STAT Modulators and Their Uses

Abstract

Compounds of formula (I): The present invention provides compounds of formula I of the present disclosure, and their pharma- ceutically acceptable salts, as well as their pharma-ceutically acceptable compositions.In one aspect, the compounds of formula I of the present disclosure and their pharma-ceutically acceptable salts inhibit STAT3 and / or STAT6, and are therefore useful in various therapeutic applications, such as in the treatment of cancer and inflammatory conditions.The pharmaceutical compositions containing these compounds and the pharma-ceutically acceptable salts of the compounds of formula I of the present disclosure, as well as the methods for their preparation, are also included.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Application No. 63 / 297,874, filed January 10, 2022, and U.S. Provisional Application No. 63 / 337,425, filed May 2, 2022. The entire contents of each of these U.S. provisional applications are incorporated herein by reference. [Background technology]

[0002] background The signal transducer and activator of transcription (STAT) family of proteins consists of transcription factors that play essential roles in regulating cellular processes such as proliferation, differentiation, apoptosis, and angiogenesis. Seven STAT genes, namely STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b, and STAT6, have been identified in the human genome.

[0003] STAT3 has received particular attention because it is strongly linked to promoting tumor growth and immune evasion and is the only STAT family member whose genetic loss results in fetal lethality. Indeed, abnormally elevated STAT3 activity is estimated to occur in over 70% of human cancers. Activated STAT3 mediates key gene expression changes and molecular events that dysregulate cell growth and apoptosis, promote the development of angiogenesis, invasion, metastasis, and resistance to apoptosis, and suppress host tumor immune surveillance, making constitutively active STAT3 a key mediator of oncogenesis and tumor progression.

[0004] Another STAT protein that has recently attracted interest is STAT6. Recent studies have shown that STAT6 signaling is essential for IL-4 and IL-13-induced epithelial-mesenchymal transition (EMT) and cellular aggressiveness of colorectal cancer cells (CRC). STAT6 is involved in several aspects of inflammatory diseases and other related conditions.

[0005] Given their role in regulating cellular processes, modulating the activity of one or more STAT proteins, particularly STAT3 and / or STAT6, represents a very important area of ​​investigation for the treatment of cancer, inflammatory conditions, and other therapeutic needs. Summary of the Invention [Means for solving the problem]

[0006] Abstract Provided herein are modulators of STAT3 and / or STAT6. Such modulators have the structural formula I: [ka] and pharmaceutically acceptable salts thereof, and compositions thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , q, t and p are as described herein.

[0007] In one aspect, the disclosed compounds of Formula I and pharmaceutically acceptable salts thereof inhibit STAT3 and / or STAT6 and are therefore useful in a variety of therapeutic applications, such as, for example, in the treatment of cancer and inflammatory conditions.

[0008] Also included are pharmaceutical compositions containing these compounds and pharmaceutically acceptable salts of the compounds of Formula I of the present disclosure, as well as methods for their preparation.

[0009] Also included are methods of treating conditions responsive to modulation of STAT3 and / or STAT6 using the disclosed compounds, their pharmaceutically acceptable salts, and compositions thereof. DETAILED DESCRIPTION OF THE INVENTION

[0010] Detailed Description 1. General description of the compound In a first embodiment, structural formula I: [ka] or a pharmaceutically acceptable salt thereof, is provided herein, in structural formula I: q is 0 or 1, and t is 0, 1, or 2, provided that at least one of q or t is 1; p is 1 or 2; Dotted lines represent single or double bonds; R 1 -CR 1a R 2a P(O)OR 1b OR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T 8- to 10-membered fused bicyclic heteroaryl substituted with -CR 1a R 2a P(O)OR 1b OR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T 8- to 10-membered fused bicyclic heterocyclyl substituted with CR 1a R 2a P(O)OR 1b OR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ], aryl substituted with -(C1-C4) alkyl (aryl) in which the aryl moiety is -CR 1a R 2a P(O)OR 1b OR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)ORT ] and the aryl portion of -(C1-C4) alkyl(aryl) and -(C2-C4) alkenyl(aryl) is -CR 1a R 2a P(O)OR 1b OR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ] substituted with -(C2-C4)alkenyl(aryl); R 1a and R 2a are each absent or independently selected from hydrogen, cyano, (C1-C4) alkyl, hydroxy(C1-C4) alkyl, and fluoro; or R 1a and R 2a and together with the carbon to which they are attached form oxo; R 1b and R 2b are each absent or hydrogen, (C1-C4) alkyl, halo(C1-C4) alkyl, -[(C1-C4) alkyl]-OC(O)-[(C1-C4) alkyl], -[(C1-C4) alkyl]-C(O)O-[(C1-C4) alkyl], -[(C1-C4) alkyl]-O-[(C1-C 20) alkyl], -[(C1-C4) alkyl]-OC(O)-[halo(C1-C4) alkyl], [(C1-C4) alkyl]-OC(O)O-[5- to 7-membered heterocyclyl], [(C1-C4) alkyl]-OC(O)-[5- to 7-membered heterocyclyl], -[(C1-C4) alkyl]-OC(O)-[(C1-C4) alkyl]-OH, -[(C1-C4) alkyl]-OC(O)-[(C1-C4) alkyl]-O-[(C1-C4) alkyl], -[(C1 ~C4) alkyl]-OC(O)O-[(C1~C4) alkyl], -[(C1~C4) alkyl]-OC(O)O-[halo(C1~C4) alkyl], -[(C1~C4) alkyl]-OC(O)O-[(C1~C4) alkyl]-OH, -[(C1~C4) alkyl]-OC(O)O-[(C1~C4) alkyl]-O-[(C1~C4) alkyl], -[(C1~C4) alkyl]-SC(O)-[(C1~C4) alkyl], -[(C1~C4) alkyl] Independently from -SC(O)-[halo(C1-C4)alkyl], -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl]-OH, -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl]-O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)NH(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)N[(C1-C4)alkyl]2, 5- to 6-membered heteroaryl, and aryl wherein the 5- to 6-membered heteroaryl and aryl are each independently optionally substituted, where valences allow, with 1 to 2 groups selected from halo, cyano, and (C1-C4)alkyl, and the 5- to 7-membered heterocyclyl of the [(C1-C4)alkyl]-OC(O)O-[5- to 7-membered heterocyclyl] and [(C1-C4)alkyl]-OC(O)-[5- to 7-membered heterocyclyl] are each independently selected, where valences allow, from C(O)OR h and optionally substituted independently with 1 to 2 groups selected from: R 2is selected from hydrogen, halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, cyano, and hydroxyl; R 3 and R 4 are each independently hydrogen, halo, (C1-C4)alkyl, halo(C1-C4)alkyl, hydroxy(C1-C4)alkyl, -(C1-C4)alkylphenyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, -(C1-C4)alkyl(C1-C4)alkoxy, hydroxyl, cyano, -NR a R b , phenyl, (C3-C6)cycloalkyl, 5- to 6-membered heteroaryl, and 4- to 6-membered heterocyclyl, wherein the phenyl, (C3-C6)cycloalkyl, 5- to 6-membered heteroaryl, and 4- to 6-membered heterocyclyl are each selected from, when valences allow, R S optionally substituted with 1 to 3 groups selected from: Or R 3 and R 4 and taken together on the same carbon atom form a (C3-C6)cycloalkyl or a 4- to 6-membered heterocyclyl, each of which is optionally substituted, if valences allow, with 1 to 3 groups selected from halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, and halo(C1-C4)alkoxy; R 5 and R 6 are each independently selected from hydrogen and (C1-C4) alkyl; R 7 is selected from (C1-C4) alkyl, phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl, where the (C1-C4) alkyl is, when valences allow, R Yand the phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl are each optionally substituted, where valences allow, with 1 to 3 groups selected from R Z or R 6 and R 7 together with the nitrogen atom to which they are attached form a 4- to 14-membered monocyclic or bicyclic heterocyclyl, or a 5- to 12-membered monocyclic or bicyclic heteroaryl, each of which, when valences allow, R Q optionally substituted with 1 to 3 groups selected from: AA is an α- or β-amino acid residue, natural or unnatural; R T is selected from (C1-C4)alkyl, benzyl, and phenyl, wherein said phenyl is optionally substituted with one or two groups selected from halo, (C1-C4)alkyl, and halo(C1-C4)alkyl; R Q is halo, (C2-C4)alkenyl, (C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, cyano, phenyl, hydroxyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C3-C6)cycloalkyl, oxo, imino, -O(phenyl), -C(O)R g , -C(O)OR e , -NHC(O)R e , -C(O)NR c R d , -NR a R b , -S(O)R e R f , -S(O)2R f , -S(O)=NH(C1-C4)alkyl, -S(O)NR e R f , and -S(O)NR e R fwherein the (C2-C4)alkenyl and (C1-C4)alkyl are each selected from, when valences allow, R M and wherein the phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C-C)cycloalkyl, and 4- to 6-membered heterocyclyl are each optionally substituted, where valences allow, with 1 to 3 groups selected from R F and optionally substituted independently with 1 to 3 groups selected from: R Y is halo, (C1-C4)alkoxy, halo(C1-C4)alkoxy, cyano, -C(O)R g , -C(O)OR e , -NHC(O)R e , -NR a R b , -S(O)R e R f , -S(O)2R f , -S(O)NR e R f , -S(O)=NH(C1-C4)alkyl, -S(O)2NR e R f , hydroxyl, phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl each, when valences allow, are selected from R X optionally substituted with 1 to 3 groups selected from: R J and R M each independently represents halo, (C1-C4)alkoxy, halo(C1-C4)alkoxy, cyano, -C(O)R g , -C(O)OR e , -NHC(O)R e , -C(O)NR c R d , -NR a R b , -S(O)R e R f , -S(O)2R f , -S(O)NR e R f, -S(O)=NH(C1-C4)alkyl, -S(O)2NR e R f , hydroxyl, phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl each, when valences allow, are selected from R X optionally substituted with 1 to 3 groups selected from: R F , R S , R X , and R Z are each independently halo, cyano, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkyl(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, -(C1-C4)alkylphenyl, (C2-C4)alkenyl, halo(C2-C4)alkenyl, (C2-C4)alkynyl, halo(C2-C4)alkynyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxyl, oxo, imino, phenyl, -S(O)R e R f , -S(O)2R f , -S(O)=NH(C1-C4)alkyl, -S(O)NR e R f , and -S(O)NR e R f , -C(O)OR e , -NR c C(O)R e , -C(O)R g , -C(O)NR c R d , and -NR a R b wherein the phenyl, and phenyl in the group -(C1-C4)alkylphenyl, are each selected from halo, cyano, (C1-C 10 ) Alkyl, (C2-C 10 ) alkenyl, (C2-C 10 ) alkynyl, halo(C1-C 10 ) alkyl, (C1-C 10 ) alkoxy, and halo(C1-C10 )alkoxy, wherein the (C1-C 10 ) Alkyl, (C2-C 10 ) alkenyl and (C2-C 10 ) each alkynyl is optionally substituted, where valences allow, by 5- to 10-membered monocyclic or bicyclic heteroaryl or 4- to 10-membered monocyclic or bicyclic heterocyclyl, wherein each of said 5- to 10-membered monocyclic and bicyclic heteroaryl or 4- to 10-membered monocyclic or bicyclic heterocyclyl is optionally substituted by oxo or 5- to 7-membered heterocyclyl optionally substituted by 1 to 2 oxo; and R a , R b , R c , R d , R e , R f , R g , and R h are each independently selected, when valences allow, from hydrogen, (C1-C4) alkyl, phenyl, (C3-C6) cycloalkyl, 4- to 6-membered heterocyclyl, and 5- to 6-membered heteroaryl, where the (C1-C4) alkyl is, when valences allow, R J and the phenyl, (C3-C6)cycloalkyl, 4- to 6-membered heterocyclyl, and 5- to 6-membered heteroaryl are each independently, where valences allow, optionally substituted with 1 to 3 groups selected from halo, cyano, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxyl, phenyl, and benzyl.

[0011] 2.Definition When used in connection with describing a chemical group that may have multiple points of attachment, the hyphen (-) depicts the point of attachment of that group to the defined variable, e.g., -NR c C(O)R e means that the point of attachment for this group is on the nitrogen atom.

[0012] The terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), and iodine (iodo, -I).

[0013] Unless otherwise specified, the term "alkyl," when used alone or as part of a larger moiety such as "haloalkyl," means a saturated, straight- or branched-chain monovalent hydrocarbon radical.

[0014] The term "haloalkyl" includes monohaloalkyl, polyhaloalkyl, and perhaloalkyl groups, where the halogens are independently selected from fluorine, chlorine, bromine, and iodine.

[0015] "Alkoxy" refers to an alkyl radical attached through an oxygen linking atom, represented by -O-alkyl. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, propoxy, and butoxy.

[0016] "Haloalkoxy" refers to a haloalkyl group that is attached to another moiety through an oxygen atom, such as, for example, -OCHF2 or -OCF3.

[0017] The term "oxo" refers to the group =O.

[0018] The term "imino" refers to the group =NH.

[0019] Unless otherwise specified, the term "heteroaryl" refers to a 5- to 12-membered aromatic radical containing 1 to 4 heteroatoms selected from N, O, and S. In some instances, the nitrogen atom in a heteroaryl can be quaternized. The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic." Heteroaryl groups can be monocyclic or bicyclic. Monocyclic heteroaryls include, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, and the like. Bicyclic heteroaryls include groups in which a monocyclic heteroaryl ring is fused to one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, benzoxazolyl, benzoxodiazolyl, indazolyl, benzimidazolyl, benzothiazolyl, benzothiopheneyl, quinolinyl, quinazolinyl, quinoxalinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrrolopyridinyl, thienopyridinyl, thienopyrimidinyl, indolizinyl, purinyl, cinnolinyl, naphthyridinyl, and pteridinyl. When specified, it is understood that optional substituents on a heteroaryl group can be located at any suitable position, and can include, for example, the position at which the heteroaryl is attached (where valences allow).

[0020] Unless otherwise specified, the term "heterocyclyl" means a 4- to 12-membered saturated or partially unsaturated heterocyclic ring containing 1 to 4 heteroatoms independently selected from N, O, and S. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic ring," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein. A heterocyclyl ring can be attached to its pendant group at any heteroatom or carbon atom that results in the appropriate structure. Heterocyclyl groups can be monocyclic or bicyclic (e.g., bridged, fused, or spiro bicyclic rings). Monocyclic saturated or partially unsaturated heterocyclic radicals include, but are not limited to, azetidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, dihydrooxadizolyl, and dihydroisoxazolyl. Bicyclic heterocyclyl groups include, for example, unsaturated heterocyclic radicals fused to another unsaturated heterocyclic radical, a cycloalkyl, aryl, or heteroaryl ring, such as benzodioxolyl, dihydrobenzodioxinyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl, 6-thia-2,7-diazaspiro[3.4]octanyl, 2,6-diazaspiro[3.3]heptanyl, spiro[indoline-3,3'-pyrrolidin]-yl, and thiochromanyl. When specified, it is understood that optional substituents on a heterocyclyl group can be located at any suitable position and can include, for example, the position at which the heterocyclyl is attached (where valences allow).

[0021] The term "spiro" refers to two rings that share one ring atom (eg, carbon).

[0022] The term "fused" refers to two rings that share two adjacent ring atoms with each other.

[0023] The term "bridged" refers to two rings which share three adjacent ring atoms with each other.

[0024] The term "aryl" refers to an aromatic carbocyclic monocyclic or fused bicyclic ring system containing 6 to 10 carbon atoms. Examples include phenyl, indanyl, tetrahydronaphthalene, and naphthyl. In one aspect, aryl is phenyl or naphthyl.

[0025] The term "cycloalkyl," as used alone or as part of a larger moiety, as used herein, unless otherwise specified, refers to a saturated cycloaliphatic monocyclic or bicyclic ring system having 3 to 10 carbon ring atoms. Monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, and cyclooctyl. When specified, it is understood that optional substituents on the cycloalkyl or alicyclic group can be located at any suitable position, and can include, for example, the position at which the cycloalkyl group is attached.

[0026] A "residue of an amino acid" is the moiety remaining after the formation of a bond between a reactive group in another compound (e.g., an amino group) and a carboxylic acid in that amino acid, after the formation of a bond between a reactive group in another compound (e.g., a carboxylic acid) and an amino group in that amino acid, or both. As a result of the bond(s) formation, the carboxylic acid in the amino acid no longer has an OH group but instead has a bond between the carbonyl group and the reactive group in the compound; the amino group has only one hydrogen atom but instead has a bond between a reactive group in another compound and the nitrogen of the amino group; or both. For example, an "α-amino acid residue" may be structurally depicted as -NH2CR'RC(O)-, -NHCR'RC(O)OH, or -NHCR'RC(O)-, and a "β-amino acid residue" may be structurally depicted as -NH2CR'RCH2-C(O)-, -NHCR'RCH2-C(O)OH, or -NHCR'RCH2-C(O)-, where R' is H or C1-C6 alkyl, and R is H, or halo, (C1-C3)alkoxy, OH, NH2, -NH(C1-C4 alkyl), -N[(C1-C4 alkyl)]2, SH, S(C1-C4 alkyl), imino, COOH, -COO(C1-C4 alkyl), -C1-C6 alkyl optionally substituted with 1 to 3 groups selected from -C1-C4 alkyl), -CO(C1-C4 alkyl), -CONH(C1-C4 alkyl)phenyl, phenyl, and 5- to 10-membered heteroaryl, where the C1-C6 alkyl may also be optionally interrupted by a sulfur or nitrogen heteroatom, and the phenyl may be optionally substituted with 1 to 3 groups selected from OH, cyano, (C1-C4 alkyl), and halo(C1-C4 alkyl); or R together with the nitrogen atom of the α- or β-amino acid residue forms a 4- to 6-membered heterocyclyl.For naturally occurring α-amino acids (i.e., naturally occurring amino acids), R' is H and R is selected from hydrogen, methyl, isopropyl, -CHCH(CH), -(CH)SCH, -CH(CH)(CHCH), CHOH, -CH(OH)(CH), CHSH, -CHC(O)NH, -(CH)C(O)NH, benzyl, p-hydroxybenzyl, -CH(indolyl), -(CH)NH, -(CH)NHC(=NH)NH, -CH(imidazolyl), -(CH)COOH, and -(CH)COOH; or R together with the nitrogen atom of the α- or β-amino acid residue forms a pyrrolidinyl ring.

[0027] Unnatural amino acids are known in the art and include, for example, α-alkyl amino acids (eg, α-methyl), α-alkylalkoxy amino acids (eg, α-CH 2 OCH 3 ), N-methyl amino acids, homo-amino acids, and the like.

[0028] Compounds with one or more chiral centers can exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Stereoisomers include all diastereomeric, enantiomeric, and epimeric forms, as well as racemates and mixtures thereof. "Geometric isomers" refer to isomers that differ in the orientation of substituents relative to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic ring system. Atoms (other than H) on each side of a carbon-carbon double bond can be in the E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents oriented on the same side) configuration. "Cis" refers to substituents oriented on the same side of the ring, while "trans" refers to substituents oriented on opposite sides of the ring.

[0029] When the stereochemical configuration at a chiral center in a compound having one or more chiral centers is indicated by its chemical name (e.g., when the configuration is indicated by "R" or "S" in the chemical name) or by its structure (e.g., when the configuration is indicated by a "wedge-shaped bond"), the enrichment of the indicated configuration over the opposite configuration is greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 99%, or greater than 99.9%. "Enrichment of the indicated configuration over the opposite configuration" is a molar percentage and is determined by dividing the number of compounds in the mixture having the indicated stereochemical configuration at its chiral center(s) by the total number of all compounds having the same or opposite stereochemical configuration.

[0030] When a geometric isomer is designated by name or structure, the enrichment of the designated isomer relative to the opposite isomer is greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 99%, or greater than 99.9%. "Enrichment of the designated isomer relative to the opposite isomer" is a molar percentage and is determined by dividing the number of compounds in the mixture having the designated geometric configuration by the total number of all compounds having the same or opposite geometric configuration.

[0031] When a compound of the present disclosure is named or depicted by a structure without indicating stereochemistry, it is understood that the name or structure encompasses one of the possible stereoisomers or geometric isomers without the other, or a mixture of the stereoisomers or geometric isomers encompassed.

[0032] In certain instances, compounds were isolated and tested as a 1:1 mixture of diastereomers. In such cases, the relative stereochemistry is indicated by the term "rel-" in the compound name and by the use of a flat bond rather than a wedge. For example, the structure: [ka] ((2-(((3S,6S,9S,10aR)-9-(azetidin-1-yl)-3-(rel-(trans)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid has the substituents around the pyrrolidine ring in trans orientation and is a mixture of both diastereomers. [ka] means to include.

[0033] The terms "subject" and "patient" can be used interchangeably and refer to a mammal in need of treatment, such as companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, a subject is a human in need of treatment.

[0034] The terms "inhibit," "inhibition," or "inhibiting" include a decrease in the baseline activity of a biological activity or process.

[0035] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of a disease or disorder described herein, or one or more symptoms thereof. In some aspects, treatment may be administered after one or more symptoms have developed, i.e., it may be a therapeutic treatment. In other aspects, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a suspected individual prior to the onset of treatment (e.g., in light of a history of symptoms and / or exposure to a particular organism or other suspected agent), i.e., it may be a preventative treatment. Treatment may also be continued after symptoms have resolved, e.g., to delay their recurrence.

[0036] The term " pharmaceutically acceptable carrier " refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound that is formulated together.The pharmaceutically acceptable carrier, adjuvant or vehicle that can be used in the compositions described herein includes but is not limited to ion exchange material, alumina, aluminum stearate, lecithin, serum protein such as human serum albumin, buffer substance such as phosphate, glycine, sorbic acid, potassium sorbate, partial glyceride mixture of saturated vegetable fatty acid, water, salt or electrolyte, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based material, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, wax, polyethylene-polyoxypropylene-block polymer, polyethylene glycol and wool fat.

[0037] For use in medicine, the salts of the compounds described herein refer to non-toxic "pharmaceutically acceptable salts." Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and organic acids (e.g., acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid). Compounds of the present disclosure having an acidic group, such as a carboxylic acid, can form pharmaceutically acceptable salts with pharmaceutically acceptable base(s). Suitable pharmaceutically acceptable base salts include, for example, ammonium salts, alkali metal salts (e.g., sodium salts and potassium salts), and alkaline earth metal salts (e.g., magnesium salts and calcium salts). Compounds having a quaternary ammonium group also include counterions such as chloride, bromide, iodide, acetate, perchlorate, etc. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids such as glutamic acid.

[0038] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound described herein sufficient to achieve the desired therapeutic effect (e.g., treatment of a condition described herein) under the conditions of administration, e.g., at a dosage of 0.01 to 100 mg / kg body weight / day.

[0039] 3.Compound In a first embodiment, structural formula I: [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above.

[0040] In a second embodiment, the compound of formula I has structural formula II: [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above with respect to Formula I.

[0041] In a third embodiment, the compound of Formula I has structural formula III, IV, V, or VII: [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above with respect to Formula I.

[0042] In a fourth embodiment, the compound of formula I has structural formula VIII, VIII', IX, X, XI, XII, or XIII: [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above with respect to Formula I.

[0043] In a sixth embodiment, the compound of formula I has the structure XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, or XXVIII: [ka] [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above with respect to Formula I.

[0044] In a seventh embodiment, the compound of Formula I has the structural formula XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXVII: [ka] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above with respect to Formula I.

[0045] In an eighth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XXI, XXII, XXIII, XXIV, XXV, XXIX, XXXI, XXXII, XXXIII, XXXIV, and XXXV 3 is selected from hydrogen, (C1-C4)alkyl, hydroxyl, (C1-C4)alkoxy, —(C1-C4)alkylphenyl, and 4- to 6-membered heterocyclyl, where the remaining variables are as described above for Formula I. Alternatively, as part of an eighth embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XXI, XXII, XXIII, XXIV, XXV, XXIX, XXXI, XXXII, XXXIII, XXXIV, and XXXV is 3 is selected from hydrogen, (C1-C2) alkyl, hydroxyl, (C1-C2) alkoxy, benzyl, and azetidinyl, where the remaining variables are as described above with respect to Formula I.

[0046] In a ninth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XVI, XXII, XXIV, XXVI, XXIX, XXXII, XXXIV, XXXV, and XXXVI 4 is selected from hydrogen, (C1-C4) alkyl, and hydroxyl, where the remaining variables are as described above with respect to Formula I.

[0047] In a tenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XVI, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXIX, XXXI, XXXII, XXXIII, XXXIV, XXXV, and XXXVI 3and R 4 is hydrogen, where the remaining variables are as described above for Formula I.

[0048] In an eleventh embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XVI, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXIX, XXXI, XXXII, XXXIII, XXXIV, XXXV, and XXXVI 3 and R 4 and taken together on the same carbon atom form a (C-C)cycloalkyl, where the remaining variables are as described above for Formula I. Alternatively, as part of an eleventh embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XVI, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXIX, XXXI, XXXII, XXXIII, XXXIV, XXXV, and XXXVI is 3 and R 4 and are joined on the same carbon atom to form cyclopropyl, where the remainder of the variables are as described above for Formula I.

[0049] In a twelfth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 2is selected from hydrogen and hydroxyl, where the remaining variables are as described above with respect to Formula I or any one of the eighth through eleventh embodiments. Alternatively, as part of the twelfth embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 2 is hydrogen, where the remaining variables are as described above with respect to Formula I or any one of the eighth through eleventh embodiments.

[0050] In a thirteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, VIII', XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 5 is hydrogen, where the remaining variables are as described above with respect to Formula I or any one of the eighth through twelfth embodiments.

[0051] In a fourteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1 is selected from 8- to 10-membered fused bicyclic heteroaryl and aryl, each of which is -CR 1a R 2a P(O)OR 1bOR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T

[0047] wherein the remaining variables are as described above with respect to Formula I or any one of the eighth through thirteenth embodiments. Alternatively, as part of the fourteenth embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII is 1 is selected from benzothiophenyl, indolyl, and naphthalenyl, each of which is selected from -CR 1a R 2a P(O)OR 1b OR 2b or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T

[0047] In another alternative, as part of the fourteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII is substituted with 1 teeth, [ka] wherein the remaining variables are as described above with respect to Formula I or any one of the eighth through thirteenth embodiments. In yet another alternative, as part of the fourteenth embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1 teeth, [ka] where the remaining variables are as described above with respect to Formula I or any one of the eighth through thirteenth embodiments.

[0052] In a fifteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1a is hydrogen and R 2a is fluoro or R 1a is fluoro and R 2ais fluoro, where the remaining variables are as described above with respect to Formula I or any one of the eighth through fourteenth embodiments. Alternatively, as part of the fifteenth embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1a is fluoro and R 2a is fluoro, where the remaining variables are as described above with respect to Formula I or any one of the eighth through fourteenth embodiments.

[0053] In a sixteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1b and R 2bare each independently selected from hydrogen, (C1-C4) alkyl, —[(C1-C4) alkyl]-OC(O)—[(C1-C4) alkyl], —[(C1-C4) alkyl]-OC(O)O—[(C1-C4) alkyl], —[(C1-C4) alkyl]-SC(O)—[(C1-C4) alkyl], —[(C1-C4) alkyl]-SC(O)—[halo(C1-C4) alkyl], —[(C1-C4) alkyl]-SC(O)—[(C1-C4) alkyl]-OH, phenyl, pyridinyl, and naphthalenyl, wherein the phenyl, pyridinyl, and naphthalenyl are each optionally independently substituted with cyano, and wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 15th embodiments. Alternatively, as part of the sixteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1b and R 2b are each -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl], where the remaining variables are as described above with respect to Formula I or any one of the 8th through 15th embodiments. In another alternative, as part of the 16th embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1b and R 2b are each hydrogen, where the remaining variables are as described above with respect to Formula I or any one of the eighth through fifteenth embodiments.

[0054] In a seventeenth embodiment, -CR in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 1a R 2a P(O)OR 1b OR 2b teeth, [ka] [ka] and wherein the remaining variables are as described above with respect to Formula I or any one of the eighth through sixteenth embodiments. Alternatively, as part of the seventeenth embodiment, -CR in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII. 1a R 2a P(O)OR 1b OR 2b teeth, [ka] where the remaining variables are as described above with respect to Formula I or any one of the 8th through 16th embodiments.

[0055] In an eighteenth embodiment, —NH[AA]C(O)OR in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII′, IX, X, XI, XII, XIII, XIV, XV, XVI, XVI′, XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII T is -NHC(R')(R)C(O)R T or -NHC(R')(R)CHC(O)R T where R ’ is hydrogen, (C1-C3)alkyl, or (C1-C3)alkyl(C1-C3)alkoxy, and R is selected from hydrogen, methyl, isopropyl, —CH2CH(CH3)2, —(CH2)2SCH3, —CH(CH3)(CH2CH3), CH2OH, —CH(OH)(CH3), CH2SH, —CH2C(O)NH2, —(CH2)2C(O)NH2, benzyl, p-hydroxybenzyl, —CH2(indolyl), —(CH2)4NH2, —(CH2)3NHC(═NH2)NH2, —CH2(imidazolyl), —(CH2)COOH, and —(CH2)2COOH; or R is —NHC(R′)(R)C(O)R T or -NHC(R')(R)CHC(O)R T together with the nitrogen atom to form a pyrrolidinyl ring, where the remainder of the variables are as described above with respect to Formula I or any one of the 8th through 17th embodiments.

[0056] In a nineteenth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 7is selected from (C1-C4)alkyl, phenyl, 5- or 6-membered monocyclic heterocyclyl, 9- or 10-membered fused bicyclic heterocyclyl, 5- or 6-membered monocyclic heteroaryl, and 9- or 10-membered fused bicyclic heteroaryl, where the (C1-C4)alkyl is, when valences allow, R Y and wherein the phenyl, 5- or 6-membered monocyclic heterocyclyl, 9- or 10-membered fused bicyclic heterocyclyl, 5- or 6-membered monocyclic heteroaryl, and 9- or 10-membered fused bicyclic heteroaryl each optionally substituted with 1 to 3 groups selected from, where valences allow, R Z and the remaining variables are as described above with respect to Formula I or any one of the 8th through 14th and 18th embodiments. Alternatively, as part of the 19th embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII is 7 is selected from (C1-C4)alkyl, phenyl, pyrrolidinyl, thiochromanyl, dihydrobenzo[b]thiophenyl, pyridinyl, indazolyl, cinnolinyl, and quinolinyl, wherein the (C1-C4)alkyl is, when valences allow, R Y and wherein phenyl, pyrrolidinyl, thiochromanyl, dihydrobenzo[b]thiophenyl, pyridinyl, indazolyl, cinnolinyl, and quinolinyl are each optionally substituted with 1 to 3 groups selected from R Z wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 18th embodiments.

[0057] In a twentieth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII Y is selected from halo, hydroxyl, phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl each, when valences allow, are selected from R X and optionally substituted with 1 to 3 groups selected from: Y is selected from halo, hydroxyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the 4- to 6-membered heterocyclyl and the 5- to 10-membered monocyclic or bicyclic heteroaryl each, when valences allow, are selected from R X wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 19th embodiments.

[0058] In a twenty-first embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII Y is selected from —C(O)NH, —C(O)N(C1-C4)alkyl[phenyl], hydroxyl, phenyl, imidazolyl, 1,2-dihydropyridinyl, pyridinyl, and pyrazolo[3,4-b]pyridinyl, wherein said phenyl, imidazolyl, 1,2-dihydropyridinyl, pyridinyl, and pyrazolo[3,4-b]pyridinyl are each, when valences allow, R X and optionally substituted with 1 to 3 groups selected from: Y is selected from hydroxyl, imidazolyl, 1,2-dihydropyridinyl, pyridinyl, and pyrazolo[3,4-b]pyridinyl, wherein the phenyl, imidazolyl, 1,2-dihydropyridinyl, pyridinyl, and pyrazolo[3,4-b]pyridinyl are each, when valences permit, R X wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 20th embodiments.

[0059] In a twenty-second embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII Z is selected from phenyl, —C(O)(C1-C4)alkyl, hydroxyl, (C1-C4)alkyl, halo, cyano, and (C1-C4)alkoxy, where the remaining variables are as described above with respect to Formula I or any one of the 8th through 21st embodiments.

[0060] In a twenty-third embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII 6 and R 7 together with the nitrogen atom to which they are attached form a 4- to 6-membered monocyclic heterocyclyl, a 7- to 13-membered spiro bicyclic heterocyclyl, or a 9- to 10-membered fused bicyclic heterocyclyl, each of which, when valences allow, is selected from R Q and optionally substituted with 1 to 3 groups selected from: 6 and R 7together with the nitrogen atom to which they are attached form pyrrolidinyl, azetidinyl, piperazinyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl, 6-thia-2,7-diazaspiro[3.4]octanyl, 2-thia-6-azaspiro[3.3]heptanyl, 4-azaspiro[2.4]heptanyl, spiro[indoline-3,3'-pyrrolidinyl], or 1,2,3,4,5,6-hexahydro-2,6-naphthyridinyl, each of which, when valences permit, is selected from the group consisting of ... R Q and optionally substituted with 1 to 3 groups selected from: 6 and R 7 together with the nitrogen atom to which they are attached form pyrrolidinyl or azetidinyl, each of which, when valences permit, may be selected from R Q wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 18th embodiments.

[0061] In a twenty-fourth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII Q is halo, (C2-C4)alkenyl, (C1-C4)alkyl, cyano, phenyl, hydroxyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C3-C6)cycloalkyl, oxo, imino, -O(phenyl), -C(O)R g , -NHC(O)R e , -S(O)=NH(C1-C4)alkyl, and -S(O)2NR eR f wherein the (C2-C4)alkenyl and (C1-C4)alkyl are each selected from, when valences allow, R M and wherein the phenyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, and (C-C)cycloalkyl are each optionally substituted, where valences allow, with 1 to 3 groups selected from R F and optionally independently substituted with 1 to 3 groups selected from: Q is halo, (C2-C4)alkenyl, (C1-C4)alkyl, cyano, phenyl, hydroxyl, morpholinyl, dihydropyridinyl, tetrahydro-2H-thiopyranyl, pyridinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, triazolyl, indazolyl, benzimidazolyl, pyrazolo[3,4-b]pyridinyl, cyclohexyl, cyclopropyl, oxo, imino, -O(phenyl), -C(O)R g , -NHC(O)R e , -S(O)=NH(C1-C4)alkyl, and -S(O)2NR e R f wherein the (C2-C4)alkenyl and (C1-C4)alkyl are each selected from, when valences allow, R Mand wherein phenyl, morpholinyl, dihydropyridinyl, tetrahydro-2H-thiopyranyl, pyridinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, triazolyl, indazolyl, benzimidazolyl, pyrazolo[3,4-b]pyridinyl, cyclohexyl, and cyclopropyl are each optionally substituted, where valences allow, with 1 to 3 groups selected from R F wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 23rd embodiments.

[0062] In a twenty-fifth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII M is selected from 4- to 6-membered heterocyclyl, 5- to 6-membered monocyclic heteroaryl, —S(O)═NH(C1-C4)alkyl, cyano, and phenyl, wherein the 4- to 6-membered heterocyclyl, 5- to 6-membered monocyclic heteroaryl, and phenyl each, when valences allow, are selected from R X and optionally substituted with 1 to 3 groups selected from: wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 24th embodiments. Alternatively, as part of the 25th embodiment, R in a compound of any one of Formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII is Mis selected from tetrahydropyranyl, pyrazolyl, —S(O)═NH(C1-C4)alkyl, cyano, and phenyl, wherein said tetrahydropyranyl, pyrazolyl, and phenyl each, when valences allow, are selected from R X wherein the remaining variables are as described above with respect to Formula I or any one of the 8th through 24th embodiments.

[0063] In a twenty-sixth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII X is selected from (C1-C4) alkyl, (C1-C4) alkoxy, and oxo, where the remaining variables are as described above with respect to Formula I or any one of the 8th through 25th embodiments.

[0064] In a twenty-seventh embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII F is selected from halo, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, (C1-C4)alkoxy, (C2-C4)alkynyl, cyano, oxo, and imino, where the remaining variables are as described above with respect to Formula I or any one of the 8th through 26th embodiments.

