STAT modulators and uses thereof

Abstract

Compounds of Formula (I), as well as pharmaceutically acceptable salts and compositions thereof, are provided, which are useful in the treatment of various conditions associated with STAT3 and / or STAT6. # imgabs0 #

Description

[0001] Related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 325,908, filed March 31, 2022, and U.S. Provisional Application No. 63 / 337,388, filed May 2, 2022, the entire contents of each of these U.S. Provisional Applications are incorporated herein by reference. Background Technology

[0003] The protein signal transducer and activator of transcription (STAT) family consists of transcription factors that play important roles in the regulation of cellular processes such as proliferation, differentiation, apoptosis, and angiogenesis. Seven STAT genes have been identified in the human genome: STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b, and STAT6.

[0004] STAT3 has received particular attention because it is closely associated with the promotion of tumor growth and immune evasion, and is the only STAT family member whose gene deletion leads to embryonic lethality. In fact, it is estimated that abnormally elevated STAT3 activity occurs in more than 70% of human cancers. Activated STAT3 mediates key gene expression changes and molecular events that dysregulate cell growth and apoptosis, promotes angiogenesis, invasion, metastasis, and the development of apoptosis resistance, and inhibits host immune surveillance of tumors, thereby making constitutively active STAT3 a key mediator of oncogenicity and tumor progression.

[0005] Another STAT protein that has recently attracted attention is STAT6. Recent studies have shown that STAT6 signaling is crucial for IL-4-induced and IL-13-induced epithelial-mesenchymal transition (EMT) and the invasiveness of colorectal cancer cells (CRC). STAT6 is involved in several aspects of inflammatory diseases and other related conditions.

[0006] Given their role in regulating cellular processes, regulating the activity of one or more STAT proteins, particularly STAT3 and / or STAT6, represents a key area of ​​research for the treatment of cancer, inflammatory conditions, and other diseases. Summary of the Invention

[0007] This document provides regulators for STAT3 and / or STAT6. These regulators comprise regulators having Formula I:

[0008]

[0009] And its pharmaceutically acceptable salts and combinations thereof, wherein R 1 R 2 R3 R 4 R 5 R 6 R 7 R 8 q, t, and p are as described in this article.

[0010] On the one hand, the disclosed Formula I compound and its pharmaceutically acceptable salt inhibit STAT3 and / or STAT6 and can be used for a variety of therapeutic applications, such as the treatment of cancer and inflammatory conditions.

[0011] It also includes pharmaceutical compositions comprising compounds of formula I disclosed and pharmaceutically acceptable salts, and methods for their preparation.

[0012] It also includes methods for treating symptoms that respond to regulation of STAT3 and / or STAT6 using the disclosed compounds, pharmaceutically acceptable salts, and combinations thereof. Detailed Implementation

[0013] 1. General description of the compound

[0014] In a first embodiment, this document provides a compound having structural formula I:

[0015]

[0016] Or its pharmaceutically acceptable salt, wherein:

[0017] q is 0 or 1, and t is 0, 1 or 2, provided that at least one of q or t is 1;

[0018] p is 1 or 2;

[0019] R 1 Selected from the following: -CR 1a R 2a P(O)OR 1b OR 2b -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b -[P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R2a P(O)[NH(AA)C(O)OR T [NH(AA)C(O)OR T or -P(O)[OR 1b [NH(AA)C(O)OR T substituted 8- to 10-membered fused bicyclic heteroaryl; substituted by -CR 1a R 2a P(O)OR 1b OR 2b 、-CR 1a R 2a P(O)[OR 1b [NH(AA)C(O)OR T 、-CR 1a R 2a P(O)[NHR Ty [NH(AA)C(O)OR T 、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty [NH(AA)C(O)OR T 、-CR 1a R 2a P(O)[NH(AA)C(O)OR T [NH(AA)C(O)OR T or -P(O)[OR 1b [NH(AA)C(O)OR T substituted 8- to 10-membered fused bicyclic heterocyclic; substituted by -CR 1a R 2a P(O)OR 1b OR[[ID=-60]] 2b 、-CR 1a R 2a P(O)[OR 1b [NH(AA)C(O)OR T 、-CR 1a R 2a P(O)[NHR Ty [NH(AA)C(O)OR T 、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty [NH(AA)C(O)OR T 、-CR 1a R 2a P(O)[NH(AA)C(O)OR T [NH(AA)C(O)ORT ] or -P(O)[OR 1b ][NH(AA)C(O)OR T Substituted aryl; -(C1-C4)alkyl (aryl), wherein the aryl moiety of the -(C1-C4)alkyl (aryl) is replaced by -CR 1a R 2a P(O)OR 1b OR 2b -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b -[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -CR 1a R 2a P(O)[NH(AA)C(O)OR T ]]

