)-2'-chloro-2-ethynyl-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (C64) To a mixture of (2S)-tert-butyl 2-ethynyl-4-oxo-piperidine-1-carboxylate S16 (3100 mg, 13.88 mmol) in DCM (57 mL) was added 2-(5-chloro-3-thienyl)ethanol S2 (2 mL) followed by methanesulfonic acid (3.5 mL, 53.94 mmol). After stirring for 40 min, the mixture was washed with NaOH (9.3 mL of a 6 M solution, 55.80 mmol), diluted with 20 mL of water and the organic layer was removed. The aqueous layer was extracted with DCM (20 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated. Purification by silica gel chromatography (gradient: 0-10% MeOH in DCM) afforded (2S)-2'-chloro-2-ethynyl-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] C64 (2.5 g, 56%) as a mixture of diastereomers. LCMS m / z 268.13 [M+H]+.

[0278] Step 2. Synthesis of (2'S,7S)-2-chloro-2'-ethynyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (S38) (2S)-2'-chloro-2-ethynyl-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] C64 (2.5 g) was separated into its constituent diastereomers by chiral SFC separation. Column: Phenomenex Lux® Cellulose-2, 20×250 mm; Mobile phase: 20% MeOH (containing 5 mM ammonia), 80% carbon dioxide. Peak A gave (2'S,7S)-2-chloro-2'-ethynyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] S38 (350 mg, 9%) as a brown oil. 1H NMR (300MHz, methanol-d4)δ 6.68(s,1H),3.91(td,J=5.6,1.5Hz,2H),3.80(dt,J=11.7,2.6Hz,1H),2.99(td,J=12.7,2.9Hz,1H),2.84(ddd,J=12.8,4.8,2.1Hz,1H ),2.70(d,J=2.3Hz,1H),2.62-2.56(m,2H),2.22(dt,J=13.8,2.8Hz,1H),2.01-1.92(m,1H),1.83-1.73(m,1H),1.74-1.63(m,1H).LCMS m / z 268.04[M+H]+. NOTE: The stereochemistry of this intermediate was confirmed by synthesizing compound 2 and using it to find the data.

[0279] compound 54 3-(4-((2S,4S)-2'-chloro-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-2-yl)-1H-1,2,3-triazol-1-yl)benzamide (54) [ka]

[0280] To a mixture of 3-aminobenzamide (33.86 mg, 0.2487 mmol) in DMSO (0.5 mL), aqueous sodium bicarbonate (174.1 μL of 12% w / v, 0.2487 mmol) was added, followed by a solution of N-diazosulfamoyl fluoride (31.11 mg, 0.2487 mmol) in MTBE (0.54 mL) (prepared according to the literature in Meng, G., Guo, T., Ma, T. et al. Modular click chemistry library for functional screening using diazotized reagents. Nature 574, 86-89 (2019)). The mixture was stirred vigorously for 5 min. At this point, (2'S,7S)-2-chloro-2'-ethynyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] S38 (20 mg, 0.07469 mmol) in DMSO (0.4 mL) was added, followed by aqueous CuSO4 (59.61 μL of 1% w / v, 0.003735 mmol) and sodium ascorbate (2.631 mg, 0.01494 mmol). The mixture was heated to 50° C. in air and stirred overnight. The mixture was evaporated to remove residual MTBE and purified directly by reverse-phase HPLC (Method: C18 Waters Sunfire column (30 × 150 mm, 5 micron). Gradient: MeCN in HO containing 5 mM HCl) to give 3-(4-((2S,4S)-2'-chloro-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-2-yl)-1H-1,2,3-triazol-1-yl)benzamide 54 (18.3 mg, 52%) as the hydrochloride salt. 1H NMR (300MHz, methanol-d4)δ 8.74(s,1H),8.37(t,J=1.9Hz,1H),8.11-7.97(m,2H),7.71(t,J=8.0Hz,1H),6.78(s,1H),4.99(dd,J=12.6,3.1Hz,1H),4.06(td,J=5.6,2.4 Hz,2H),3.68-3.55(m,1H),3.51-3.40(m,1H),2.70(t,J=5.4Hz,2H),2.66(s,3H),2.50-2.33(m,2H),2.13(td,J=14.6,14.2,4.8Hz,1H).LCMS m / z 430.19[M+H]+.