[0065] In a twenty-eighth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII g is selected from (C1-C4) alkyl, morpholinyl, imidazolyl, benzyl, and cyclopropyl, where the remaining variables are as described above with respect to Formula I or any one of the 8th through 27th embodiments.

[0066] In a twenty-ninth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII e and R f are each independently selected from (C1-C4) alkyl and halo(C1-C4) alkyl, where the remaining variables are as described above with respect to Formula I or any one of the 8th through 28th embodiments.

[0067] In a thirtieth embodiment, R in a compound of any one of formulas I, II, III, IV, V, VII, VIII, VIII', IX, X, XI, XII, XIII, XIV, XV, XVI, XVI', XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXXVII c and R dare each independently selected from hydrogen and (C1-C4) alkyl, where the remaining variables are as described above with respect to Formula I or any one of the 8th through 29th embodiments. Compounds having Formula I are further described in the examples and are included in this disclosure, including pharmaceutically acceptable salts and neutral forms thereof.

[0068] 4. Use, Formulation and Administration The compounds and compositions described herein are generally useful for regulating the activity of STAT proteins, particularly STAT3 and / or STAT6. In some aspects, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein inhibit STAT3 and / or STAT6 activity.

[0069] In some aspects, the compounds and pharmaceutical compositions described herein are useful for conditions responsive to modulation of STAT3 and / or STAT6. Accordingly, provided herein is a method of treating a condition responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6 in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0070] Also provided is the use of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a condition responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6. Also provided is a pharmaceutical composition containing a compound described herein, or a pharmaceutically acceptable salt thereof, or a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in treating a condition responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6.

[0071] In one aspect, conditions responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6 include, but are not limited to, cancer, a neurodegenerative disorder, a viral disease, an autoimmune disease, an inflammatory disorder, a genetic disorder, a hormone-related disease, a metabolic disorder, a condition associated with organ transplantation, an immunodeficiency disorder, a destructive bone disorder, a proliferative disorder, an infectious disease, a condition associated with cell death, thrombin-induced platelet aggregation, a liver disease, a pathological immune condition involving T-cell activation, a cardiovascular disorder, or a CNS disorder.

[0072] In another aspect, conditions responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6 include cancer (see, e.g., Turkson & Jove, Oncogene 2000, 19:6613-6626), diabetes (see, e.g., Gurzov et al., FEBS 2016, 283:3002), cardiovascular disease (see, e.g., Grote et al., Vasc. Pharmacol. 2005, 43:2005), viral diseases (see, e.g., Gao et al., J Hepatol. 2012, 57(2):430), autoimmune diseases such as lupus (see, e.g., Goropevsek et al., Clin. Rev. Alleg. & Immun. 2017, 52(2):164), and rheumatoid arthritis (see, e.g., Walker & Smith, J. Rheumat. 2005, 32(9):1650), autoinflammatory syndromes (see, e.g., Rauch et al., Jak-Stat 2013, 2(l):e23820), atherosclerosis (see, e.g., Ortiz-Munoz et al., Arterio., Thromho., Vase. Bio. 2009, 29:525), psoriasis (see, e.g., Andres et al., Exp. Derm. 2013, 22(5):323), allergic diseases (see, e.g., Oh et al., Eur. Respir. Rev. 2019, 19(115):46), inflammatory bowel disease (see, e.g., Sugimoto, World J Gastroenterol. 2008, 14(33):5110), inflammation (see, e.g., Tamiya et al. al., Arierio. Thrombo., Vasc. Bio. 2011, 31:980), acute and chronic gout and gouty arthritis, neuropathy (see, e.g., Campbell, Brain Res. Rev. 2005, 48(2):166), metabolic syndrome, immunodeficiency disorders such as AIDS and HIV (see, e.g., O'Shea et al., N. Engl. J. Med. 2013, 368:161), destructive bone disorders (see, e.g., Jatiani et al., Genes & Can.2011,1(10):979), osteoarthritis, proliferative disorders, Waldenstrom's macroglobulinemia (see, e.g., Hodge et al., Blood 2014,123(7):1055), infectious diseases, conditions associated with cell death, pathologic immune conditions involving T cell activation, and CNS disorders.

[0073] Proliferative disorders include benign or malignant tumors, solid tumors, liquid tumors, tumors of the brain, kidney, liver, adrenal gland, bladder, breast, stomach, and gastric tumors. tumor), carcinoma of the ovary, colon, rectum, prostate, pancreas, lung, vagina, cervix, testis, genitourinary tract, esophagus, larynx, skin, bone or thyroid gland, sarcoma, glioblastoma, neuroblastoma, multiple myeloma, gastrointestinal cancer, especially colon cancer or colorectal adenoma, tumors of the head and neck, epidermal hyperproliferation, psoriasis, benign prostatic hyperplasia, neoplasia, epithelial neoplasia, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, lymphoma, Hodgkin's and non-Hodgkin's, breast cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, IL-I-driven disorders, MyD88-driven disorders, smoldering asymptomatic multiple myeloma, or hematological malignancies (leukemia, diffuse large B-cell lymphoma (DLBCL), ABC These include, but are not limited to, DLBCL, chronic lymphocytic leukemia (CLL), chronic lymphocytic lymphoma, primary effusion lymphoma, Burkitt's lymphoma / leukemia, acute lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenstrom's macroglobulinemia (WM), splenic marginal zone lymphoma, multiple myeloma, plasmacytoma, and intravascular large B-cell lymphoma.

[0074] In some embodiments, the cancer to be treated is selected from glioma, breast cancer, prostate cancer, squamous cell carcinoma of the head and neck, cutaneous melanoma, ovarian cancer, malignant neurilemmoma (MPNST), and pancreatic cancer. In other embodiments, the cancer to be treated is selected from glioma, breast cancer, prostate cancer, squamous cell carcinoma of the head and neck, cutaneous melanoma, ovarian cancer, malignant neurilemmoma (MPNST), pancreatic cancer, non-small cell lung cancer (NSCLC) including EGFR-mutated NSCLC, urothelial cancer, liver cancer, bile duct cancer, kidney cancer, colon cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, and hematological malignancies include lymphoma, leukemia, myeloma, myeloproliferative neoplasm, and myelodysplastic syndrome. In other embodiments, the cancer is selected from solid tumors (e.g., prostate cancer, renal cancer, hepatic cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, Kaposi's sarcoma, Castleman's disease, uterine leiomyosarcoma, melanoma, etc.), hematological cancers (e.g., lymphoma, leukemia, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), or multiple myeloma), and skin cancers, such as cutaneous T-cell lymphoma (CTCL) and cutaneous B-cell lymphoma. Exemplary CTCLs include Sézary syndrome and mycosis fungoides.

[0075] The compounds, salts, and compositions described herein are also useful in the treatment of inflammatory or obstructive airway diseases, for example, resulting in a reduction in tissue damage, airway inflammation, bronchial hyperresponsiveness, remodeling, or disease progression. Inflammatory or obstructive airway diseases include asthma of any type or onset, including both intrinsic (non-allergic) and extrinsic (allergic) asthma, mild asthma, moderate asthma, severe asthma, bronchitis asthma, exercise-induced asthma, occupational asthma, and asthma induced after bacterial infection. It should be understood that the treatment of asthma also includes the treatment of subjects who exhibit symptoms of wheezing, for example, under 4 or 5 years of age, and who have been diagnosed or can be diagnosed as "wheezing infants." Wheezing infants are an established patient category of major medical concern and are often currently identified as having early or early asthma.

[0076] The compounds, salts, and compositions described herein are also useful in treating xenoimmune diseases, including, but not limited to, graft-versus-host disease, transplants, blood transfusions, anaphylaxis, allergies (e.g., allergies to plant pollen, latex, drugs, foods, insect venom, animal hair, animal dander, house dust mites, or cockroach calyx), type I hypersensitivity, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

[0077] The compounds, salts, and compositions described herein are also suitable for the present invention and are useful in treating other inflammatory or obstructive airway diseases and conditions, including acute lung injury (ALI), adult / acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary, airway, or lung disease (COPD, COAD, or COLD), including chronic bronchitis or related dyspnea, emphysema, and exacerbation of airway hyperactivity as a result of other medications, particularly other inhaled medications. The compounds, salts, and compositions described herein are also useful in treating bronchitis, including, but not limited to, acute, arachidonic, catarrhal, croupus, chronic, or tuberculous bronchitis. The compounds, salts, and compositions described herein are also useful in the treatment of any type or occurrence of pneumoconiosis (an inflammatory, generally occupational, lung disease that may be chronic or acute, is frequently associated with airway obstruction, and is caused by repeated inhalation of dust), including, for example, aluminum lung disease, anthracosis, asbestosis, stone pneumoconiosis, ostrich pneumoconiosis, siderosis, silicosis, tobacco poisoning, and byssinosis.

[0078] The compounds, salts, and compositions described herein are also useful in the treatment of inflammatory or allergic conditions of the skin, such as psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforma, dermatitis herpetiformis, scleroderma, vitiligo, hypersensitivity vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, systemic lupus erythematosus, pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, epidermolysis bullosa acquisita, acne vulgaris, and other inflammatory or allergic conditions of the skin.

[0079] The compounds, salts, and compositions described herein are also useful in the treatment of other diseases or conditions, e.g., diseases or conditions that have an inflammatory component, e.g., diseases and conditions of the eye such as ocular allergies, conjunctivitis, keratoconjunctivitis sicca, and vernal conjunctivitis, diseases affecting the nose, including allergic rhinitis, and inflammatory diseases that involve an autoimmune response or have an autoimmune component or etiology, such as autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell aplasia, and erythroblastic anemia). and idiopathic thrombocytopenia), systemic lupus erythematosus, rheumatoid arthritis, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease), irritable bowel syndrome, celiac disease, periodontitis, pulmonary hyaline membrane disease, kidney disease, glomerular disease, alcoholic liver disease, multiple sclerosis, endocrine ophthalmopathy opthalmopathy, Graves' disease, sarcoidosis, alveolitis, alveolar osteitis, chronic hypersensitivity pneumonitis, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), Sjögren's syndrome, keratoconjunctivitis sicca, and vernal conjunctivitis), interstitial pulmonary fibrosis, psoriatic arthritis, systemic juvenile idiopathic arthritis, cryopyrin-associated periodic syndromes, nephritis, vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis (with and without nephrotic syndrome, e.g., idiopathic nephrotic syndrome or minimal change nephropathy), chronic granulomatous disease, endometriosis, leptospirosis renal disease, glaucoma, retinal disease, aging, headache, pain, complex regional pain syndrome, cardiac hypertrophy, muscle wasting, catabolic disordersdisorder), obesity, fetal growth retardation, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, ahidrotic ectodermal dysplasia, Behçet's disease, incontinentia pigmenti, Paget's disease, pancreatitis, hereditary periodic fever syndromes, asthma (allergic and non-allergic, mild, moderate, severe, bronchitis, and exercise-induced), acute lung injury, acute respiratory distress syndrome, eosinophilia, hypersensitivity, anaphylaxis, sinusitis, ocular allergies, silica-induced disease, COPD (injury, airway inflammation, bronchial hyperresponsiveness, reduced remodeling or disease progression), pulmonary disease, cystic fibrosis, acid-induced lung injury, pulmonary hypertension, polyneuropathy cataracts, muscle inflammation associated with systemic sclerosis, inclusion body myositis, myasthenia gravis, thyroiditis, Addison's disease, lichen planus, type 1 or type 2 diabetes, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic graft rejection, colitis, conjunctivitis, Crohn's disease, cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, encephalomyelitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis, Henoch-Schönlein purpura, hepatitis, hidradenitis abscess, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, osteomyelitis It is useful in treating myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis, pleuritis, phlebitis, pneumonitis, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, ulcerative colitis, uveitis, vaginitis, vasculitis, or vulvitis.

[0080] In some embodiments, cardiovascular diseases that may be treated by the present methods include, but are not limited to, restenosis, cardiomegaly, atherosclerosis, myocardial infarction, ischemic stroke, congestive heart failure, angina pectoris, reocclusion after angioplasty, restenosis after angioplasty, reocclusion after aortocoronary bypass, restenosis after aortocoronary bypass, stroke, transient ischemia, peripheral arterial occlusive disorder, pulmonary embolism, and deep vein thrombosis.

[0081] In some embodiments, neurodegenerative diseases that may be treated by the present methods include, but are not limited to, treatment of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, cerebral ischemia, and neurodegenerative diseases caused by trauma, glutamate neurotoxicity, hypoxia, epilepsy, diabetes, metabolic syndrome, obesity, organ transplantation, and graft-versus-host disease.

[0082] In certain aspects, the pharmaceutical compositions described herein are formulated for administration to a patient in need of such a composition. The pharmaceutical compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarterial, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection and infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the pharmaceutical compositions described herein can be aqueous or oily suspensions. These suspensions can be formulated using suitable dispersing or wetting agents and suspending agents according to techniques known in the art.

[0083] In some aspects, the pharmaceutical composition is administered orally.

[0084] The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, excretion rate, drug combination, as well as the judgment of the treating physician and the severity of the particular disease being treated. The amount of the compound described herein in the composition will also depend on the specific compound in the pharmaceutical composition. [Example]

[0085] Example Preparation of compounds The compounds claimed herein were prepared according to the procedures outlined in the following schemes. Compound names were generated using software built into ChemDraw. In the event of a discrepancy between the name of a compound and its depicted structure, the depicted chemical structure shall be deemed to be the appropriate compound.

[0086] Steps for synthesis of the core: Synthesis of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0087] Step 1: Preparation of methyl (2S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)pyrrolidine-2-carboxylate

[0088] To a cooled (0 °C) solution of (2S)-methyl 5-allylpyrrolidine-2-carboxylate hydrochloride (200 g, 972 mmol, 1.00 equiv.) and (S)-2-((tert-butoxycarbonyl)amino)pent-4-enoic acid (209 g, 972 mmol, 1.00 equiv.) in CHCl (1.60 L) was added EtN (406 mL, 2.92 mol, 3.00 equiv.) and 2-chloro-1-methylpyridinium iodide (CMPI) (273 g, 1.07 mol, 1.10 equiv.). The solution was warmed to 25 °C and stirred for 1 h. The mixture was poured into water (5.0 L) and extracted with CHCl (2.00 L × 3). The combined organic layers were washed with brine (2.0 L), dried over NaSO, filtered, and concentrated under reduced pressure. Six equal-scale batches were run in parallel and combined during workup. The residue was purified by column chromatography (petroleum ether: EtOAc = 100:1 to 10:1) to give methyl (2S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)pyrrolidine-2-carboxylate (1.18 kg, 3.22 mol, 55.2% yield) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.84 - 5.78 (m, 2H), 5.17 - 4.99 (m, 5H), 4.50 - 4.34 (m, 3H), 3.76 - 3.70 (m, 3H), 2.50 - 2.15 (m, 6H), 1.96 - 1.91 (m, 2H), 1.41 (s, 9H).

[0089] The intermediates shown in Table 1 were prepared according to the protocol outlined in Step 1 above using (S)-5-oxopyrrolidine-2-carboxylic acid, the appropriate N-protected amino acid [(S)-2-((tert-butoxycarbonyl)amino)but-3-enoic acid) or (S)-2-((tert-butoxycarbonyl)amino)-2-methylpent-4-enoic acid], CMPI, triethylamine, and CHCl. ​​The intermediate(s) were purified using standard methods. [Table 1]

[0090] Step 2: Preparation of methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0091] To a solution of methyl (2S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)pyrrolidine-2-carboxylate (200 g, 546 mmol, 1.00 equiv.) in CHCl (2.00 L) was added first-generation Grubbs' catalyst (44.9 g, 54.6 mmol, 0.10 equiv.) at 25 °C. The solution was then heated to 50 °C and stirred for 36 h. Six equal-scale batches were run in parallel and combined during workup. The combined reaction mixture was concentrated to give a residue. The residue was purified twice by column chromatography (petroleum ether: EtOAc = 100:1 to 0:1) to give the crude product. The crude product was triturated with petroleum ether (2.00 L) for 12 h and filtered. The filter cake was dried under reduced pressure to give methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (510 g, 1.40 mol, 51.3% yield, 92.9% purity) as a solid. LCMS (ESI) m / z = 361.2 [M+Na] + ; 1H NMR (400 MHz, CDCl3) δ 5.82 - 5.80 (m, 1H), 5.73 - 5.71 (m, 1H), 5.58 -5.56 (m, 1H), 4.88 - 4.85 (m, 1H), 4.52 - 4.49 (m, 1H), 4.16 - 4.14 (m, 1H), 3.71 (s, 3H), 2.81 - 2.74 (m, 2H), 2.45 - 2.38 (m, 1H), 2.33 - 2.24 (m, 1H), 2.14 - 2.05 (m, 2H), 1.98 - 1.93 (m, 2H), 1.43 (s, 9H).

[0092] The intermediates shown in Table 2 were prepared according to the representative protocol described above in Step 2, with appropriate modifications. [Table 2]

[0093] Step 3: Preparation of methyl (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0094] To a solution of methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (47.9 g, 137 mmol, 96.8% purity, 1.00 equiv) in EtOAc (500 mL) was added 10% Pd / C (9.58 g) under N2(g). The suspension was degassed under reduced pressure and purged with H2(g) several times. The mixture was stirred at 25 °C under H2(g) (50 psi) for 16 h. The mixture was filtered, and the filtrate was concentrated to give methyl (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (48.0 g) as a brown oil. LCMS (ESI) m / z = 341.0 [M+H] + ;1 H NMR (400 MHz, CDCl3) δ 5.43 (d, J = 8.4 Hz, 1H), 4.69 - 4.57 (m, 1H), 4.50 - 4.41 (m, 1H), 4.27 - 4.18 (m, 1H), 3.76 (s, 3H), 2.34 - 2.25 (m, 1H), 2.21 - 2.11 (m, 1H), 2.04 - 1.68 (m, 8H), 1.64 - 1.57 (m, 2H), 1.43 (s, 9H).

[0095] The intermediates shown in Table 3 were prepared according to the protocol described above in step 3, with appropriate modifications. [Table 3]

[0096] Step 4: Preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0097] To a cooled (0 °C) solution of methyl (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (17.0 g, 49.9 mmol, 1.00 equiv.) in 1,4-dioxane (85.0 mL) was added a 2 M aqueous solution of LiOH·HO (74.9 mL, 3.00 equiv.). The mixture was gradually warmed to room temperature and stirred at 25 °C for 16 h. The reaction mixture was acidified with 1.0 N HCl to pH 4.0. The mixture was extracted with EtOAc (3×). The combined organic layers were dried over NaSO and concentrated to give a residue. The residue was dissolved in water (150 mL) and acetonitrile (30 mL). The mixture was lyophilized to give (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (15.5 g, 44.8 mmol, 92.3% yield over two steps, 94.3% purity) as a gray solid. LCMS (ESI) m / z = 327.0 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 7.01 (s, 1H), 5.53 (d, J = 8.0 Hz, 1H), 4.72 - 4.62 (m, 1H), 4.54 (t, J = 8.6 Hz, 1H), 4.32 - 4.21 (m, 1H), 2.32 - 2.12 (m, 3H), 2.05 - 1.95 (m, 1H), 1.85 - 1.55 (m, 8H), 1.43 (s, 9H).

[0098] Step 5: Preparation of (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0099] (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid was prepared using the hydrolysis conditions described above in step 4 for the preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid and starting from methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate. Starting from (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid (35.0 g, 103 mmol, 1.00 equiv.), LiOH·HO (8.64 g, 3.00 equiv.), and THF (150 mL) / water (50 mL), (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid (31.1 g, 99.1 mmol, 96.1% yield) was obtained as a white solid. LCMS (ESI) m / z = 325.2 [M+H] + ;

[0100] The intermediate(s) shown in Table 4 were prepared using the hydrolysis conditions described above in Step 4 for the preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid and starting from the appropriate starting materials and conditions. [Table 4] Synthesis of (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate [ka]

[0101] Step 1: Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-5-oxo-7-(trimethylsilyl)hept-6-ynoate

[0102] To a cooled (0 °C) solution of isopropylmagnesium chloride (2.0 M, 9.04 L, 1.10 equiv.) and THF (4.00 L) was added ethynyltrimethylsilane (1.86 kg, 18.9 mol, 2.62 L, 1.15 equiv.). The reaction mixture was stirred for 1 hour, after which a solution of 1-(tert-butyl)2-methyl (S)-5-oxopyrrolidine-1,2-dicarboxylate (4.00 kg, 16.4 mol, 1.00 equiv.) in THF (8.0 L) was added slowly over 1.5 hours. After stirring for an additional 30 minutes, the reaction mixture was transferred to a stirred, cooled (0 °C) biphasic mixture of iPAc (5.00 L) and 20% aqueous NH4Cl (16.0 L). The biphasic mixture was separated, and the aqueous layer was extracted with iPAc (8.00 L). The combined organic layers were washed with 20% aqueous NH4Cl (8.00 L), brine (8.00 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to give methyl (S)-2-((tert-butoxycarbonyl)amino)-5-oxo-7-(trimethylsilyl)hept-6-ynoate (5.50 kg) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.11 - 5.09 (m, 1H), 4.32 - 4.30 (m, 1H), 3.77 - 3.72 (m, 3H), 2.72 - 2.64 (m, 2H), 2.04 - 2.01 (m, 1H), 1.73 - 1.71 (m, 1H), 1.41 (s, 9H), 0.24 (s, 9H).

[0103] Step 2: Preparation of 1-(tert-butyl)2-methyl (2S,5R)-5-((trimethylsilyl)ethynyl)pyrrolidine-1,2-dicarboxylate

[0104] To a suspension of NaBH(OAc) (4.03 kg, 19.0 mol, 1.30 equiv.) in iPAc (5.00 L) was added a solution of methyl (S)-2-((tert-butoxycarbonyl)amino)-5-oxo-7-(trimethylsilyl)hept-6-ynoate in iPAc (20.0 L). The batch was cooled to -10 °C, and then TFA (7.18 kg, 62.9 mol, 4.66 L, 4.30 equiv.) was added slowly over 1.5 h. The mixture was warmed to 10 °C and stirred for an additional 2 h. The reaction mixture was transferred to a cooled (0 °C) 25% aqueous KHPO solution (40.0 L). The pH of the suspension was adjusted to pH 6-7 with aqueous saturated NaHCO. The biphasic mixture was separated, and the aqueous layer was extracted with iPAc (8.00 L). The combined organic layers were washed with 25% KHPO solution (8.00 L), brine (5.00 L), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 30 / 1 to 3 / 1) to give 1-(tert-butyl)2-methyl (2S,5R)-5-((trimethylsilyl)ethynyl)pyrrolidine-1,2-dicarboxylate (2.27 kg, 6.97 mol, 42.4% yield) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ 4.49 - 4.32 (m, 1H), 4.16 (s, 1H), 3.63 (s, 3H), 2.16 (s, 2H), 1.95 - 1.88 (m, 2H), 1.40 - 1.32 (m, 9H), 0.13 (s, 9H).

[0105] Step 3: Preparation of (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0106] To a cooled (0 °C) solution of 1-(tert-butyl)2-methyl (2S,5R)-5-((trimethylsilyl)ethynyl)pyrrolidine-1,2-dicarboxylate (500 g, 1.54 mol, 1.00 equiv.) in THF (2.00 L) was added a 1.0 M solution of TBAF in THF (1.84 L, 1.20 equiv.), and the mixture was stirred for 1 h. Four equal-scale batches were run in parallel, and the reaction mixtures were combined and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 1 / 0 to 0 / 1) to give 1-(tert-butyl)2-methyl (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylate (I-14) (920 g, 2.95 mol, 48.0% yield, 81.2% purity) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 4.62 - 4.50 (m, 1H), 4.30 - 4.20 (m, 1H), 3.74 - 3.70 (m, 3H), 2.31 - 2.14 (m, 5H), 1.47 - 1.40 (m, 9H).

[0107] Step 4: Preparation of (2S,5R)-5-vinylpyrrolidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0108] To a suspension of 1-(tert-butyl)2-methyl (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylate (150 g, 592 mmol, 1.00 equiv.) in EtOAc (1.50 L) under N2(g) was added Lindlar's catalyst (7.50 g, 1.82 mmol, 5.0% wt.) and quinoline (163 g, 1.27 mol, 150 mL, 2.14 equiv.). The suspension was degassed under reduced pressure and purged with H2(g) (three times). The mixture was stirred under H2(g) (50 psi) for 1 h. Six equal-scale batches were run in parallel, filtered, and the filtrates were combined during workup. The mixture was filtered, and the filtrate was washed with 1 N aqueous HCl (9.00 L), separated, and concentrated. The combined residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 1 / 0 to 0 / 1) to give 1-(tert-butyl)2-methyl (2S,5R)-5-vinylpyrrolidine-1,2-dicarboxylate (745 g, 2.23 mol, yield 62.8%, purity 76.5%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ 5.84 - 5.76 (m, 1H), 5.34 - 5.27 (m, 1H), 5.06 - 5.04 (m, 1H), 4.29 - 4.17 (m, 2H), 3.65 - 3.62 (m, 3H), 2.16 - 2.14 (m, 2H), 1.79 - 1.78 (m, 1H), 1.69 - 1.65 (m, 1H), 1.34 - 1.32 (m, 9H).

[0109] Step 5: Preparation of methyl (2S,5R)-5-vinylpyrrolidine-2-carboxylate

[0110] To a solution of 1-(tert-butyl)2-methyl (2S,5R)-5-vinylpyrrolidine-1,2-dicarboxylate (245 g, 959 mmol, 1.00 equiv.) in EtOAc (1.25 L) was added a solution of 4.0 M HCl in EtOAc (959 mL, 4.00 equiv.). Three equal-scale batches were run in parallel, and the mixture was stirred at 25° C. for 2 h. The reaction mixtures were combined and concentrated under reduced pressure to give the product, methyl (2S,5R)-5-vinylpyrrolidine-2-carboxylate (520 g, HCl) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 6.03 - 5.95 (m, 1H), 5.44 - 5.33 (m, 2H), 4.49 - 4.45 (m, 1H), 4.11 - 4.05 (m, 1H), 3.76 (s, 3H), 2.31 - 2.28 (m, 1H), 2.16 - 2.10 (m, 1H), 1.81 - 1.76 (m, 1H).

[0111] Step 6: Preparation of methyl (2S,5R)-1-((S)-2-((tert-butoxycarbonyl)amino)hex-5-enoyl)-5-vinylpyrrolidine-2-carboxylate

[0112] To a solution of (S)-2-((tert-butoxycarbonyl)amino)hex-5-enoic acid (80.6 g, 352 mmol, 1.00 equiv.) in CHCl (674 mL) was added (2S,5R)-methyl 5-vinylpyrrolidine-2-carboxylate (67.4 g, 352 mmol, 1.00 equiv., HCl) and EtN (147 mL, 1.06 mol, 3.00 equiv.). The mixture was cooled to 0 °C, and CMPI (98.8 g, 387 mmol, 1.10 equiv.) was added. The reaction mixture was then warmed to 25 °C and stirred for 3 h. The reaction mixture was poured into water (500 mL) and extracted with CHCl (200 mL × 3). The combined organic layers were washed with saturated aqueous NH4Cl (300 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 50 / 1 to 5 / 1) to give methyl (2S,5R)-1-((S)-2-((tert-butoxycarbonyl)amino)hex-5-enoyl)-5-vinylpyrrolidine-2-carboxylate (115 g, 247 mmol, 70.2% yield, 78.9% purity) as a yellow oil. LCMS (ESI) m / z = 367.2 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 6.03 - 5.94 (m, 1H), 5.53 (d, J = 17.2 Hz, 1H), 5.23 (d, J = 10.4 Hz, 1H), 5.03 - 4.94 (m, 3H), 4.81 (t, J = 12.0 Hz, 1H), 4.53 - 4.40 (m, 2H), 3.75 (s, 3H), 2.25 - 2.18 (m, 2H), 2.16 - 2.12 (m, 1H), 2.10 - 2.07 (m, 1H), 2.05 (s, 1H), 2.00-1.97 (m, 1H), 1.89 - 1.84 (m, 1H), 1.80 - 1.73 (m, 1H), 1.43 (s, 9H).

[0113] The intermediates shown in Table 5 were synthesized using methyl (2S,5R)-5-vinylpyrrolidine-2-carboxylate, the appropriate N-protected amino acid, CMPI, triethylamine, and CHCl under the reaction conditions described above. The intermediate(s) were purified using standard methods. [Table 5]

[0114] Step 7: Preparation of methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0115] To a solution of methyl (2S,5R)-1-((S)-2-((tert-butoxycarbonyl)amino)hex-5-enoyl)-5-vinylpyrrolidine-2-carboxylate (38.2 g, 104 mmol, 1.00 equiv.) in CHCl (1.90 L) was added Grubb's catalyst 1 (8.58 g, 10.4 mmol, 0.10 equiv.). The mixture was heated to 50 °C and stirred for 12 h. Three equal-scale batches were run in parallel. The reaction mixtures were then combined and concentrated to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 100 / 1 to 7 / 3) to give the crude product as a gray solid. The crude product (62.0 g) was triturated with petroleum ether (122 mL) for 1.5 h. The mixture was filtered, and the filter cake was dried under reduced pressure to give methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (50.0 g, 50.3% yield) as a gray solid. LCMS (ESI) m / z = 339.3 [M+H] + ; 1H NMR (400 MHz, CDCl3) δ 5.78 - 5.71 (m, 1H), 5.48 (d, J = 12.0 Hz, 1H), 5.28 (d, J = 8.8 Hz, 1H), 4.75 - 4.68 (m, 2H), 4.62 (t, J = 6.4 Hz, 1H), 3.74 (s, 3H), 3.10 - 3.07 (m, 1H), 2.30 - 2.24 (m, 1H), 2.15 - 2.07 (m, 2H), 2.00 - 1.88 (m, 2H), 1.64 - 1.57 (m, 1H), 1.42 (s, 9H).

[0116] The intermediates shown in Table 6 were synthesized under the conditions described above for the preparation of methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate using the appropriate starting materials and reagents. [Table 6]

[0117] Step 8: Preparation of (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0118] (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid was prepared using the hydrolysis conditions previously described for the preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid in step 4. Methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate, LiOH (185 mg, 4.41 mmol, 3 equiv.), and THF / water (9 mL / 3 mL) were used to give (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid (488 mg, 1.50 mmol) as a white solid. LCMS (ESI) m / z = 339.1 [M+H] + . [ka]

[0119] The intermediates shown in Table 7 were synthesized under the hydrogenation conditions described for the preparation of methyl (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate in Step 3 using the appropriate starting materials and reagents. [Table 7] [ka]

[0120] The intermediates shown in Table 8 were synthesized under the hydrolysis conditions described for the preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid in Step 4 using the appropriate starting materials and reagents. [Table 8] Synthesis of (4S,7S,11aS)-7-((tert-butoxycarbonyl)amino)-6-oxodecahydro-2H-pyrido[1,2-a]azocine-4-carboxylic acid [ka]

[0121] Step 1: Preparation of (S)-6-oxopiperidine-2-carboxylate methyl

[0122] To a cooled (0 °C) solution of MeOH (1.50 L), SOCl (137 g, 1.15 mol, 83.6 mL, 1.10 equiv.) and (S)-6-oxopiperidine-2-carboxylic acid (150 g, 1.05 mol, 1.00 equiv.) were added sequentially. The mixture was warmed to 25 °C and stirred for 12 h. Two equal-scale batches were run in parallel. The reaction mixtures were combined and concentrated in vacuo. The resulting residue was dissolved in a mixture of toluene (1.15 L) and EtN (292 mL, 2.10 mol, 2.00 equiv.). The mixture was stirred at 25 °C for 30 min, then filtered, and the filtrate was concentrated to give (S)-methyl 6-oxopiperidine-2-carboxylate (282 g) as a yellow oil. LCMS (ESI) m / z = 158.2 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 6.53 (s, 1H), 4.10 (t, J = 5.60 Hz, 1H), 3.77 (s, 3H), 2.43 - 2.33 (m, 2H), 2.20 - 2.15 (m, 1H), 1.89 - 1.77 (m, 3H).

[0123] Step 2: Preparation of (S)-6-oxopiperidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0124] To a solution of (S)-methyl 6-oxopiperidine-2-carboxylate (141 g, 897 mmol, 1.00 equiv) in acetonitrile (1.41 L) was added BocO (235 g, 1.08 mol, 247 mL, 1.20 equiv), DMAP (21.9 g, 179 mmol, 0.20 equiv), and EtN (136 g, 1.35 mol, 187 mL, 1.50 equiv) at 25 °C. The mixture was stirred at 25 °C for 3 h. Two equal-scale batches were run in parallel, and the reaction mixtures were combined and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 100 / 1 to 80 / 1) to give (S)-1-(tert-butyl)2-methyl 6-oxopiperidine-1,2-dicarboxylate (431 g, 1.67 mol, 79.5% yield over two steps, 99.4% purity) as a yellow solid. LCMS (ESI) m / z = 158.2 [(M-Boc)+H] + ; 1 H NMR: (400 MHz, CDCl3) δ 4.71 - 4.69 (m, 1H), 3.76 (s, 3H), 2.60 - 2.43 (m, 2H), 2.18 - 2.14 (m, 1H), 2.09 - 2.00 (m, 1H), 1.81 - 1.72 (m, 2H), 1.49 (s, 9H).

[0125] Step 3: Preparation of (2S)-6-hydroxypiperidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0126] To a cooled (−78° C.) solution of 1-(tert-butyl)2-methyl (S)-6-oxopiperidine-1,2-dicarboxylate (120 g, 464 mmol, 99.4% purity, 1.00 equiv.) in THF (960 mL) was added LiEtBH (1 M, 510 mL, 1.10 equiv.). The mixture was stirred under N2 (g) for 1 h. The reaction mixture was poured into saturated aqueous NH4Cl (4.50 L) and extracted with EtOAc (2.00 L × 2). Three equal-scale batches were prepared and worked up in the same way. The combined organic layers were washed with brine (1.50 L × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 1-(tert-butyl)2-methyl (2S)-6-hydroxypiperidine-1,2-dicarboxylate (487 g) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ 5.55 (s, 1H), 5.34 (s, 1H), 4.58 (s, 1H), 3.58 (m, 3H), 2.11 (d, J = 13.2 Hz, 1H), 1.83 - 1.73 (m, 1H), 1.65 (d, J = 1.2 Hz, 2H), 1.57 - 1.51 (m, 2H), 1.41 (s, 9H).

[0127] Step 4: Preparation of (2S)-6-methoxypiperidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0128] To a solution of 1-(tert-butyl)2-methyl (2S)-6-hydroxypiperidine-1,2-dicarboxylate (266 g, 1.03 mol, 1.00 equiv.) in MeOH (1.33 L) was added TsOH·HO (39.1 g, 205 mmol, 0.20 equiv.). The reaction mixture was stirred at 25 °C for 12 h. Two equal-scale batches were performed. The reaction batches were combined and concentrated in vacuo. The crude residue was dissolved in EtOAc (3.00 L) and washed with saturated aqueous NaHCO (2.00 L × 2). The combined organic layers were washed with brine (1.50 L × 2), dried over NaSO, filtered, and concentrated in vacuo. The resulting residue was purified by column chromatography (petroleum ether / EtOAc=100 / 0 to 20 / 1) to give 1-(tert-butyl)2-methyl (2S)-6-methoxypiperidine-1,2-dicarboxylate (427 g) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.33 (m, 1H), 4.74 (m, 1H), 3.71 (s, 3H), 3.30 (s, 3H), 2.33 - 2.25 (m, 1H), 1.92 - 1.81 (m, 2H), 1.69 - 1.58 (m, 2H), 1.50 (s, 9H), 1.47 (s, 1H).