[0020] [NH(AA)C(O)OR T ]、-P(O)[OR 1b ][NH(AA)C(O)OR T Substitution; and -(C2-C4)alkenyl (aryl), wherein the aryl moiety of -(C2-C4)alkenyl (aryl) is replaced by -CR 1a R 2a P(O)OR 1b OR 2b –CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b -[P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T]][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]replace;

[0021] R 1a and R 2a Each is independently selected from hydrogen, cyano, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, and fluorine; or R 1a and R 2a It forms an oxygen with the carbon it is attached to;

[0022] R 1b and R 2b Each is independently selected from 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 [(C1-C4)alkyl], [(C1-C4)alkyl]-OC(O)-[halogenated(C1-C4)alkyl], [(C1-C4)alkyl]-OC(O)O-[5- to 7-membered heterocyclic], [(C1-C4)alkyl]-OC(O)-[5- to 7-membered heterocyclic], [(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-[halogenated(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)alkylphenyl]-C(O)O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)-[NH(AA)C(O)OR T], -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl], -[(C1-C4)alkyl]-SC(O)-[halogenated(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 The 5- to 6-membered heteroaryl group and the aryl group are optionally and independently substituted by 1 to 2 groups selected from halogenated, cyanoated, and (C1-C4)alkyl groups, wherein, where the valence allows, the 5- to 7-membered heteroaryl group of [(C1-C4)alkyl]-OC(O)O-[5- to 7-membered heterocyclic group] and [(C1-C4)alkyl]-OC(O)-[5- to 7-membered heterocyclic group] are optionally and independently substituted by 1 to 2 groups selected from C(O)OR h Substitution of groups;

[0023] R 2 Selected from hydrogen, halogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, cyano and hydroxy;

[0024] R 3 and R 4 Each is independently selected from hydrogen, halogenated, and (C1-C4)alkyl groups;

[0025] R 5 and R 6 Each is independently selected from hydrogen, phenyl, and (C1-C4) alkyl;

[0026] R 7 The group is selected from (C1-C4) alkyl, phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic groups and 5- to 10-membered monocyclic or bicyclic heteroaryl groups, wherein, where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 groups selected from R Y The phenyl group is substituted, and where the valence allows, the phenyl group, the 4- to 9-membered monocyclic or bicyclic heterocyclic group, and the 5- to 10-membered monocyclic or bicyclic heteroaryl group are each optionally replaced by 1 to 3 groups selected from R Z Substitution of groups; or

[0027] R 6 and R 7Together with the nitrogen atom to which it is attached, it forms a 4- to 14-membered monocyclic or bicyclic heterocyclic group or a 5- to 12-membered monocyclic or bicyclic heteroaryl group, wherein, where the valence allows, each of the 4- to 14-membered monocyclic or bicyclic heterocyclic group or the 5- to 12-membered monocyclic or bicyclic heteroaryl group is optionally surrounded by 1 to 3 atoms selected from R Q Substitution of groups;

[0028] R 8 It is hydrogen or (C1-C4) alkyl;

[0029] AA is a residue of α or β natural or non-natural amino acids;

[0030] R T and R Ty Each group is independently selected from (C1-C4)alkyl, (C1-C4)alkyl-C(O)O(C1-C4)alkyl, benzyl and phenyl, wherein the phenyl group is optionally substituted by one or two groups selected from halogenated, (C1-C4)alkyl and halogenated (C1-C4)alkyl;