[0281] compound 55 (2S,4S)-2'-Chloro-2-(1-phenyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (55) [ka]

[0282] Preparation of (2S,4S)-2'-chloro-2-(1-phenyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] (55) To azidobenzene (16 mg, 0.1343 mmol) in methanol (800.0 μL) was added aqueous copper(II) sulfate (100 μL of 1% w / v, 0.006265 mmol) and to the pale blue solution was added (2S)-tert-butyl 2-ethynyl-4-oxo-piperidine-1-carboxylate S16 (20 mg, 0.08958 mmol) followed by sodium ascorbate (2 mg, 0.01136 mmol). After stirring overnight, the mixture was concentrated and the residue was diluted with DCM and saturated aqueous sodium bicarbonate. The organic layer was collected through a phase separator to give a C65 solution. To this was added MsOH (25 μL, 0.3853 mmol) followed by 2-(5-chloro-3-thienyl)ethanol S2 (25 mg, 0.1537 mmol) and the mixture was stirred overnight. At this point, the mixture was pH adjusted with saturated aqueous sodium bicarbonate, and the organics were collected via a phase separator and blown off with nitrogen. Purification by reverse-phase HPLC (Method: Purification by C18 Waters Sunfire column (30×150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) gave (2S,4S)-2'-chloro-2-(1-phenyl-1H-1,2,3-triazol-4-yl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran] 55 (24.9 mg, 55%) as the trifluoroacetate salt. 1H NMR (300 MHz, methanol-d4) δ 8.67(d,J=0.5Hz,1H),7.90-7.76(m,2H),7.65-7.55(m,2H),7.55-7.42(m,1H),6 .76(s,1H),4.97(dd,J=12.4,3.2Hz,1H),4.05(td,J=5.6,2.2Hz,2H),3.61(td,J= 13.1,3.2Hz,1H),3.46(ddd,J=12.9,4.8,2.1Hz,1H),2.73-2.61(m,3H),2.45(dd, LCMS m / z 387.25[M+H]+.

[0283] Compounds 56~77 Compounds 56-77 (see Table 5) were prepared by methods similar to compounds 54 or 55, with modifications that would be apparent to one of skill in the art. The amines or azides were obtained from commercial sources. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7]

[0284] compound 78 2-Ethyl-2'-(4-fluorophenyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] (78) [ka]

[0285] Step 1. Synthesis of benzyl 2-(4-fluorophenyl)-4-oxo-piperidine-1-carboxylate (C65) A solution of copper(I) bromide dimethylsulfide complex (1.5 g, 7.296 mmol) in THF (25 mL) was cooled to -78 °C. 4-Fluorophenylmagnesium bromide (7.3 mL of a 1 M solution in THF, 7.300 mmol) was added slowly via an addition funnel. After stirring at -78 °C for 1 h, diethyloxonio(trifluoro)boranuide (896 μL, 7.260 mmol) was added and stirred for 5 min. To the newly formed complex was then added a solution of benzyl 4-oxo-2,3-dihydropyridine-1-carboxylate C34 (999.0 mg, 4.32 mmol) in THF (15 mL) slowly over 1 h. After stirring at -78 °C for 2 h, 16 mL of 20% NH4Cl / conc. NH4OH (1:1) was added and the mixture was allowed to warm to room temperature. The mixture was extracted with EtOAc (3×100 mL) and the combined organic layers were washed with brine, dried over MgSO4, and concentrated. Purification by silica gel chromatography afforded the product 2-(4-fluorophenyl)-4-oxo-piperidine-1-carboxylate benzyl C66 (883 mg, 33%). LCMS m / z 327.93 [M+H]+.

[0286] Step 2. Synthesis of 2-ethyl-2'-(4-fluorophenyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] (C67) A solution of 2-(4-fluorophenyl)-4-oxo-piperidine-1-carboxylate benzyl C66 (115 mg, 0.3513 mmol) and 2-(5-ethyl-2-thienyl)ethanol S6 (55 mg, 0.3520 mmol) in dioxane (2 mL) was cooled to 0 °C and treated dropwise with trifluoromethanesulfonic acid (95 μL, 1.074 mmol). The mixture was stirred at 0 °C for 30 min and allowed to warm to room temperature. After an additional 30 min, saturated sodium bicarbonate was added and the mixture was extracted with DCM (3 × 3 mL). The combined organic layers were dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0-30% EtOAc in hexanes) afforded the CBz-protected intermediate, which was immediately treated with Pd / C (38 mg of 10 wt%, 0.03571 mmol) and dissolved in MeOH (10 mL). The reaction was purged and evacuated (3 times) and stirred under a hydrogen balloon atmosphere. After 1 h, the mixture was filtered through a pad of Celite and the filtrate was concentrated. Purification by silica gel chromatography (gradient: 0-10% 0.7 M ammonia in MeOH in DCM) provided 2-ethyl-2'-(4-fluorophenyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]C67 (32 mg, 26%). LCMS m / z 332.03[M+H]+.