[0129] Step 5: Preparation of (2S)-6-allylpiperidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0130] To a solution of 1-(tert-butyl)2-methyl (2S)-6-methoxypiperidine-1,2-dicarboxylate (136 g, 496 mmol, 1.00 equiv.) in CHCl (1.35 L) was added allyltrimethylsilane (113 g, 992 mmol, 158 mL, 2.00 equiv.). The reaction mixture was cooled to -78 °C, and then BF .EtO (76.1 g, 536 mmol, 66.1 mL, 1.08 equiv) was added dropwise and stirred for 1 h. Three equal-scale batches were performed. The reaction mixtures were combined and quenched by pouring into HO (10.0 L). The reaction mixture was extracted with CHCl (1.00 L × 2). The combined organic layers were washed with brine (4.00 L × 2), dried over NaSO, filtered, and concentrated to give 1-(tert-butyl)2-methyl (2S)-6-allylpiperidine-1,2-dicarboxylate (334 g) as a yellow oil.

[0131] Step 6: Preparation of (2S)-6-allylpiperidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl

[0132] To a cooled (0 °C) solution of 1-(tert-butyl)2-methyl (2S)-6-allylpiperidine-1,2-dicarboxylate (188 g, 662 mmol, 1.00 equiv.) in MeOH (938 mL) was added a solution of 4 M HCl in 1,4-dioxane (827 mL, 5.00 equiv.). The mixture was warmed to 25 °C and stirred for 2 h. Two equal batches were run, and the mixtures were combined and concentrated under reduced pressure. The resulting residue was suspended in EtOAc (5.00 L) and HO (2.50 L). The aqueous layer was removed, and the organic layer was washed with saturated aqueous NaHCO (5.00 L × 2). The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give 1-(tert-butyl)2-methyl (2S)-6-allylpiperidine-1,2-dicarboxylate (76.6 g) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 5.81 - 5.72 (m, 1H), 5.15 - 5.07 (m, 2H), 3.71 (s, 3H), 3.35 - 3.32 (m, 1H), 2.58 - 2.53 (m, 1H), 2.24 - 2.14 (m, 3H), 2.12 - 2.00 (m, 1H), 1.91 - 1.84 (m, 1H), 1.64 (d, J = 13.2 Hz, 1H), 1.45 - 1.34 (m, 2H), 1.14 - 1.07 (m, 1H).

[0133] Step 7: Preparation of methyl (2S)-6-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)piperidine-2-carboxylate

[0134] To a solution of 1-(tert-butyl)2-methyl (2S)-6-allylpiperidine-1,2-dicarboxylate (56.5 g, 308 mmol, crude purity, 1.00 equiv.) in THF (1.41 L) was added (S)-2-((tert-butoxycarbonyl)amino)pent-4-enoic acid (67.7 g, 314 mmol, 1.02 equiv.) and IIDQ (100 g, 330 mmol, 1.07 equiv.). The mixture was stirred for 12 h and then diluted with EtOAc (2.80 L). The organic layer was washed with 1 N aqueous HCl (2.80 L) and then saturated aqueous NaHCO (2.80 L). The combined organic layers were washed with brine (2.80 L), dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / EtOAc = 100 / 0 to 5 / 1) to give methyl (2S)-6-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)piperidine-2-carboxylate (22.4 g, 54.4 mmol, 4.7% yield over 5 steps, 92.6% purity) as a pale yellow oil. LCMS (ESI) m / z = 381.3 [M+H] + .

[0135] The intermediates shown in Table 9 were synthesized using the procedures described above and starting from 1-(tert-butyl)2-methyl (2S)-6-allylpiperidine-1,2-dicarboxylate, the appropriate N-protected amino acid [(S)-2-((tert-butoxycarbonyl)amino)pent-4-enoic acid], IIDQ, and THF. [Table 9]

[0136] Step 8: Preparation of methyl (4S,7S,11aR,Z)-7-((tert-butoxycarbonyl)amino)-6-oxo-1,3,4,6,7,8,11,11a-octahydro-2H-pyrido[1,2-a]azocine-4-carboxylate

[0137] To a solution of methyl (2S)-6-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)piperidine-2-carboxylate (21.8 g, 53.0 mmol, 92.6% purity, 1.00 equiv.) in CHCl (1.09 L) was added Grubb's catalyst 1 (8.72 g, 10.6 mmol, 0.20 equiv.). The mixture was heated to 50° C. After 5 h, LCMS analysis indicated the reaction contained approximately 14% starting material. Additional Grubb's catalyst 1 (8.72 g, 10.6 mmol, 0.20 equiv.) was introduced to the mixture, and the mixture was aged for an additional 2 h. The reaction mixture was cooled to room temperature and concentrated to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 100 / 1 to 20 / 1) to give methyl (4S,7S,11aR,Z)-7-((tert-butoxycarbonyl)amino)-6-oxo-1,3,4,6,7,8,11,11a-octahydro-2H-pyrido[1,2-a]azocine-4-carboxylate (10.0 g) as a brown oil. LCMS (ESI) m / z = 353.3 [M+H] + .

[0138] The intermediates shown in Table 10 were synthesized by ring-closing metathesis using the conditions described above for the preparation of methyl (4S,7S,11aR,Z)-7-((tert-butoxycarbonyl)amino)-6-oxo-1,3,4,6,7,8,11,11a-octahydro-2H-pyrido[1,2-a]azocine-4-carboxylate and starting from methyl (2S)-6-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)but-3-enoyl)piperidine-2-carboxylate and using first generation Grubb's catalyst. [Table 10]

[0139] Step 9: Preparation of methyl (4S,7S,11aS)-7-((tert-butoxycarbonyl)amino)-6-oxodecahydro-2H-pyrido[1,2-a]azocine-4-carboxylate

[0140] To a solution of methyl (4S,7S,11aR,Z)-7-((tert-butoxycarbonyl)amino)-6-oxo-1,3,4,6,7,8,11,11a-octahydro-2H-pyrido[1,2-a]azocine-4-carboxylate (10.4 g, 29.5 mmol, 1.00 equiv) in EtOAc (100 mL) was added 10% Pd / C (2.08 g) under N2(g). The suspension was subjected to three cycles of degassing under reduced pressure and purging with H2(g). The mixture was stirred at 25 °C under H2(g) (50 psi). After 12 h, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (Phenomenex luna C18 250 × 80 mm × 10 μm; mobile phase: [water (TFA)-acetonitrile]; B%: 50%-80%). The pH of the mixture was adjusted to 7-8 with saturated aqueous NaHCO3 and carefully concentrated under reduced pressure to remove acetonitrile. The aqueous layer was extracted with EtOAc (250 mL × 2). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give methyl (4S,7S,11aS)-7-((tert-butoxycarbonyl)amino)-6-oxodecahydro-2H-pyrido[1,2-a]azocine-4-carboxylate (6.46 g, 18.1 mmol, 31.2% yield over two steps) as a dark brown gum. LCMS (ESI) m / z = 355.2 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 5.82 (d, J = 7.2 Hz, 1H), 5.20 (t, J = 2.4 Hz, 1H), 4.67 - 4.62 (m, 1H), 4.24 - 4.21 (m, 1H), 3.72 (s, 3H), 3.31 - 3.28 (m, 1H), 2.14 - 2.10 (m, 1H), 2.03 - 1.65 (m, 6H), 1.60 - 1.36 (m, 5H), 1.44 (s, 9H), 1.27 - 1.21 (m, 1H).

[0141] The intermediates shown in Table 11 were synthesized using the hydrogenation conditions described above for the preparation of methyl (4S,7S,11aS)-7-((tert-butoxycarbonyl)amino)-6-oxodecahydro-2H-pyrido[1,2-a]azocine-4-carboxylate and starting from methyl (4S,7S,10aR)-7-((tert-butoxycarbonyl)amino)-6-oxo-1,2,3,4,6,7,10,10a-octahydropyrido[1,2-a]azepine-4-carboxylate. [Table 11]

[0142] Step 10: Preparation of (4S,7S,11aS)-7-((tert-butoxycarbonyl)amino)-6-oxodecahydro-2H-pyrido[1,2-a]azocine-4-carboxylic acid

[0143] (4S,7S,11aS)-7-((tert-butoxycarbonyl)amino)-6-oxodecahydro-2H-pyrido[1,2-a]azocine-4-carboxylic acid was prepared using the hydrolysis conditions previously described in step 4 for the preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid.

[0144] The intermediates shown in Table 12 were synthesized using the conditions described in Step 4 for the preparation of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid. [Table 12]

[0145] Synthesis of (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylic acid and (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylic acid [ka]

[0146] Preparation of (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate 1-(tert-butyl) 2-methyl

[0147] (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl was prepared according to the protocol reported in WO2015010626.

[0148] Step 1: Preparation of 1-(tert-butyl)2-methyl (2S,4R)-5-(but-3-en-1-yl)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate

[0149] To a cooled (−78° C.) solution of 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate (165 g, 442 mmol, 1.00 equiv.) in THF (1.50 L) was added a 1 M solution of but-3-en-1-ylmagnesium bromide in THF (552 mL, 1.25 equiv.). The reaction mixture was stirred for 1 h. Two equal-scale batches were run in parallel and combined for workup. The reaction mixture was poured into saturated aqueous NH4Cl (6.00 L) and extracted with EtOAc (5.00 L × 3). The combined organic layers were washed with brine (5.00 L), dried over Na2SO4, filtered, and concentrated to give 1-(tert-butyl)2-methyl (2S,4R)-5-(but-3-en-1-yl)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate (357 g) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.95 - 5.72 (m, 1H), 5.13 - 4.92 (m, 2H), 4.53 - 4.39 (m, 1H), 4.18 (t, J = 5.2 Hz, 1H), 3.79 - 3.71 (m, 3H), 2.84 - 2.70 (m, 1H), 2.63 - 2.54 (m, 1H), 2.38 - 2.24 (m, 2H), 2.22 - 2.08 (m, 2H), 1.47 - 1.39 (m, 9H), 0.99 - 0.90 (m, 9H), 0.18 - 0.12 (m, 3H), 0.11 - 0.06 (m, 3H).

[0150] Step 2: Preparation of methyl (2S,4R,5S)-5-(but-3-en-1-yl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate

[0151] To a cooled (0 °C) solution of 1-(tert-butyl)2-methyl (2S,4R)-5-(but-3-en-1-yl)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate (178 g, 414 mmol, 1.00 equiv.) in CHCl (1.80 L) was added TFA (472 g, 4.14 mol, 307 mL, 10.0 equiv.) followed by EtSiH (241 g, 2.07 mol, 331 mL, 5.00 equiv.). The mixture was stirred for 3 h while maintaining the reaction temperature between 0 °C and 25 °C. Two equal-scale batches were run in parallel and combined during workup. The combined reaction mixture was concentrated to give a residue. The resulting residue was dissolved in EtOAc (2.00 L), and saturated aqueous NaHCO was added until the aqueous layer was basic (pH ∼8). The mixture was extracted with EtOAc (2.00 L × 3). The combined organic layers were washed with brine (2.00 L), dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / EtOAc = 1 / 0 to 10 / 1) to give methyl (2S,4R,5S)-5-(but-3-en-1-yl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (102 g, 325 mmol, 39.3% over 4 steps) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 5.89 - 5.79 (m, 1H), 5.08 - 5.03 (m, 1H), 5.00 - 4.97 (m, 1H), 3.97 (t, J = 8.0 Hz, 1H), 3.89 - 3.84 (m, 1H), 3.74 (s, 3H), 2.94 - 2.90 (m, 1H), 2.24 - 2.11 (m, 3H), 2.04 - 1.96 (m, 2H), 1.73 - 1.61 (m, 1H), 1.51 - 1.42 (m, 1H), 0.89 (s, 9H), 0.06 (d, J = 1.0 Hz, 6H).

[0152] Step 3: Preparation of methyl (2S,4R,5S)-5-(but-3-en-1-yl)-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate

[0153] To a solution of methyl (2S,4R,5S)-5-(but-3-en-1-yl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate in DMF (1.00 L), N,N-diisopropylethylamine (283 mL, 1.63 mol, 5.00 equiv.), (S)-2-((tert-butoxycarbonyl)amino)pent-4-enoic acid (77.0 g, 358 mmol, 1.10 equiv.), and HATU (247 g, 651 mmol, 2.00 equiv.) were added sequentially. The mixture was stirred at ambient temperature for 3 hours and then poured into water (2.00 L). The mixture was extracted with EtOAc (1.00 L × 3). The combined organic layers were washed with saturated aqueous NaHCO (1.00 L × 3), saturated aqueous NH Cl (1.00 L), brine (1.00 L), dried over Na SO , filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / EtOAc = 1 / 0 to 10 / 1) to give methyl (2S,4R,5S)-5-(but-3-en-1-yl)-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (121 g, 220 mmol, 67.5% yield, 92.7% purity) as a yellow oil. LCMS (ESI) m / z = 511.2 [M+H] + ; 1H NMR (400 MHz, CDCl3) δ 5.91 - 5.66 (m, 2H), 5.18 - 5.00 (m, 4H), 4.66 - 4.56 (m, 1H), 4.50 - 4.41 (m, 1H), 4.24 (d, J = 3.2 Hz, 1H), 4.16 - 4.04 (m, 1H), 3.98 - 3.95 (m, 1H), 3.80 - 3.71 (m, 3H), 2.58 - 2.48 (m, 1H), 2.43 - 2.27 (m, 2H), 2.24 - 2.13 (m, 3H), 2.09 - 2.01 (m, 1H), 1.85 - 1.75 (m, 1H), 1.44 - 1.39 (m, 9H), 0.89 - 0.82 (m, 9H), 0.09 - 0.04 (m, 6H).

[0154] Step 4: Preparation of (1R,3S,6S,11aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-2,3,5,6,7,10,11,11a-octahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate

[0155] To a solution of methyl (2S,4R,5S)-5-(but-3-en-1-yl)-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (60.5 g, 118 mmol, 1.00 equiv.) in CHCl (1.20 L) was added Grubb's catalyst 1 (9.75 g, 11.9 mmol, 0.10 equiv.) under N. The mixture was heated to 55° C. and stirred for 12 h. Two equal-scale batches were run in parallel and combined for workup. The combined reaction mixture was cooled to ambient temperature and concentrated to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 1 / 0 to 10 / 1) to give (1R,3S,6S,11aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-2,3,5,6,7,10,11,11a-octahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (70.8 g, 130 mmol, 54.9% yield, 88.6% purity) as a brown oil. LCMS (ESI) m / z = 483.2 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 6.02 - 5.90 (m, 1H), 5.83 (d, J = 6.8 Hz, 1H), 5.71 - 5.49 (m, 1H), 4.78 (t, J = 9.0 Hz, 1H), 4.34 (t, J = 6.8 Hz, 1H), 4.10 - 4.07 (m, 1H), 3.88 - 3.82 (m, 1H), 3.75 (s, 3H), 2.66 - 2.58 (m, 1H), 2.51 - 2.38 (m, 1H), 2.35 - 2.30 (m, 1H), 2.27 - 2.18 (m, 1H), 2.16 - 2.09 (m, 2H), 1.54 - 1.46 (m, 2H), 1.42 (s, 9H), 0.80 (s, 9H), 0.09 - 0.03 (m, 6H).

[0156] Step 5: Preparation of (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate

[0157] To a solution of (1R,3S,6S,11aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-2,3,5,6,7,10,11,11a-octahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (70.8 g, 147 mmol, 1.00 equiv) in EtOH (700 mL) was added Pd / C (7.00 g, 10% wt) under a constant stream of N2(g). The reaction mixture was degassed under reduced pressure and purged with H2(g) (3 times). The reaction mixture was stirred under H2(g) (50 psi) for 12 h. The reaction mixture was filtered, and the filter cake was washed with EtOH (200 mL × 3). The filtrate was concentrated to give methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (65.0 g, 125 mmol, 85.5% yield, 93.5% purity) as an off-white solid. LCMS (ESI) m / z = 485.1 [M+H] + ; 1H NMR (400 MHz, CDCl3) δ 5.17 (d, J = 8.8 Hz, 1H), 4.86 - 4.76 (m, 1H), 4.70 - 4.65 (m, 1H), 4.10 (d, J = 2.0 Hz, 1H), 3.86 (d, J = 10.0 Hz, 1H), 3.77 (s, 3H), 2.20 - 2.15 (m, 1H), 2.13 - 2.03 (m, 1H), 2.02 - 1.94 (m, 1H), 1.88 - 1.73 (m, 5H), 1.59 (s, 2H), 1.54 - 1.44 (m, 2H), 1.41 (s, 9H), 0.85 (s, 9H), 0.08 (d, J = 5.0 Hz, 6H).

[0158] Step 6: Preparation of methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate

[0159] A cooled (−10° C.) solution of methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (65.0 g, 134 mmol, 1.00 equiv.) in THF (650 mL) was added dropwise to a 1 M solution of TBAF in THF (268 mL, 2.00 equiv.). The reaction mixture was stirred at 0° C. for 1 h and then poured into water (1.50 L). The mixture was extracted with EtOAc (1.00 L × 2). The combined organic layers were washed with brine (1.00 L), dried over NaSO, filtered, and concentrated to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 1 / 0 to 3 / 2) to give methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (40.8 g, 107 mmol, 79.9% yield, 97.3% purity) as an off-white solid. LCMS (ESI) m / z = 371.4 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 5.22 (d, J = 8.0 Hz, 1H), 4.74 - 4.70 (m, 2H), 4.19 (d, J = 3.2 Hz, 1H), 4.01 (d, J = 10.0 Hz, 1H), 3.76 (s, 3H), 2.78 (s, 1H), 2.34 - 2.28 (m, 1H), 2.12 - 1.97 (m, 2H), 1.89 - 1.83 (m, 2H), 1.79 - 1.74 (m, 3H), 1.61 - 1.57 (m, 4H), 1.42 (s, 9H).

[0160] Step 7: Preparation of methyl (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxo-5,6,7,8,9,10,11,11a-octahydro-3H-pyrrolo[1,2-a]azonine-3-carboxylate

[0161] To a cooled (−78 °C) solution of methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (12.0 g, 32.4 mmol, 1.00 equiv.) in CHCl (120 mL) was added DAST (25.7 mL, 194 mmol, 6.00 equiv.) slowly under N (g). After complete addition of DAST, the mixture was warmed to 25 °C and stirred for 8 h. The reaction mixture was cooled to 0 °C and then quenched by the addition of saturated aqueous NaHCO (300 mL). The biphasic mixture was extracted with CHCl (200 mL × 3). The combined organic layers were washed with brine (200 mL), dried over NaSO, filtered, and concentrated to give a residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 1 / 0 to 3 / 1) to give methyl (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxo-5,6,7,8,9,10,11,11a-octahydro-3H-pyrrolo[1,2-a]azonine-3-carboxylate (3.00 g, 8.36 mmol, 25.8% yield, 98.2% purity) as a white solid. LCMS (ESI) m / z = 353.1 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 5.87 - 5.78 (m, 2H), 5.43 (s, 1H), 5.18 (d, J = 8.0 Hz, 1H), 4.88 - 4.74 (m, 2H), 3.77 (s, 3H), 2.04 - 1.93 (m, 2H), 1.89 - 1.76 (m, 4H), 1.63 - 1.55 (m, 4H), 1.43 (s, 9H).

[0162] Step 8: Preparation of methyl (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate

[0163] To a solution of methyl (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxo-5,6,7,8,9,10,11,11a-octahydro-3H-pyrrolo[1,2-a]azonine-3-carboxylate (3.00 g, 8.51 mmol, 1.00 equiv) in MeOH (30.0 mL) was added Pd / C (0.60 g, 10% wt) under a constant stream of N2(g). The mixture was degassed under reduced pressure and purged with H2(g) (15 psi) (3 times). The mixture was heated to 45 °C and stirred under H2(g) (15 psi) for 22 h. The mixture was filtered, and the filter cake was washed with MeOH (50.0 mL × 3). The filtrate was concentrated to give a residue. The residue was dissolved in acetonitrile (5.00 mL) and HO (30.0 mL) and then lyophilized to give methyl (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (5.72 g, 15.9 mmol, 94.2% yield, 98.2% purity) as an off-white solid. LCMS (ESI) m / z = 355.1 [M+H[ + ; 1 H NMR (400 MHz, CDCl3) δ 5.16 (d, J = 8.8 Hz, 1H), 4.89 - 4.78 (m, 1H), 4.60 - 4.51 (m, 1H), 4.22 - 4.13 (m, 1H), 3.76 (s, 3H), 2.30 - 2.18 (m, 2H), 2.01 (s, 2H), 1.83 - 1.73 (m, 7H), 1.60 - 1.55 (m, 3H), 1.43 (s, 9H).

[0164] Step 9: Preparation of (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylic acid

[0165] To a cooled (0 °C) solution of methyl (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (1.00 g, 2.82 mmol, 1.00 equiv) in THF (10 mL) was added LiOH HO (153 mg, 3.66 mmol, 1.3 equiv) and water (10 mL). The mixture was stirred for 16 h and then acidified with NaHSO. The aqueous layer was extracted with EtOAc (3 times). The combined organic layers were dried over NaSO and concentrated to give (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylic acid (855 mg, 2.51 mmol, 89.1% yield) as a solid. LCMS (ESI) m / z = 341.4 [M+H] + .

[0166] Step 10: Preparation of methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate

[0167] Methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate was prepared using the hydrolysis procedure described above for the preparation of (3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylic acid and starting from methyl (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate. Starting with (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (500 mg, 1.34 mmol, 1.00 equiv.), LiOH·HO (107 mg, 2.68 mmol, 2.0 equiv.), THF (12 mL), and water (4 mL), (1R,3S,6S,11aS)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxodecahydro-1H-pyrrolo[1,2-a]azonine-3-carboxylate (340 mg, 0.953 mmol, 71.2% yield) was obtained as a solid. LCMS (ESI) m / z = 357.3 [M+H] + . (3S,6S,9R,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid, (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid Synthesis of (3S,6S,8S,10aR)-6-((tert-butoxycarbonyl)amino)-8-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid and (3S,6S,8R,10aR)-6-((tert-butoxycarbonyl)amino)-8-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0168] Step 1: Preparation of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-8-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate [ka]

[0169] Methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-8-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate were prepared starting from methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate and synthesized in the Journal of Medicinal Chemistry. It was prepared using a similar protocol established in Chemistry (2010), 53(17), 6361-6367.

[0170] The mixture of alcohol diastereomers and regioisomers was separated by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5-40% MeOH in water (containing 0.1% formic acid)), followed by chiral SFC separation (Lux i-Cellulose-5 21.2 x 250 mm 5 μm column, column temperature = 40 °C, flow rate 75 mL / min, 20% MeOH, cycle time: 5 min). A representative reaction starting with methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (4.3 g, 12.7 mmol) gave the following products:

[0171] Peak 1: (3S,6S,9R,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (760 mg, 2.13 mmol, 16.8% yield) as a white solid. LCMS (ESI) m / z = 357.2 [M+H] + .

[0172] Peak 2: (3S,6S,9S,10aR)-methyl 6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (375 mg, 19% yield) as a clear thick oil. LCMS (ESI) m / z = 357.2 [M+H] + .

[0173] Peak 3: (3S,6S,8R,10aR)-6-{[(tert-butoxy)carbonyl]amino}-8-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (795 mg, 40% yield) as a white foam. LCMS (ESI) m / z = 357.2 [M+H] + .

[0174] Peak 4: (3S,6S,8S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-8-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (600 mg, 30% yield) as a white solid. LCMS (ESI) m / z = 357.2 [M+H] + .

[0175] Step 2: Preparation of (3S,6S,9R,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid. [ka]

[0176] To a solution of (3S,6S,9R,10aR)-methyl 6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (peak 1) (1.08 g, 3.03 mmol, 1 equiv.) in a mixture of THF (24 mL) and water (8 mL) was added LiOH·HO (380 mg, 9.08 mmol, 3.0 equiv.). The reaction mixture was stirred at room temperature for 2 h. The reaction was then concentrated under reduced pressure to remove tetrahydrofuran. The crude residue was purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5-60% acetonitrile in water (containing 0.1% formic acid)) to give (3S,6S,9R,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid (780 mg, 76% yield) as a white solid. LCMS (ESI) m / z = 343.2 [M+H] + .

[0177] The following intermediates in Table 13 were prepared following the general procedures described above, starting from the appropriate starting materials. [Table 13] Synthesis of (3S,6S,9S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-9-methyl-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid and (3S,6S,8R,10aR)-6-((tert-butoxycarbonyl)amino)-8-hydroxy-8-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0178] Step 1: Preparation of methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate [ka]

[0179] Methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate were prepared using the protocol described in US 2009 / 0123480. The crude reaction mixture was purified by reverse-phase chromatography using a C18 cartridge eluted with a gradient of 5 to 100% acetonitrile in water (containing 0.1% FA) to give a mixture of isomeric ketones (1.2 g) as a beige solid. The resulting ketone isomers were separated by chiral SFC (Lux i-cellulose-5 21.2 × 250 mm 5 μm column, flow rate 75 mL / min, 15% MeOH) to give methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (9) (424 mg, 35.6% yield) as a white solid and methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (433 mg, 36.3% yield) as a white solid.

[0180] (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate methyl: LCMS (ESI) m / z = 299.1 [M+H] + ; 1H NMR: (400 MHz, CDCl3) δ 5.27 (d, J = 8.1 Hz, 1H), 4.59 (t, J = 8.7 Hz, 1H), 4.45 - 4.31 (m, 2H), 3.80 (s, 3H), 3.20 (td, J = 12.5, 4.9 Hz, 1H), 3.06 (t, J = 12.0 Hz, 1H), 2.47 - 2.15 (m, 5H), 2.13 - 2.00 (m, 1H), 1.87 (dd, J = 12.1, 7.0 Hz, 1H), 1.68 - 1.62 (m, 1H), 1.41 (s, 9H).

[0181] (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate methyl: LCMS (ESI) m / z = 299.1 [M+H] + ; 1 H NMR: (400 MHz, CDCl3) δ 5.56 (d, J = 7.6 Hz, 1H), 5.13 (ddd, J = 12.2, 7.6, 4.9 Hz, 1H), 4.51 (t, J = 8.8 Hz, 1H), 4.33 - 4.23 (m, 1H), 3.73 (s, 3H), 3.07 (dd, J = 13.8, 4.5 Hz, 1H), 2.96 (td, J = 11.9, 2.9 Hz, 1H), 2.65 - 2.50 (m, 2H), 2.36 - 1.91 (m, 4H), 1.90 - 1.76 (m, 2H), 1.45 (s, 9H).

[0182] Step 2: Preparation of (3S,6S,9S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-9-methyl-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0183] To a cooled (-78 °C) solution of methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (250 mg, 705 μmol, 1 equiv.) in THF (15 mL) was added a 3 M solution of MeMgCl in THF (480 μL, 1.44 mmol, 2.05 equiv.). The reaction mixture was allowed to warm slowly to 0 °C with stirring for 2 h and then quenched with saturated aqueous NH Cl. The product was extracted with EtOAc (twice), and the combined organic layers were dried over Na SO , filtered, and concentrated in vacuo. The crude residue was purified by reverse phase chromatography [C18 cartridge eluted with a gradient of 5-60% acetonitrile in water (containing 0.1% formic acid)] to give the resulting tertiary alcohol (240 mg, 647 μmol, 93.3%) as a white solid [ 1 Based on H NMR, the product was obtained as a 1:1 mixture of ketone and tertiary alcohol. This mixture was dissolved in tetrahydrofuran (6 mL) and water (2 mL), and then LiOH·HO (81.4 mg, 1.94 mmol, 3 equiv.) was added. The reaction mixture was stirred at room temperature for 4 h. The reaction mixture was concentrated under reduced pressure and purified by reverse-phase chromatography (C18 cartridge, elution gradient: 5–60% acetonitrile in water (containing 0.1% FA)). Pure fractions were combined and evaporated to dryness to give (3S,6S,9S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-9-methyl-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid (82.0 mg, 230 μmol, 35.6% yield) as a white solid. LCMS (ESI) m / z = 357.2 [M+H] + .

[0184] The following intermediates in Table 14 were prepared according to the protocol outlined above and starting from methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate. [Table 14] Synthesis of (3S,6S,9S,10aR)-9-(azetidin-1-yl)-6-{[(tert-butoxy)carbonyl]amino}-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0185] Step 1: Preparation of methyl (3S,6S,9S,10aR)-9-(azetidin-1-yl)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0186] To a solution of methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (150 mg, 0.4232 mmol, 1 equiv.) and AcOH (26.5 μL, 465 μmol, 1.1 equiv.) in CHCl (10 mL) was added azetidine (55.7 μL, 846 μmol, 2 equiv.). The mixture was stirred at room temperature for 2 hours. To the mixture was added NaBH(OAc) (134 mg, 634 μmol, 1.5 equiv.), and the reaction was stirred at room temperature for 1.5 hours. The reaction was quenched by the addition of a saturated aqueous solution of NaHCO (10 mL). The product was extracted with CHCl (3 times). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure to give crude (3S,6S,9S,10aR)-9-(azetidin-1-yl)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate as a white solid. LCMS (ESI) m / z = 396.3 [M+H] + .

[0187] Step 2: Preparation of (3S,6S,9S,10aR)-9-(azetidin-1-yl)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0188] To a solution of (3S,6S,9S,10aR)-9-(azetidin-1-yl)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (180 mg, 0.4551 mmol) in THF (24 mL) and water (8 mL) was added LiOH·HO (57.0 mg, 1.36 mmol). The reaction was stirred at room temperature for 4 h. The reaction was then concentrated under reduced pressure. The crude residue was purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 0–40% acetonitrile in water). Pure fractions were concentrated to dryness, and the product was then lyophilized to give (3S,6S,9S,10aR)-9-(azetidin-1-yl)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (140 mg, 0.3669 mmol, 80.9% yield) as a white solid. LCMS (ESI) m / z = 382.4 [M+H] + .

[0189] The following intermediates in Table 15 were prepared according to the procedures outlined above and starting from methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate. [Table 15] Synthesis of (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid [ka]

[0190] Step 1: Preparation of methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methylidene-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate

[0191] To a cooled (0°C) solution of methyltriphenylphosphonium bromide (628 mg, 1.76 mmol, 2.5 equiv) in THF (10 mL) was added a solution of 1.0 M NaHMDS in THF (1.76 mL, 1.76 mmol, 2.5 equiv). The resulting yellow suspension was warmed to room temperature and stirred for 30 minutes. The yellow suspension was cooled to 0°C, after which a solution of methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,9-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (250 mg, 0.7054 mmol, 1 equiv) in THF (10 mL) was added. The resulting reaction mixture was stirred for 10 minutes and then allowed to warm to ambient temperature over 1.5 hours. The reaction mixture was quenched by the addition of brine (10 mL), and the product was extracted with EtOAc (30 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5 to 60% acetonitrile containing 10 mM ammonium bicarbonate / ammonium hydroxide buffer pH = 10) to afford methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methylidene-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (210 mg, 0.5958 mmol, 84.6% yield) as a white solid. LCMS (ESI) m / z = 353.2 [M+H] + .

[0192] Step 2: Preparation of (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methylidene-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0193] To a solution of methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methylidene-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate (210 mg, 0.5958 mmol, 1 equiv.) in THF (6 mL) and water (2 mL) was added LiOH HO (74.6 mg, 1.78 mmol, 3 equiv.). The reaction was stirred at room temperature for 20 h. The reaction was then concentrated under reduced pressure and purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5-60% acetonitrile in water (containing 0.1% FA)) to afford (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methylidene-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid (186 mg, 0.5496 mmol, 92.5% yield) as a white solid. LCMS (ESI) m / z = 339.2 [M+H] + .

[0194] Step 3: Preparation of (3S,6S,10aR)-2,2-dibromo-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid

[0195] To a solution of (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methylidene-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carboxylic acid (186 mg, 0.5496 mmol, 1 equiv.) in CHCl (2 mL) was added bromoform (384 μL, 4.35 mmol, 7.9 equiv.) and TEBAC (15.0 mg, 65.9 μmol, 0.12 equiv.), followed by a 50 wt.% aqueous solution of NaOH (0.56 mL, 7.0 mmol, 12.7 equiv.). The reaction was heated to reflux for 21 h. The reaction mixture was then diluted with 1 N aqueous NaOH, and the product was extracted with CHCl. ​​The organic layer was concentrated under reduced pressure and dried under high vacuum. The resulting residue was purified by reverse-phase chromatography [C18 cartridge using a gradient of 5-80% acetonitrile in water (containing 0.1% FA)] to give (3S,6S,10aR)-2,2-dibromo-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid (175 mg, 0.3429 mmol, 62.5% yield) as a white solid. LCMS (ESI) m / z = 509.0 [M+H] + .

[0196] Step 4: Preparation of (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid

[0197] To a solution of (3S,6S,10aR)-2,2-dibromo-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid (175 mg, 0.3429 mmol, 1 equiv.) in 2-propanol (6 mL) under a N2(g) atmosphere was added KOH (115 mg, 2.05 mmol, 6 equiv.) and 10% Pd / C (50% wet, 170 mg, 0.1597 mmol, 0.466 equiv.). The reaction mixture was heated to 70 °C under H2(g) (40 psi) for 21 h. The reaction mixture was then diluted with MeOH and filtered through a pad of Celite®. The filtrate was concentrated under reduced pressure and then purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5-70% acetonitrile in water (containing 0.1% FA)) to give (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid (104 mg, 0.2950 mmol, 86.6% yield) as a white solid. LCMS (ESI) m / z = 353.2 [M+H] + .

[0198] The following intermediates in Table 16 were prepared according to the steps (steps 1-4) outlined above for the synthesis of (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-spiro[cyclopropane-1,9-pyrrolo[1,2-a]azocine]-3-carboxylic acid and starting from methyl (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-5,8-dioxo-decahydropyrrolo[1,2-a]azocine-3-carboxylate. [Table 16] Synthesis of (3S,6S,10S,10aR)-6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0199] Step 1: Preparation of (2S,5R)-methyl 5-ethynylpyrrolidine-2-carboxylate hydrochloride

[0200] To a solution of 1-(tert-butyl)2-methyl (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylate (52.0 g, 205 mmol, 1.00 equiv.) in EtOAc (260 mL) was added a solution of 4.0 M HCl in EtOAc (154 mL, 3.00 equiv.). After 3.5 h, the mixture was concentrated under reduced pressure to give methyl (2S,5R)-5-ethynylpyrrolidine-2-carboxylate hydrochloride (44.0 g, 232 mmol, HCl salt) as a brown oil. 1 H NMR (400 MHz CDCl3) δ 4.74 - 4.71 (m, 1H), 4.62 (t, J = 7.2 Hz, 1H), 3.91 (s, 3H), 2.69 (d, J = 1.6 Hz, 1H), 2.61 - 2.35 (m, 3H), 2.21 - 2.14 (m, 1H).

[0201] Step 2: Preparation of methyl (2S,5R)-1-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)pent-4-enoyl)-5-ethynylpyrrolidine-2-carboxylate

[0202] To a mixture of (2S,5R)-methyl 5-ethynylpyrrolidine-2-carboxylate hydrochloride (44.0 g, 232 mmol, 1.00 equiv, HCl) and (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)pent-4-enoic acid (62.6 g, 186 mmol, 0.80 equiv) in CHCl (880 mL) was added BOPCl (59.1 g, 232 mmol, 1.00 equiv) and NaHCO (78.0 g, 928 mmol, 36.1 mL, 4.00 equiv) under N (g). The mixture was heated to 40 °C and stirred. After 2 h, the mixture was cooled to ambient temperature and concentrated in vacuo. The residue was purified by column chromatography (petroleum ether / EtOAc = 100 / 1 to 4 / 1) to give methyl (2S,5R)-1-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)pent-4-enoyl)-5-ethynylpyrrolidine-2-carboxylate (85.0 g, 169 mmol, purity 93.9%) as a yellow oil. LCMS (ESI) m / z = 473.0 [M+H] + ; 1 H NMR (400 MHz CDCl3) δ 7.76 (d, J = 7.6 Hz, 2H), 7.60 - 7.56 (m, 2H), 7.40 (t, J = 7.6 Hz, 2H), 7.33 - 7.30 (m, 2H), 5.92 - 5.81 (m, 1H), 5.50 (d, J = 8.4 Hz, 1H), 5.30 - 5.10 (m, 2H), 5.00 - 4.92 (m, 1H), 4.73 - 4.69 (m, 1H), 4.54 - 4.50 (m, 1H), 4.40 - 4.35 (m, 1H), 4.31 - 4.27 (m, 1H), 4.23 - 4.19 (m, 1H), 3.76 - 3.70 (m, 2H), 2.80 - 2.75 (m, 1H), 2.58 - 2.53 (m, 1H), 2.50 - 2.49 (m, 1H), 2.37 - 2.31 (m, 1H), 2.27 - 2.18 (m, 3H).