[0031] R Q Selected from halogenated, (C2-C4)alkenyl, (C1-C4)alkyl, (C1-C4)alkoxy, halogenated (C1-C4)alkoxy, cyano, phenyl, hydroxyl, 4- to 9-membered monocyclic or bicyclic heterocyclic groups, 5- to 10-membered monocyclic or bicyclic heteroaryl groups, (C3-C6)cycloalkyl, oxo, imino, -OR e -C(O)R g -C(O)OR e -NR c C(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)2NR e R f Where the valence allows, the (C2-C4) alkenyl and the (C1-C4) alkyl are each optionally and independently composed of 1 to 3 molecules selected from R M The group is substituted, and wherein, where the valence allows, the phenyl group, the 5- to 10-membered monocyclic or bicyclic heteroaryl group, the (C3-C6) cycloalkyl group, and the 4- to 9-membered monocyclic or bicyclic heterocyclic group are each optionally and independently replaced by 1 to 3 groups selected from R F Substitution of groups;

[0032] RY Selected from halogenated, (C1-C4)alkoxy, halogenated (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 heterocyclic group and 5 to 10-membered monocyclic or bicyclic heteroaryl, wherein, where the valence allows, the phenyl, the 4 to 6-membered heterocyclic group and the 5 to 10-membered monocyclic or bicyclic heteroaryl group are each optionally composed of 1 to 3 groups selected from R X Substitution of groups;

[0033] R M and R J Each is independently selected from halogenated, (C1-C4)alkyl, (C1-C4)alkoxy, halogenated (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)=NR e (C1-C4)alkyl, -S(O)2NR e R f , hydroxyl, phenyl, 4 to 6-membered heterocyclic group and 5 to 10-membered monocyclic or bicyclic heteroaryl, wherein, where the valence allows, the phenyl, the 4 to 6-membered heterocyclic group and the 5 to 10-membered monocyclic or bicyclic heteroaryl group are each optionally composed of 1 to 3 groups selected from R X Substitution of groups;

[0034] R F R X and R ZEach is independently selected from halogen, cyano, (C1-C4)alkyl, cyano(C1-C4)alkyl, C3-C6 cycloalkyl, halogen(C1-C4)alkyl, -(C1-C4)alkylC(O)NR c R d -(C1-C4)alkyl(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, -(C1-C4)alkylphenyl, -(C1-C4)alkylheteroaryl, (C2-C4)alkenyl, halo(C2-C4)alkenyl, (C2-C4)ynyl, halo(C2-C4)ynyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, -OR e , oxo, imino, phenyl, 4- to 6-membered heterocyclic groups, 5- to 6-membered monocyclic heteroaryl groups, -S(O)R e R f -S(O)2R f -S(O)=NH(C1-C4)alkyl, -S(O)NR e R f -S(O)2NR e R f -C(O)OR e -NR c C(O)R e -(C1-C4 alkyl)C(O)R g -C(O)R g -C(O)NR c R d NO2 and -NR a R b Where the valence allows, the phenyl group, the 4- to 6-membered heterocyclic group, and the phenyl group of the -(C1-C4)alkylphenyl group are each optionally and independently substituted by 1 to 3 groups selected from the following: halogenated, cyanoated, oxoated, (C1-C4)alkylphenyl groups. 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 ) alkynyl, halogenated (C1-C 10 )alkyl, (C1-C 10 )alkoxy, -(C1-C4)alkyl (C1-C4)alkoxy and halogen (C1-C4)alkoxy 10 )alkoxy, wherein, where the valence allows, the (C1-C 10 )alkyl, the (C2-C 10 )alkenyl and the (C2-C 10Each of the alkynyl groups is optionally substituted with a 5- to 10-membered monocyclic or bicyclic heteroaryl group or a 4- to 10-membered monocyclic or bicyclic heterocyclic group, each of the 5- to 10-membered monocyclic and bicyclic heteroaryl groups or the 4- to 10-membered monocyclic or bicyclic heterocyclic groups being optionally substituted with an oxo group or a 5- to 7-membered heterocyclic group being optionally substituted with one or two oxo groups; and

[0035] When valence is permissible, R a R b R c R d R e R f R g and R h Each is independently selected from hydrogen, (C1-C4)alkyl, (C 2- C4) alkynyl, -(C1-C4) alkylphenyl, phenyl, (C3-C6) cycloalkyl, 4- to 6-membered heterocyclic and 5- to 6-membered heteroaryl, wherein, where the valence permits, the (C1-C4) alkyl is optionally surrounded by 1 to 3 radicals selected from R J The groups are substituted, and where the valence allows, the phenyl, the (C3-C6)cycloalkyl, the 4- to 6-membered heterocyclic and the 5- to 6-membered heteroaryl are each independently and optionally substituted by 1 to 3 groups selected from the group consisting of: halogen, cyano, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxy, phenyl and benzyl.