[0287] Step 3. Synthesis of 2-ethyl-2'-(4-fluorophenyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] (78) Racemic 2-ethyl-2'-(4-fluorophenyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]C67 (26 mg, 0.07292 mmol) was separated into its four constituent stereoisomers by chiral SFC separation. Column: Daicel Chiralpak® AD-H, 10×250 mm; Mobile phase: 15% EtOH (5 mM ammonia), 85% carbon dioxide. Peak A gave 2-ethyl-2'-(4-fluorophenyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]78 (3.2 mg, 42%). 1H NMR (300MHz, chloroform-d)δ 7.53-7.31(m,2H),7.10-6.90(m,2H),6.50(d,J=1.1Hz,1H),4.09(dd,J=11.5,2.6Hz,1H),4.05-3.91(m,2H),3.37-3.17 LCMS m / z 331.53[M+H]+. EXAMPLES

[0288] Assays for detecting and measuring the APOL1 inhibitor properties of compounds MultiTox-Fluor multiplex cytotoxicity assay The MultiTox-Fluor multiplex cytotoxicity assay is a single-reagent-addition, homogeneous fluorescent assay that simultaneously measures the number of live and dead cells in a culture well. The assay measures cell viability and cytotoxicity by detecting two distinct protease activities. Live cell protease activity is restricted to intact live cells and measured using a fluorogenic cell-permeant peptide glycyl-phenylalanylaminofluorocoumarin (GF-AFC) substrate. The substrate enters intact cells where it is cleaved to generate a fluorescent signal proportional to the number of live cells. This live cell protease activity marker becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium. A second cell-impermeant fluorescent peptide substrate (bis-AAF-R110 substrate) is used to measure dead cell proteases released from cells that have lost membrane integrity. The ratio of dead to live cells is used to normalize the data.

[0289] Briefly, tet-inducible transgenic APOL1 T-REx-HEK293 cell lines were incubated in duplicate with 50ng / mL tet in the presence of 3-(2-(4-fluorophenyl)-1H-indol-3-yl)-N-((3S,4R)-4-hydroxy-2-oxopyrrolidin-3-yl)propenamide 10.03, 3.24, 1.13, 0.356, 0.129, 0.042, 0.129, 0.0045, 0.0015, 0.0005μM in a humidified 37°C incubator for 24 hours to induce APOL1. MultiTox reagent was added to each well and placed back in the incubator for an additional 30 minutes. Plates were read on an EnVision plate reader. Dead to live cell ratio was used for normalization and data was imported, analyzed, and fitted using Genedata Screener (Basel, Switzerland) software. Data was normalized using percentages of control, no tet (100% viability), and 50 ng / mL tet treatment (0% viability) and fitted using Smart Fit. Reagents, methods, and a complete protocol for the MultiTox assay are described below. [Table 6-1] [Table 6-2] [Table 7]

[0290] Multiple toxicity assay protocols Human embryonic kidney (HEK293) cell lines G0 DC2.13, G1 DC3.25, and G2 DC4.44 containing the tet-inducible expression system (T-REx™, Invitrogen, Carlsbad, CA) and the adeno-associated virus site 1 pAAVS1-Puro-APOL1 G0 or pAAVS1-Puro-APOL1 G1 or pAAVS1-Puro-APOL1 G2 clones were grown at approximately 90% confluency in T-225 flasks in cell growth medium (DMEM solution, 10% Tet-free FBS, 2 mM L-glutamine, 100 units / mL penicillin-streptomycin, 5 μg / mL blasticidin S HCl, 1 μg / mL puromycin dihydrochloride). The cells were washed with DPBS and then trypsinized to dissociate from the flasks. The trypsin was quenched using medium, then the cells were pelleted at 200 g and resuspended in fresh cell assay medium (DMEM solution, 2% Tet-free FBS, 2 mM L-glutamine, 100 units / mL penicillin-streptomycin). The cells were counted and diluted to 1.17×106 cells / mL. 20 μL of cells (23,400 / well) were dispensed into each well of a 384-well poly-D-lysine coated plate using a Multidrop dispenser. The plate was then incubated at room temperature for one hour.