[0203] Step 3: Preparation of methyl (3S,6S,10aR,Z)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0204] A solution of methyl (2S,5R)-1-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)pent-4-enoyl)-5-ethynylpyrrolidine-2-carboxylate in toluene (2.80 L) was sparged with ethylene for 10 minutes. To this solution was added Grubbs Generation I catalyst (3.65 g, 5.82 mmol, 0.05 equiv.), and the mixture was stirred under an atmosphere of ethylene. Three equal-scale batches were run in parallel and combined for workup. After 3 hours, the reaction mixture was concentrated in vacuo. The resulting product was purified by reverse-phase HPLC (0.10% formic acid) to give methyl (3S,6S,10aR,Z)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (43.0 g, 87.1 mmol, purity 95.7%) as a dark brown oil. LCMS (ESI) m / z = 473.0 [M+H] + ; 1H NMR (400 MHz CDCl3) δ 7.77 (d, J = 7.6 Hz, 2H), 7.62 - 7.58 (m, 2H), 7.41 (t, J = 7.2 Hz, 2H), 7.33 - 7.29 (m, 2H), 6.24 - 6.15 (m, 1H), 5.94 - 5.81 (m, 1H),5.58 (d, J = 8.8 Hz, 1H), 5.27 - 5.16 (m, 1H), 5.04 - 5.01 (m, 1H), 4.81 - 4.69 (m, 1H), .4.54 - 4.50 (m, 1H), 4.40 - 4.20 (m, 3H), 3.77 - 3.70 (m, 3H), 2.79 - 2.66 (m, 1H), 2.57 - 2.49 (m, 1H), 2.38 - 2.16 (m, 3H), 2.11 - 2.07 (m, 1H).

[0205] Step 4: Preparation of methyl (3S,6S,10aR,Z)-6-amino-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0206] To a solution of methyl (3S,6S,10aR,Z)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (13.0 g, 27.5 mmol, 1.00 equiv.) in THF (260 mL) was added piperidine (7.03 g, 82.5 mmol, 8.15 mL, 3.00 equiv.). After stirring for 2 h, the mixture was poured into an aqueous solution of 2 N HCl (200 mL), and the biphasic mixture was extracted with EtOAc (200 mL × 2). The combined organic layers were discarded, and the aqueous phase was made basic with NaHCO3 (adjusted to pH = 8-9). The resulting basic aqueous layer was extracted with EtOAc (200 mL × 2). The combined organic layers were discarded, and the aqueous phase was collected to give the crude product. This crude product was purified by prep-HPLC (Phenomenex Luna C18 250 mm × 100 mm × 10 μm; mobile phase: [water (TFA)-acetonitrile]; B%: 10% to 30%, 20 min) and lyophilized to give methyl (3S,6S,10aR,Z)-6-amino-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (2.65 g, 10.6 mmol, 38.5% yield) as a white solid. LCMS (ESI) m / z = 250.9 [M+H] + ; 1 H NMR (400 MHz CDCl3) δ 8.30 (s, 3H), 6.30 - 6.23 (m, 1H), 5.89 (t, J = 4.0 Hz, 1H), 5.29 - 5.25 (m, 1H), 5.04 - 5.01 (m, 1H), 4.88 (s, 1H), 4.63 - 4.61 (m, 1H), 4.46 (s, 1H), 3.61 (s, 3H), .2.49 - 2.43 (m, 1H), 2.39 - 2.03 (m, 1H), 1.93 - 1.92 (m, 1H), 1.91 - 1.82 (m, 1H).

[0207] Step 5: Preparation of methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0208] To a solution of (3S,6S,10aR,Z)-6-amino-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (2.55 g, 10.2 mmol, 1.00 equiv.) and EtN (2.06 g, 20.4 mmol, 2.84 mL, 2.00 equiv.) in CHCl (25.5 mL) was added BocO (2.45 g, 11.2 mmol, 2.57 mL, 1.10 equiv.). The reaction was stirred for 2 h and then poured into saturated aqueous NHCl (50.0 mL). The biphasic mixture was extracted with EtOAc (30.0 mL × 2). The combined organic layers were dried over NaSO and concentrated to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 100 / 1 to 10 / 1) to give methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (1.52 g, 4.26 mmol, 98.1% purity, 41.8% yield) as a colorless oil. LCMS (ESI) m / z = 251.2 [(M-100)+H] + ; 1H NMR (400 MHz CDCl3) δ 6.21 - 6.14 (m, 1H), 5.90 (d, J = 4.0 Hz, 1H), 5.76 (d, J = 3.2 Hz, 1H), 5.20 (d, J = 8.4 Hz, 1H), 4.99 (d, J = 5.4 Hz, 1H),4.79 - 4.72 (m, 3H), 4.65 - 4.61 (m, 1H), 3.71 (s, 3H), .2.61 - 2.59 (m, 1H), 2.53 - 2.38 (m, 2H), 2.07 - 2.04 (m, 2H), 1.43 (s, 9H).

[0209] Step 6: Preparation of methyl (3S,6S,10S,10aR)-6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0210] To a solution of methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-10-methylene-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (1.52 g, 4.34 mmol, 1.00 equiv) in EtOAc (15.2 mL) under N2(g) was carefully added 10% Pd / C (0.30 g, 868 μmol, 0.20 equiv) at 25 °C. The suspension was subjected to three cycles of evacuation and purging with H2(g). The mixture was stirred under H2(g) (50 Psi) at 25 °C. After stirring for 3 h, the reaction vessel was evacuated, and the reaction mixture was subjected to three cycles of evacuation and purging with N2(g). The reaction mixture was filtered through Celite®, and the filtrate was concentrated to give methyl (3S,6S,10S,10aR)-6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (1.14 g, crude) as a colorless oil. LCMS (ESI) m / z = 355.0 [M+H] + .

[0211] Step 7: Preparation of (3S,6S,10S,10aR)-6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0212] To a solution of (3S,6S,10S,10aR)-methyl 6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (1.16 g, 3.26 mmol, 1.00 equiv.) in THF (11.0 mL) was added a solution of LiOH.HO (411 mg, 9.79 mmol, 3.00 equiv.) in HO (2.20 mL). After 12 h, HO (20.0 mL) was added to the reaction mixture, and the solution was acidified to pH 2 with 1 N aqueous HCl. The mixture was extracted with EtOAc (20.0 mL × 3). The combined organic layers were dried over NaSO and concentrated to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18 150 × 40 mm × 15 μm; mobile phase: [water (TFA)-ACN]; B%: 31% to 61%, 10 min) and lyophilized to give (3S,6S,10S,10aR)-6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (0.80 g, 2.35 mmol, 77.9% over two steps) as a white solid.

[0213] Two equal batches were combined to give (3S,6S,10S,10aR)-6-amino-10-methyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (1.12 g, 4.66 mmol, 99.1% purity) as a white solid. LCMS (ESI) m / z = 241.0 [(M-100)+H] + ; 1H NMR (400 MHz CDCl3) δ 5.69 (d, J = 3.0 Hz, 1H), 4.75 - 4.66 (m, 1H), 4.35 - 4.32 (m, 1H), 4.05 (t, J = 8.8 Hz, 1H), 3.66 - 3.51 (m, 1H), 2.55 - 2.47 (m, 1H), 2.41 - 2.07 (m, 3H), 1.97 - 1.67 (m, 3H), 1.69 - 1.55 (m, 2H), 1.46 (s, 9H), 1.36 - 1.20 (m, 1H), 1.10 - 0.95 (m, 1H), 0.89 - 0.84 (m, 3H). Synthesis of (3S,6S,9R,10aR)-6-amino-9-(benzyloxy)-3-(3-(morpholine-4-carbonyl)azetidine-1-carbonyl)octahydropyrrolo[1,2-a]azocin-5(1H)-one [ka]

[0214] Step 1: Preparation of tert-butyl N-[(3S,6S,9R,10aR)-9-hydroxy-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-5-oxo-decahydropyrrolo[1,2-a]azocin-6-yl]carbamate

[0215] To a solution of (3S,6S,9R,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (0.780 g, 2.27 mmol, 1 equiv.), 4-(azetidine-3-carbonyl)morpholine trifluoroacetate (645 mg, 2.27 mmol, 1 equiv.), and N,N-diisopropylethylamine (2.35 mL, 13.6 mmol, 6 equiv.) in DMF (10 mL) was added HATU (1.03 g, 2.72 mmol, 1.2 equiv.). The reaction was stirred at room temperature for 2 hours. The reaction mixture was then concentrated in vacuo and subsequently purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5-75% acetonitrile in water (containing 0.1% FA)) to afford tert-butyl N-[(3S,6S,9R,10aR)-9-hydroxy-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-5-oxo-decahydropyrrolo[1,2-a]azocin-6-yl]carbamate (1.16 g, 2.34 mmol, 103% yield) as a beige foam. LCMS (ESI) m / z = 495.3 [M+H] + .

[0216] Step 2: Preparation of tert-butyl ((3S,6S,9R,10aR)-9-(benzyloxy)-3-(3-(morpholine-4-carbonyl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate

[0217] To a solution of tert-butyl N-[(3S,6S,9R,10aR)-9-hydroxy-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-5-oxo-decahydropyrrolo[1,2-a]azocin-6-yl]carbamate (75 mg, 0.1516 mmol, 1 equiv.) in CHCl (3 mL) was added EtN (63.2 μL, 454 μmol, 3 equiv.) and TMSCl (28.7 μL, 227 μmol, 1.5 equiv.), and the mixture was stirred at room temperature. After 30 min, brine (10 mL) was added, and the organic material was extracted with CHCl (10 mL × 2). The combined organic extracts were dried over sodium sulfate, filtered, and concentrated in vacuo to give crude tert-butyl N-[(3S,6S,9R,10aR)-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-5-oxo-9-[(trimethylsilyl)oxy]-decahydropyrrolo[1,2-a]azocin-6-yl]carbamate (85.9 mg, 0.1516 mmol, 100% yield) as a white solid.

[0218] A solution of tert-butyl N-[(3S,6S,9R,10aR)-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-5-oxo-9-[(trimethylsilyl)oxy]-decahydropyrrolo[1,2-a]azocin-6-yl]carbamate (85.9 mg, 0.1516 mmol, 1 equiv) in CHCl (5 mL) was cooled to −78° C., followed by the addition of benzaldehyde (22.9 μL, 227 μmol, 1.5 equiv), EtSiH (36.1 μL, 227 μmol, 1.5 equiv), and trimethylsilyl triflate (20.4 μL, 113 μmol, 0.75 equiv). The reaction was stirred at −78° C. for 10 min and then at 0° C. for 90 min. The reaction was quenched by the addition of saturated aqueous NaHCO (10 mL), and the product was extracted with CHCl (10 mL × 3). The combined organic extracts were dried over sodium sulfate and filtered to give a solution of tert-butyl ((3S,6S,9R,10aR)-9-(benzyloxy)-3-(3-(morpholine-4-carbonyl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate.

[0219] Step 3: Preparation of (3S,6S,9R,10aR)-6-amino-9-(benzyloxy)-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-decahydropyrrolo[1,2-a]azocin-5-one

[0220] To this solution was added trifluoroacetic acid (0.5 mL), and the reaction was stirred at room temperature for 30 minutes, then concentrated under reduced pressure. The reaction was purified by reverse-phase chromatography (C18 cartridge eluted with a gradient of 5-40% acetonitrile in water (containing 0.1% FA)) to afford (3S,6S,9R,10aR)-6-amino-9-(benzyloxy)-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-decahydropyrrolo[1,2-a]azocin-5-one (41.0 mg, 0.08460 mmol, 56.0% yield) as a white solid. LCMS (ESI) m / z = 485.4 [M+H] +.

[0221] The following intermediates in Table 17 were prepared following the protocol outlined for the synthesis of (3S,6S,9R,10aR)-6-amino-9-(benzyloxy)-3-[3-(morpholine-4-carbonyl)azetidine-1-carbonyl]-decahydropyrrolo[1,2-a]azocin-5-one and using the appropriate advanced intermediate(s) as starting materials. Where appropriate, the final treatment with TFA (Step 3) was omitted and the N-Boc-protected amine was isolated. [Table 17] Synthesis of tert-butyl ((3R,6R,8R,9R,10aS)-8,9-dihydroxy-5-oxo-3-(3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate, tert-butyl ((3S,6S,8R,9R,10aR)-9-hydroxy-8-methyl-5-oxo-3-((S)-3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate, and tert-butyl ((3S,6S,8S,9S,10aR)-8-hydroxy-9-methyl-5-oxo-3-((S)-3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate [ka]

[0222] Step 1: Preparation of methyl (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxylene[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate and methyl (1aR,3S,6S,8aR,9aS)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxylene[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate

[0223] To a cooled (0 °C) solution of 1H-pyrazole (40.2 mg, 591 μmol, 0.1 equiv.), methyltrioxorhenium (14.7 mg, 59.1 μmol, 0.01 equiv.), and HO (6.66 g, 59.1 mmol, 10 equiv.) in CFCHOH (3 mL) was slowly added methyl (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (2.00 g, 5.91 mmol, 1 equiv.). The reaction mixture was allowed to warm to room temperature and stirred for 4 h. To the mixture was added a solution of aqueous saturated NaHSO, and the mixture was extracted with CHCl (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting residue was purified by C18 column (gradient 15% to 60% acetonitrile in water) to give methyl (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate (1.50 g, 4.23 mmol, 72% yield) as the major product and methyl (1aR,3S,6S,8aR,9aS)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate (minor adduct) (190 mg, 0.54 mmol, 9% yield) as the minor adduct.

[0224] (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate methyl ester (major diastereomer): LCMS (ESI) m / z = 355 [M+H] + ; 1H NMR (400 MHz, CDCl3) δ 5.51 (d, J = 6.9 Hz, 1H), 4.68 (dd, J = 14.9, 7.6 Hz, 1H), 4.60 - 4.48 (m, 1H), 4.27 - 4.13 (m, 1H), 3.74 (s, 3H), 3.29 - 3.20 (m, 1H), 3.18 - 3.08 (m, 1H), 2.55 - 2.36 (m, 2H), 2.34 - 2.13 (m, 3H), 2.09 - 1.91 (m, 3H), 1.42 (s, 9H).

[0225] (1aR,3S,6S,8aR,9aS)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate methyl ester (minor diastereomer): LCMS (ESI) m / z = 355 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 5.90 (d, J = 6.4 Hz, 1H), 4.53 - 4.42 (m, 2H), 4.14 (t, J = 6.3 Hz, 1H), 3.75 (s, 3H), 3.31 - 3.19 (m, 1H), 3.11 - 3.06 (m, 1H), 2.37 - 2.27 (m, 2H), 2.25 - 2.11 (m, 3H), 2.03 - 1.87 (m, 3H), 1.44 (s, 9H).

[0226] Step 2: Preparation of (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylic acid

[0227] To a solution of methyl (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylate (390 mg, 1.10 mmol, 1.0 equiv.) in a mixture of THF (8 mL) and HO (2 mL) was added LiOH·HO (138 mg, 3.30 mmol, 3.0 equiv.). The reaction mixture was stirred at room temperature for 4 h and then acidified to pH = 5–6 with a 1 N aqueous solution of HCl. The resulting acidic mixture was extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The crude product was purified on a C18 column (gradient 15% to 60% acetonitrile in water) to give (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylic acid (300 mg, 0.88 mmol, 80% yield) as a white solid. LCMS (ESI) m / z = 341 [M+H] + .

[0228] Step 3: Preparation of tert-butyl ((1aS,3S,6S,8aR,9aR)-4-oxo-6-(3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate

[0229] To a solution of (1aS,3S,6S,8aR,9aR)-3-((tert-butoxycarbonyl)amino)-4-oxodecahydroxyleno[2,3-d]pyrrolo[1,2-a]azocine-6-carboxylic acid (300 mg, 0.88 mmol, 1 equiv.), EtN (445 mg, 4.40 mmol, 5 equiv.), and rac-3-phenylpyrrolidine (129 mg, 0.88 mmol, 1 equiv.) in DMF (10 mL) was added HATU (399 mg, 1.05 mmol, 1.2 equiv.) in one portion. The resulting mixture was stirred at room temperature for an additional 12 h and then diluted with water (5 mL). The biphasic mixture was extracted with CHCl (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel to give tert-butyl ((1aS,3S,6S,8aR,9aR)-4-oxo-6-(3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate (200 mg, 0.43 mmol, 48% yield) as a clear oil. LCMS (ESI) m / z = 470.3 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 7.37 - 7.30 (m, 2H), 7.27 - 7.19 (m, 3H), 5.87 - 5.37 (m, 1H), 4.74 - 4.41 (m, 2H), 4.36 - 4.08 (m, 2H), 4.02 - 3.65 (m, 2H), 3.61 - 3.31 (m, 4H), 2.50 - 2.26 (m, 3H), 2.25 - 1.92 (m, 6H), 1.90 - 1.82 (m, 1H), 1.43 (s, 9H).

[0230] Step 4: Preparation of tert-butyl ((1aS,3S,6S,8aR,9aR)-4-oxo-6-((S)-3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate and tert-butyl ((1aS,3S,6S,8aR,9aR)-4-oxo-6-((R)-3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate

[0231] tert-Butyl rac-((1aR,3R,6R,8aS,9aS)-4-oxo-6-(3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate (600 mg, 1.27 mmol, 1 equiv.), triethylamine (976 mg, 9.65 mmol, 5 equiv.), and (3S)-3-phenylpyrrolidine (284 mg, 1.93 mmol, 1 equiv.) were added to a 250 × 21.2 mm SFC (ChiralCel OX) column. ID, 5 μm, A is CO, B is MeOH + 0.1% NHOH, 40 mL / min) gave peak 1: ((1aS,3S,6S,8aR,9aR)-4-oxo-6-((R)- or (S)-3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)tert-butyl carbamate (400 mg, 0.8518 mmol, 67.1% yield). LCMS (ESI) m / z = 470.3 [M+H] + and peak 2: ((1aS,3S,6S,8aR,9aR)-4-oxo-6-((S)- or (R)-3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate tert-butyl ester (130 mg, 0.2768 mmol, 21.8% yield) was obtained as a clear oil. LCMS (ESI) m / z = 470.3 [M+H] + .

[0232] Step 5: Preparation of tert-butyl ((3S,6S,8R,9R,10aR)-9-hydroxy-8-methyl-5-oxo-3-((S)-3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate and tert-butyl ((3S,6S,8S,9S,10aR)-8-hydroxy-9-methyl-5-oxo-3-((S)-3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate

[0233] To a cooled (0 °C) solution of tert-butyl ((1aS,3S,6S,8aR,9aR)-4-oxo-6-((R)- or (S)-3-phenylpyrrolidine-1-carbonyl)decahydroxyleno[2,3-d]pyrrolo[1,2-a]azocin-3-yl)carbamate (peak 1) (200 mg, 0.43 mmol, 1 equiv.) in toluene (5 mL) was added AlMe3 (613 mg, 8.51 mmol, 20 equiv.) slowly in a dropwise manner. The reaction mixture was then allowed to warm to room temperature. After 30 min, the reaction mixture was cooled to 0 °C, and then MeOH (10 mL) was added to quench the excess AlMe3. The resulting mixture was concentrated under reduced pressure and purified on a C18 column (gradient 20% to 70% acetonitrile in water) to give an inseparable mixture of regioisomeric epoxide ring-opened products (160 mg, 0.33 mmol, 77% yield). LCMS (ESI) m / z = 486.3 [M+H] + ; 1 H NMR (400 MHz, mixture of isomers, CDCl3) δ 7.40 - 7.28 (m, 3H), 7.26 - 7.21 (m, 2H), 5.78 - 5.50 (m, 1H), 4.73 - 4.49 (m, 2H), 4.36 - 4.09 (m, 2H), 4.04 - 3.30 (m, 6H), 2.44 - 1.97 (m, 10H), 1.43 (s, 9H), 1.13 - 0.83 (m, 3H).

[0234] This mixture of regioisomeric ring-opened products (60 mg, 0.124 mmol) was purified by SFC (ChiralPak®,®-WHELK, 250 × 21.2 mm ID, 5 μm; mobile phase: A is CO, B is MeOH + 0.1% NHOH. Gradient: B 25%; flow rate: 40 mL / min) to give:

[0235] Peak 1: tert-butyl ((3S,6S,8R,9R,10aR)-9-hydroxy-8-methyl-5-oxo-3-((R)- or (S)-3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate (3.00 mg, 0.006177 mmol). LCMS (ESI) m / z = 486.3 [M+H] + .

[0236] Peak 2: tert-butyl ((3S,6S,8S,9S,10aR)-8-hydroxy-9-methyl-5-oxo-3-((S)- or (R)-3-phenylpyrrolidine-1-carbonyl)decahydropyrrolo[1,2-a]azocin-6-yl)carbamate (5.00 mg, 0.01029 mmol) as a white solid. LCMS (ESI) m / z = 486.3 [M+H] + . Procedure for the synthesis of the linker:

[0237] The following intermediates in Table 18 were prepared according to the protocols described in WO 2020 / 205467. [Table 18] Synthesis of (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylic acid [ka]

[0238] (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylic acid was prepared according to the protocol described in US 2004 / 0225146. Representative Procedure for the Preparation of Activated Ester Phosphonic Acids Synthesis of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid [ka]

[0239] Step 1: Preparation of 4-nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0240] A mixture of 5-[(diethoxyphosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylic acid (10.0 g, 27.4 mmol), EDCI (7.85 g, 41.0 mmol), and DMAP (836 mg, 6.85 mmol) in CHCl (80 mL) was stirred at room temperature. After 15 min, 4-nitrophenol (4.75 g, 34.2 mmol) was added, and the resulting yellow mixture was stirred at room temperature for 18 h. The reaction was quenched with water (30 mL), and the product was extracted with CHCl (10 mL × 2). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by reverse phase chromatography (C18 cartridge eluted with a gradient of 5-100% acetonitrile in water) and the appropriate fractions were concentrated to give 4-nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (7.80 g, 16.0 mmol, 59.0% yield) as a yellow solid. LCMS m / z = 486.2 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 8.32 - 8.37 (m, 3H), 8.22 (s, 1H), 8.01 (d, J = 9.1 Hz, 1H), 7.77 (d, J = 7.8 Hz, 1 H), 7.51 - 7.45 (m, 2 H), 4.14 - 4.32 (m, 4H), 1.34 (t, J = 7.8 Hz, 6H).

[0241] Step 2: Preparation of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0242] To a cooled (0 °C) solution of 4-nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (4.47 g, 9.20 mmol) in CHCl (39 mL) was added N,O-bis(trimethylsilyl)trifluoroacetamide (12.1 mL, 46.0 mmol) and iodotrimethylsilane (5.23 mL, 36.8 mmol) as a solution in CHCl (10 mL). The reaction mixture was gradually warmed to ambient temperature. To the reaction mixture was added a mixture (50 mL) of 2:1 HO / acetonitrile (containing 0.1% TFA), and precipitation of the product was observed. The volatiles were removed in vacuo, and the crude residue was suspended in a mixture of acetonitrile / water (1:1 v / v, 100 mL). The suspension was filtered, the solid washed with a 2:1 mixture of acetonitrile / water solution, and the solid dried under reduced pressure to give [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid (6.5 g, 94%) as a beige solid. The filtrate was concentrated to 50% of the solvent volume, and the resulting suspension was filtered and washed with a 1:2 solution of acetonitrile / water. The solid was dried under reduced pressure to give additional [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid (0.4 g) as a beige solid. Both products were lyophilized to give (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (6.90 g, 16.0 mmol, 98.0% yield). 1 H NMR (400 MHz, DMSO-d6) δ 7.66 - 7.74 (m, 3H), 8.27 (d, J = 8.3 Hz, 1H), 8.30 (s, 1H), 8.36 - 8.41 (m, 2H), 8.66 (s, 1H).

[0243] The following intermediates in Table 19 were prepared using a similar protocol as outlined above for the synthesis of [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid and utilizing the appropriate advanced intermediate(s) as the starting material(s). [Table 19] Synthesis of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid [ka]

[0244] Preparation of 5-methylbenzo[b]thiophene-2-carboxylic acid

[0245] 5-Methylbenzo[b]thiophene-2-carboxylic acid was prepared according to the procedure described in WO 2016 / 100184.

[0246] Step 1: Preparation of benzyl 5-methylbenzo[b]thiophene-2-carboxylate

[0247] To a solution of 5-methylbenzo[b]thiophene-2-carboxylic acid (21.2 g, 110.0 mmol, 1.0 equiv.) and KCO (30.4 g, 220.0 mmol, 2.0 equiv.) in DMF (200 mL) was added benzyl bromide (20.6 g, 121.0 mmol, 1.1 equiv.). The mixture was stirred at room temperature for 14 hours. The reaction mixture was poured into ice water (400 mL) and stirred for 5 minutes. The resulting solid was filtered, and the filter cake was washed with water (50 mL) and dried in vacuo to give benzyl 5-methylbenzo[b]thiophene-2-carboxylate (30.1 g, 106.0 mmol, 97% yield) as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.01 (s, 1H), 7.71 (t, J = 12.2 Hz, 1H), 7.65 (s, 1H), 7.46 (d, J = 6.8 Hz, 2H), 7.42 - 7.35 (m, 3H), 7.29 - 7.26 (m, 1H), 5.38 (s, 2H), 2.47 (s, 3H).

[0248] Step 2: Preparation of benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate

[0249] To a solution of benzyl 5-methylbenzo[b]thiophene-2-carboxylate (15.0 g, 53.1 mmol, 1.0 equiv.) and NBS (10.3 g, 58.4 mmol, 1.1 equiv.) in CCl4 (30 mL) was added benzoyl peroxide (1.3 g, 5.31 mmol, 0.1 equiv.). The reaction flask was subjected to three cycles of evacuation and back-filling with N2(g). The mixture was stirred at 80 °C for 16 h under a constant atmosphere of N2(g). The reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to afford benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (6.80 g, 18.8 mmol, 36% yield) as a yellow solid.

[0250] Step 3: Preparation of benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0251] A solution of benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (10.3 g, 28.5 mmol, 1.0 equiv) in triethyl phosphite (30.0 g, 180.0 mmol, 6.3 equiv) was stirred at 100° C. for 5 h. The reaction mixture was directly concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica gel to give benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (6.5 g, 15.5 mmol, 55% yield) as a colorless oil. LCMS (ESI) m / z = 419 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.80 (d, J = 8.3 Hz, 2H), 7.49 - 7.33 (m, 6H), 5.39 (s, 2H), 4.08 - 3.93 (m, 4H), 3.26 (d, J = 21.5 Hz, 2H), 1.24 (t, J = 7.1 Hz, 6H).

[0252] Step 4: Preparation of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0253] To a solution of benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (5.6 g, 13.3 mmol, 1.0 equiv.) in a mixture of THF (80 mL) and HO (10 mL) was added LiOH (1.10 g, 26.6 mmol, 2.0 equiv.). The mixture was stirred at room temperature for 3 hours and then acidified with 1N aqueous HCl (adjusted to pH 3-4). Upon acidification, the product precipitated from the solution. The resulting solid was filtered, the filter cake was washed with water (20 mL × 2), and the solid was dried under reduced pressure to give 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (3.9 g, 11.8 mmol, 88.9% yield) as a white solid. LCMS (ESI) m / z = 329 [M+H] + .

[0254] Step 5: Preparation of perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0255] To a cooled (0 °C) solution of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (3.9 g, 11.8 mmol, 1.0 equiv.) in CHCl (50 mL) was added oxalyl chloride (2.2 g, 17.7 mmol, 1.5 equiv.), followed by two drops of DMF. The mixture was stirred at 0 °C for 30 min, after which the reaction mixture was evaporated to dryness. The resulting solid was dissolved in CHCl (50 mL), followed by the addition of EtN (3.6 g, 35.4 mmol, 3.0 equiv.) and pentafluorophenol (2.6 g, 14.1 mmol, 1.2 equiv.). The resulting mixture was stirred at room temperature for an additional 2 h and then poured onto HO (30 mL). The biphasic solution was extracted with EtOAc (30 mL × 3). The combined organic layers were dried over MgSO, filtered, and concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel to give perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (4.7 g, 9.5 mmol, 81% yield) as a white solid. LCMS (ESI) m / z = 419 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 8.29 (s, 1H), 7.88 (d, J = 9.1 Hz, 2H), 7.51 (d, J = 8.4 Hz, 1H), 4.14 - 3.94 (m, 4H), 3.29 (d, J = 21.5 Hz, 2H), 1.26 (t, J = 7.0 Hz, 6H).

[0256] Step 6: Preparation of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0257] To a solution of perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (4.7 g, 9.5 mmol, 1.0 equiv.) in CHCl (60 mL) was added bromotrimethylsilane (12 mL). The mixture was stirred at room temperature for 14 hours and then concentrated under reduced pressure. The residue was purified by C column chromatography to give ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (3.7 g, 8.4 mmol, 89% yield) as a white solid. LCMS (ESI) m / z = 439 [M+H] + . Synthesis of (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid and (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid [ka]

[0258] Step 1: benzyl rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylate

[0259] To a cooled (−78° C.) solution of benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (2.4 g, 5.73 mmol, 1 equiv.) in THF (75 mL) and 2-(benzenesulfonyl)-3-phenyloxaziridine (2.97 g, 11.4 mmol, 2 equiv.) was added a solution of 1 M NaHMDS in THF (11.4 mL, 11.4 mmol, 2 equiv.). Upon addition of the base, a deep purple solution was observed, which turned orange after complete addition of the base. The mixture was stirred for an additional 10 min, after which aqueous saturated NH4Cl (50 mL) was added. The mixture was warmed to ambient temperature, and EtOAc (75 mL) and water (25 mL) were added. After stirring for an additional 30 min, the phases were separated. The aqueous layer was extracted with EtOAc (125 mL × 2). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Separate batches of equal size were run and combined for purification. The combined material (6.42 mmol, 12.15 mmol total) was purified by flash chromatography (20% to 100% EtOAc:heptane) to afford benzyl rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylate (3.69 g, 8.49 mmol, 70%) as a white sticky solid. LCMS (ESI) m / z = 869.4 [2M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 8.10 (s, 1H), 8.04 - 8.00 (m, 1H), 7.88 (d, J = 8.6 Hz, 1H), 7.61 (d, J = 8.6 Hz, 1H), 7.51 - 7.47 (m, 2H), 7.46 - 7.35 (m, 3H), 5.42 (s, 2H), 5.17 (dd, J = 10.4, 4.5 Hz, 1H), 4.18 - 3.95 (m, 4H), 3.10 - 2.99 (m, 1H), 1.33 - 1.20 (m, 6H).

[0260] Step 2: benzyl rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate

[0261] To a cooled (−78 °C) solution (under N(g)) of benzyl rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylate (cc) (1.56 g, 3.59 mmol, 1 equiv.) in CHCl (30 mL) was added (diethylamino)sulfur trifluoride (568 μL, 4.30 mmol, 1.2 equiv.). The reaction was stirred for 15 min, after which aqueous saturated NaHCO (50 mL) was added. After warming to room temperature, the product was extracted with CHCl (50 mL × 3). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by reverse-phase chromatography on a C18 cartridge (eluting with 5-80% acetonitrile in water) to give benzyl rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (650 mg, 1.48 mmol, 41.6%) as a thick clear oil. LCMS (ESI) m / z = 873.2 [2M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 8.12 (s, 1H), 8.02 - 7.99 (m, 1H), 7.92 (d, J = 8.7 Hz, 1H), 7.61 (d, J = 8.7 Hz, 1H), 7.51 - 7.51 (m, 2H), 7.46 - 7.36 (m, 3H), 5.82 (dd, J = 44.4, 7.5 Hz, 1H), 5.42 (s, 2H), 4.21 - 4.02 (m, 4H), 1.34 - 1.26 (m, 6H).

[0262] Step 3: Preparation of (R)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate and (S)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate

[0263] Benzyl rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (650 mg, 1.48 mmol) was subjected to chiral SFC separation (Lux i-amylose 3, 21.2 × 250 mm 5 μm column, 75 mL / min, 40% MeOH) to give (R)- or (S)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (304 mg, 0.70 mmol, 46.8% recovery, 99.9% ee) as a thick, clear oil (peak 1) and (R)- or (S)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (317 mg, 0.73 mmol, 49% recovery, 99.9% ee) as a thick, clear oil (peak 2). Note: The fastest eluting enantiomer by SFC was arbitrarily assigned as (R)-5-(fluoro(phosphono)methyl)benzo[b]thiophene-2-carboxylic acid, and the slowest eluting enantiomer by SFC was assigned as (S)-5-(fluoro(phosphono)methyl)benzo[b]thiophene-2-carboxylic acid. HPLC method for enantiomeric excess analysis: Lux Cellulose-3 150 mm 45% HO + 0.05% TFA / 55% MeCN 1 mL / min for 8 min.

[0264] Step 4: Preparation of (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0265] A mixture of 10% Pd / C (60 mg, 50% wet) and (R)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (peak 1) (60 mg, 0.1374 mmol, 1 equiv.) in THF (5 mL) was degassed with N(g) for 5 min. H(g) was bubbled through the mixture for 5 min, and then the reaction was stirred at room temperature under H(g) (1 atm). The reaction mixture was stirred until consumption of the starting material was detected by LCMS. The reaction mixture was then sparged with N(g) for 15 min and filtered through a pad of Celite®. The filter cake was washed with 2-MeTHF, and the filtrate was concentrated to give (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid (47.4 mg, 0.0137 mmol, 99%) as a thick clear oil. LCMS (ESI) m / z = 347.2 [M+H] + .

[0266] The following intermediates in Table 20 were prepared using the procedure outlined above (Step 4), starting from (S)- or (R)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (Peak 2) and using the appropriate reagents. [Table 20] Representative Methods for the Synthesis of Activated Linkers

[0267] Method 1: Stepwise acid chloride approach for the synthesis of mixed linkers

[0268] Representative Procedure for the Synthesis of 4-Nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate [ka]

[0269] Step 1: Preparation of 4-nitrophenyl 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0270] To a cooled (0 °C) heterogeneous solution of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (100 mg, 0.2329 mmol, 1 equiv.) in CHCl (4 mL) was added a catalytic amount of DMF (2 drops), followed by the dropwise addition of oxalyl chloride (198 μL, 2.32 mmol, 10 equiv.). The homogeneous reaction mixture was allowed to warm to room temperature and stirred for 2 h. The reaction was concentrated in vacuo and further dried under high vacuum for 30 min to give a yellow solid. The yellow solid was diluted with CHCl (4 mL) and cooled to −78 °C. A solution of pyridin-3-ol (22.0 mg, 232 μmol, 1 equiv.) and triethylamine (64.7 μL, 465 μmol, 2 equiv.) in CHCl (1 mL) [sonicated for 1 min to allow solubilization] was slowly added. The homogeneous reaction mixture was stirred at −78° C. for 2 min, then allowed to warm to ambient temperature and stirred overnight. After 24 h, the reaction mixture became heterogeneous, and the reaction was concentrated under reduced pressure. The crude product, 4-nitrophenyl 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate, was used directly in the next step without further purification or manipulation. LCMS m / z = 507.2 [M+H] + .