[0036] 2. definition

[0037] When used to describe a chemical group that may have multiple connection points, the hyphen (-) indicates the connection point between the group and the variable that defines it. For example, -NR c C(O)R e This means that the bonding point of this group appears on the nitrogen atom.

[0038] The terms “halogen” and “halogen” refer to atoms selected from fluorine (fluorinated, -F), chlorine (chlorinated, -Cl), bromine (brominated, -Br), and iodine (iodinated, -I).

[0039] Unless otherwise stated, the term "alkyl" when used alone or as part of a larger part such as "halogenated alkyl" means a saturated straight-chain or branched monovalent hydrocarbon group.

[0040] The term "haloalkyl" includes monohaloalkyl, polyhaloalkyl, and perhaloalkyl, wherein the halogen is independently selected from fluorine, chlorine, bromine, and iodine.

[0041] "Alkoxy" refers to an alkyl group represented by -O-alkyl linked by an oxygen atom. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, propoxy, and butoxy.

[0042] "Haloalkoxy" is a haloalkyl group that is connected to another part by an oxygen atom, such as -OCHF2 or -OCF3.

[0043] The term "oxo" refers to a group with the radical =O.

[0044] The term "imino" refers to the group =NH.

[0045] Unless otherwise stated, the term "heteroaryl" refers to a 5- to 12-membered aromatic group containing 1 to 4 heteroatoms selected from N, O, and S. In some cases, the nitrogen atom in the heteroaryl group may be quaternized. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl," or "heteroaromatic." Heteroaryl groups can be monocyclic or bicyclic. Monocyclic heteroaryl groups include, for example, thienyl, furanyl, pyrroleyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrazinyl, etc. Bicyclic heteroaryl groups include groups in which a monocyclic heteroaryl ring is fused with one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, benzoxazolyl, benzooxadiazolyl, indazolyl, benzimidazolyl, benzothiazolyl, benzothiophene, quinolinyl, quinazolinyl, quinoxalolinyl, pyrrolopyridyl, pyrrolopyridyl, thienopyridyl, thienopyridyl, indoleyl, purinyl, cenolinyl, naphridyl, and pteridyl. It should be understood that, when specified, optional substituents on the heteroaryl group may be present at any substituted position and include, for example, positions connecting to the heteroaryl group (if the valence allows).

[0046] Unless otherwise stated, the term "heterocyclic group" means a 4- to 12-membered saturated or partially unsaturated heterocycle containing 1 to 4 heteroatoms independently selected from N, O, and S. The terms "heterocycle," "heterocyclyl group," "heterocyclic ring," "heterocyclic radical," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein. A heterocyclic ring can be attached to any heteroatom or carbon atom forming a stable structure, with its side groups attached. Heterocyclic groups can be monocyclic or bicyclic (e.g., bridged, fused, or spirobicyclic). Examples of monocyclic saturated or partially unsaturated heterocyclic groups include, but are not limited to, aza-butyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, pyrrolyl, pyrrolidone, piperidinyl, oxazolyl, piperazine, dioxacycloalkyl, dioxopentyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridyl, tetrahydropyridyl, dihydropyrimidinyl, tetrahydropyrimidinyl, dihydrooxadiazolyl, and dihydroisooxazolyl. Bicyclic heterocyclic groups include, for example, unsaturated heterocyclic groups fused with another unsaturated heterocyclic group, cycloalkyl group, aryl group, or heteroaryl ring, such as benzodioxacyclopentenyl, dihydrobenzodioxinyl, dihydrobenzofuranyl, dihydrobenzothiopheneyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl, 6-thia-2,7-diazaspiro[3.4]octyl, 2,6-diazaspiro[3.3]heptyl, spiro[indoline 3,3'-pyrrolidine]-yl, thiobenzodihydropyranyl, etc. It should be understood that, when specified, optional substituents on the heterocyclic group may be present at any substituted position and include, for example, positions connecting to the heterocyclic group (if the valence allows).