[0291] Tetracycline is required to induce APOL1 expression. 1 mg / mL tet stock in water was diluted to 250 ng / mL (5x) in cell assay medium. 60 μL of cell assay medium (no tet control) was dispensed into columns 1 and 24, and 60 μL of 5Xtet in a 384-PP-round bottom plate was dispensed into columns 2-23 with a Multidrop dispenser.

[0292] Assay ready plates from the Global Compound Archive were ordered using the template 384_APOL1Cell_DR10n2_50uM_v3. Compounds were dispensed at 200nL in DMSO. The final top concentration was 10μM and for the MultiTox assay, 10-point 3-fold dilutions were done in duplicate.

[0293] 20 μL was transferred from the 5Xtet plate to the ARP and mixed, then 5 μL of 5Xtet plus compound was transferred to the cell plate and mixed using a Bravo. The cell plate was placed in a humidified 37° C. 5% CO2 incubator for 24 hours.

[0294] The MultiTox-Fluor Multiplex Cytotoxicity Assay was performed according to the manufacturer's protocol. After incubating cells with tet and compounds for 24 hours, 25 μL of 1× MultiTox reagent was added to each well using a Multidrop dispenser and the plate was placed on a plate shaker (600 rpm) for 2 minutes, then briefly centrifuged and returned to a 37° C. incubator for 30 minutes. Cell viability (excitation: 400 nm, emission: 486 nm) and cytotoxicity (excitation: 485 nm, emission: 535 nm) were read using an EnVision plate reader. The ratio of dead cells (cytotoxicity) to live cells (viability) was reported. Data was exported and analyzed in Genedata. Data was normalized using percentages of control, no tet (100% viability), and 50 ng / mL tet treatment (0% viability) and fitted using the Smart Fit setting in Genedata.

[0295] Efficacy data for compounds 1-78 Compounds of formula I are useful as inhibitors of APOL1 activity. Table 8 below illustrates the IC50s of compounds 1-78 using the above procedure. The potency of any compound of formula I may be determined using the above procedure. In Table 8 below, the following meanings apply: For IP50 (i.e., IC50 for cell proliferation), "+++" means <0.1 μM, "++" means 0.1-0.5 μM, and "+" means >0.5-1.0 μM. [Table 8]

[0296] Other embodiments The present disclosure provides only non-limiting, exemplary embodiments of the disclosed subject matter. Those skilled in the art will readily recognize from this disclosure and the following claims that various changes, modifications, and variations can be made without departing from the spirit and scope of the present disclosure, as defined in the following claims.