[0271] Step 2: Preparation of 4-nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0272] To a cooled (0 °C) solution of 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoryl)methyl)benzo[b]thiophene-2-nitrophenyl carboxylate (117 mg, 0.2329 mmol, 1 equiv.) in CHCl (5 mL) was added two drops of DMF, followed by the dropwise addition of oxalyl chloride (198 μL, 2.32 mmol, 10 equiv.). The reaction mixture was warmed to room temperature and stirred for 1.5 h. LCMS analysis indicated partial conversion to the desired activated intermediate. Additional oxalyl chloride (198 μL, 2.32 mmol, 10 equiv.) was introduced to the reaction mixture, and the mixture was stirred for an additional 1 h. The reaction was concentrated in vacuo and further dried under high vacuum for 30 min to give a yellow solid. The yellow solid was diluted with CHCl (5 mL) and cooled to −78 °C. To this cooled solution, a solution of 1-[(2-hydroxyethyl)sulfanyl]butan-1-one (103 mg, 698 μmol, 3 equiv.) diluted in CHCl (1 mL) [pre-dried by passage through anhydrous NaSO] was slowly added, followed by triethylamine (134 μL, 967 μmol, 2 equiv.). After stirring for 2 minutes, the resulting mixture was allowed to warm to ambient temperature and stirred overnight. Celite® was added to the mixture, and the mixture was carefully concentrated in vacuo. The crude residue was purified by flash chromatography (gradient elution 0 to 60% EtOAc in heptane) to afford 4-nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (22.0 mg, 0.03455 mmol, 14.9% yield) as a clear oil. LCMS m / z = 637.2 [M+H] + ; 1H NMR: (400 MHz, DMSO-d6) δ 8.50 - 8.44 (m, 2H), 8.33 - 8.38 (m, 3H), 8.25 (s, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.79 (d, J = 8.6 Hz, 1H), 7.57 - 7.53 (m, 1 H), 7.50 - 7.45 (m, 2H), 7.31 - 7.26 (m, 1H), 4.37 - 4.24 (m, 2 H), 3.20 - 3.08 (m, 2H), 2.52 (t, J = 7.6 Hz, 2H), 1.67 (sextet, J = 7.3 Hz, 2H), 0.94 (t, J = 7.6 Hz, 3H).

[0273] The following intermediates in Table 21 were prepared using a similar protocol as described above for the synthesis of 4-nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediate(s) as the starting material(s). [Table 21-1] [Table 21-2]

[0274] Method 2: One-pot acid chloride method for the synthesis of activated linkers

[0275] Representative procedure for the synthesis of perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate [ka]

[0276] To a cooled (0 °C) solution of 3 (200 mg, 0.42 mmol, 1.0 equiv) in dry CHCl (15 mL) and a catalytic amount of DMF (3.2 μL, 42.1 μmol, 0.1 equiv) was added oxalyl chloride (266 mg, 2.10 mmol, 5.0 equiv) dropwise. The reaction mixture was allowed to warm to 40 °C. After stirring for 2 h, the reaction mixture was concentrated in vacuo and dried (to remove excess oxalyl chloride). The resulting solid was redissolved in anhydrous CHCl (5 mL) and cooled to 0 °C. To this cooled solution was added a solution of S-(4-hydroxybutyl) 3-methylbutanethioate (239 mg, 1.26 mmol, 3.0 equiv), DMAP (5.14 mg, 42.1 μmol, 0.1 equiv), and N,N-diisopropylethylamine (217 mg, 1.68 mmol, 4.0 equiv) in anhydrous CHCl (10 mL). The reaction mixture was allowed to warm to room temperature and stirred for an additional 18 h. The reaction was quenched by the addition of HO (10 mL) and extracted with CHCl (3 × 10 mL). The organic layers were combined and washed with brine (20 mL), dried over anhydrous NaSO, and concentrated in vacuo. The residue was purified by flash column chromatography to give perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (15.0 mg, 18.3 μmol, 4.4% yield). LCMS (ESI) m / z = 819 [M+H] + .

[0277] The following intermediates in Table 22 were prepared using a similar protocol as described above for the synthesis of perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediate(s) as the starting material(s). [Table 22]

[0278] Method 3: One-pot silver salt method for the synthesis of activated linkers

[0279] Representative procedure for the synthesis of perfluorophenyl 5-((bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate [ka]

[0280] Step 1: Preparation of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate silver(I)

[0281] To a solution of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (300 mg, 684 μmol, 1.0 equiv.) in a mixture of deionized HO (4 mL) and THF (2 mL) was added Amberlite IR120® resin (Na + To the filtrate was added AgNO (463 mg, 2.73 mmol, 4.0 equiv.) in deionized HO (2 mL), and the mixture was stirred at room temperature for an additional 1 h. A white precipitate was observed to form, and the solid was collected by filtration. The filter cake was then washed with cold HO (3 × 2 mL), and the solid was dried under reduced pressure to give silver(I) ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate as a dry powder. This silver salt was used without further purification.

[0282] Step 2: Preparation of perfluorophenyl 5-((bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0283] A suspension of silver(I) ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate was suspended in anhydrous toluene (10 mL), and iodomethyl 2-methylpropanoate (500 mg, 2.05 mmol, 3.0 equiv.) was added dropwise. After the addition, the resulting mixture was stirred at room temperature for an additional 12 hours. The progress of the reaction was monitored by LCMS, and upon completion, the unreacted silver salt was collected by filtration. The filtrate solution was concentrated in vacuo, and the resulting residue was purified by reverse-phase chromatography [C18 column, gradient elution: water / acetonitrile = 90% to 1%] to afford perfluorophenyl 5-((bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (165 mg, 246 μmol, 36% yield) as a white solid. LCMS (ESI) m / z = 671 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 8.28 (s, 1H), 7.96 - 7.80 (m, 2H), 7.48 (d, J = 8.5 Hz, 1H), 5.70 - 5.53 (m, 4H), 4.90 (dt, J = 12.6, 6.2 Hz, 2H), 3.42 (d, J = 22.2 Hz, 2H), 1.31 (d, J = 6.2 Hz, 12H).

[0284] The following intermediates in Table 23 were prepared using a similar protocol as described above for the synthesis of perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediate(s) as the starting material(s). [Table 23-1] [Table 23-2] [Table 23-3] (R)- or (S)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid [ka]

[0285] Step 1: Preparation of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid

[0286] To a cooled (0 °C) solution of (R)- or (S)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (peak 1) (205 mg, 0.4697 mmol) in CHCl (8.0 mL) was added BSTFA (746 μL, 2.81 mmol), followed by a 1.0 M solution of trimethylsilyl iodide in CHCl (1.87 mL, 1.87 mmol). After stirring for 1 h, a mixture of acetonitrile (0.66 mL), water (0.33 mL), and 0.1% TFA was added. The solvent was removed under reduced pressure at 0 °C. The crude residue was purified by reverse-phase chromatography (C18 cartridge eluting with 5–40% acetonitrile in water) to afford (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid (163 mg, 0.4285 mmol) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 1H), 8.11 - 8.06 (m, 2H), 7.58 (d, J = 8.6 Hz, 1H), 7.51 - 7.47 (m, 2H), 7.45 - 7.33 (m, 3H), 5.84 (dd, J = 44.6, 8.3 Hz, 1H), 5.40 (s, 2H).

[0287] Step 2: Preparation of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid

[0288] [The following reaction was carried out in a foil-covered vessel in the absence of ambient light]: To a suspension of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid (163 mg, 0.4285 mmol, 1 equiv.) in water (5 mL) was added a solution of sodium hydroxide (34.2 mg, 857 μmol, 2 equiv.) in water (2 mL). To this yellow solution was added silver(I) nitrate (181 mg, 1.07 mmol, 2.5 equiv.), and the resulting off-white suspension was stirred at room temperature for 1.5 h. The suspension was cooled to 0° C., filtered, and dried under high vacuum. The solid was resuspended in acetonitrile, concentrated under reduced pressure, and further dried under high vacuum (3 h). The resulting dark yellow powder was suspended in toluene (10 mL), and iodomethyl 2,2-dimethylpropanoate (191 μL, 1.28 mmol, 3 equiv.) was added. After stirring for 20 h, the reaction mixture was stirred at room temperature for 20 h. The reaction mixture was filtered and rinsed with toluene. The filtrate was concentrated under reduced pressure. The crude residue was purified (C18 cartridge eluting with 5–100% acetonitrile in water) to afford (R)- or (S)-((((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphoryl)bis(oxy))bis(methylene) bis(2,2-dimethylpropanoate) (122 mg, 0.2004 mmol, 46.9%) as a clear oil. 1 H NMR (400 MHz, CDCl3) δ 8.11 (s, 1 H), 8.01 - 7.99 (m, 1 H), 7.92 (d, J = 8.3 Hz, 1H), 7.59 (d, J = 8.3 Hz, 1H), 7.52 - 7.47 (m, 2 H), 7.46 - 7.36 (m, 3H), 8.87 (dd, J = 44.3, 7.5 Hz, 1H), 5.72 - 5.62 (m, 4H), 5.43 (s, 2H), 1.21 (s, 18H).

[0289] Step 3: Preparation of (R)- or (S)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0290] To a solution of (R)-((((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphoryl)bis(oxy))bis(methylene) bis(2,2-dimethylpropanoate) (122 mg, 0.200 mmol, 1 equiv) in THF (10 mL) under N2(g) was added 10% Pd / C (50% wet, 120 mg, 0.1127 mmol, 0.56 equiv). H2(g) was bubbled through the suspension for 5 min. The reaction mixture was stirred at room temperature under H2(g) (1 atm). After stirring for 22 h, the reaction mixture was purged with N2(g) and filtered through Celite®. The filter pad was washed with THF and concentrated under reduced pressure to give (R)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid (0.200 mmol, 99.9%) as a thick clear oil. LC-MS (ESI) m / z [M+H] + = 519.1.

[0291] The following intermediates in Table 24 were prepared using a protocol similar to that described above for the synthesis of (R)- or (S)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid and utilizing the appropriate advanced intermediate(s) as the starting material(s). The absolute configuration of the starting material (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid was not determined, but the elution peak ("Peak 1" or Peak 2") of the starting material utilized is indicated in the table. [Table 24]

[0292] Method 4: Stepwise silver salt method for the synthesis of mixed linkers

[0293] Representative Procedure for Perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate [ka]

[0294] Step 1: Preparation of perfluorophenyl 5-(difluoro(hydroxy(2-(pivaloylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0295] Silver(I) (difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate was synthesized starting from (difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid using the method described in Step 1, Method 3 for the synthesis of silver(I) ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate.

[0296] To a suspension of silver(I) ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate (647 mg, 940 μmol, 1.0 equiv.) in anhydrous toluene (10 mL) was added S-(2-iodoethyl) 2,2-dimethylpropanethioate (310 mg, 1.14 mmol, 1.2 equiv.) in a dropwise manner. After complete addition of the alcohol, the resulting mixture was stirred at room temperature for an additional 12 h. The heterogeneous mixture was filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by reverse-phase chromatography to afford perfluorophenyl 5-(difluoro(hydroxy(2-(pivaloylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (240 mg, 388 μmol, 41% yield). LCMS (ESI) m / z = 617 [MH] - .

[0297] Step 2: Preparation of perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0298] Perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate was synthesized using a similar protocol outlined above. Starting with perfluorophenyl 5-(difluoro(hydroxy(2-(pivaloylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (190 mg, 307 μmol, 1.0 equiv.), AgNO (207 mg, 1.22 mmol, 4.0 equiv.), and S-(2-iodoethyl)butanethioate (94.9 mg, 368 μmol, 1.2 equiv.), afforded perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (55.0 mg, 73.4 μmol, 24%) as a white solid. LCMS (ESI) m / z = 749 [M+H] + .

[0299] The following intermediates in Table 25 were prepared using a similar protocol as described above for the synthesis of perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediate(s) as the starting material(s). [Table 25] Preparation of 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid [ka]

[0300] Step 1: Preparation of allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0301] To a suspension of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (1.0 g, 3.0 mmol, 1.0 equiv.) and KCO (839 mg, 6.1 mmol, 2.0 equiv.) in DMF (20 mL) was added 3-bromoprop-1-ene (440 mg, 3.6 mmol, 1.2 equiv.). The mixture was stirred at room temperature for 14 hours and poured onto water (30 mL). The mixture was extracted with EtOAc (25 mL × 3). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure, and the residue was purified by column chromatography to give allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (0.980 g, 2.7 mmol, 88% yield) as a pale yellow solid. LCMS (ESI) m / z = 369 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.84 - 7.77 (m, 2H), 7.45 - 7.37 (m, 1H), 6.18 - 5.94 (m, 1H), 5.49 - 5.39 (m, 1H), 5.36 - 5.28 (m, 1H), 4.87 - 4.83 (m, 2H), 4.08 - 3.97 (m, 4H), 3.27 (d, J = 21.4 Hz, 2H), 1.25 (t, J = 7.1 Hz, 6H).

[0302] Step 2: Preparation of ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0303] To a solution of allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (980 mg, 2.7 mmol, 1.0 equiv) in CHCl (15 mL) was added bromotrimethylsilane (3 mL). The mixture was stirred at room temperature for 14 h and then concentrated under reduced pressure. The resulting residue was triturated with HO (5 mL) and the resulting precipitate was filtered. The filter cake was washed with HO (5 mL × 2) and dried under reduced pressure to give ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (0.710 g, 2.3 mmol, 86% yield) as a white solid. LCMS (ESI) m / z = 313 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.20 (s, 1H), 7.96 (d, J = 8.4 Hz, 1H), 7.92 - 7.83 (m, 1H), 7.50 - 7.38 (m, 1H), 6.12 - 5.98 (m, 1H), 5.46 - 5.38 (m, 1H), 5.32 - 5.27 (m, 1H), 4.85 - 4.80 (m, 2H), 3.08 (d, J = 21.2 Hz, 2H).

[0304] Step 3: Preparation of allyl 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0305] To a cooled (0 °C) solution (under a constant stream of N (g)) of ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (2.80 g, 8.96 mmol, 1 equiv.) and a catalytic amount of DMF (1 drop) in dry CHCl (50 mL) was added oxalyl chloride (3.40 g, 26.8 mmol, 3 equiv.). After gas effervescence ceased, the mixture was warmed to 40 °C. After 2 h, the mixture was cooled to room temperature and concentrated in vacuo to give a yellow solid. The solid was then diluted with CHCl (50 mL) and cooled to 0 °C. To this cooled solution was added phenol (0.843 g, 8.96 mmol, 1 equiv.) and EtN (4.53 g, 44.8 mmol, 5 equiv.). After complete addition, the mixture was warmed to room temperature and stirred for 1 hour, after which (2S)-2-aminopropanoic acid propan-2-yl (1.75 g, 13.4 mmol, 1.5 equiv.) was introduced to the mixture. After stirring for an additional 2 hours, the mixture was concentrated to dryness. The residue was purified by C18 column (elution: 50% to 80% acetonitrile in water) to give allyl 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (2.23 g, 4.44 mmol, 49.6% yield) as a white solid. LCMS (ESI) m / z = 502.0 [M+H] + ; 1H NMR (400 MHz, CDCl3) δ 8.05 (d, J = 6.9 Hz, 1H), 7.91 - 7.80 (m, 2H), 7.53 - 7.43 (m, 1H), 7.29 (d, J = 8.1 Hz, 2H), 7.18 - 7.09 (m, 3H), 6.05 (ddd, J = 16.1, 10.9, 5.6 Hz, 1H), 5.48 - 5.40 (m, 1H), 5.32 (dd, J = 10.4, 1.2 Hz, 1H), 4.98 - 4.87 (m, 1H), 4.85 (d, J = 5.7 Hz, 2H), 4.04 - 3.85 (m, 1H), 3.44 (dd, J = 20.7, 14.1 Hz, 2H), 3.12 (t, J = 10.9 Hz, 1H), 1.21 - 1.10 (m, 9H).

[0306] Step 4: Preparation of 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0307] A solution of allyl 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (90 mg, 0.1794 mmol, 1 equiv), pyrrolidine (12.7 mg, 179 μmol, 1 equiv), Pd(PPh) (10.3 mg, 8.97 μmol, 0.05 equiv) in CHCl (5 mL) was stirred under N(g). After 2 h, the reaction was concentrated in vacuo. The residue was purified by C18 column (elution 30% to 70% acetonitrile in water) to give 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (64.0 mg, 0.1386 mmol, 77.4% yield) as a white solid. LCMS (ESI) m / z = 462.1 [M+H] + .

[0308] The following intermediates in Table 26 were prepared using that described above for the synthesis of 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid, utilizing the appropriate starting materials and modifications. [Table 26] Synthesis of phosphonate analogues:

[0309] Method 1: Representative procedure for the synthesis of phosphonate analogs via amino acid coupling of a carboxylic acid linker with an amino acid core.

[0310] (Difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (1) [ka]

[0311] Step 1: Preparation of tert-butyl ((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate

[0312] To a solution of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (400 mg, 1.22 mmol, 1.0 equiv.), N-methylaniline (261 mg, 2.44 mmol, 2.0 equiv.), and EtN (246 mg, 2.44 mmol, 2.0 equiv.) in CHCl (10 mL) was added TP (1.55 g, 2.44 mmol, 2 equiv.). The resulting mixture was stirred at 40 °C for 12 h. The reaction mixture was diluted with water (5 mL) and extracted with CHCl (10 mL × 3). The organic layers were combined and washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography to give tert-butyl ((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate (240 mg, 578 μmol, 47% yield) as a white solid. LCMS (ESI) m / z = 416 [M+H] + .

[0313] Step 2: Preparation of (3S,6S,10aS)-6-amino-N-methyl-5-oxo-N-phenyldecahydropyrrolo[1,2-a]azocine-3-carboxamide

[0314] To a solution of tert-butyl ((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate (100 mg, 240 μmol, 1 equiv.) in CHCl (6 mL) was added trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 1 h, then cooled to 0 °C and carefully neutralized with aqueous NaHCO until basic (pH adjusted to 8–9). The resulting mixture was then extracted with CHCl (10 mL × 3 mL), and the combined organic layers were washed with brine (2 × 10 mL), dried over anhydrous NaSO, and concentrated under reduced pressure to give crude (3S,6S,10aS)-6-amino-N-methyl-5-oxo-N-phenyldecahydropyrrolo[1,2-a]azocine-3-carboxamide (76 mg, 240 μmol) as a white solid, which was used directly in the next step without further purification. LCMS (ESI) m / z = 316 [M+H] + .

[0315] Step 3: Preparation of diethyl (difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonate

[0316] To a solution of (3S,6S,10aS)-6-amino-N-methyl-5-oxo-N-phenyldecahydropyrrolo[1,2-a]azocine-3-carboxamide (76 mg, 240 μmol, 1.0 equiv.) and 5-[(diethoxyphosphoryl)difluoromethyl]-1H-indole-2-carboxylic acid (83.3 mg, 240 μmol, 1.0 equiv.) in DMF (3 mL) was added HATU (118 mg, 312 μmol, 1.3 equiv.) and EtN (72.8 mg, 720 μmol, 3.0 equiv.). The resulting mixture was stirred at room temperature for an additional 2 h. The reaction mixture was diluted with HO (10 mL) and extracted with CHCl (10 mL × 3). The organic layers were combined and washed with brine (20 mL), dried over anhydrous NaSO, and concentrated in vacuo. The resulting residue was purified by flash column chromatography to give diethyl (difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonate (100 mg, 155 μmol, 65% yield) as a white solid. LCMS (ESI) m / z = 645 [M+H] + .

[0317] Step 4: Preparation of (difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (1)

[0318] To a cooled (0 °C) solution of diethyl (difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonate (100 mg, 155 μmol, 1.0 equiv) in CHCl (10 mL) was added bromotrimethylsilane (355 mg, 2.32 mmol, 15.0 equiv) in a dropwise manner. The reaction was allowed to warm to room temperature and stirred. After 12 h, the reaction mixture was quenched by the addition of HO (5 mL). The biphasic mixture was extracted with CHCl (3 × 10 mL). The organic layers were combined and washed with brine (20 mL), dried over anhydrous NaSO, and concentrated in vacuo. The resulting residue was purified by reverse-phase HPLC to give (difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (1) (8.70 mg, 14.7 μmol, 9.5% yield) as a white solid. LCMS (ESI) m / z = 589 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 11.74 (s, 1H), 8.51 (d, J = 7.1 Hz, 1H), 7.82 (s, 1H), 7.41 (m, 8H), 4.96 (d, J = 5.4 Hz, 1H), 4.17 (m, 4H), 3.16 (s, 3H), 1.93 (m, 8H), 1.57 (m, 4H).

[0319] The following compounds in Table 27 were prepared according to the representative procedure described above for the synthesis of (difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (1) and utilizing the appropriate starting materials and modifications. [Table 27-1] [Table 27-2] [Table 27-3] [Table 27-4] [Table 27-5] [Table 27-6] [Table 27-7] [Table 27-8] [Table 27-9] [Table 27-10] [Table 27-11] [Table 27-12]

[0320] Method 2: Representative procedure for the synthesis of phosphonate analogs by direct coupling of perfluorophenyl or p-nitrophenyl activated linker esters with amino acid cores

[0321] Synthesis of difluoro(2-(((3S,6S,10aR,Z)-3-(methyl(phenyl)carbamoyl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (30) [ka]

[0322] Step 1: Preparation of 4-nitrophenyl 5-((bis((trimethylsilyl)oxy)phosphoryl)difluoromethyl)-1H-indole-2-carboxylate

[0323] To a solution of 4-nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)-1H-indole-2-carboxylate (70 mg, 149 μmol, 1.0 equiv.) in CHCl (4 mL) was added TMSBr (228 mg, 1.49 mmol, 10.0 equiv.) dropwise at room temperature. The mixture was stirred at room temperature for 5 hours. Upon completion, the reaction was concentrated under reduced pressure to afford 4-nitrophenyl 5-((bis((trimethylsilyl)oxy)phosphoryl)difluoromethyl)-1H-indole-2-carboxylate (77.0 mg, 146 μmol, 99%) as a yellow solid, which was used directly in the next step without further purification. LCMS (ESI) m / z = 413 [(M-144)+H] + (The TMS ester was hydrolyzed during LCMS, and only the mass of the parent acid was detected.)

[0324] Step 2: Preparation of (3S,6S,10aR,Z)-6-amino-N-methyl-5-oxo-N-phenyl-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxamide

[0325] (3S,6S,10aR,Z)-6-amino-N-methyl-5-oxo-N-phenyl-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxamide was synthesized starting from (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylic acid and using a similar protocol described above for the preparation of (3S,6S,10aS)-6-amino-N-methyl-5-oxo-N-phenyldecahydropyrrolo[1,2-a]azocine-3-carboxamide.

[0326] Step 1: Preparation of difluoro(2-(((3S,6S,10aR,Z)-3-(methyl(phenyl)carbamoyl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (30)

[0327] (3S,6S,10aR,Z)-6-amino-N-methyl-5-oxo-N-phenyl-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxamide (137 mg, 440 μmol, 1.1 equiv.) [difluoro(2-(((3S,6S,10aS)-3-(methyl(phenyl)carbamoyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl]-1H-isothiazolinone To a solution of 4-nitrophenyl 5-({bis[(trimethylsilyl)oxy]phosphoryl}difluoromethyl)-1H-indole-2-carboxylate (223 mg, 400 μmol, 1 equiv.) in DMF (3 mL) was added 4-nitrophenyl 5-({bis[(trimethylsilyl)oxy]phosphoryl}difluoromethyl)-1H-indole-2-carboxylate (1), which was previously synthesized using a protocol similar to that detailed in Steps 1 and 2 of Method 1 for the preparation of (indol-5-yl)methyl)phosphonic acid (1). The resulting mixture was stirred at 40 °C for 12 h. The reaction mixture was cooled to ambient temperature, and then HO (10 mL) was introduced. The biphasic mixture was extracted with CHCl (10 mL × 3). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and concentrated in vacuo. The residue was purified by reverse-phase HPLC to give (difluoro(2-(((3S,6S,10aR,Z)-3-(methyl(phenyl)carbamoyl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (30) (77 mg, 132 μmol, 33% yield) as a white solid. LCMS (ESI) m / z = 587 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 11.80 (s, 1H), 8.55 (d, J = 7.6 Hz, 1H), 7.83 (s, 1H), 7.52-7.34 (m, 8H), 5.88-5.73 (m, 2H), 5.03-4.93 (m, 1H), 4.26-4.11 (m, 1H), 3.18-3.11 (m, 4H), 2.67-2.56 (m, 2H), 2.54-2.40 (m, 2H), 1.97-1.73 (m, 4H).

[0328] The following compounds in Table 28 were prepared according to the representative procedure described above for the synthesis of (difluoro(2-(((3S,6S,10aR,Z)-3-(methyl(phenyl)carbamoyl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid and utilizing the appropriate starting materials and modifications. [Table 28-1] [Table 28-2] [Table 28-3] [Table 28-4] [Table 28-5] [Table 28-6] [Table 28-7] [Table 28-8] [Table 28-9] [Table 28-10] [Table 28-11] [Table 28-12] [Table 28-13]

Table 28-14

Table 28-15

Table 28-16

Table 28-17

Table 28-18

Table 28-19

Table 28-20

Table 28-21

Table 28-22

Table 28-23

Table 28-24

Table 28-25

Table 28-26

Table 28-27

Table 28-28

Table 28-29

Table 28-30

[0329] Step 1: Preparation of tert-butyl ((3S,6S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate

[0330] To a solution of 3-(azetidin-3-yl)pyridine (0.16 g, 1.2 mmol, 1 equiv.) and N,N-diisopropylethylamine (0.47 g, 3.7 mmol, 3.0 equiv.) in DMF (1.0 mL) was sequentially added a solution of (3S,6S,9aR)-6-{[(tert-butoxy)carbonyl]amino}-8-methyl-5-oxo-octahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylic acid (0.40 g, 1.2 mmol, 1 equiv.) in DMF (1.0 mL). To this mixture was added HATU (0.7 g, 1.8 mmol, 1.5 equiv.) and N,N-diisopropylethylamine (0.47 g, 3.7 mmol, 3.0 equiv.). The yellow solution was stirred for 1 h and then diluted with water (50 mL). The aqueous phase was extracted with EtOAc (50 mL × 2). The combined organic layers were washed with saturated aqueous brine (50 mL × 2), dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by column chromatography (eluted with CHCl / MeOH) to afford tert-butyl ((3S,6S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate (0.29 mg, 53% yield) as a yellow oil. LCMS (ESI) m / z = 443.1 [M+H] + .

[0331] Step 2: Preparation of tert-butyl ((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate and tert-butyl ((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate

[0332] tert-Butyl ((3S,6S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate (0.29 mg) was purified by SFC (Column: DAICEL CHIRALPAK AD 250 mm × 30 mm, 10 μm, Mobile phase: Phase A is CO and Phase B is Neu-IPA; Gradient elution: 30% B in A) to give:

[0333] Peak 1: tert-butyl ((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate or tert-butyl ((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate (0.14 g, 48% yield, tR = 1.536 min) as a white solid. LCMS (ESI) m / z = 443.1 [M+H] + .

[0334] Peak 2: tert-butyl ((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate or tert-butyl ((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate (80 mg, 28% yield, tR = 1.662 min) as a white solid. LCMS (ESI) m / z = 443.1 [M+H] + .

[0335] Step 3: Preparation of (3S,6S,8R,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one or (3S,6S,8S,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one

[0336] To a solution of tert-butyl ((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate or tert-butyl ((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate [peak 1] (1 equivalent) in CHCl (0.3 mL) was added trifluoroacetic acid (0.1 mL). The resulting yellow reaction mixture was stirred for 15 minutes, then concentrated in vacuo and dried to give (3S,6S,8R,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one or (3S,6S,8S,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one (140 mg, TFA salt) as a yellow oil. LCMS (ESI) m / z = 343.1 [M+H] + .

[0337] The intermediates in Table 29 were prepared according to the methods described above, starting from tert-butyl ((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate or tert-butyl ((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamate [peak 2] and using the appropriate conditions. [Table 29]

[0338] Step 4: Preparation of (difluoro(2-(((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid or (difluoro(2-(((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (109 / 110)

[0339] To a solution of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (1 equiv.) in DMF (1 mL) was added HOBt (1.5 equiv.) and N,N-diisopropylethylamine (3 equiv.). After stirring for 30 minutes, a solution of (3S,6S,8R,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one or (3S,6S,8S,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one [from peak 1 in step 3] (1.2 equivalents) in DMF (1 mL) and N,N-diisopropylethylamine (3 equivalents) was added to the reaction mixture. The resulting yellow reaction mixture was stirred for 30 minutes and then purified by prep-HPLC (column: Phenomenex Luna C18 The sample was purified using a 150×25mm×10um mobile phase: water (NH4HCO3)-acetonitrile; B%: 4% to 34%, 8 minutes, to obtain (difluoro(2-(((3S,6S,8R,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thio (difluoro(2-(((3S,6S,8S,9aR)-8-methyl-5-oxo-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (109) (21 mg, 14% yield). LCMS (ESI) m / z = 633.3 [M+H] + ; 1H NMR (400 MHz, methanol-d4) δ 8.69 - 8.40 (m, 2H), 8.22 - 8.06 (m, 3H), 7.94 (d, J = 8.4 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.48 (s, 1H), 4.77 (d, J = 10.0 Hz, 1H), 4.69 (s, 1H), 4.62 - 4.47 (m, 2H), 4.45 - 4.24 (m, 1H), 4.14 - 3.93 (m, 3H), 2.31 (s, 1H), 2.18 - 2.03 (m, 2H), 1.96 (d, J = 9.6 Hz, 3H), 1.86 - 1.72 (m, 1H), 1.70 - 1.52 (m, 2H), 1.05 (s, 3H).

[0340] The compounds in Table 30 were prepared according to the methods described above, starting from (3S,6S,8S,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one or (3S,6S,8R,9aR)-6-amino-8-methyl-3-(3-(pyridin-3-yl)azetidine-1-carbonyl)octahydro-5H-pyrrolo[1,2-a]azepin-5-one [from peak 2] and using the appropriate conditions. [Table 30]

[0341] Method 3: Representative procedure for the synthesis of phosphonate analogs by amino acid coupling of core carboxylic acids with amines

[0342] Synthesis of (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (111) [ka]

[0343] Step 1: Preparation of benzyl (3S,6S,9aS)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate

[0344] To a solution of (3S,6S,9aS)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylic acid (590 mg, 1.88 mmol, 1 equiv.) and CsCO (1.22 g, 3.76 mmol, 2 equiv.) in THF (15 mL) was added benzyl bromide (352 mg, 2.06 mmol, 1.1 equiv.). The resulting mixture was stirred at room temperature for an additional 12 h, after which HO (10 mL) was added. The resulting suspension was extracted with CHCl (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by flash column chromatography to give benzyl (3S,6S,9aS)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (633 mg, 1.57 mmol, 84% yield) as a white solid. LCMS (ESI) m / z = 403 [M+H] + .

[0345] Step 2: Preparation of benzyl (3S,6S,9aS)-6-amino-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate

[0346] To a solution of benzyl (3S,6S,9aS)-6-((tert-butoxycarbonyl)amino)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (663 mg, 1.64 mmol, 1.0 equiv.) in CHCl (16 mL) was added trifluoroacetic acid (8 mL), and the resulting mixture was stirred at room temperature for 2 h. The reaction mixture was then cooled (0 °C), and saturated aqueous NaHCO was carefully added to make the mixture basic (adjust the pH to 8-9). The resulting mixture was extracted with CHCl (10 mL×3), and the combined organic layers were washed with brine (10 mL×2), dried over anhydrous NaSO, and then concentrated under reduced pressure to give crude (3S,6S,9aS)-6-amino-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate benzyl (498 mg, 1.64 mmol, quantitative) as a white solid, which was used directly in the next step without further purification. LCMS (ESI): m / z = 303 [M+H] + .

[0347] Step 3: Preparation of benzyl (3S,6S,9aS)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate

[0348] A solution of 5-[(diethoxyphosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylic acid (597 mg, 1.64 mmol, 1 equiv.), EDCI (412 mg, 2.13 mmol, 1.3 equiv.), HOBt (287 mg, 2.13 mmol, 1.3 equiv.), and N,N-diisopropylethylamine (635 mg, 4.92 mmol, 3 equiv.) in CHCl (15 mL) was stirred for 30 minutes, after which benzyl (3S,6S,9aS)-6-amino-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (498 mg, 1.64 mmol, 1 equiv.) was added. The mixture was stirred at room temperature for 12 hours, after which HO (10 mL) was added. The biphasic mixture was extracted with CHCl (10 mL × 3). The organic layers were combined and washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by flash column chromatography to give benzyl (3S,6S,9aS)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (530 mg, 817 μmol, 50%) as a white solid. LCMS (ESI): m / z = 649 [M+H] + .

[0349] Step 4: Preparation of (3S,6S,9aS)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylic acid

[0350] A mixture of (3S,6S,9aS)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (510 mg, 786 μmol, 1 equiv.) and 10% wt wet Pd / C (50 mg) in MeOH (15 mL) was stirred under an H(g) atmosphere. After complete consumption of the starting material (as judged by LCMS), the reaction mixture was filtered through a pad of Celite®. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography to give (3S,6S,9aS)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylic acid (376 mg, 673 μmol, 86% yield) as an off-white solid. LCMS (ESI): m / z = 559 [M+H] + .

[0351] Step 5: Preparation of diethyl (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonate

[0352] A solution of (3S,6S,9aS)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylic acid (120 mg, 214 μmol, 1 equiv.), 2-chloro-1-methylpyridinium iodide (164 mg, 642 μmol, 3.0 equiv.), and N,N-diisopropylethylamine (82.9 mg, 642 μmol, 3.0 equiv.) in CHCl (15 mL) was stirred at room temperature for 30 min, after which N-methylaniline (22.9 mg, 214 μmol, 1.0 equiv.) was added. After stirring for 12 h, the reaction mixture was diluted with HO (10 mL), and the resulting biphasic mixture was extracted with CHCl (10 mL × 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by flash column chromatography to give diethyl (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonate (90 mg, 138 μmol, 65% yield) as a white solid. LCMS (ESI) m / z = 648 [M+H] + .

[0353] Step 6: Preparation of (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (111)

[0354] To a cooled (0 °C) solution of diethyl (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonate (50 mg, 77.1 μmol, 1.0 equiv) in CHCl (15 mL) was added bromotrimethylsilane (235 mg, 1.54 mmol, 20.0 equiv) in a dropwise manner. After the addition, the reaction mixture was allowed to warm to room temperature and stirred for an additional 12 h. After complete consumption of the starting material (as judged by LCMS), the reaction mixture was quenched by the addition of HO (5 mL), and the resulting biphasic mixture was extracted with CHCl (10 mL × 3). The organic layers were combined and washed with brine (20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by reverse-phase HPLC to give (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (111) (2.1 mg, 3.5 μmol, 4.5% yield) as a white solid. LCMS (ESI) m / z = 592 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.74 (d, J = 6.7 Hz, 1H), 8.29 (s, 1H), 8.13 - 7.96 (m, 2H), 7.65 (d, J = 7.3 Hz, 1H), 7.53 - 7.25 (m, 5H), 4.67 - 4.55 (m, 1H), 4.41 - 4.27 (m, 1H), 3.99 - 3.85 (m, 1H), 3.15 (s, 3H), 2.16 - 1.62 (m, 10H).

[0355] The following compounds in Table 31 were prepared according to the representative procedure described above for the synthesis of (difluoro(2-(((3S,6S,9aS)-3-(methyl(phenyl)carbamoyl)-5-oxooctahydro-1H-pyrrolo[1,2-a]azepin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid and utilizing the appropriate starting materials and modifications. [Table 31-1] [Table 31-2] [Table 31-3] [Table 31-4]

[0356] Synthesis of azetidine building blocks:

[0357] Synthesis of 1-phenyl-2-(2,6-diazaspiro[3.3]heptan-2-yl)ethan-1-one [ka]

[0358] Step 1: Preparation of tert-butyl 6-(2-oxo-2-phenylethyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0359] To a solution of 2-phenylacetic acid (500 mg, 3.67 mmol, 1 equiv.) in DMF (4 mL) was added N,N-diisopropylethylamine (1.42 g, 11.0 mmol, 3 equiv.) and HATU (2.09 g, 5.50 mmol, 1.5 equiv.). After stirring for 10 minutes, a solution of tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (945 mg, 4.77 mmol, 1.3 equiv.) in DMF (4 mL) and N,N-diisopropylethylamine (1.42 g, 11.0 mmol, 3 equiv.) was added. The mixture was stirred for an additional 1 hour and then diluted with water (20 mL). The mixture was extracted with EtOAc (10 mL × 2). The combined organic layers were washed with saturated aqueous brine (10 mL × 2), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0% to 5% CH₂Cl₂:MeOH) to give tert-butyl 6-(2-phenylacetyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (1.00 g, 3.16 mmol, 86.2% yield) as a white solid. LCMS (ESI) m / z = 317.0 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 7.36 - 7.29 (m, 2H), 7.28 - 7.22 (m, 3H), 4.18 (s, 2H), 4.11 (s, 2H), 3.47 (s, 2H), 2.81 (s, 4H), 1.72 (s, 4H), 1.43 (s, 9H).