[0047] The term "spirocycle" refers to two rings that share a single ring atom (e.g., carbon).

[0048] The term "fusion" refers to two rings that share two adjacent ring atoms.

[0049] The term "bridging" refers to two rings that share three ring atoms with each other.

[0050] The term "aryl" refers to an aromatic carbocyclic monocyclic or two-ring system containing 6 to 10 carbon atoms. Examples include phenyl, indanyl, tetrahydronaphthalene, and naphthyl. On the one hand, aryl is either phenyl or naphthyl.

[0051] The term "cycloalkyl" used alone or as part of a larger body refers to a saturated cyclic aliphatic monocyclic or bicyclic system as described herein, having 3 to 10 carbon ring atoms unless otherwise specified. Monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, and cyclooctyl. It should be understood that, when specified, optional substituents on the cycloalkyl or alicyclic group may be present at any substituted position and include, for example, positions connecting to the cycloalkyl group.

[0052] "Amino acid residues" refers to the portion remaining after a bond is formed between a reactive group (e.g., an amino group) in another compound and a carboxylic acid in the amino acid, or both. As a result of bond formation, the carboxylic acid in the amino acid no longer has an OH group, but instead has a bond between a carbonyl group and a reactive group in the compound; the amino group has only one hydrogen atom, and a bond between a reactive group in another compound and the nitrogen atom of the amino group; or both. For example, an "α-amino acid residue" can be structurally described as NH2CR'RC(O)-, -NHCR'RC(O)OH, or -NHCR'RC(O)-; and a "β-amino acid residue" can be structurally described as NH2CR'RCH2-C(O)-, -NHCR'RCH2-C(O)OH, or -NHCR'RCH2-C(O)-, where R' is H or a C1-C6 alkyl group, and R is H or a C1-C6 alkyl group optionally substituted with 1 to 3 groups selected from: halogenated, (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), -CO(C1-C4 alkyl), -CONH(C1-C4 alkyl)phenyl, phenyl and 5 to 10-membered heteroaryl, wherein the C1-C6 alkyl may optionally be interrupted by a sulfur or nitrogen heteroatom, and wherein the phenyl may optionally be substituted by 1 to 3 groups selected from OH, cyano, (C1-C4 alkyl) and halogenated (C1-C4 alkyl); or R together with a nitrogen atom from an α or β amino acid residue to form a 4 to 6-membered heterocyclic group. For naturally occurring α-amino acids (i.e., naturally occurring amino acids), R' is H, 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 (indole), -(CH2)4NH2, -(CH2)3NHC(=NH2)NH2, -CH2 (imidazolyl), -(CH2)COOH and -(CH2)2COOH; or R forms a pyrrolidinyl ring together with the nitrogen atom of the α or β amino acid residue.

[0053] Non-natural amino acids are known in the art and include, for example, α-alkyl amino acids (e.g., α-methyl), α-alkylalkoxy amino acids (e.g., α-CH2OCH3), N-methyl amino acids, high amino acids, etc.

[0054] Compounds with one or more chiral centers can exist in a variety of stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Stereoisomers encompass all diastereomers, enantiomers, and epimers, as well as racemates and mixtures thereof. "Geometric isomers" refer to isomers with different orientations of substituent groups associated with a carbon-carbon double bond, cycloalkyl ring, or bridging bicyclic system. Atoms on each side of the carbon-carbon double bond (except H) can have E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents oriented on the same side) configurations. "cis" refers to substituents oriented on the same side of the ring, while "trans" refers to substituents oriented on opposite sides of the ring.

[0055] When the stereochemical configuration at the chiral center in a compound having one or more chiral centers is described by its chemical name (e.g., where the configuration is indicated by "R" or "S" in the chemical name) or structure (e.g., the configuration is indicated by a "wedge" bond), the enrichment of the indicated configuration relative to the opposite configuration is greater than 50%, 60%, 70%, 80%, 90%, 99%, or 99.9%. "Enrichment of the indicated configuration relative to the opposite configuration" is a molar percentage and is determined by dividing the number of compounds having the indicated stereochemical configuration at the chiral center by the total number of all compounds in the mixture having the same or opposite stereochemical configurations.