Claims

1. A compound represented by the following structural formula: 【Transformation 62】 tautomers thereof, deuterated derivatives of these compounds or tautomers, or pharmaceutically acceptable salts of any of the foregoing, wherein: X １ S and -CR ２ａ is selected from X ２ S and -CR ２ｂ is selected from: X １ and X ２ One of them is S, X １ If S, then X ２ Ha-CR ２ｂ and X ２ If S, then X １ Ha-CR ２ａ and R １ is hydrogen, halogen, cyano, -OH, C １ -C ６ Alkyl, C １ -C ６ Alkoxy, and C ３ -C ６ cycloalkyl; R １ The above C1-C ６ alkyl is independently selected from halogen, cyano, —OH, —NH ２ , and C １ -C ４ optionally substituted with 1 to 3 groups selected from alkoxy (optionally substituted with 1 to 3 halogen groups); R １ The above C １ -C ６ the alkoxy is optionally substituted with 1 to 3 independently selected halogen; R １ The above C ３ -C ６ Cycloalkyl is halogen, cyano, —OH, —NH ２ , C １ -C ４ Alkyl, and C １ -C ４ optionally substituted with 1 to 3 groups independently selected from alkoxy; R ２ａ is hydrogen, halogen, cyano, —OH, and C １ -C ６ alkyl, wherein R ２ａ C １ -C ６ The alkyl is selected from halogen, cyano, —OH, and C １ -C ４ optionally substituted with 1 to 3 groups independently selected from alkoxy; R ２ｂ is hydrogen, halogen, cyano, —OH, and C １ -C ６ alkyl, Each R ３ａ are independently selected from halogen, cyano, —OH, C １ -C ６ Alkoxy, and C １ -C ６ alkyl (optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and —OH); or Two R's ３ａ together form an oxo group, Each R ３ｂ But independently, C １ -C ２ alkyl (optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and —OH); or Two R's ３ｂ together form an oxo group, R ４ and R ５ One of them is hydrogen and the other is C １ -C ６ Alkyl, C ２ -C ６ alkynyl, and 【Transformation 63】 is selected from: R ４ or R ５ C １ -C ６ Alkyl is halogen, cyano, —OH, —NH ２ , and C １ -C ４ optionally substituted with 1 to 3 groups independently selected from alkoxy; Ring A is C ３ -C １２ cycloalkyl, 3- to 12-membered heterocyclyl, C ６ and C １０ aryl, and 5- to 10-membered heteroaryl, wherein ring A is selected from 1, 2, 3, 4, or 5 R ａ optionally substituted with a group, R ａ For each occurrence, halogen, cyano, C １ -C ６ Alkyl, C ２ -C ６ Alkenyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkyl, C １ -C ６ Haloalkenyl, C １ -C ６ Haloalkoxy, —C(═O)NR ｈ R ｉ , -NR ｈ R ｉ , -NR ｈ C(=O)R ｋ , -NR ｈ C(=O)OR ｋ , -NR ｈ C(=O)NR ｉ R ｊ , -NR ｈ S(=O)pR ｋ , -OR ｋ , —OC(═O)R ｋ , -OC(=O)OR ｋ , -OC(=O)NR ｈ R ｉ , -[O(CH ２ ) ｑ ] ｒ O (C １ -C ６ alkyl), -S(=O) ｐ R ｋ , -S(=O) ｐ NR ｈ R ｉ , -C(=O)OR ｋ , C ３ -C １２ cycloalkyl, 3- to 12-membered heterocyclyl, C ６ and C １０ aryl, and 5- to 10-membered heteroaryl; R ａ The above C １ -C ６ Alkyl, C １ -C ６ Alkoxy, C １ -C ６ haloalkyl, and the C ２ -C ６ Each alkenyl is C ６ -C １０ Aryl (1 to 3 R ｍ group), 5- to 10-membered heterocyclyl (1-3 R ｍ aryl (optionally substituted with 1 to 3 R ｍ group), cyano, —C(═O)R ｋ , -C(=O)OR ｋ , —C(═O)NR ｈ R ｉ , -NR ｈ R ｉ , -NRhC(=O)R ｋ , -NR ｈ C(=O)OR ｋ , -NR ｈ C(=O)NR ｉ R ｊ , -NR ｈ S (= O) ｐ R ｋ , -OR ｋ , —OC(═O)R ｋ , -OC(=O)OR ｋ , -OC(=O)NR ｈ R ｉ , -S(=O) ｐ R ｋ , -S(=O) ｐ NR ｈ R ｉ , and C ３ -C ６ Cycloalkyl group (1 to 3 R ｍ optionally substituted with 1 to 3 groups independently selected from R ａ The above C ３ -C １２ cycloalkyl, the 3- to 12-membered heterocyclyl, the C ６ and C １０ The aryl and the 5- to 10-membered heteroaryl are each selected from halogen, cyano, C １ -C ４ Alkyl, —(C═O)NR ｈ R ｉ , -NR ｈ R ｉ , -OR ｋ optionally substituted with 1 to 3 groups independently selected from , , and oxo; R ｈ , R ｉ , and R ｊ are hydrogen, C, １ -C ４ Alkyl, C ６ -C １０ Aryl, and C ３ -C ６ cycloalkyl; R ｈ , R ｉ , and R ｊ Any one of the C １ -C ４ the alkyl is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and —OH; R ｋ For each occurrence, hydrogen, C １ -C ４ Alkyl, 5- to 10-membered heterocyclyl, and C ３ -C ６ independently selected from carbocycles, wherein R ｋ Any one of the C １ -C ４ the alkyl is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and —OH; R ｍ For each occurrence, halogen, cyano, oxo, C １ -C ６ Alkyl, C １ -C ６ Alkoxy, —S(═O) ｐ R ｋ , and -OR ｋ are independently selected from R ｍ The above C １ -C ６ the alkyl is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and —OH; k is an integer selected from 0, 1, and 2, where R ３ａ is oxo, k is 1; m is an integer selected from 0, 1, and 2, where R ３ｂ is oxo, m is 1; p, for each occurrence, is an integer selected from 1 and 2; A compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt, wherein q and r are integers independently selected from 1, 2, 3, and 4, respectively, for each occurrence.