[0360] Step 2: Preparation of 1-phenyl-2-(2,6-diazaspiro[3.3]heptan-2-yl)ethan-1-one

[0361] To a solution of tert-butyl 6-(2-phenylacetyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (1.00 g, 3.16 mmol) in CHCl (6 mL) was added trifluoroacetic acid (2 mL, 0.0175 mmol, 0.006 equiv.), and the reaction mixture was stirred for 1 h. The resulting yellow reaction mixture was concentrated under reduced pressure to give a residue. The product, 1-phenyl-2-(2,6-diazaspiro[3.3]heptan-2-yl)ethan-1-one (approximately 1 g, TFA salt), was obtained as a yellow oil and used without further purification. Synthesis of 1-((2R,3S)-2-methylazetidin-3-yl)-1H-imidazole [ka]

[0362] Step 1: Preparation of tert-butyl (2R,3R)-2-methyl-3-((methylsulfonyl)oxy)azetidine-1-carboxylate

[0363] To a solution of tert-butyl (2R,3R)-3-hydroxy-2-methylazetidine-1-carboxylate (500 mg, 2.67 mmol, 1.0 equiv.) and EtN (810 mg, 8.01 mmol, 3.0 equiv.) in CHCl (20 mL) was added methanesulfonyl chloride (335 mg, 2.93 mmol, 1.1 equiv.). The reaction mixture was stirred at room temperature for 14 h, after which HO (10 mL) was added. The resulting biphasic mixture was extracted with CHCl (20 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by Biotage® C18 column to give tert-butyl (2R,3R)-2-methyl-3-((methylsulfonyl)oxy)azetidine-1-carboxylate (710 mg, 2.67 mmol, 100% yield) as a colorless oil. LCMS (ESI) m / z = 210 [(M-56)+H] + .

[0364] Step 2: Preparation of tert-butyl (2R,3S)-3-(1H-imidazol-1-yl)-2-methylazetidine-1-carboxylate

[0365] To a cooled (0 °C) solution of 1H-imidazole (599 mg, 8.81 mmol, 3.3 equiv) in DMF (10 mL) was added NaH (352 mg, 8.81 mmol, 3.3 equiv) in portions under a constant stream of N2 (g). The mixture was stirred until gas evolution ceased (approximately 30 min). To this mixture was added a solution of tert-butyl (2R,3R)-2-methyl-3-((methylsulfonyl)oxy)azetidine-1-carboxylate (710 mg, 2.67 mmol, 1.0 equiv) in DMF (3 mL). The reaction mixture was then heated to 80 °C. After stirring for 72 h, the reaction mixture was cooled to 0 °C, and saturated aqueous NH4Cl (10 mL) was added. The mixture was extracted with EtOAc (20 mL × 3). The organic layers were combined, washed with brine (10 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by Biotage C18 column to give tert-butyl (2R,3S)-3-(1H-imidazol-1-yl)-2-methylazetidine-1-carboxylate (73.0 mg, 0.31 mmol, 12% yield) as a colorless oil. LCMS (ESI) m / z = 238 [M+H] + .

[0366] Step 3: Preparation of 1-((2R,3S)-2-methylazetidin-3-yl)-1H-imidazole

[0367] To a solution of tert-butyl (2R,3S)-3-(1H-imidazol-1-yl)-2-methylazetidine-1-carboxylate (73.0 mg, 0.31 mmol, 1.0 equiv.) in CHCl (3 mL) was added TFA (1 mL), and the resulting mixture was stirred at room temperature. After stirring for 16 h, the reaction mixture was concentrated under reduced pressure to give 1-((2R,3S)-2-methylazetidin-3-yl)-1H-imidazole (40 mg, TFA salt) as a white solid, which was used without further purification. LCMS (ESI) m / z = 138 [M+H] + .

[0368] ((2-(((3S,6S,10aS)-3-(3-butylamido-3-(pyridin-2-yl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid (122) [ka]

[0369] Step 1: Preparation of tert-butyl 3-butylamido-3-(pyridin-2-yl)azetidine-1-carboxylate

[0370] To a solution of butyric acid (95 mg, 1.1 mmol, 0.9 equiv.) in DMF (3 mL) was added HATU (0.69 g, 1.8 mmol, 1.5 equiv.) and N,N-diisopropylethylamine (0.46 g, 3.6 mmol, 3 equiv.). The mixture was stirred for 10 minutes, after which tert-butyl 3-amino-3-(pyridin-2-yl)azetidine-1-carboxylate (0.30 g, 1.2 mmol, 1 equiv.) was added. After stirring for an additional hour, the reaction mixture turned brown. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with saturated aqueous brine (30 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (SepaFlash® column, 0% to 100% EtOAc / petroleum ether gradient) to afford tert-butyl 3-butylamido-3-(pyridin-2-yl)azetidine-1-carboxylate (0.26 g, 68% yield) as a white solid. LCMS (ESI) m / z = 320.1 [M+H] + .

[0371] Step 2: Preparation of N-(3-(pyridin-2-yl)azetidin-3-yl)butyramide

[0372] A solution of tert-butyl 3-butylamido-3-(pyridin-2-yl)azetidine-1-carboxylate (0.24 g, 0.75 mmol, 1 equiv.) in CHCl (2 mL) and TFA (1 mL) was stirred at room temperature. After 1 h, the resulting yellow solution was concentrated under reduced pressure to give N-(3-(pyridin-2-yl)azetidin-3-yl)butyramide (0.24 g, TFA salt), which was used in the next step without further purification.

[0373] The following intermediates in Table 32 were prepared according to the representative procedures described above (Step 1 and Step 2), starting from tert-butyl 3-butylamido-3-(pyridin-2-yl)azetidine-1-carboxylate and utilizing the appropriate starting materials and modifications. [Table 32]

[0374] Step 3: Preparation of tert-butyl ((3S,6S,10aS)-3-(3-butylamido-3-(pyridin-2-yl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate

[0375] To a solution of (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (0.36 g, 1.1 mmol, 1 equiv.) in DMF (3 mL) was added N,N-diisopropylethylamine (0.43 g, 3.3 mmol, 3 equiv.) and HATU (0.63 g, 1.6 mmol, 1.5 equiv.). After 10 min, N-(3-(pyridin-2-yl)azetidin-3-yl)butyramide (0.24 g, 1.1 mmol, 1 equiv.) was added. After 1 h, the brown solution was diluted with water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with 60 mL (20 mL × 3) of saturated aqueous brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (SepaFlash® silica flash column, 0% to 100% EtOAc / petroleum ether) to give tert-butyl ((3S,6S,10aS)-3-(3-butylamido-3-(pyridin-2-yl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate (0.29 g, 51% yield) as a white solid. LCMS (ESI) m / z = 528.2 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 9.11 - 8.95 (m, 1H), 8.61 (d, J = 4.4 Hz, 1H), 7.82 - 7.73 (m, 1H), 7.43 - 7.26 (m, 2H), 6.76 - 6.67 (m, 1H), 4.66 - 4.53 (m, 1H), 4.32 (td, J = 8.8, 9.2 Hz, 2H), 4.19 - 4.08 (m, 2H), 4.07 - 3.99 (m, 1H), 2.25 - 2.13 (m, 3H), 1.99 (s, 1H), 1.87 - 1.65 (m, 6H), 1.54 (dt, J = 7.8, 11.2 Hz, 6H), 1.38 - 1.30 (m, 9H), 1.17 (t, J = 7.2 Hz, 1H), 0.88 (td, J = 5.2, 5.2 Hz, 3H).

[0376] Step 4: Preparation of N-(1-((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-3-(pyridin-2-yl)azetidin-3-yl)butyramide

[0377] A solution of tert-butyl ((3S,6S,10aS)-3-(3-butylamido-3-(pyridin-2-yl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamate (0.22 g, 0.42 mmol, 1 equiv.) in CHCl (1 mL) and TFA (0.5 mL) was stirred at room temperature. After 1 h, the resulting yellow solution was concentrated under reduced pressure to give N-(1-((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-3-(pyridin-2-yl)azetidin-3-yl)butyramide (0.22 g, TFA salt), which was used in the next step without further purification.

[0378] Step 5: Preparation of ((2-(((3S,6S,10aS)-3-(3-butylamido-3-(pyridin-2-yl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid (122)

[0379] To a solution of N-(1-((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-3-(pyridin-2-yl)azetidin-3-yl)butyramide (80 mg, 0.19 mmol, 1 equiv.) in DMF (1 mL) was added (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (80 mg, 0.19 mmol, 1 equiv.), N,N-diisopropylethylamine (73 mg, 0.57 mmol, 3 equiv.), and HOBt (25 mg, 0.19 mmol, 1 equiv.). The mixture was stirred at 25° C. for 10 minutes, and then the yellow mixture was filtered. The filtrate was purified by prep-HPLC (Phenomenex Luna C18 150 × 25 mm × 10 μm; water (0.1% TFA)-ACN; B%: 32% to 62% in 10 min) to give ((2-(((3S,6S,10aS)-3-(3-butylamido-3-(pyridin-2-yl)azetidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid (122) (40 mg, 5.6% yield) as a white solid. LCMS (ESI) m / z = 718.4 [M+H] + ; 1H NMR (400 MHz, methanol-d4) δ 8.67 - 8.33 (m, 1H), 8.21 - 8.12 (m, 1H), 8.03 (s, 1H), 8.02 - 7.93 (m, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.67 (dd, J = 8.4, 6.4 Hz, 1H), 7.58 - 7.51 (m, 1H), 5.06 - 4.93 (m, 2H), 4.84 - 4.77 (m, 1H), 4.68 - 4.60 (m, 1H), 4.55 - 4.47 (m, 1H), 4.55 - 4.47 (m, 1H), 4.45 (d, J = 10.0 Hz, 1H), 4.36 - 4.26 (m, 1H), 2.34 - 2.21 (m, 4H), 2.14 - 1.92 (m, 6H), 1.91 - 1.79 (m, 2H), 1.75 - 1.68 (m, 1H), 1.68 - 1.51 (m, 3H), 0.90 (td, J = 7.2, 12.0 Hz, 3H).

[0380] The following compounds in Table 33 were prepared according to the representative procedures described above, starting from (3S,6S,10aS)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid and utilizing the appropriate starting materials and modifications. [Table 33]

[0381] Synthesis of pyrrolidine building blocks: Synthesis of rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile [ka]

[0382] Step 1: Preparation of (E)-3-cyclohexylacrylonitrile

[0383] To a solution of cyclohexanecarbaldehyde (500 mg, 4.45 mmol, 1.0 equiv.) in THF (15 mL) was added t-BuOK (998 mg, 8.90 mmol, 2.0 equiv.) and diethyl (cyanomethyl)phosphonate (788 mg, 4.45 mmol, 1.0 equiv.) at room temperature. The solution was stirred at room temperature for 1 h. After completion, the reaction mixture was quenched by adding HO (10 mL) and then extracted with EtOAc (10 mL × 3). The organic layers were combined and washed with brine (10 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give (E)-3-cyclohexylacrylonitrile (300 mg, 2.21 mmol, 50% yield) as a colorless oil. LC-MS (ESI) m / z = 136 [M+H] + .

[0384] Step 2: Preparation of rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carbonitrile

[0385] To a solution of (E)-3-cyclohexylacrylonitrile (300 mg, 2.21 mmol, 1.0 equiv) in CHCl (5 mL) was added N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methanamine (524 mg, 2.21 mmol, 1.0 equiv) and TFA (25.1 mg, 221 μmol, 0.1 equiv). The reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was diluted with CHCl (10 mL) and washed with aqueous saturated NaHCO (5 mL). The organic layer was separated and concentrated under reduced pressure to give crude rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carbonitrile (700 mg) as a colorless oil. LC-MS (ESI) m / z = 269 [M+H] + .

[0386] Step 3: Preparation of rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile

[0387] To a solution of (trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carbonitrile (700 mg) in dry 1,2-dichloroethane (15 mL) was added 1-chloroethyl carbonochloridate (3.71 g, 26.0 mmol, 10.0 equiv.). The resulting mixture was stirred at 70° C. for 12 hours, then concentrated under reduced pressure. The crude product was dissolved in MeOH (5 mL) and stirred at 70° C. for 1 hour. Upon completion, the reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to afford rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile (211 mg, 1.18 mmol, 46% yield) as a colorless oil. LC-MS (ESI) m / z = 179 [M+H] + .

[0388] The following intermediates in Table 34 were prepared according to the representative procedure (Steps 1-3) described for rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile, utilizing the appropriate starting materials and modifications. The compounds were prepared as racemates with trans stereochemical configurations about the C3 and C4 stereocenters in the pyrrolidine ring. [Table 34]

[0389] tert-Butyl rel-(trans)-3-cyano-4-phenylpyrrolidine-1-carboxylate and tert-butyl (trans)-3-cyano-4-(2-oxo-1,2-dihydropyridin-4-yl)pyrrolidine-1-carboxylate were prepared according to the method described above for the synthesis of rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile. The racemic mixture of trans isomers was purified under SFC conditions, and its absolute stereochemistry was arbitrarily assigned as shown. [ka]

[0390] Preparative separation method:

[0391] Instrument: Waters Thar 80 Preparative SFC; Column: ChiralPak C-IG, 100 × 4.6 mm ID, 5 μm; Mobile phase: A is CO2, B is methanol (0.05% diethylamine); Gradient: 10% to 40% B in 8 min; Flow rate: 2.5 mL / min; Back pressure: 100 bar; Column temperature: 40 °C; Wavelength: 210 nm; Cycle time: 2 min. [Table 35] Isolation of 2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile [ka]

[0392] 2-Oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile (1.00 g, single diastereomer of unknown relative stereochemistry, CAS# 1423027-48-8) was purchased and separated under SFC conditions (Lux-Cellulose-5 21.2 x 250 mm, 5 um column, 3.7 mg / inj, concentration 36.9 mg / mL, column T = 40°C, flow rate 75 mL / min, 15% MeOH (with 0.1% diethylamine), cycle time: 4.9 min) to give peak 1 or peak 2.

[0393] Peak 1: (3S,4'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile and (3R,4'R)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile (230 mg of each enantiomer) or (3S,4'R)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile or (3R,4'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile (220 mg of each enantiomer)

[0394] Peak 2: (3S,4'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile and (3R,4'R)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile (220 mg of each enantiomer) or (3S,4'R)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile or (3R,4'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carbonitrile (220 mg of each enantiomer) Synthesis of 6-phenyl-4-azaspiro[2.4]heptane [ka]

[0395] Step 1: Preparation of methyl 3-cyano-2-phenylpropanoate

[0396] To a cooled (−78° C.) solution of methyl 2-phenylacetate (5.0 g, 33.3 mmol, 1.0 equiv.) in dry THF (50 mL) was slowly added a 2 M solution of LDA in THF (20 mL, 40.0 mmol, 1.2 equiv.). After 1 h, 2-bromoacetonitrile (4.2 g, 35.0 mmol, 1.1 equiv.) was slowly added dropwise, and the reaction was further aged at −78° C. for an additional 1 h. Saturated aqueous NH₄Cl (5 mL) was added to the mixture, and the mixture was allowed to warm to room temperature. The mixture was then washed with EtOAc (20 mL × 3). The organic layers were combined, washed with brine (150 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give methyl 3-cyano-2-phenylpropanoate (5.0 g, 26.5 mmol, 79%) as a white solid. LCMS (ESI) m / z = 190.1 [M+H] + .

[0397] Step 2: Preparation of 6-phenyl-4-azaspiro[2.4]heptan-5-one

[0398] To a cooled (0 °C) solution of methyl 3-cyano-2-phenylpropanoate (5.0 g, 26.5 mmol, 1.0 equiv.) and Ti(OiPr)4 (9.0 g, 31.7 mmol, 1.2 equiv.) in dry THF (100 mL) was slowly added 3 M EtMgBr solution (20 mL, 59.6 mmol, 2.25 equiv.) while maintaining the internal temperature between -5 °C and 0 °C. After complete addition of EtMgBr, the mixture was stirred at 0 °C for an additional 1 h. The reaction mixture was quenched by the addition of 2 N aqueous HCl (60 mL). The resulting acidic solution was extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (20 mL × 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by flash column chromatography to give 6-phenyl-4-azaspiro[2.4]heptan-5-one (2.1 g, 11.2 mmol, 42% yield) as a white solid. LCMS (ESI) m / z = 188 [M+H] + ; 1 H NMR (400 MHz, CDCl3) δ 7.39 - 7.24 (m, 6H), 3.86 (dd, J = 9.4, 7.6 Hz, 1H), 2.51 (dd, J = 12.9, 9.4 Hz, 1H), 2.29 (dd, J = 12.9, 7.6 Hz, 1H), 0.85 - 0.88 (m, 1H), 0.87 - 0.81 (m, 1H), 0.76 - 0.64 (m, 2H).

[0399] Step 3: Preparation of 6-phenyl-4-azaspiro[2.4]heptane

[0400] To a cooled (10 °C) solution of 6-phenyl-4-azaspiro[2.4]heptan-5-one (2.1 g, 11.2 mmol, 1.0 equiv.) in dry THF (50 mL) was added NaBH (2.1 g, 56.0 mmol, 5.0 equiv.) in several portions. To this mixture was added BF·EtO (6.7 mL, 56.0 mmol, 5.0 equiv.) in a dropwise manner. The reaction mixture was heated at 60 °C for 16 h. The mixture was cooled to ambient temperature, and 2 N aqueous HCl (30 mL) was slowly introduced. The acidic mixture was extracted with EtOAc (30 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by Biotage® C18 column to give 6-phenyl-4-azaspiro[2.4]heptane (1.5 g, 8.7 mmol, 78%) as a colorless oil. LCMS (ESI) m / z = 174 [M+H] + ; 1 H NMR (400 MHz, CDCl3) 8.59 (s, 1H), δ 7.38 - 7.27 (m, 5H), 3.85 - 3.57 (m, 2H), 3.40 - 3.17 (m, 1H), 2.43 - 2.09 (m, 2H), 1.46 - 1.18 (m, 2H), 0.98 - 0.66 (m, 2H). Synthesis of 3-(2-oxo-1,2-dihydropyridin-4-yl)pyrrolidin-1-ium chloride [ka]

[0401] Step 1: Preparation of benzyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,3-dihydro-1H-pyrrole-1-carboxylate

[0402] To a cooled (−78° C.) solution of benzyl 3-oxopyrrolidine-1-carboxylate (5.0 g, 22.8 mmol, 1 equiv.) in dry THF (80 mL) was slowly added a solution of LDA (25.0 mmol, 1.1 equiv.) in THF. The mixture was stirred for an additional 30 minutes, after which a solution of 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (8.93 g, 25.0 mmol, 1.1 equiv.) in THF (70 mL) was added. After complete addition of 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide, the reaction mixture was allowed to warm to room temperature and stirred for 14 hours. The reaction mixture was quenched by the addition of HO (20 mL) and extracted with EtOAc (150 mL×3). The organic layers were combined, washed with brine (150 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give benzyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,3-dihydro-1H-pyrrole-1-carboxylate (3.00 g, 8.53 mmol, 37.5% yield) as a yellow oil. LCMS (ESI) m / z = 352 [M+H] + .

[0403] Step 2: Preparation of benzyl 4-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate

[0404] To a solution of benzyl 4-(((trifluoromethyl)sulfonyl)oxy)-2,3-dihydro-1H-pyrrole-1-carboxylate (3.0 g, 8.53 mmol, 1.0 equiv), (2-methoxypyridin-4-yl)boronic acid (1.55 g, 10.2 mmol, 1.2 equiv), and KCO (2.94 g, 21.3 mmol, 2.5 equiv) in a mixture of toluene (60.0 mL), EtOH (15.0 mL), and HO (15.0 mL) was added Pd(PPh) (492 mg, 426 μmol, 0.05 equiv). The reaction mixture was then heated at 100 °C under N for 12 h. The reaction mixture was cooled to room temperature, diluted with HO (40 mL), and extracted with EtOAc (100 mL × 3). The organic layers were combined, washed with brine (100 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give benzyl 4-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate (750 mg, 2.41 mmol, 28.4% yield) as a white solid. LCMS (ESI) m / z = 311 [M+H] + .

[0405] Step 3: Preparation of 2-methoxy-4-(pyrrolidin-3-yl)pyridine

[0406] To a solution of benzyl 4-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-pyrrole-1-carboxylate (750 mg, 2.41 mmol, 1 equiv.) in MeOH (50 mL) under N2(g) was added 10% Pd / C (100 mg). The suspension was subjected to three cycles of evacuation and purging with H2(g). The resulting mixture was stirred at room temperature under H2(g) (1 atm) for 14 h. The reaction mixture was then subjected to three cycles of evacuation and purging with N2(g), and the suspension was filtered through a pad of Celite®. The filter cake was carefully washed with MeOH (20 mL), and the combined filtrates were concentrated to dryness to give 2-methoxy-4-(pyrrolidin-3-yl)pyridine (375 mg, 2.10 mmol, 87.4% yield) as an off-white solid. LCMS (ESI) m / z = 179 [M+H] + .

[0407] Step 4: Preparation of 3-(2-oxo-1,2-dihydropyridin-4-yl)pyrrolidin-1-ium chloride

[0408] A solution of 2-methoxy-4-(pyrrolidin-3-yl)pyridine (375 mg, 2.10 mmol, 1 equiv) in a solution of 1N HCl in 1,4-dioxane (4 mL) was heated and stirred at 100° C. in a sealed tube. After 24 h, the reaction mixture was concentrated under reduced pressure to give 3-(2-oxo-1,2-dihydropyridin-4-yl)pyrrolidin-1-ium chloride (400 mg, HCl salt) as a white solid, which was used directly without further purification. LCMS (ESI) m / z = 165 [M+H] + .

[0409] Representative Procedure for Direct Coupling of Perfluorophenyl or p-Nitrophenyl Activated Linker Esters to Amino Acid Cores for the Construction of Analogues S,S'-(((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl) bis(oxy)) bis(ethane-2,1-diyl)) dibutanethioate (125) [ka]

[0410] Step 1: Preparation of S,S'-(((((2-(((3S,6S,10aS)-3-((3S,4R)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl) bis(oxy)) bis(ethane-2,1-diyl)) dibutanethioate

[0411] To a solution of perfluorophenyl 5-((bis(2-(butyrylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (30 mg, 42.9 μmol, 1.0 equiv.) and (3S,4R or 3R,4S)-1-((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-4-phenylpyrrolidine-3-carbonitrile (16 mg, 42.0 μmol, 1.0 equiv.) (using pyrrolidine SFC peak 2) in CHCl (2 mL) was added N,N-diisopropylethylamine (10.8 mg, 84 μmol, 2.0 equiv.) at room temperature. After stirring for 48 h, the reaction mixture was concentrated under reduced pressure. The residue was purified by reverse-phase HPLC to give S,S'-(((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) dibutanethioate (35 mg, 39 μmol, 94% yield) as a white solid. LCMS (ESI) m / z =895 [M+H]+; 1H NMR (400 MHz, CDCl3) δ 7.86-7.71 (m, 3H), 7.45-7.29 (m, 6H), 7.22-7.13 (m, 1H), 5.19-5.08 (m, 1H), 4.60-4.49 (m, 1H), 4.39-4.23 (m, 2H), 4.11-3.93 (m, 6H), 3.81-3.60 (m, 2H), 3.28 (d, JHP = 21.2 Hz, 2H), 3.22-3.12 (m, 1H), 3.06 (t, J = 6.3 Hz, 4H), 2.52 (t, J = 7.4 Hz, 4H), 2.29-1.58 (m, 16H), 0.94 (t, J= 8.0 Hz, 6H).

[0412] The following compounds in Table 36 were prepared using the representative protocol described above for the synthesis of S,S'-(((((2-(((3S,6S,10aS)-3-((3S,4R)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) dibutanethioate and utilizing the appropriate starting materials and modifications. [Table 36-1] [Table 36-2] [Table 36-3] [Table 36-4] [Table 36-5] [Table 36-6] [Table 36-7] [Table 36-8] [Table 36-9] [Table 36-10] [Table 36-11] [Table 36-12] [Table 36-13]

Table 36-14

Table 36-15

Table 36-16

Table 36-17

Table 36-18

Table 36-19

Table 36-20

Table 36-21

Table 36-22

Table 36-23

Table 36-24

Table 36-25

Table 36-26

Table 36-27

Table 36-28

Table 36-29

Table 36-30

[0413] Representative Procedure for Amino Acid Coupling Conditions Between the Acid Ester of the Linker and the Amino Acid Core for Construction of Analogs

[0414] Preparation of (((R)- or (S)-(2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphoryl)bis(oxy))bis(methylene) bis(2,2-dimethylpropanoate)) (183) [ka]

[0415] To a solution of (3S,4R or 3R,4S)-1-[(3S,6S,10aS)-6-amino-5-oxo-decahydropyrrolo[1,2-a]azocine-3-carbonyl]-4-phenylpyrrolidine-3-carbonitrile (pyrrolidine peak 2) (45.6 mg, 120 μmol, 1.2 equiv.), 5-[(S)-[bis({[(2,2-dimethylpropanoyl)oxy]methoxy})phosphoryl](fluoro)methyl]-1-benzothiophene-2-carboxylic acid (phosphonate peak 2) (51.8 mg, 0.10 mmol, eq.) and N,N-diisopropylethylamine (104 μL, 600 μmol, 6 equiv.) in DMF (3 mL) was added HATU (45.6 mg, 120 μmol, 1.2 equiv.). The reaction mixture was stirred at room temperature for 15 minutes, after which the solution was purified by reverse-phase chromatography using a C18 column (gradient 5% to 100% acetonitrile in water) to give ({[(R)- or (S)-(2-{[(3S,6S,10aS)-3-[(3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl]-5-oxo-decahydropyrrolo[1,2-a]azocin-6-yl]carbamoyl}-1-benzothiophen-5-yl)(fluoro)methyl]({[(2,2-dimethylpropanoyl)oxy]methoxy})phosphoryl}oxy)methyl 2,2-dimethylpropanoate (182) (36.4 mg, 0.04136 mmol, 41.3%) as a white solid. LCMS (ESI) m / z = 881.2 [M+H] + ; 1H NMR (400 MHz, acetonitrile-d3) δ 8.03 - 7.91 (m, 3H), 7.56 - 7.46 (m, 2H), 7.46 - 7.29 (m, 5H), 5.99 (dd, J = 43.0, 7.8 Hz, 1H), 5.67 - 5.52 (m, 4H), 5.10 - 4.96 (m, 1H), 4.63 - 4.53 (m, 1H), 4.42 - 4.23 (m, 2H), 4.13 - 3.95 (m, 1H), 3.85 - 3.68 (m, 1H), 3.65 - 3.31 (m, 3H), 2.35 - 2.21 (m, 1H). 2.12 - 1.96 (m, 5H), 1.91 - 1.66 (m, 4H), 1.65 - 1.54 (m, 2H), 1.12 (s, 18H).

[0416] The following compounds in Table 37 were prepared according to the representative procedure described above for the synthesis of ({[(R)- or (S)-(2-{[(3S,6S,10aS)-3-[(3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl]-5-oxo-decahydropyrrolo[1,2-a]azocin-6-yl]carbamoyl}-1-benzothiophen-5-yl)(fluoro)methyl]({[(2,2-dimethylpropanoyl)oxy]methoxy})phosphoryl}oxy)methyl 2,2-dimethylpropanoate (182) and using the appropriate starting materials and modifications. [Table 37-1] [Table 37-2] [Table 37-3] [Table 37-4] [Table 37-5] [Table 37-6] [Table 37-7] [Table 37-8] [Table 37-9] [Table 37-10] S,S'-(((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl))=bis(3-hydroxy-2,2-dimethylpropanethioate)trifluoroacetate (199) [ka]

[0417] Step 1: Preparation of S,S'-((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanethioate)

[0418] S,S'-((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanethioate) was reacted with S,S'-((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanethioate). The procedure described for the synthesis of 5-((bis(2-((3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanoyl)thio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate was used, starting from perfluorophenyl 5-((bis(2-((3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanoyl)thio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate and (3S,4R or 3R,4S)-1-((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-4-phenylpyrrolidine-3-carbonitrile (SFC peak 2 was used). The product was purified using standard chromatographic techniques.

[0419] S,S'-((((2-(((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-4-phenylpyrrolidine-3-carbonitrile) was obtained by reaction of 5-((bis(2-(butyrylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-perfluorophenyl carboxylate (25.0 mg, 25.3 μmol, 1.0 equiv.) with (3S,4R or 3R,4S)-1-((3S,6S,10aS)-6-amino-5-oxodecahydropyrrolo[1,2-a]azocine-3-carbonyl)-4-phenylpyrrolidine-3-carbonitrile (9.62 mg, 25.3 μmol, 1.0 equiv.). aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanethioate) (14.0 mg, 11.8 μmol, 46.8% yield) was obtained as a colorless oil after prep-HPLC purification.

[0420] Step 2: Preparation of S,S'-((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-hydroxy-2,2-dimethylpropanethioate) trifluoroacetate (199)

[0421] To a solution of S,S'-((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpropanethioate) (14 mg, 11.8 μmol, 1.0 equiv) in CHCl (0.5 mL) was added TFA (13.4 mg, 118 μmol, 10.0 equiv). The reaction mixture was stirred at 25 °C for 0.5 h and then concentrated in vacuo. The crude product was pre-purified by column chromatography, followed by prep-HPLC purification to give S,S'-((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-hydroxy-2,2-dimethylpropanethioate) trifluoroacetate (199) (4.90 mg, 5.13 μmol, 43.7% yield). LCMS ESI (m / z) = 955.0 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.77 (m, 1H), 8.21 (m, 1H), 7.98 - 7.91 (m, 1H), 7.84 (s, 1H), 7.41 (m, 5H), 7.34 - 7.26 (m, 1H), 5.08 - 4.91 (m, 1H), 4.54 (m, 1H), 4.29 (m, 1H), 3.98 (m, 4H), 3.84 - 3.62 (m, 3H), 3.47 - 3.38 (m, 8H), 3.04 (m, 4H), 2.30 - 2.22 (m, 1H), 1.84 (m, 6H), 1.59 (m, 3H), 1.24 (s, 2H), 1.10 (s, 12H).

[0422] The following compounds in Table 38 were prepared using the representative protocol described above for the synthesis of S,S'-(((((2-(((3S,6S,10aS)-3-((3S,4R or 3R,4S)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydropyrrolo[1,2-a]azocin-6-yl)carbamoyl)benzo[b]thiophen-5-yl)methyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl)) bis(3-hydroxy-2,2-dimethylpropanethioate) trifluoroacetate (199) and utilizing the appropriate starting materials and modifications. [Table 38-1] [Table 38-2] List of SFC methods:

[0423] A list of specific SFC conditions is provided in Table 39. [Table 39-1] [Table 39-2] Further Compounds and Methods Steps for synthesis of the core: Synthesis of methyl (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate [ka]

[0424] Step 1: (2S,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl)2-methyl

[0425] To a solution of 1-(tert-butyl)2-methyl (2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (330 g, 1.35 mol, 1.00 equiv.) in DMF (2.50 L) was added imidazole (183 g, 2.69 mol, 2.00 equiv.) at 25° C. under N2. The mixture was cooled to 0° C. and stirred for 0.50 h. To the mixture was added TBSCl (243 g, 1.61 mol, 198 mL, 1.20 equiv.) at 0° C., and the mixture was stirred at 0° C. for 0.5 h. The mixture was warmed to 25° C. and stirred at 25° C. for 12 h. TLC (petroleum ether / ethyl acetate=1 / 3) showed the presence of 1-(tert-butyl)2-methyl (2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (R f =0.50) was completely consumed, and the major new spot (R f =0.80). Three batches were combined. The mixture was poured into 15.0 L of water and extracted with ethyl acetate (5.00 L × 2). The organic layer was washed with saturated NaHCO3 (5.00 L × 2), saturated NH4Cl (5.00 L × 2), brine (5.00 L), dried over Na2SO4, filtered, and concentrated to give 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (1.54 kg, crude) as a colorless oil. 1 H NMR: (400 MHz, CDCl3) δ 4.49 - 4.29 (m, 2H), 3.80 - 3.69 (m, 3H), 3.67 - 3.54 (m, 1H), 3.47 - 3.28 (m, 1H), 2.25 - 2.12 (m, 1H), 2.08 - 1.96 (m, 1H), 1.52 - 1.38 (m, 9H), 0.88 (s, 9H), 0.07 (s, 6H).

[0426] Step 2: Synthesis of 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate

[0427] To a solution of NaIO (250 g, 1.17 mol, 64.7 mL, 3.00 equiv.) in HO (2.20 L) was added RuO.HO (11.8 g, 77.9 mmol, 0.20 equiv.) at 25 °C under N. The mixture was stirred at 25 °C for 0.50 h. To this mixture was added a solution of 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (140 g, 389 mmol, 1.00 equiv.) in ethyl acetate (1.20 L) at 25 °C. The mixture was stirred at 25 °C for 12 h. TLC (petroleum ether / ethyl acetate=10 / 1) showed that (2S,4R)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate 1-(tert-butyl)2-methyl (R f =0.40) was consumed, and the major new spot (R f =0.45). Six batches were combined. The mixture was filtered. The filtrate was extracted with ethyl acetate (10.0 L × 3). The combined organic layers were washed with NaSO (10.0 L × 2), brine (10.0 L), dried over NaSO, filtered, and concentrated to give 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate (822 g, crude) as a yellow oil. 1 H NMR: (400 MHz, CDCl3) δ 4.60 - 4.57 (m, 1H), 4.45 - 4.40 (m, 1H), 3.79 (s, 3H), 2.39 - 2.34 (m, 1H), 2.25 - 2.17 (m, 1H), 1.51 (s, 9H), 0.90 (s, 9H), 0.18 (s, 3H), 0.13 (s, 3H).

[0428] Step 3: Synthesis of 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate

[0429] To a solution of 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate (450 g, 1.20 mol, 1.00 equiv.) in THF (1.35 L) was added LiBHEt (1 M, 1.33 L, 1.10 equiv.) at −78° C., followed by stirring at −78° C. for 3 h. TLC (petroleum ether / ethyl acetate=3 / 1) showed that 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-1,2-dicarboxylate (R f =0.60) indicates that the desired spot (R f =0.45, 0.50) was detected. The resulting mixture was quenched with saturated NaHCO3 (1000 mL) and then extracted with ethyl acetate (1500 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate (450 g, crude) was obtained as a pale yellow oil. 1 H NMR: (400 MHz, CDCl3) δ 5.50 - 5.15 (m, 1H), 4.56 - 4.18 (m, 2H), 3.76 - 3.68 (m, 3H), 2.28 - 2.09 (m, 2H), 1.46 - 1.38 (m, 9H), 0.87 (d, J = 3.0 Hz, 9H), 0.10 - 0.04 (m, 6H).

[0430] Step 4: Synthesis of 1-(tert-butyl)2-methyl (2S,4R)-5-acetoxy-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate

[0431] To a solution of 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate (384 g, 1.02 mol, 1.00 equiv.), AcO (156 g, 1.53 mol, 143 mL, 1.50 equiv.), and TEA (156 g, 1.54 mol, 215 mL, 1.51 equiv.) in DCM (2.00 L) was added DMAP (25.0 g, 204 mmol, 0.20 equiv.) at 25° C., followed by stirring at 25° C. for 2 h. TLC (petroleum ether / ethyl acetate=4 / 1) showed that 1-(tert-butyl)2-methyl (2S,4R)-4-((tert-butyldimethylsilyl)oxy)-5-hydroxypyrrolidine-1,2-dicarboxylate (R f = 0.40, 0.45) was consumed, and the desired spot (R f =0.45, 0.55) was detected. The combined reaction mixture was poured into saturated NaHCO3 (2000 mL) and extracted with DCM (1000 mL x 2). The combined organic phase was concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 10 / 1, R f =0.45) to give 1-(tert-butyl)2-methyl (2S,4R)-5-acetoxy-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (220 g, 527 mmol, 51.5% yield, 100% purity) as a pale yellow oil. 1 H NMR: EW28523-2-P1A1 (400 MHz, CDCl3) δ 6.17 (s, 1H), 4.56 - 4.37 (m, 1H), 4.14 (d, J = 7.2 Hz, 1H), 3.77 (s, 3H), 2.18 - 2.07 (m, 5H), 1.45 (d, J = 6.8 Hz, 9H), 0.87 (s, 9H), 0.16 - 0.07 (m, 6H).