[0056] When geometric isomers are described by name or structure, the enrichment of the indicated isomer relative to the opposite isomer is greater than 50%, 60%, 70%, 80%, 90%, 99%, or 99.9%. "Enrichment of the indicated isomer relative to the opposite isomer" is a molar percentage and is determined by dividing the number of compounds having the indicated geometry by the total number of all compounds in the mixture having the same or opposite geometry.

[0057] When a disclosed compound is named or described by structure without indicating stereochemistry, it should be understood that the name or structure covers one stereoisomer of the possible stereoisomers or a geometric isomer that does not contain another stereoisomer, or a mixture of the covered stereoisomers or geometric isomers.

[0058] In some cases, compounds are isolated and tested as a 1:1 mixture of diastereomers. In such cases, relative stereochemistry is indicated by the term "rel-", and flat bonds are used instead of wedge bonds. For example,

[0059]

[0060] ((((2-(((3S,6S,7aS,8aR,9aR)-3-((rel-trans)-3-cyano-4-phenylpyrrolidine-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionate)

[0061] This means that the substituents around the pyrrolidine ring are trans-substituted, and cover two diastereomers.

[0062]

[0063] The terms "subject" and "patient" are used interchangeably and refer to mammals in need of treatment, such as companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cattle, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, a subject is a person in need of treatment.

[0064] The term "inhibit / inhibition / inhibiting" refers to a reduction in the baseline activity of a biological activity or process.

[0065] As used herein, the term "treatment / treat / treating" refers to reversing, alleviating, delaying the onset of a disease or condition or one or more symptoms thereof as described herein, or inhibiting its progression. In some respects, treatment may be administered after the onset of one or more symptoms, i.e., therapeutic treatment. In other respects, treatment may be administered in the absence of symptoms. For example, treatment may be administered to susceptible individuals before the onset of symptoms (e.g., based on symptom history and / or based on exposure to a particular organism or other susceptibility factors), i.e., preventative treatment. Treatment may also continue after symptoms have subsided, for example, to delay their recurrence.

[0066] The term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant, or catalyst that does not impair the pharmacological activity of the compound formulated with it. Pharmaceutically acceptable carriers, adjuvants, or catalysts that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of saturated vegetable fatty acids in the form of glycerides, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene glycol-polyoxypropylene block polymers, polyethylene glycol, and lanolin.

[0067] For use in pharmaceuticals, 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 salts of, for example, inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and organic acids (such as acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid). Compounds of the teachings of this invention having acidic groups such as carboxylic acids can form pharmaceutically acceptable salts with pharmaceutically acceptable bases. Suitable pharmaceutically acceptable basic salts include, for example, ammonium salts, alkali metal salts (such as sodium and potassium salts), and alkaline earth metal salts (such as magnesium and calcium salts). Compounds having quaternary ammonium groups also contain counter anions such as chloride, bromide, iodide, acetate, perchlorate, etc. Other examples of such salts include hydrochlorides, hydrobromic acids, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids such as glutamic acid.

[0068] The term “effective amount” or “therapeutic effective amount” refers to the amount of the compound described herein that is sufficient to achieve the desired therapeutic effect (treatment of the symptoms described herein) under administration conditions (e.g., a dose of 0.01-100 mg / kg body weight / day).

[0069] 3. compound

[0070] In the first embodiment, a compound having Formula I is provided:

[0071]