2. X １ S and -CR ２ａ is selected from X ２ S and -CR ２ｂ is selected from: X １ and X ２ One of them is S, X １ If S, then X ２ Ha-CR ２ｂ and X ２ If S, then X １ Ha-CR ２ａ and R １ But halogen, C １ -C ６ Alkyl, and C ３ -C ６ cycloalkyl, wherein R １ The above C １ -C ６ the alkyl is optionally substituted with 1 to 3 groups independently selected from halogen; R １ The above C ３ -C ６ cycloalkyl is optionally substituted with 1 to 3 groups independently selected from halogen; R ２ａ is hydrogen and C １ -C ６ alkyl, R ２ａ The above C １ -C ６ the alkyl is optionally substituted with 1 to 3 —OH groups; R ２ｂ is hydrogen, Each R ３ａ are independently —OH, C １ -C ６ Alkoxy, and C １ -C ６ alkyl (optionally substituted with 1 to 3 groups independently selected from halogen); or Two R's ３ａ together form an oxo group, Each R ３ｂ But independently, C １ -C ２ alkyl (optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and —OH); or Two R's ３ｂ together form an oxo group, R ４ and R ５ one of which is hydrogen and the other is 【Chemistry 64】 is selected from: Ring A is C ３ -C １２ Cycloalkyl, C ６ aryl, and 5- to 10-membered heteroaryl, and ring A is selected from 1, 2, or 3 R ａ optionally substituted with a group, R ａ may, in each occurrence, independently be a halogen, C １ -C ６ Alkyl, cycloalkyl-C(=O)NR ｈ R ｉ , -OR ｋ , 3- to 12-membered heterocyclyl, C ６ aryl, and 5- to 10-membered heteroaryl; R ａ The above C １ -C ６ The alkyl is —OR ｋ , and -S(=O) ｐ NR ｈ R ｉ and optionally substituted with 1 to 3 groups independently selected from R ａ C ３ -C １２ cycloalkyl, 3- to 12-membered heterocyclyl, C ６ The aryl and the 5- to 10-membered heteroaryl are each selected from halogen, C １ -C ４ Alkyl, —C(═O)NR ｈ R ｉ , -OR ｋ and oxo; R ｈ , R ｉ and R ｊ are hydrogen and C for each occurrence, respectively. １ -C ４ independently selected from alkyl, R ｋ are, for each occurrence, hydrogen and C, respectively. １ -C ４ independently selected from alkyl, k is an integer selected from 0, 1, and 2, where R ３ａ is oxo, k is 1; m is an integer selected from 0, 1, and 2; 2. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt of claim 1, wherein p, for each occurrence, is an integer selected from 1 and 2.