[0432] Step 5: Synthesis of 1-(tert-butyl)2-methyl (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate

[0433] To a solution of 1-(tert-butyl)2-methyl (2S,4R)-5-acetoxy-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (100 g, 239 mmol, 1.00 equiv) in DCM (1.00 L) was added BF EtO (85.1 g, 599 mmol, 74.0 mL, 2.50 equiv) and allyltrimethylsilane (123 g, 1.08 mol, 172 mL, 4.50 equiv) at −70 °C under N. The reaction mixture was stirred at −70 °C for 3 h. LCMS (EW28523-3-P1A) showed that 1-(tert-butyl)2-methyl (2S,4R)-5-acetoxy-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate had been consumed, and the desired mass was detected (RT = 1.153 min). The reaction was quenched with saturated NaHCO3 (1000 mL) and then extracted with DCM (1000 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. 1-(tert-butyl)2-methyl (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (200 g, crude) was obtained as a yellow oil. The crude product was used in the next step without purification. LCMS: RT=1.153 min, m / z=300.1 [M-Boc] of product + . 1 H NMR: (400 MHz, CDCl3) δ 5.97 - 5.74 (m, 1H), 5.17 - 4.95 (m, 1H), 4.67 - 3.94 (m, 3H), 3.81 - 3.70 (m, 3H), 2.65 - 2.42 (m, 1H), 2.37 - 2.15 (m, 1H), 2.12 - 1.92 (m, 2H), 1.60 - 1.36 (m, 8H), 0.92 - 0.83 (m, 9H), 0.09 - 0.03 (m, 6H).

[0434] Step 6: Synthesis of methyl (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate

[0435] To a solution of 1-(tert-butyl)2-methyl (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (200 g, 500 mmol, crude purity, 1.00 equiv.) in DCM (1.50 mL) was added TFA (571 g, 5.01 mol, 370 mL, 10.0 equiv.) at 0° C., followed by stirring at 25° C. for 16 h. TLC (petroleum ether / ethyl acetate=2 / 1) showed that 1-(tert-butyl)2-methyl (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-1,2-dicarboxylate (R f = 0.50, 0.55) has been consumed, and the desired spot (R f =0.35) was detected. The resulting mixture was quenched with saturated NaHCO3 (3000 mL) and then extracted with DCM (2000 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by fast silica gel chromatography (petroleum ether / ethyl acetate = 10 / 1, R f =0.35) to give methyl (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (135 g, crude) as a yellow oil. 1 H NMR: (400 MHz, CDCl3) δ 5.89 - 5.76 (m, 1H), 5.19 - 5.03 (m, 3H), 4.30 - 4.21 (m, 1H), 4.07 (t, J = 8.2 Hz, 1H), 3.73 (s, 3H), 3.18 (dt, J = 3.4, 7.0 Hz, 1H), 2.35 - 2.27 (m, 2H), 2.21 - 2.11 (m, 2H), 2.08 - 1.99 (m, 2H), 0.91 - 0.89 (m, 9H), 0.08 (d, J = 5.4 Hz, 6H).

[0436] Step 7: Synthesis of methyl (2S,4R,5S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate and methyl (2S,4R,5R)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate

[0437] To a solution of (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (135 g, 451 mmol, crude purity, 1.00 equiv.) and (S)-2-((tert-butoxycarbonyl)amino)pent-4-enoic acid (111 g, 518 mmol, 1.15 equiv.) in DMF (1.30 L), HATU (343 g, 902 mmol, 2.00 equiv.) and DIEA (291 g, 2.25 mol, 393 mL, 5.00 equiv.) were added at 0° C., followed by stirring at 25° C. for 16 h. TLC (petroleum ether / ethyl acetate=3 / 1) showed that (2S,4R)-5-allyl-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (R f = 0.40, 0.45) was consumed, and the desired spot (R f=0.50) was detected. The reaction was poured into water (4.00 L) and then extracted with ethyl acetate (2.00 L × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by pre-HPLC (column: Phenomenex Luna C18 (250 × 80 mm × 15 um); mobile phase: [water (0.1% TFA)-ACN]; B%: 80%-100%, 35 min]). The aqueous was concentrated under reduced pressure to give the product. Methyl (2S,4R,5S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (84.0 g, 163 mmol, 32.2% yield over three steps, 96.5% purity) was obtained as a pale yellow oil. Methyl (2S,4R,5R)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (24.0 g, 48.3 mmol, 10.7% yield over three steps) was obtained as a pale yellow oil. HPLC (2S,4R,5S)-methyl 5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate: product RT=4.394 min, purity 96.5%, 220 nm. 1H NMR methyl (2S,4R,5S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate: (400 MHz, CDCl3) δ 5.98 - 5.76 (m, 2H), 5.28 - 5.05 (m, 5H), 4.63 - 4.48 (m, 2H), 4.43 - 4.31 (m, 1H), 4.14 - 4.01 (m, 1H), 3.74 (s, 3H), 2.71 - 2.59 (m, 1H), 2.48 - 2.35 (m, 2H), 2.31 (s, 1H), 2.24 - 2.14 (m, 1H), 2.08 - 2.00 (m, 1H), 1.48 - 1.40 (m, 9H), 0.94 - 0.88 (m, 9H), 0.09 (d, J = 3.2 Hz, 6H).

[0438] Step 8a: Synthesis of methyl (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0439] To a solution of methyl (2S,4R,5S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (30.0 g, 58.3 mmol, 96.5% purity, 1.00 equiv.) in DCM (1.50 L) was added Grubbs 1st (9.59 g, 11.7 mmol, 0.20 equiv.) at 25° C., followed by heating to reflux (55° C.) for 16 h. LCMS (EW28523-6-P1A) showed that methyl (2S,4R,5S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate remained (RT=1.138 min), and the desired mass was detected (RT=1.065 min). The mixture was concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate=10 / 1, petroleum ether / ethyl acetate=3 / 1, R f =0.40). (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (23.0 g, crude) was obtained as a brown oil. LCMS: RT=1.065 min, m / z=469.1 (M+H) for the product. + .

[0440] Step 8b: Synthesis of methyl (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0441] To a solution of methyl (2S,4R,5R)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate (16.3 g, 32.8 mmol, 1.00 equiv) in DCM (800 mL) was added Grubb's 1st (2.70 g, 3.28 mmol, 0.10 equiv) at 25° C., followed by heating to reflux (60° C.) for 16 h. LCMS (EW21491-265-P1A) showed that methyl (2S,4R,5R)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-4-((tert-butyldimethylsilyl)oxy)pyrrolidine-2-carboxylate was consumed, and the desired mass was detected (RT = 1.077 min). The resulting mixture was concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1, R f = 0.50, plate: petroleum ether / ethyl acetate = 1 / 1). (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (18.0 g, crude) was obtained as a brown oil. LCMS: RT = 1.077 min, m / z = 469.1 (M+H) of the product. + .

[0442] Step 9a: Synthesis of methyl (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0443] To a solution of methyl (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (23.0 g, 49.1 mmol, crude purity, 1.00 equiv.) in THF (220 mL) was added TBAF (1 M, 98.1 mL, 2.00 equiv.) at −10° C., followed by stirring at 0° C. for 1 h. TLC (petroleum ether / ethyl acetate = 3 / 1, plate 1) revealed the presence of (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate methyl ester (R f =0.30) was consumed, and the desired spot (R f = 0.00) was detected. The reaction mixture was poured into water (50.0 mL) and then extracted with ethyl acetate (50.0 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with petroleum ether / ethyl acetate = 2 / 1 (80.0 mL) at 20 °C for 0.5 h and then filtered. The filter cake was dried under reduced pressure. Methyl (1R,3S,6S,10aS,Z)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (5.20 g, 14.2 mmol, 12.1% yield for two steps, 96.7% purity) was obtained as a pale yellow solid. LCMS: RT=0.774 min, m / z=299.0 [M-55] of product + HPLC product RT = 1.910 min, purity 96.7%, 220 nm SFC: RT = 1.218 min, ee% = 100% (220 nm) 1H NMR: (400 MHz, CDCl3) δ 6.05 - 5.71 (m, 3H), 4.77 - 4.62 (m, 2H), 4.41 (d, J = 3.2 Hz, 1H), 4.30 - 4.22 (m, 1H), 3.75 (s, 3H), 2.89 - 2.78 (m, 1H), 2.71 - 2.55 (m, 2H), 2.46 (ddd, J = 3.0, 8.2, 16.8 Hz, 1H), 2.26 (ddd, J = 3.8, 9.2, 13.4 Hz, 1H), 2.16 - 1.95 (m, 2H), 1.44 (s, 9H).

[0444] Step 9b: Synthesis of methyl (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0445] To a solution of methyl (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (18.0 g, 38.4 mmol, crude purity, 1.00 equiv.) in THF (100 mL) was added TBAF (1 M, 76.8 mL, 2.00 equiv.) at −10° C., followed by stirring at 0° C. for 1 h. TLC (petroleum ether / ethyl acetate = 3 / 1, plate 1) revealed the presence of (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-((tert-butyldimethylsilyl)oxy)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate methyl ester (R f =0.30) was consumed, and the desired spot (R f=0.00) was detected. The reaction was poured into water (200 mL) and then extracted with ethyl acetate (100 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (TFA condition), then the pH was adjusted to about 7 with saturated aqueous NaHCO3 (50.0 mL) and extracted with ethyl acetate (500 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. Methyl (1R,3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-1-hydroxy-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (4.20 g, 11.4 mmol, 36.0% yield over two steps, 96.2% purity) was obtained as a yellow solid. LCMS: RT = 0.776 min, m / z = 299.0 (M-55) for the product. + HPLC: Product RT = 1.960 min, purity 96.2%, 220 nm SFC: RT = 1.483 min, ee% = 100% (220 nm) 1 H NMR: (400 MHz, CDCl3) δ 5.96 - 5.81 (m, 1H), 5.78 - 5.63 (m, 2H), 4.84 - 4.65 (m, 1H), 4.58 (t, J = 7.2 Hz, 1H), 4.16 (d, J = 4.6 Hz, 1H), 3.89 (d, J = 4.6 Hz, 1H), 3.72 (s, 3H), 3.40 - 2.82 (m, 1H), 2.65 (t, J = 16.8 Hz, 2H), 2.55 - 2.34 (m, 2H), 2.27 - 2.07 (m, 2H), 1.42 (s, 9H). Synthesis of (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid and (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0446] Step 1: Preparation of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0447] To a solution of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-5,9-dioxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (250 mg, 705 μmol, 1 equiv.) in anhydrous tetrahydrofuran (10 mL) was added ethylmagnesium chloride solution (2 M in THF) (720 μL, 1.44 mmol, 2.05 equiv.) at 0 °C. The reaction mixture was stirred at 0 °C for 2 h and then quenched with saturated aqueous NH4Cl. The mixture was extracted with EtOAc (2 × 15 mL), and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was obtained by eluting 30 g of C with 5–60% MeCN in water (containing 0.1% formic acid). 18 Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure and then freeze-dried to give methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (197 mg, 72.6%) as a white solid. LCMS (ESI): m / z = 385.2 [M+H] + . 1H NMR (400 MHz, CDCl3) δ 5.38 (d, J = 7.8 Hz, 1H), 5.08 - 4.96 (m, 1H), 4.58 (dd, J = 11.7, 7.6 Hz, 1H), 4.53 - 4.43 (m, 1H), 3.78 - 3.71 (m, 3H), 2.41 - 2.29 (m, 1H), 2.26 - 2.12 (m, 3H), 2.06 - 1.91 (m, 2H), 1.73 (dd, J = 12.1, 6.7 Hz, 1H), 1.68 - 1.57 (m, 2H), 1.54 - 1.46 (m, 4H), 1.44 - 1.38 (m, 9H), 0.91 (t, J = 7.5 Hz, 3H).

[0448] The intermediates shown in Table 40 were prepared according to the protocol described above in Step 1, with appropriate modifications. [Table 40]

[0449] Step 2: Preparation of methyl (3S,6S,10aR,E)-6-((tert-butoxycarbonyl)amino)-9-ethylidene-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0450] To a solution of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (197 mg, 512 μmol, 1 equiv.) in anhydrous toluene (5 mL) was added Burgess reagent (486 mg, 2.04 mmol, 4 equiv.). The reaction mixture was warmed to 70 °C and stirred for 1 h. The solvent was evaporated. The crude product was purified by elution with 50 g of C4H4O, eluting with 5-100% MeCN in water (containing 0.1% formic acid). 18Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure to give methyl (3S,6S,10aR,E)-6-((tert-butoxycarbonyl)amino)-9-ethylidene-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (158 mg, 84.4%) as a yellow oil. LCMS (ESI): m / z = 367.4 [M+H] + .

[0451] The intermediates shown in Table 41 were prepared according to the protocol described above in Step 2, with appropriate modifications. [Table 41]

[0452] Step 3: Preparation of methyl (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0453] To a solution of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethylidene-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (158 mg, 431 μmol, 1 equiv.) in methanol (5 mL) was added 10% palladium on carbon (50% wet) (136 mg, 129 μmol, 0.3 equiv.) while bubbling with nitrogen. Hydrogen gas was bubbled through the suspension for 5 minutes, and the reaction mixture was then stirred at room temperature under a hydrogen atmosphere (1 atm) for 20 hours. Nitrogen was bubbled through the suspension for 5 minutes. The reaction mixture was filtered through Celite (rinsing with MeOH), and the filtrate was concentrated under reduced pressure. The crude product was obtained by eluting 100 g of C with 5-100% MeCN in water (containing 0.1% formic acid). 18Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure and then freeze-dried to give methyl (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (86.0 mg, 54.4%, major isomer) as a white solid. LCMS (ESI): m / z = 369.2 [M+H] + . 1 H NMR (400 MHz, C6D6) δ 5.69 (d, J = 8.3 Hz, 1H), 4.91 - 4.84 (m, 1H), 4.45 (t, J = 8.7 Hz, 1H), 3.66 - 3.62 (m, 1H), 3.37 - 3.29 (m, 3H), 2.18 - 2.15 (m, 1H), 1.96 - 1.68 (m, 1H), 1.72 - 1.65 (m, 1H), 1.65 - 1.50 (m, 3H), 1.44 (s, 9H), 1.33 - 1.30 (m, 1H), 1.25 - 1.14 (m, 2H), 1.11 - 1.02 (m, 2H), 1.00 - 0.90 (m, 2H), 0.74 (t, J = 7.3 Hz, 3H). (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (10.0 mg, 6.32%, minor isomer) was also isolated as a white solid. LCMS (ESI): m / z = 369.2 [M+H] + . 1H NMR (400 MHz, C6D6) δ 6.20 (d, J = 6.8 Hz, 1H), 4.57 - 4.45 (m, 1H), 4.36 (t, J = 9.0 Hz, 1H), 3.62 - 3.50 (m, 1H), 3.31 (s, 3H), 2.27 - 2.22 (m, 1H), 1.77 - 1.65 (m, 2H), 1.63 - 1.47 (m, 3H), 1.45 (s, 9H), 1.37 - 1.29 (m, 1H), 1.25 - 1.12 (m, 3H), 1.11 - 1.01 (m, 1H), 0.95 - 0.88 (m, 1H), 0.83 (t, J = 7.5 Hz, 3H), 0.75 - 0.64 (m, 1H). Note: The absolute stereochemistry at the carbon atom bearing the ethyl group is unknown for both isomers.

[0454] The intermediates shown in Table 42 were prepared according to the protocol described above in Step 3, with appropriate modifications. Note: The absolute stereochemistry at the carbon atom bearing the isopropyl group is unknown for both isomers. [Table 42]

[0455] Step 4: Preparation of (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0456] To a solution of methyl (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (40 mg, 108 μmol, 1 equiv.) in tetrahydrofuran (6 mL) and water (2 mL) was added lithium hydroxide monohydrate (13.5 mg, 324 μmol, 3 equiv.). The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure (to remove THF). The crude product was purified by elution with 50 g of C using 5-60% MeCN in water (containing 0.1% formic acid). 18 Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure and then freeze-dried to give (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (32.0 mg, 83.7%) as a white solid. LCMS (ESI): m / z = 355.2 [M+H] + .

[0457] The intermediates shown in Table 43 were prepared according to the protocol described above in step 4, with appropriate modifications. [Table 43]

[0458] Step 4': Preparation of (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0459] To a solution of methyl (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (10 mg, 27.1 μmol, 1 equiv.) in tetrahydrofuran (1 mL) and water (0.2 mL) was added lithium hydroxide monohydrate (3.41 mg, 81.3 μmol, 3 equiv.). The reaction mixture was stirred at room temperature for 18 hours. 1 N HCl (1 mL) was added. The reaction mixture was concentrated under reduced pressure to give crude (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-9-ethyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (9.6 mg, 100%) as a white solid. LCMS (ESI): m / z = 355.2 [M+H] + .

[0460] The intermediates shown in Table 44 were prepared according to the protocols described above in Step 4 and Step 4', with appropriate modifications. [Table 44-1] Synthesis of (3S,6S,7S,10aS)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0461] Step 1: Preparation of (E)-ethyl 3-cyclopropylacrylate

[0462] To a suspension of 60% sodium hydride in mineral oil (658 mg, 16.5 mmol, 1.1 equiv.) in tetrahydrofuran (25 mL) at 0 °C, ethyl 2-(diethoxyphosphoryl)acetate (3.69 g, 16.5 mmol, 1.1 equiv.) was slowly added. The reaction mixture was warmed to room temperature and stirred for 30 minutes. A solution was observed. Cyclopropanecarbaldehyde (1.05 g, 15 mmol, 1 equiv.) was added dropwise at room temperature. An exotherm was observed, and a thick paste formed. Stirring was difficult, and a thick paste precipitated at the bottom of the flask. The reaction was stirred at room temperature for 20 hours. The reaction was quenched with saturated aqueous NaHCO3 (50 mL) and diluted with EtOAc (100 mL) and water (50 mL). The phases were separated, and the aqueous layer was re-extracted with EtOAc (2 × 75 mL). The combined organic layers were dried over sodium sulfate, filtered, adsorbed onto silica gel, and carefully concentrated under reduced pressure (care should be taken during concentration in vacuo as the product is volatile). The crude residue was purified by flash-chromatography on silica gel eluting with 1-25% DCM in pentane to give (£)-ethyl 3-cyclopropylacrylate (1.25 g, 59.5%) as a clear liquid. 1 H NMR (400 MHz, CDCl3) δ 6.44 (dd, J = 15.3, 9.7 Hz, 1H), 5.91 (d, J = 15.7 Hz, 1H), 4.15 (q, J = 7.2 Hz, 2H), 1.64 - 1.53 (m, 1H), 1.29 (t, J = 7.2 Hz, 3H), 0.99 - 0.93 (m, 2H), 0.69 - 0.62 (m, 2H).

[0463] Step 2: Preparation of (E)-3-cyclopropylprop-2-en-1-ol

[0464] To a solution of ethyl (E)-3-cyclopropylacrylate (1.59 g, 11.3 mmol, 1 equiv.) in diethyl ether (50 mL) at −78° C. was slowly added 1 M DIBAL-H in n-hexane (24.0 mL, 24.0 mmol, 2.13 equiv.). The reaction was stirred at −78° C. for 1 h and at room temperature for 2.25 h. The reaction was cooled to 0° C. and quenched by the addition of MeOH (15 mL), followed by 10% aqueous Rochelle's salt solution (70 mL). The mixture was stirred vigorously at room temperature for 1.5 h, and then the phases were separated. The organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure to give (E)-3-cyclopropylprop-2-en-1-ol (986 mg, 89.6%) as a clear liquid. 1 H NMR (400 MHz, CDCl3) δ 5.75 (dt, J = 15.2, 6.1 Hz, 1H), 5.29 - 5.21 (m, 1H), 4.09 (d, J = 5.8 Hz, 2H), 1.48 - 1.38 (m, 1H), 0.78 - 0.72 (m, 2H), 0.43 - 0.38 (m, 2H).

[0465] Step 3: Preparation of (E)-(tert-butoxycarbonyl)glycine 3-cyclopropylallyl

[0466] To a solution of (£)-3-cyclopropylprop-2-en-1-ol (1.05 g, 10.6 mmol, 1 equiv.) and (tert-butoxycarbonyl)glycine (1.85 g, 10.6 mmol, 1 equiv.) in methylene chloride (50 mL) was added dicyclohexylcarbodiimide (2.18 g, 10.6 mmol, 1.0 equiv.) followed by DMAP (403 mg, 1.05 mmol, 0.1 equiv.) at 0° C. The reaction mixture was allowed to warm to room temperature and stirred overnight.

[0467] The white precipitate was filtered off, and the filtrate was washed with 1 N HCl (50 mL) followed by saturated aqueous NaHCO3 (50 mL). The organic layer was adsorbed onto silica gel and concentrated under reduced pressure. The crude residue was purified by flash-chromatography on silica gel eluting with 0 to 50% EtOAc in heptane to give (E)-(tert-butoxycarbonyl)glycine 3-cyclopropylallyl (1.49 g, 55.1%) as a sticky off-white solid. 1 H NMR (400 MHz, CDCl3) δ 5.66 (dt, J = 15.4, 7.1 Hz, 1H), 5.34 - 5.29 (m, 1H), 5.05 - 4.97 (m, 1H), 4.61 - 4.57 (m, 2H), 3.98 - 3.82 (m, 2H), 1.47 (s, 9H), 1.44 - 1.39 (m, 1H), 0.80 - 0.74 (m, 2H), 0.46 - 0.40 (m, 2H).

[0468] Step 4: Preparation of racemic (2S,3S)-2-((tert-butoxycarbonyl)amino)-3-cyclopropylpent-4-enoic acid

[0469] To a solution of (E)-(tert-butoxycarbonyl)glycine 3-cyclopropylallyl (1 g, 3.91 mmol, 1 equiv.) in tetrahydrofuran (40 mL) at -78 °C, chlorotrimethylsilane (1.48 mL, 11.7 mmol, 3 equiv.) was added, followed by the dropwise addition of a 1 M solution of LiHMDS in THF (11.7 mL, 11.7 mmol, 3 equiv.). The reaction was maintained in an ice bath and stirred overnight, slowly warming to room temperature. The reaction was quenched with 1 N HCl (3 mL) and concentrated under reduced pressure. The crude residue was purified by elution with 5-80% MeCN in water (containing 0.1% formic acid). 18 Purification by reverse phase chromatography on a cartridge gave racemic (2S,3S)-2-((tert-butoxycarbonyl)amino)-3-cyclopropylpent-4-enoic acid (730 mg, 73.1%) as a yellowish viscous oil.1 H NMR (400 MHz, CDCl3) δ 5.83 - 5.68 (m, 1H), 5.28 - 5.10 (m, 3H), 4.51 - 4.43 (m, 1H), 1.93 - 1.82 (m, 1H), 1.45 (s, 9H), 0.94 - 0.82 (m, 1H), 0.62 - 0.49 (m, 2H), 0.34 - 0.24 (m, 1H), 0.19 - 0.08 (m, 1H).

[0470] Step 5: Preparation of methyl (2S,5R)-5-allyl-1-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-cyclopropylpent-4-enoyl)pyrrolidine-2-carboxylate

[0471] HATU (977 mg, 2.57 mmol, 1.2 equiv.) was added to a solution of crude methyl (2S)-5-allylpyrrolidine-2-carboxylate (434 mg, 2.57 mmol, eq.) HCl salt, racemic (2S,3S)-2-((tert-butoxycarbonyl)amino)-3-cyclopropylpent-4-enoic acid (550 mg, 2.15 mmol, eq.) and DIPEA (2.23 mL, 12.9 mmol, 6 equiv.) in N,N-dimethylformamide (2 mL). The reaction was stirred at room temperature for 15 min. The product was obtained by eluting with 5–80% MeCN in water (containing 0.1% formic acid). 18 Direct purification by reverse-phase chromatography on a cartridge gave methyl (2S,5R)-5-allyl-1-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-cyclopropylpent-4-enoyl)pyrrolidine-2-carboxylate (525 mg, 60.0%) as a thick clear oil. LCMS (ESI) m / z = 407.2 [M+H] + .

[0472] Step 6: Preparation of methyl (3S,6S,7R,10aR,Z)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0473] To a solution of methyl (2S,5R)-5-allyl-1-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-cyclopropylpent-4-enoyl)pyrrolidine-2-carboxylate (525 mg, 1.29 mmol, 1 equiv.) in methylene chloride (75 mL) under nitrogen was added Grubbs II catalyst (109 mg, 0.129 mmol, 0.1 equiv.). The reaction was heated to reflux for 48 hours. The reaction was concentrated under reduced pressure. The crude residue was purified by elution with 5-80% MeCN in water (containing 0.1% formic acid) to afford 100 g of C. 18 Purification by reverse-phase chromatography on a cartridge gave methyl (3S,6S,7R,10aR,Z)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (218 mg, 44.6%) as a brownish oil (complex mixture of diastereomers). LCMS (ESI) m / z = 379.2 [M+H] + .

[0474] Step 7: Preparation of methyl (3S,6S,7R,10aS)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0475] To a solution of methyl (3S,6S,7R,10aR,Z)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (218 mg, 0.5760 mmol, 1 equiv.) in ethanol (25 mL) under nitrogen was added 10% palladium on carbon (50% wet) (120 mg, 0.05638 mmol, 0.10 equiv.). Hydrogen gas was bubbled through the suspension for 2 minutes, and then the reaction was stirred at room temperature under a hydrogen atmosphere (1 atm) for 40 hours. Nitrogen was bubbled through the suspension, and the reaction was filtered through Celite (rinsing with MeOH). The filtrate was concentrated under reduced pressure. The crude residue was purified by eluting with 5-80% MeCN in water (0.1% formic acid). 18 Purification by reverse-phase chromatography on a cartridge gave methyl (3S,6S,7R,10aS)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (122 mg, 56%) as a white solid. LCMS (ESI) m / z = 381.4 [M+H] + . 1 H NMR (400 MHz, C6D6) δ 5.56 (d, J = 9.1 Hz, 1H), 4.95 (t, J = 9.5 Hz, 1H), 4.39 (t, J = 8.6 Hz, 1H), 3.74 - 3.65 (m, 1H), 3.36 (s, 3H), 2.39 - 2.29 (m, 1H), 1.80 - 1.70 (m, 1H), 1.68 - 1.50 (m, 5H), 1.46 (s, 9H), 1.20 - 1.06 (m, 1H), 1.05 - 0.78 (m, 4H), 0.74 - 0.61 (m, 1H), 0.27 - 0.16 (m, 2H), -0.23 - -0.31 (m, 1H).

[0476] Step 8: Preparation of (3S,6S,7S,10aS)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0477] To a solution of methyl (3S,6S,7R,10aS)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (122 mg, 0.3206 mmol, 1 equiv.) in tetrahydrofuran (3 mL) and water (1 mL) was added lithium hydroxide monohydrate (26.9 mg, 0.6412 mmol, 2 equiv.). The reaction was stirred for 20 h. The reaction was quenched with the addition of 1 N HCl (1 mL) and concentrated under reduced pressure to remove THF. The crude product was purified by elution with 5-80% MeCN in water (containing 0.1% formic acid). 18 Purification by reversed-phase chromatography on a cartridge gave (3S,6S,7S,10aS)-6-((tert-butoxycarbonyl)amino)-7-cyclopropyl-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (100 mg, 85.4%) as a white solid. LCMS (ESI) m / z = 367.1 [M+H] + . 1 H NMR (400 MHz, C6D6) δ 5.72 - 5.59 (m, 1H), 4.92 - 4.81 (m, 1H), 4.35 - 4.25 (m, 1H), 3.73 - 3.57 (m, 1H), 2.01 - 1.83 (m, 2H), 1.67 - 1.49 (m, 3H), 1.46 (s, 9H), 1.42 - 1.28 (m, 3H), 1.19 - 1.04 (m, 1H), 0.99 - 0.87 (m, 2H), 0.87 - 0.75 (m, 2H), 0.71 - 0.57 (m, 1H), 0.30 - 0.12 (m, 2H), -0.19 - -0.31 (m, 1H). Synthesis of (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylic acid and (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylic acid [ka]

[0478] Step 1: Preparation of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-cyclopropyl-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0479] To a solution of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-5,9-dioxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (490 mg, 1.38 mmol, 1 equiv.) in anhydrous tetrahydrofuran (10 mL) was added cyclopropylmagnesium bromide solution (0.5 M in THF) (5.52 mL, 2.76 mmol, 2 equiv.) at 0° C. The reaction mixture was stirred at 0° C. for 2 h. Cyclopropylmagnesium bromide solution (0.5 M in THF) (5.52 mL, 2.76 mmol, 2 equiv.) was added. The reaction mixture was stirred at 0° C. for 2 h and then quenched with saturated aq. NH4Cl. The product was extracted with EtOAc (2×15 mL), and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by eluting 50 g of C with 5-60% MeCN in water (containing 0.1% formic acid). 18Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure and then freeze-dried to give methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-cyclopropyl-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (420 mg, 76.7%) as a beige solid. LCMS (ESI): m / z = 397.2 [M+H] + .

[0480] Step 2: Preparation of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-cyclopropyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate

[0481] To a solution of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-cyclopropyl-9-hydroxy-5-oxodecahydropyrrolo[1,2-a]azocine-3-carboxylate (420 mg, 1.05 mmol, 1 equiv.) in anhydrous toluene (15 mL) was added Burgess reagent (1 g, 4.20 mmol, 4 equiv.). The reaction mixture was warmed to 70 °C and stirred for 1 h. The solvent was evaporated. The crude product was purified by elution with 100 g of C4H4O, eluting with 5-100% MeCN in water (containing 0.1% formic acid). 18 Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure to give methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-cyclopropyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (285 mg, 71.7%) as a beige solid. LCMS (ESI): m / z = 379.2 [M+H] + .

[0482] Step 3: Preparation of methyl (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylate and methyl (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylate

[0483] To a solution of methyl (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-cyclopropyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azocine-3-carboxylate (190 mg, 502 μmol, 1 equiv.) in ethanol (10 mL) was added 10% palladium on carbon (50% wet) (159 mg, 150 μmol, eq.) while bubbling with nitrogen. Hydrogen was bubbled through the suspension for 5 minutes, and then the reaction was stirred at 70° C. under hydrogen pressure (60 psi) for 18 hours. Nitrogen was bubbled through the suspension for 5 minutes. The reaction mixture was filtered through Celite (rinsing with EtOH), and the filtrate was concentrated under reduced pressure. The crude product was purified by eluting 100 g of C with 5-100% MeCN in water (containing 0.1% formic acid). 18 Purification was carried out by reverse-phase chromatography on a cartridge. The combined fractions were concentrated under reduced pressure and then freeze-dried to give methyl (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylate (107 mg, 55.7%, major isomer) as a white solid. LCMS (ESI): m / z = 383.2 [M+H] + . 1H NMR (400 MHz, C6D6) δ 5.71 (d, J = 8.3 Hz, 1H), 4.94 - 4.82 (m, 1H), 4.45 (t, J = 8.7 Hz, 1H), 3.69 - 3.60 (m, 1H), 3.34 (s, 3H), 2.22 - 2.11 (m, 1H), 2.02 - 1.91 (m, 1H), 1.71 - 1.52 (m, 3H), 1.44 - 1.42 (m, 9H), 1.43 (s, 9H), 1.35 - 1.25 (m, 2H), 1.22 - 1.16 (m, 1H), 1.15 - 1.05 (m, 2H), 1.04 - 0.89 (m, 5H), 0.83 (t, J = 7.1 Hz, 3H). (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylate (16.0 mg, 8.33%, minor isomer) was also isolated as a white solid. LCMS (ESI): m / z = 383.2 [M+H] + . 1 H NMR (400 MHz, C6D6) δ 6.21 (d, J = 7.1 Hz, 1H), 4.57 - 4.46 (m, 1H), 4.36 (t, J = 9.0 Hz, 1H), 3.63 - 3.53 (m, 1H), 3.31 (s, 3H), 2.30 - 2.21 (m, 1H), 1.85 - 1.75 (m, 1H), 1.74 - 1.64 (m, 1H), 1.62 - 1.53 (m, 2H), 1.52 - 1.47 (m, 1H), 1.45 (s, 9H), 1.29 - 1.19 (m, 3H), 1.13 - 1.05 (m, 3H), 0.95 - 0.85 (m, 5H), 0.71 - 0.64 (m, 1H). Note: The absolute stereochemistry at the carbon atom bearing the n-propyl group is unknown for both isomers.

[0484] Step 4: Preparation of (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0485] To a solution of methyl (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylate (90 mg, 235 μmol, 1 equiv.) in tetrahydrofuran (6 mL) and water (2 mL) was added lithium hydroxide monohydrate (17 mg, 405 μmol, 1.7 equiv.). The reaction mixture was stirred overnight at room temperature. 1 N HCl (1 mL) was added. The reaction was concentrated under reduced pressure (to remove THF). The crude product was purified by elution with 5-100% MeCN in water (containing 0.1% formic acid) to afford 30 g of C. 18 Purification by reverse-phase chromatography on a cartridge was performed. Pure fractions were evaporated and lyophilized to give (3S,6S,9R,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (65.0 mg, 75.1%) as a white solid. LCMS (ESI): m / z = 369.2 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 6.79 (d, J = 7.6 Hz, 1H), 4.55 - 4.28 (m, 1H), 4.25 - 4.15 (m, 2H), 2.25 - 1.99 (m, 2H), 1.94 - 1.78 (m, 2H), 2.28 - 1.78 (m, 4H), 1.76 - 1.44 (m, 6H), 1.40 - 1.28 (m, 11H), 1.22 - 1.14 (m, 2H), 0.86 (t, J = 7.2 Hz, 3H).

[0486] Step 4': Preparation of (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylic acid

[0487] To a solution of methyl (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylate (15 mg, 39.2 μmol, 1 equiv.) in tetrahydrofuran (1 mL) and water (0.2 mL) was added lithium hydroxide monohydrate (4.90 mg, 117 μmol, 3 equiv.). The reaction mixture was stirred at room temperature for 18 hours. 1 N HCl (1 mL) was added. The reaction was concentrated under reduced pressure (to remove THF). The crude product was purified by elution with 5-60% MeCN in water (containing 0.1% formic acid) to afford 50 g of C. 18 Purification by reversed-phase chromatography on a cartridge was carried out. Pure fractions were evaporated and lyophilized to give (3S,6S,9S,10aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-9-propyldecahydropyrrolo[1,2-a]azocine-3-carboxylic acid (13.0 mg, 90.2%) as a beige solid. LCMS (ESI): m / z = 369.2 [M+H] + . Synthesis of methyl (3S,6S,8S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate and methyl (3S,6S,8R,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate [ka]

[0488] Step 1: Preparation of methyl (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate

[0489] To a solution of Pd / C (wet) (0.50 g, 4.85 mmol, 0.3 equiv) in MeOH (50 mL) was added methyl (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (5.5 g, 16 mmol, 1 equiv) and the mixture was stirred at 25 °C under H (15 Psi) for 2 h. The reaction mixture was filtered to obtain the filtrate and concentrated under reduced pressure to give methyl (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (5.70 g, crude) as a yellow oil. 1 H NMR (400 MHz, CD3OD) δ 4.57 - 4.46 (m, 1H), 4.24 - 4.10 (m, 1H), 4.00 - 3.89 (m, 1H), 3.68 (d, J = 1.2 Hz, 3H), 2.32 - 2.18 (m, 2H), 2.14 - 2.06 (m, 1H), 1.99 - 1.93 (m, 1H), 1.84 - 1.65 (m, 4H), 1.60 - 1.53 (m, 1H), 1.40 (s, 9H), 1.15 (d, J = 7.2 Hz, 1H), 0.96 (d, J = 6.4 Hz, 2H).