[0072] Or a pharmaceutically acceptable salt thereof, wherein the variables are as described above. Alternatively, as part of the first embodiment, R F R X and R Z Each is independently selected from halogen, cyano, (C1-C4)alkyl, cyano(C1-C4)alkyl, (C3-C6)cycloalkyl, halogen(C1-C4)alkyl, -(C1-C4)alkylC(O)NR c R d -(C1-C4)alkyl(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, -(C1-C4)alkylphenyl, -(C1-C4)alkylheteroaryl, (C2-C4)alkenyl, halo(C2-C4)alkenyl, (C2-C4)ynyl, halo(C2-C4)ynyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, -OR e , oxo, imino, phenyl, 4- to 6-membered heterocyclic groups, 5- to 6-membered monocyclic heteroaryl groups, -S(O)R e R f-S(O)2R f -S(O)=NH(C1-C4)alkyl, -S(O)NR e R f -S(O)2NR e R f -C(O)OR e -NR c C(O)R e -(C1-C4 alkyl)C(O)R g -C(O)R g -C(O)NR c R d NO2 and -NR a R b Where the valence allows, the phenyl group, the 4- to 6-membered heterocyclic group, and the phenyl group of the -(C1-C4)alkylphenyl group are each optionally and independently substituted by 1 to 3 groups selected from the following: halogenated, cyanoated, oxoated, (C1-C4)alkylphenyl groups. 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 ) alkynyl, halogenated (C1-C 10 )alkyl, (C1-C 10 )alkoxy and halogen (C1-C 10 )alkoxy, wherein, where the valence allows, the (C1-C 10 )alkyl, the (C2-C 10 )alkenyl and the (C2-C 10 Each of the alkynyl groups is optionally substituted with a 5- to 10-membered monocyclic or bicyclic heteroaryl or a 4- to 10-membered monocyclic or bicyclic heterocyclic group, each of which is optionally substituted with an oxo-substituted 5- to 7-membered heterocyclic group, and the remaining variables are as described above.

[0073] In the second embodiment, the compound of formula I has formula II:

[0074]

[0075] Or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

[0076] In the third embodiment, q in the compound of formula I or II or its pharmaceutically acceptable salt is q, wherein the remaining variables are as described above for formula I.

[0077] In the fourth embodiment, R in the compound of formula I or II or its pharmaceutically acceptable salt 2The value is hydrogen, and the other variables are as described above for Formula I or the third embodiment.

[0078] In the fifth embodiment, the compound of formula I has formula III or IV.

[0079] Or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

[0080] In the sixth embodiment, R in the compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 5 The value is hydrogen, wherein the remaining variables are as described for any of the embodiments in Formula I or the third or fourth embodiment.

[0081] In the seventh embodiment, R in the compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 3 and R 4 Each is independently selected from hydrogen and halogenated, wherein the remaining variables are as described for any of the embodiments of Formula I or the third, fourth or sixth embodiments. Alternatively, as part of the seventh embodiment, R in the compounds of Formula I, II, III or IV or their pharmaceutically acceptable salts. 3 and R 4 Each is hydrogen, wherein the remaining variables are as described for any of the embodiments of Formula I or the third, fourth or sixth embodiments. Alternatively, as part of the seventh embodiment, R in the compounds of Formula I, II, III or IV or their pharmaceutically acceptable salts. 3 and R 4 Each is fluorine, wherein the remaining variables are as described for any of the embodiments of Formula I or the third, fourth or sixth embodiments.

[0082] In the seventh embodiment, R in the compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 1 Selected from 8 to 10 fused bicyclic heteroaryl and aryl groups, each of the 8 to 10 fused bicyclic heteroaryl and the aryl group being -CR 1a R 2a P(O)OR 1b OR 2b -CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T ]][NH(AA)C(O)OR T ] or -CR 1a R2a P(O)[OR 1b ][NH(AA)C(O)OR T [Replace, wherein the remaining variables are as described for any of the embodiments of Formula I or the third, fourth, sixth or seventh embodiments. Alternatively, as part of the eighth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt.] 1 Selected from benzothiophene and naphthyl, each of the benzothiophene and naphthyl groups being -CR 1a R 2a P(O)OR 1b OR 2b -CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T ]][NH(AA)C(O)OR T ] or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T [Replace, wherein the remaining variables are as described for any of the embodiments of Formula I or the third, fourth, sixth or seventh embodiments. Alternatively, as part of the eighth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt.] 1 Selected from

[0083]

[0084] The remaining variables are as described for any of the embodiments of Formula I or the third, fourth, sixth, or seventh embodiments. Alternatively, as part of the eighth embodiment, R in the compound of Formula I, II, III, or IV or its pharmaceutically acceptable salt. 1 Selected from The remaining variables are as described for any of the embodiments of Formula I or the third, fourth, sixth, or seventh embodiments. Alternatively, as part of the seventh embodiment, R in the compound of Formula I, II, III, or IV or its pharmaceutically acceptable salt. 1 for The remaining variables are as described for any of the embodiments in Equation I or the third, fourth or sixth embodiments.