3. X １ S and -CR ２ａ is selected from X ２ S and -CR ２ｂ is selected from: X １ and X ２ One of them is S, X １ If S, then X ２ Ha-CR ２ｂ and X ２ If S, then X １ Ha-CR ２ａ and R １ But halogen, C １ -C ６ Alkyl, and C ３ -C ６ cycloalkyl, wherein R １ The above C １ -C ６ the alkyl is optionally substituted with 1 to 3 groups independently selected from halogen; R １ The above C ３ -C ６ cycloalkyl is optionally substituted with 1 or 2 groups independently selected from halogen; R ２ａ is hydrogen and C １ -C ６ alkyl, R ２ａ The above C １ -C ６ the alkyl is optionally substituted with one —OH; R ２ｂ is hydrogen, R ３ａ are independently —OH, C １ -C ６ Alkyl, and C １ -C ６ or two R 3a together form an oxo group, wherein: R ３ａ The above C １ -C ６ the alkyl is optionally substituted with 1 to 3 groups independently selected from halogen; R ３ｂ But C １ -C ２ alkyl, 【Transformation 65】 But in each occurrence, R ３ａ is —OH and optionally substituted C １ -C ６ alkyl, or R ３ｂ is C １ -C ２ alkyl is a single bond, or alternatively, 【Transformation 6】 But in each occurrence, R ３ａ is an oxo, then it is a double bond, R ４ and R ５ one of which is hydrogen and the other is 【Transformation 67】 is selected from: Ring A is C ３ -C １２ Cycloalkyl, C ６ aryl, and 5- to 10-membered heteroaryl, and ring A is selected from 1, 2, or 3 R ａ optionally substituted with a group, R ａ may, in each occurrence, independently be a halogen, C １ -C ６ Alkyl, C ６ aryl, and 5- to 10-membered heteroaryl; R ａ C １ -C ６ Each alkyl is —OR ｋ and -S(=O) ｐ R ｋ and optionally substituted with 1 to 3 groups independently selected from R ａ The above C ６ The aryl and the 5- to 10-membered heteroaryl are each independently selected from halogen, C １ -C ４ Alkyl, —C(═O)NR ｈ R ｉ , -OR ｋ and oxo, R ｈ , R ｉ and R ｊ are hydrogen and C for each occurrence, respectively. １ -C ４ independently selected from alkyl, R ｋ For each occurrence, hydrogen and C １ -C ４ independently selected from alkyl, k is an integer selected from 0, 1, and 2, where R ３ａ is oxo, k is 1; m is an integer selected from 0, 1, and 2; 2. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt of claim 1, wherein p, for each occurrence, is an integer selected from 1 and 2.

4. The compound is represented by one of the following structural formulas: 【Transformation 68】 tautomers thereof, deuterated derivatives of these compounds or tautomers, or pharmaceutically acceptable salts of any of the foregoing, wherein: For each occurrence of ring A, C ３ -C ６ cycloalkyl, phenyl, and 5- to 10-membered heteroaryl, each of which is selected from 1, 2, or 3 R ａ optionally substituted with a group, 10. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt of claim 1.

5. The compound is represented by one of the following structural formulas: 【Transformation 69】 tautomers thereof, deuterated derivatives of these compounds or tautomers, or pharmaceutically acceptable salts of any of the foregoing, wherein: For each occurrence of ring A, C ３ -C ６ cycloalkyl, phenyl, and 5- to 10-membered heteroaryl, each of which is selected from 1, 2, or 3 R ａ 10. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt of claim 1, optionally substituted with a group.

6. Each ring A is selected from the group consisting of 1, 2 or 3 R ａ optionally substituted with a group, 【Transformation 70】 2. The compound, tautomer, deuterated derivative or pharmaceutically acceptable salt of claim 1 selected from:

7. Each ring A is selected from the group consisting of 1, 2 or 3 R ａ optionally substituted with a group, 【Chemistry 71】 2. The compound, tautomer, deuterated derivative or pharmaceutically acceptable salt of claim 1 selected from: 【Request Item 8】 【Table 11-1】 Table 11-2 Table 11-3 Table 11-4 Table 11-5 Table 11-6 A compound, tautomer, deuterated derivative or pharmaceutically acceptable salt selected from tautomers thereof, deuterated derivatives of those compounds or tautomers, and pharmaceutically acceptable salts of any of the foregoing.

9. A pharmaceutical composition comprising a compound, tautomer, deuterated derivative or pharmaceutically acceptable salt according to any one of claims 1 to 8.

10. 10. Use of a compound, tautomer, deuterated derivative or pharmaceutically acceptable salt according to any one of claims 1 to 8 in the manufacture of a medicament for treating an APOL1-mediated kidney disease.

11. Use of a compound, tautomer, deuterated derivative or pharmaceutically acceptable salt according to any one of claims 1 to 8 in the manufacture of a pharmaceutical for treating focal segmental glomerulosclerosis (FSGS) and / or non-diabetic kidney disease (NDKD).

12. 10. A composition for use in treating an APOL1 mediated disease, comprising a compound, tautomer, deuterated derivative or pharmaceutically acceptable salt according to any one of claims 1 to 8.

13. A composition for use in the treatment of focal segmental glomerulosclerosis (FSGS) and / or non-diabetic kidney disease (NDKD), comprising a compound, tautomer, deuterated derivative or pharmaceutically acceptable salt according to any one of claims 1 to 8.

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