[0490] Step 2: Preparation of methyl (3S,6S,8S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate and methyl (3S,6S,8R,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate

[0491] Methyl (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (5.70 g, 16.70 mmol, 1 equiv.) was purified by SFC (Method / IC-3-MeOH(DEA)-5-40-3 mL-35 T.1 cm) to give methyl (3S,6S,8S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (1.2 g, 3.5 mmol, 21% yield, peak 2) as a pale yellow oil. SFC (Rt = 1.342 min). 1 H NMR (400 MHz, CD3OD) δ 4.53 (d, J = 4.0 Hz, 1H), 4.44 (d, J = 3.2 Hz, 1H), 4.20 - 4.13 (m, 1H), 3.77 - 3.67 (m, 3H), 2.33 - 2.19 (m, 2H), 2.18 - 2.09 (m, 1H), 2.05 - 1.97 (m, 1H), 1.92 - 1.79 (m, 2H), 1.77 - 1.66 (m, 2H), 1.65 - 1.57 (m, 1H), 1.44 (s, 9H), 1.23 - 1.15 (m, 3H) and (3S,6S,8R,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxooctahydro-1H-pyrrolo[1,2-a]azepine-3-carboxylate (3.0 g, 8.8 mmol, 53% yield, peak 1) were obtained as a pale yellow oil. SFC (Rt = 1.138 min). 1 H NMR (400 MHz, CD3OD) δ 4.58 (d, J = 3.6 Hz, 1H), 4.23 (d, J = 11.2 Hz, 1H), 3.98 (d, J = 10.4 Hz, 1H), 3.72 (s, 3H), 2.35 - 2.25 (m, 1H), 2.18 - 2.08 (m, 1H), 2.06 - 1.91 (m, 3H), 1.89 - 1.79 (m, 2H), 1.79 - 1.69 (m, 2H), 1.44 (s, 9H), 1.00 (d, J = 6.4 Hz, 3H). [Table 44-2] * The individual isomers were separated by chiral SFC using the following conditions: Column: Chiralpak IC-3 50 x 4.6 mm, ID 3 um. Mobile phase: CO2 / MeOH / 0.05% DEA). Peak 1 rt = 1 min, Peak 2 rt = 1.135 min. Procedure for the synthesis of the linker: Synthesis of 4-nitrophenyl 5-((bis(2-(benzoylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate) [ka]

[0492] Step 1: Preparation of S-benzoic acid (2-hydroxyethyl)

[0493] To a solution of 2-mercaptoethanol (5.0 g, 64 mmol) and triethylamine (6.7 g, 64 mmol) in DCM (50 mL) was added benzoyl chloride (8.1 g, 58 mmol) in DCM (10 mL) at −70° C. The mixture was stirred at 25° C. for 12 hours to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give S-benzoate (2-hydroxyethyl) (5.60 g, 30.73 mmol, 48% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 8.09 - 8.03 (m, 2H), 7.63 - 7.55 (m, 1H), 7.50 - 7.42 (m, 2H), 4.46 (t, J = 6.4 Hz, 2H), 2.90 (td, J = 6.8, 8.4 Hz, 2H).

[0494] The following intermediates in Table 45 were prepared using those described above in Step 1 for the preparation of S-benzoate (2-hydroxyethyl) and utilizing the appropriate starting materials and modifications. [Table 45]

[0495] Step 2: Preparation of 4-nitrophenyl 5-((dichlorophosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0496] To a solution of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (0.50 g, 1.2 mmol) and DMF (42 mg, 0.58 mmol) in DCM (5 mL) was added oxalyl dichloride (0.44 g, 3.5 mmol) in DCM (5 mL) at 0° C., and the mixture was stirred at 40° C. for 1 hour to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give 4-nitrophenyl 5-((dichlorophosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (0.54 g, crude) as a yellow oil.

[0497] Step 3: Preparation of 4-nitrophenyl 5-((bis(2-(benzoylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0498] To a solution of 4-nitrophenyl 5-((dichlorophosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (0.54 g, 1.1 mmol) in DCM (5 mL) was added 2-hydroxyethyl S-benzoate (0.35 g, 1.9 mmol) dropwise at −78° C., and then triethylamine (0.27 g, 2.7 mmol) in DCM (5 mL) was added dropwise at −78° C. and stirred at −78° C. for 5 minutes to give a yellow solution. The mixture was purified by column chromatography to give 4-nitrophenyl 5-((bis(2-(benzoylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (0.20 g, 0.26 mmol, 25% yield) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 8.39 - 8.26 (m, 3H), 8.23 ​​- 8.06 (m, 1H), 8.06 - 7.95 (m, 5H), 7.85 - 7.73 (m, 1H), 7.62 - 7.54 (m, 2H), 7.53 - 7.40 (m, 6H), 4.52 - 4.40 (m, 4H), 3.39 - 3.15 (m, 4H).

[0499] The following intermediates in Table 46 were prepared using the method described above for the preparation of 4-nitrophenyl 5-((bis(2-(benzoylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate in Step 1 and utilizing the appropriate starting materials and modifications. [Table 46] Synthesis of 4-nitrophenyl 5-((bis(2-((2-methoxyacetyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate [ka]

[0500] Step 1: Preparation of S-2-Methoxyethanethioic acid (2-hydroxyethyl)

[0501] A solution of 2-methoxyacetyl chloride (1.5 g, 13.8 mmol, 1 equiv) and triethylamine (1.91 mL, 13.8 mmol, 1.0 equiv) in DCM (20 mL) was cooled to −78° C. under nitrogen, then 2-sulfanylethan-1-ol (1.07 g, 13.8 mmol, 1 equiv) was added dropwise. The reaction was stirred at −78° C. for 1 h, then allowed to warm to room temperature and stirred for 1 h. The reaction was filtered and rinsed with DCM (20 mL), then the filtrate was washed with water (30 mL). The organic layer was washed with DCM (2×20 mL), and the combined organic extracts were adsorbed onto silica gel and concentrated under reduced pressure. The residue was purified by flash-chromatography on silica gel eluting with 0-60% EtOAc in heptane to give S-2-hydroxyethyl 2-methoxyethanethioate (1.27 g, 61.3%) as a clear oil. 1 H NMR (400 MHz, CDCl3) d 4.11 (s, 2H), 3.78 (t, J = 6.2 Hz, 2H), 3.49 (s, 3H), 3.12 (t, J = 6.2 Hz, 2H).

[0502] Step 2: Preparation of S-2-Methoxyethanethioic acid (2-iodoethyl)

[0503] To a solution of 2-hydroxyethyl S-2-methoxyethanethioate (500 mg, 3.32 mmol, 1 equiv.) in 10 mL of THF at 0 °C was added triphenylphosphine (870 mg, 3.32 mmol, 1.0 equiv.), imidazole (452 ​​mg, 6.64 mmol, 2.0 equiv.), and iodine (842 mg, 3.32 mmol, 1.0 equiv.). The resulting brown solution was stirred at 0 °C for 1 h. The solution was adsorbed onto silica gel and concentrated under reduced pressure. The crude product was separated by flash chromatography on silica gel eluting with 0–30% EtOAc in heptane to give 2-iodoethyl S-2-methoxyethanethioate (757 mg, 87.7%) as a clear oil. 1H NMR (400 MHz, CDCl3) d 4.09 (s, 2H), 3.49 (s, 3H), 3.40 - 3.34 (m, 2H), 3.29 - 3.23 (m, 2H).

[0504] Step 3: Preparation of 4-nitrophenyl 5-((bis(2-((2-methoxyacetyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0505] Sodium hydroxide (74.3 mg, 1.86 mmol) in water (1 mL) was added dropwise to a stirred suspension of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (400 mg, 0.9317 mmol) in water (10 mL). When the mixture reached a pH of approximately 8, a clear yellowish solution was obtained, and silver nitrate (473 mg, 2.79 mmol) was added. A white precipitate was observed. After 2 h at room temperature, the white mixture was cooled to 0 °C, and the white precipitate was collected by filtration, washed with water, and transferred to a 100 mL flask with MeCN. The solvent was removed under reduced pressure, and the resulting beige solid was dried under reduced pressure. The powder was suspended in dry toluene (10 mL), and S-2-methoxyethanethiolate (2-iodoethyl) (787 mg, 3.02 mmol, 3.24 equiv.) was added. The mixture was stirred at 50° C. for 16 h. The reaction was adsorbed onto Celite and concentrated under reduced pressure. The crude residue was purified by flash-chromatography on silica gel eluting with a gradient of EtOAc (0 to 60%) in heptane to afford 4-nitrophenyl 5-((bis(2-((2-methoxyacetyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (178 mg, 27.5%) as a thick, clear oil. 1H NMR (400 MHz, CDCl3) d 8.39 - 8.34 (m, 3H), 8.23 ​​(s, 1H), 8.02 (d, J = 8.7 Hz, 1H), 7.76 (d, J = 8.7 Hz, 1H), 7.48 (d, J = 9.1 Hz, 2H), 4.35 - 4.17 (m, 4H), 4.09 (s, 4H), 3.49 (s, 6H), 3.26 - 3.11 (m, 4H). Synthesis of perfluorophenyl 5-((bis(2-((3-methoxypropanoyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate [ka]

[0506] Step 1: Preparation of 2-((tetrahydro-2H-pyran-2-yl)oxy)ethane-1-thiol

[0507] To a solution of 2-mercaptoethan-1-ol (9 g, 115 mmol, 1 equiv.) and 3,4-dihydro-2H-pyran (14.5 g, 173 mmol, 1.5 equiv.) in EtOH (200 mL) was added PPTS (2.9 g, 11.5 mmol, 0.1 equiv.). The reaction solution was stirred at 70 °C under N for 2 hours. Upon completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give 2-((tetrahydro-2H-pyran-2-yl)oxy)ethane-1-thiol (12 g, 74.0 mmol, 64%) as a colorless oil. LCMS (ESI) m / z = 163 [M+H] + .

[0508] Step 2: Preparation of S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 3-methoxypropanethioate

[0509] To a solution of 3-methoxypropanoic acid (5.5 g, 52.8 mmol, 1 equiv.) in DCM (30 mL) was added CDI (9.39 g, 58.0 mmol, 1.1 equiv.) at 0° C. The solution was stirred at room temperature for 30 minutes, and then 2-((tetrahydro-2H-pyran-2-yl)oxy)ethane-1-thiol (8.56 g, 52.8 mmol, 1 equiv.) was added to the solution. The solution was stirred at room temperature for 2 hours. After completion, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 3-methoxypropanethioate (8.00 g, 32.2 mmol, 61%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.65-4.60 (m, 1H), 3.92-3.74 (m, 3H), 3.71-3.64 (m, 2H), 3.62-3.46 (m, 2H), 3.38-3.30 (m, 3H), 3.21-3.08 (m, 2H), 2.88-2.78 (m, 2H), 1.82 (ddd, J = 15.1, 10.1, 3.4 Hz, 2H), 1.76-1.65 (m, 1H), 1.64-1.46 (m, 4H).

[0510] The following intermediates in Table 47 were prepared using the method described above for the preparation of S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 3-methoxypropanethioate in Step 2, utilizing the appropriate starting materials and modifications. [Table 47]

[0511] Step 3: Preparation of S-(2-hydroxyethyl) 3-methoxypropanethioate

[0512] To a solution of S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 3-methoxypropanethioate (8 g, 32.2 mmol, 1 equiv.) in EtOH (40 mL) was added PPTS (809 mg, 3.22 mmol, 0.1 equiv.). The reaction solution was stirred at 50° C. under N2 for 2 h. Upon completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to afford S-(2-hydroxyethyl) 3-methoxypropanethioate (4.00 g, 24.3 mmol, 76%) as a colorless oil. LCMS (ESI) m / z = 165 [M+H] + .

[0513] The following intermediates in Table 48 were prepared using the method described above for the preparation of S-(2-hydroxyethyl) 3-methoxypropanethioate in Step 3, utilizing the appropriate starting materials and modifications. [Table 48]

[0514] Step 4: Preparation of S-(2-iodoethyl) 3-methoxypropanethioate

[0515] To a solution of S-(2-hydroxyethyl) 3-methoxypropanethioate (1.4 g, 8.54 mmol, 1 equiv.) and triphenylphosphine (2.2 g, 8.54 mmol, 1.0 equiv.) in DCM (50 mL) was added 1-iodopyrrolidine-2,5-dione (1.92 g, 8.54 mmol, 1.0 equiv.) at 0° C. The solution was stirred at room temperature for 1 hour. Upon completion, the reaction solution was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give S-(2-iodoethyl) 3-methoxypropanethioate (1.3 g, 4.74 mmol, 56%) as a yellow oil. LCMS (ESI) m / z = 163 [M+H] + .

[0516] The following intermediates in Table 49 were prepared using the method described above for the preparation of S-(2-iodoethyl) 3-methoxypropanethioate in Step 4, utilizing the appropriate starting materials and modifications. [Table 49]

[0517] Step 5: Preparation of perfluorophenyl 5-((bis(2-((3-methoxypropanoyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0518] Amberlite IR120® (2.92 g, 127 mmol, 10 equiv.) in water (2 mL) was added dropwise to a stirred solution of (difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (6 g, 12.7 mmol, 1 equiv.) in HO (60 mL). When the mixture became clear (pH ∼9), silver(I) nitrate (4.75 g, 27.9 mmol, 2.2 equiv.) was added. After stirring at 0 °C for 2 h, the gray precipitate was collected by filtration and dried under reduced pressure. To a solution of the gray precipitate (2.2 g, 3.19 mmol, 1 equiv.) in MeCN (10 mL) was added 1S-(2-iodoethyl) 3-methoxypropanethioate (5, 2.00 g, 7.33 mmol, 2.3 equiv.). The solution was stirred at room temperature for 48 hours. After completion, the mixture was filtered, and the filtrate was purified by Prep-HPLC to give perfluorophenyl 5-((bis(2-((3-methoxypropanoyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (500 mg, 652 μmol, 20%) as a yellow oil. LCMS (ESI) m / z = 767 [M+H] + .

[0519] The following intermediates in Table 50 were prepared using the method described above for the preparation of perfluorophenyl 5-((bis(2-((3-methoxypropanoyl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate in Step 5 and utilizing the appropriate starting materials and modifications. [Table 50] Synthesis of perfluorophenyl 7-((bis(2-((2-(oxetan-3-yl)acetyl)thio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoate: [ka]

[0520] Step 1: Preparation of S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 2-(oxetan-3-yl)ethanethioate

[0521] To a solution of 2-(oxetan-3-yl)acetic acid (1.0 g, 8.62 mmol, 1.0 equiv.) and 2-((tetrahydro-2H-pyran-2-yl)oxy)ethane-1-thiol (1.5 g, 9.50 mmol, 1.1 equiv.) in DCM (30 mL) was added CDI (2.79 g, 17.2 mmol, 2.0 equiv.). The mixture was stirred at room temperature for 1 hour. After completion, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to afford S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 2-(oxetan-3-yl)ethanethioate (1.1 g, 4.23 mmol, 49%) as a white solid. LCMS (ESI) m / z = 261 [M+H] + .

[0522] Step 2: Preparation of S-(2-hydroxyethyl) 2-(oxetan-3-yl)ethanethioate

[0523] To a solution of S-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl) 2-(oxetan-3-yl)ethanethioate (1.1 g, 4.23 mmol, 1 equiv.) in EtOH (15 mL), 4-methylbenzene-1-sulfonic acid; pyridine (106 mg) was added at room temperature. The reaction solution was stirred at 50° C. under N2 for 5 hours. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give S-(2-hydroxyethyl) 2-(oxetan-3-yl)ethanethioate (600 mg, 3.40 mmol, 81%) as an oil. LCMS (ESI) m / z = 177 [M+H] + .

[0524] Step 3: Preparation of perfluorophenyl 7-((bis(2-((2-(oxetan-3-yl)acetyl)thio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoate

[0525] Oxalyl chloride (243 mg, 1.92 mmol, 3 equiv.) was added dropwise to a solution of (difluoro(7-((perfluorophenoxy)carbonyl)naphthalen-2-yl)methyl)phosphonic acid (300 mg, 0.64 mmol, 1 equiv.) in dry DCM (15 mL) and DMF (4.68 mg, 64 μmol, 0.1 equiv.) at 20° C. The reaction mixture was stirred at 40° C. for an additional 1 h. The reaction was monitored by pipetting a small sample and quenching it with MeOH to ensure complete formation of the bis-Cl phosphoryl chloride (bis-methoxyphosphonate was observed by LCMS). Upon completion, excess oxalyl chloride and solvent were removed under reduced pressure. The residue was redissolved in anhydrous DCM (2 mL) and then added to a mixture of S-(2-hydroxyethyl) 2-(oxetan-3-yl)ethanethioate (3, 281 mg, 1.60 mmol, 2.5 equiv) and DIEA (330 mg, 2.56 mmol, 4 equiv) in anhydrous DCM (2 mL) at 0 °C. The reaction was allowed to warm to 25 °C and stirred for an additional 2 h. Upon completion, the reaction mixture was quenched with HCl (0.3 mL, 2N) and concentrated under reduced pressure. The residue was purified by Biotage® C18 column chromatography to give perfluorophenyl 7-((bis(2-((2-(oxetan-3-yl)acetyl)thio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoate (50.0 mg, 63 μmol, 10%). LCMS (ESI) m / z = 785.0 [M+H] + . Synthesis of (E)-(((difluoro(4-(3-(4-nitrophenoxy)-3-oxoprop-1-en-1-yl)phenyl)methyl)phosphoryl)bis(oxy))bis(methylene) bis(2,2-dimethylpropanoate) [ka]

[0526] Step 1: Preparation of (E)-4-nitrophenyl 3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylate

[0527] Oxalyl chloride (1.13 g, 8.98 mmol, 3 equiv) was added dropwise to a solution of (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylic acid (1 g, 2.99 mmol, 1 equiv) in dry DCM (20 mL) and DMF (0.1 mL) at 0° C. The reaction mixture was stirred at 40° C. for an additional 1 h. Upon completion, excess oxalyl chloride and solvent were removed under reduced pressure. The residue was redissolved in anhydrous DCM (10 mL) and then added to a mixture of 4-nitrophenol (457 mg, 3.29 mmol, 1.1 equiv) and TEA (1.5 g, 15.0 mmol, 5 equiv) in anhydrous DCM (10 mL) at 0° C. The reaction was allowed to warm to 25° C. and stirred for an additional 2 h. Upon completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel to give (E)-4-nitrophenyl 3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylate (1 g, 2.20 mmol, 74%) as a white solid. LCMS (ESI): m / z = 456 [M+H] + .

[0528] Step 2: Preparation of (E)-(difluoro(4-(3-(4-nitrophenoxy)-3-oxoprop-1-en-1-yl)phenyl)methyl)phosphonic acid

[0529] To a solution of (E)-4-nitrophenyl 3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylate (1 g, 2.20 mmol, 1.0 equiv) in DCM (15 mL) was added TMSBr (6.7 g, 44 mmol, 20.0 equiv) dropwise at 0 °C. After the addition, the reaction mixture was allowed to warm to room temperature and stirred for an additional 12 h. The progress of the ...

Claims

1. Structural formula I: 【Chemistry 197】 or a pharmaceutically acceptable salt thereof, wherein in structural formula I: q is 0 or 1 and t is 0, 1, or 2, provided that at least one of q or t is 1; p is 1 or 2; The dotted lines represent single or double bonds; R １ is -CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ 8- to 10-membered fused bicyclic heteroaryl substituted with -CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ 8- to 10-membered fused bicyclic heterocyclyl substituted with CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ aryl substituted with -(C １ ~C ４ ) The aryl portion of alkyl(aryl) is —CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ ] is substituted with -(C １ ~C ４ ) alkyl (aryl), and -(C ２ ~C ４ ) The aryl portion of alkenyl (aryl) is —CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ ] is substituted with -(C ２ ~C ４ ) alkenyl(aryl); R １ａ and R ２ａ are each absent, hydrogen, cyano, (C １ ~C ４ ) alkyl, hydroxy (C １ ~C ４ ) independently selected from alkyl and fluoro; or R １ａ and R ２ａ together with the carbon to which they are attached form oxo; R １ｂ and R ２ｂ are each absent or hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -[(C １ ~C ４ ) alkyl]-OC(O)-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-C(O)O-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-O-[(C １ ~C ２０ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)-[halo(C １ ~C ４ ) alkyl], [(C １ ~C ４ ) alkyl]-OC(O)O-[5- to 7-membered heterocyclyl], [(C １ ~C ４ ) alkyl]-OC(O)-[5- to 7-membered heterocyclyl], -[(C １ ~C ４ ) alkyl]-OC(O)-[(C １ ~C ４ ) alkyl]-OH, -[(C １ ~C ４ ) alkyl]-OC(O)-[(C １ ~C ４ ) alkyl]-O-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)O-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)O-[halo(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)O-[(C １ ~C ４ ) alkyl]-OH, -[(C １ ~C ４ ) alkyl]-OC(O)O-[(C １ ~C ４ ) alkyl]-O-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-SC(O)-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-SC(O)-[halo(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-SC(O)-[(C １ ~C ４ ) alkyl]-OH, -[(C １ ~C ４ ) alkyl]-SC(O)-[(C １ ~C ４ ) alkyl]-O-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)NH(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)N[(C １ ~C ４ ) alkyl] ２ , 5- to 6-membered heteroaryl, and aryl, wherein said 5- to 6-membered heteroaryl and aryl each, when valences allow, are selected from halo, cyano, and (C １ ~C ４ ) alkyl, and the [(C １ ~C ４ ) alkyl]-OC(O)O-[5- to 7-membered heterocyclyl] and [(C １ ~C ４ ) alkyl]-OC(O)-[5- to 7-membered heterocyclyl], where valence allows, each 5- to 7-membered heterocyclyl is C(O)OR ｈ and optionally substituted independently with 1 to 2 groups selected from: R ２ is hydrogen, halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, hydroxy (C １ ~C ４ ) selected from alkyl, cyano, and hydroxyl; R ３ and R ４ are each independently hydrogen, halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, hydroxy (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkylphenyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, -(C １ ~C ４ ) alkyl(C １ ~C ４ ) alkoxy, hydroxyl, cyano, -NR ａ R ｂ , phenyl, (C ３ ~C ６ ) cycloalkyl, 5- to 6-membered heteroaryl, and 4- to 6-membered heterocyclyl, wherein said phenyl, (C ３ ~C ６ ) Cycloalkyl, 5- to 6-membered heteroaryl, and 4- to 6-membered heterocyclyl each, when valence allows, R Ｓ optionally substituted with 1 to 3 groups selected from Or R ３ and R ４ and are together on the same carbon atom, (C ３ ~C ６ ) cycloalkyl or 4- to 6-membered heterocyclyl, each of which, when valences permit, is selected from halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, and halo (C １ ~C ４ ) optionally substituted with 1 to 3 groups selected from alkoxy; Each R5 is independently selected from hydrogen and (C １ ~C ４ ) alkyl; R ６ and R ７ together with the nitrogen atom to which they are attached form a 4- to 14-membered monocyclic or bicyclic heterocyclyl or a 5- to 12-membered monocyclic or bicyclic heteroaryl, each of which, when valences allow, R Ｑ optionally substituted with 1 to 3 groups selected from AA is an α- or β-, natural or unnatural, amino acid residue; R Ｔ is (C １ ~C ４ ) alkyl, benzyl, and phenyl, wherein said phenyl is selected from halo, (C １ ~C ４ ) alkyl, and halo(C １ ~C ４ ) alkyl; R Ｑ Halo, (C ２ ~C ４ ) alkenyl, (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, cyano, phenyl, hydroxyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C ３ ~C ６ ) cycloalkyl, oxo, imino, —O(phenyl), —C(O)R ｇ , -C(O)OR ｅ , -NHC(O)R ｅ , —C(O)NR ｃ R ｄ , -NR ａ R ｂ , -S(O)R ｅ R ｆ , -S(O) ２ R ｆ , -S(O)=NH(C １ ~C ４ ) alkyl, —S(O)NR ｅ R ｆ , and -S(O) ２ NR ｅ R ｆ wherein the (C ２ ~C ４ ) alkenyl and (C １ ~C ４ ) alkyl, when valence allows, R Ｍ and the phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C ３ ~C ６ ) cycloalkyl, and 4- to 6-membered heterocyclyl each, when valence allows, R Ｆ and optionally substituted with 1 to 3 groups independently selected from: R Ｊ and R Ｍ are each independently halo, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, cyano, —C(O)R ｇ , -C(O)OR ｅ , -NHC(O)R ｅ , —C(O)NR ｃ R ｄ , -NR ａ R ｂ , -S(O)R ｅ R ｆ , -S(O) ２ R ｆ , -S(O)NR ｅ R ｆ , -S(O)=NH(C １ ~C ４ ) alkyl, —S(O) ２ NR ｅ R ｆ , hydroxyl, phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein said phenyl, 4- to 6-membered heterocyclyl, and 5- to 10-membered monocyclic or bicyclic heteroaryl each, when valences allow, are selected from R Ｘ optionally substituted with 1 to 3 groups selected from R Ｆ , R Ｓ , and RX each independently represent halo, cyano, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl(C １ ~C ４ ) alkoxy, hydroxy (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkylphenyl, (C ２ ~C ４ ) alkenyl, halo(C ２ ~C ４ ) alkenyl, (C ２ ~C ４ ) alkynyl, halo(C ２ ~C ４ ) alkynyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, hydroxyl, oxo, imino, phenyl, -S(O)R ｅ R ｆ , -S(O) ２ R ｆ , -S(O)=NH(C １ ~C ４ ) alkyl, —S(O)NR ｅ R ｆ , and -S(O) ２ NR ｅ R ｆ , -C(O)OR ｅ , -NR ｃ C(O)R ｅ , -C(O)R ｇ , —C(O)NR ｃ R ｄ , and -NR ａ R ｂ wherein the phenyl and the group -(C １ ~C ４ ) alkylphenyl, each phenyl, when valences permit, may be halo, cyano, (C １ ~C １０ ) alkyl, (C ２ ~C １０ ) alkenyl, (C ２ ~C １０ ) alkynyl, halo(C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, and halo (C １ ~C １０ ) alkoxy, wherein the (C １ ~C １０ ) alkyl, (C ２ ~C １０ ) alkenyl and (C ２ ~C １０ ) each alkynyl is optionally substituted, where valences allow, with a 5- to 10-membered monocyclic or bicyclic heteroaryl or a 4- to 10-membered monocyclic or bicyclic heterocyclyl, wherein each of said 5- to 10-membered monocyclic and bicyclic heteroaryl or 4- to 10-membered monocyclic or bicyclic heterocyclyl is optionally substituted with oxo or a 5- to 7-membered heterocyclyl optionally substituted with 1 to 2 oxo; and R ａ , R ｂ , R ｃ , R ｄ , R ｅ , R ｆ , R ｇ , and R ｈ are each independently, when valence allows, hydrogen, (C １ ~C ４ ) alkyl, phenyl, (C ３ ~C ６ ) cycloalkyl, 4- to 6-membered heterocyclyl, and 5- to 6-membered heteroaryl, wherein said (C １ ~C ４ ) alkyl, when valence allows, R Ｊ and the phenyl, (C ３ ~C ６ ) cycloalkyl, 4- to 6-membered heterocyclyl, and 5- to 6-membered heteroaryl are each independently selected from halo, cyano, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) optionally substituted with 1 to 3 groups selected from alkoxy, hydroxyl, phenyl, and benzyl; The compound or a pharmaceutically acceptable salt thereof.

2. The compound has structural formula II: 【Chemistry 198】 2. The compound of claim 1, wherein:

3. The compound has the structure III, IV, V, or VII: 【Chemistry 199】 2. The compound of claim 1, wherein:

4. The compound has the structure VIII, VIII', IX, X, XI, XII, or XIII: 【Chemistry 200】 2. The compound of claim 1, wherein:

5. 2. The compound of claim 1, wherein the compound has the structure XIV, XV, XVI, XVI′, XVII, XVIII, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, or XXVIII: 【Chemical Engineering 201】 【Chemical Engineering 202】 2. The compound of claim 1, wherein:

6. 10. The compound of claim 1, wherein the compound has structural formula XXX, XXXI, XXXII, XXXIII XXXIV, XXXV, XXXVI, or XXVII: 【Chemical 203】 2. The compound of claim 1, wherein:

7. R ３ is hydrogen, (C １ ~C ４ ) alkyl, hydroxyl, (C １ ~C ４ ) alkoxy, -(C １ ~C ４ ) alkylphenyl, and 4- to 6-membered heterocyclyl; or R ３ and R ４ and are together on the same carbon atom, (C ３ ~C ６ ) forming a cycloalkyl; or a pharmaceutically acceptable salt thereof.

8. R ４ But hydrogen (C １ ~C ４ ) alkyl, and hydroxyl; or R ３ and R ４ and are together on the same carbon atom, (C ３ ~C ６ ) forming a cycloalkyl; or a pharmaceutically acceptable salt thereof.

9. R ３ is hydrogen, (C １ ~C ２ ) alkyl, hydroxyl, (C １ ~C ２ ) selected from alkoxy, benzyl, and azetidinyl; or R ３ and R ４ and are taken together on the same carbon atom to form cyclopropyl, or a pharmaceutically acceptable salt thereof.

10. R ３ and R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

11. R ２ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is selected from hydrogen and hydroxyl.

12. R ２ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

13. R ５ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

14. R １ is selected from 8- to 10-membered fused bicyclic heteroaryl and aryl, each of which is —CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

15. R １ is selected from benzothiophenyl, indolyl, and naphthalenyl, each of which is —CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ or -CR １ａ R ２ａ P(O) [OR １ｂ ][NH(AA)C(O)OR Ｔ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

16. R １ but, 【Chemical 204】 【Chemical 205】 2. The compound of claim 1 selected from: or a pharmaceutically acceptable salt thereof.

17. R １ but, 【Chemical 206】 2. The compound of claim 1, wherein:

18. R １ａ is hydrogen and R ２ａ is fluoro or R １ａ is fluoro and R ２ａ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is fluoro.

19. R １ａ and R ２ａ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is fluoro.

20. R １ｂ and R ２ｂ are each independently hydrogen, (C １ ~C ４ ) alkyl, -[(C １ ~C ４ ) alkyl]-OC(O)-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-OC(O)O-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-SC(O)-[(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-SC(O)-[halo(C １ ~C ４ ) alkyl], -[(C １ ~C ４ ) alkyl]-SC(O)-[(C １ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the phenyl, pyridinyl, and naphthalenyl are each independently optionally substituted with cyano.

21. R １ｂ and R ２ｂ are respectively -[(C １ ~C ４ ) alkyl]-OC(O)-[(C １ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

22. R １ｂ and R ２ｂ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

23. -CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ が、 【Chemical 207】 【Chemical 208】 21. The compound of claim 1 or 20, selected from: or a pharmaceutically acceptable salt thereof.

24. -CR １ａ R ２ａ P(O)OR １ｂ OR ２ｂ が、 【Chemical Engineering 209】 21. The compound of claim 1 or 20, wherein:

25. -NH[AA]C(O)OR Ｔ is -NHC(R')(R)C(O)R Ｔ or -NHC(R')(R)CH ２ C(O)R Ｔ where R ’ is hydrogen, (C １ ~C ３ ) alkyl, or (C １ ~C ３ ) alkyl(C １ ~C ３ ) alkoxy, and R is hydrogen, methyl, isopropyl, —CH ２ CH (CH ３ ) ２ , -(CH ２ ) ２ SCH ３ , -CH(CH ３ ) (CH ２ CH ３ ), C.H. ２ OH, -CH(OH)(CH ３ ), C.H. ２ SH, -CH ２ C(O)NH ２ , -(CH ２ ) ２ C(O)NH ２ , benzyl, p-hydroxybenzyl, -CH ２ (indolyl), -(CH ２ ) ４ NH ２ , -(CH ２ ) ３ NHC (=NH ２ ) NH ２ , -CH ２ (imidazolyl), -(CH ２ ) COOH, and -(CH ２ ) ２ COOH; or R is selected from -NHC(R')(R)C(O)R Ｔ or -NHC(R')(R)CH ２ C(O)R Ｔ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R is a substituted or unsubstituted aryl group, ...

26. R ６ and R ７ and together with the nitrogen atom to which they are attached form pyrrolidinyl, azetidinyl, piperazinyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl, 6-thia-2,7-diazaspiro[3.4]octanyl, 2-thia-6-azaspiro[3.3]heptanyl, 4-azaspiro[2.4]heptanyl, spiro[indoline-3,3′-pyrrolidinyl], or 1,2,3,4,5,6-hexahydro-2,6-naphthyridinyl, each of which, when valences allow, is selected from the group consisting of R Ｑ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

27. R ６ and R ７ and together with the nitrogen atom to which they are attached form pyrrolidinyl or azetidinyl, each of which, when valences permit, may be selected from R Ｑ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

28. R Ｑ But, Halo, (C ２ ~C ４ ) alkenyl, (C １ ~C ４ ) alkyl, cyano, phenyl, hydroxyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C ３ ~C ６ ) cycloalkyl, oxo, imino, —O(phenyl), —C(O)R ｇ , -NHC(O)R ｅ , -S(O)=NH(C １ ~C ４ ) alkyl, and —S(O) ２ NR ｅ R ｆ wherein the (C ２ ~C ４ ) alkenyl and (C １ ~C ４ ) alkyl, when valence allows, R Ｍ and wherein the phenyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, and (C ３ ~C ６ ) Each cycloalkyl, when valence allows, is R Ｆ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups independently selected from:

29. R Ｑ But, Halo, (C ２ ~C ４ ) alkenyl, (C １ ~C ４ ) alkyl, cyano, phenyl, hydroxyl, morpholinyl, dihydropyridinyl, tetrahydro-2H-thiopyranyl, pyridinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, triazolyl, indazolyl, benzimidazolyl, pyrazolo[3,4-b]pyridinyl, cyclohexyl, cyclopropyl, oxo, imino, —O(phenyl), —C(O)R ｇ , -NHC(O)R ｅ , -S(O)=NH(C １ ~C ４ ) alkyl, and —S(O) ２ NR ｅ R ｆ wherein the (C ２ ~C ４ ) alkenyl and (C １ ~C ４ ) alkyl, when valence allows, R Ｍ and said phenyl, morpholinyl, dihydropyridinyl, tetrahydro-2H-thiopyranyl, pyridinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, triazolyl, indazolyl, benzimidazolyl, pyrazolo[3,4-b]pyridinyl, cyclohexyl, and cyclopropyl are each optionally substituted, where valences allow, with 1 to 3 groups selected from R Ｆ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups independently selected from:

30. R Ｍ is a 4- to 6-membered heterocyclyl, a 5- to 6-membered monocyclic heteroaryl, —S(O)═NH(C １ ~C ４ ) alkyl, cyano, and phenyl, wherein said 4- to 6-membered heterocyclyl, 5- to 6-membered monocyclic heteroaryl, and phenyl each, when valences allow, are selected from R Ｘ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

31. R Ｍ tetrahydropyranyl, pyrazolyl, -S(O)=NH(C １ ~C ４ ) alkyl, cyano, and phenyl, wherein said tetrahydropyranyl, pyrazolyl, and phenyl are each selected from R Ｘ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

32. R Ｘ However, (C １ ~C ４ ) alkyl, (C １ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R is selected from: alkoxy;

33. R Ｆ But, Halo, (C １ ~C ４ ) alkyl, hydroxy (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, (C ２ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: R is selected from alkynyl, cyano, oxo, and imino.

34. R ｇ However, (C １ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R is selected from alkyl, morpholinyl, imidazolyl, benzyl, and cyclopropyl.

35. R ｅ and R ｆ are each independently １ ~C ４ ) alkyl and halo(C １ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

36. R ｃ and R ｄ are each independently hydrogen and (C １ ~C ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

37. 2. The compound of claim 1, wherein the compound is selected from any one of compounds 1 to 1127, or a pharmaceutically acceptable salt thereof.

38. 10. A pharmaceutically acceptable composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Images