[0085] In the ninth embodiment, R in the compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 1a and R 2a Each is independently selected from hydrogen and fluorine, wherein the remaining variables are as described for formula I or any of the embodiments in the third, fourth, sixth to eighth examples. Alternatively, as part of the ninth embodiment, R in the compounds of formula I, II, III or IV or their pharmaceutically acceptable salts is selected. 1a and R 2a Each is hydrogen, wherein the remaining variables are as described for Formula I or any of the embodiments in the third, fourth, and sixth through eighth embodiments. Alternatively, as part of the ninth embodiment, R in the compounds of Formula I, II, III, or IV or their pharmaceutically acceptable salts. 1a It is hydrogen and R 2a The variable is fluorine, wherein the remaining variables are as described for formula I or any of the embodiments in the third, fourth, and sixth through eighth embodiments. Alternatively, as part of the ninth embodiment, R in the compound of formula I, II, III, or IV or a pharmaceutically acceptable salt thereof 1a and R 2a Each is fluorine, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third, fourth and sixth to eighth embodiments.

[0086] In the tenth embodiment, R in the compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 1b and R 2b Each is independently selected from 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)alkylphenyl]-C(O)O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)-[NH(AA)C(O)OR T], -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl]-OH, -[(C1-C4)alkyl]-OC(O)O-[5 to 7-membered heterocyclic], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl]-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)-[(C1-C4)alkyl]-OH and phenyl, wherein the remaining variables are as described for Formula I or any of the embodiments of the third, fourth and sixth to ninth examples. Alternatively, as part of the tenth example, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. 1b and R 2b Each is independently selected from hydrogen, [(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl]-O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl], and -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl], wherein the remaining variables are as described for Formula I or any of the embodiments in the third, fourth, and sixth through ninth examples. Alternatively, as part of the tenth example, R in the compound of Formula I, II, III, or IV or a pharmaceutically acceptable salt thereof. 1b and R 2b Each is -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl], wherein the remaining variables are as described for Formula I or any of the embodiments in the third, fourth, and sixth through ninth examples. Alternatively, as part of the tenth example, R in the compound of Formula I, II, III, or IV or its pharmaceutically acceptable salt. 1b and R 2b Each is hydrogen, wherein the remaining variables are as described for Formula I or any of the embodiments in the third, fourth and sixth through ninth embodiments.

[0087] In the tenth embodiment, the -CR in the compound of formula I, II, III or IV or its pharmaceutically acceptable salt 1a R 2a P(O)OR 1b OR 2b Selected from

[0088]

[0089] The remaining variables are as described for Formula I or any of the embodiments in the third, fourth, and sixth through eighth embodiments. Alternatively, as part of the tenth embodiment, the -CR in the compound of Formula I, II, III, or IV or its pharmaceutically acceptable salt is... 1a R 2a P(O)OR 1b OR 2b for The remaining variables are as described for Equation I or any of the embodiments in the third, fourth, and sixth through eighth embodiments.

[0090] In the eleventh embodiment, the -(AA)C(O)OR in the compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof T -C(R')(R)C(O)R T or -C(R')(R)CH2C(O)R T , where R ’ R is hydrogen and is selected from hydrogen, methyl, -CH2CH(CH3)2, benzyl and -CH2CH2-phenyl, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third, fourth and sixth to eighth embodiments.

[0091] In the twelfth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof T Selected from (C1-C4)alkyl, (C1-C4)alkyl-C(O)OC 1-4 Alkyl and benzyl, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third, fourth, sixth through eighth and eleventh embodiments.

[0092] In the thirteenth embodiment, the -CR in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T [Selected from]

[0093]

[0094] The remaining variables are as described for Formula I or any of the embodiments in the third, fourth, sixth through eighth, eleventh and twelfth embodiments. In the alternative thirteenth embodiment, the -CR in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt is... 1a R 2aP(O)[OR 1b ][NH(AA)C(O)OR T [Selected from]

[0095]

[0096] The remaining variables are as described for Equation I or any of the embodiments in the third, fourth, sixth to eighth, eleventh and twelfth embodiments.

[0097] In the fourteenth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 6 The value is hydrogen, wherein the remaining variables are as described for Formula I or any of the embodiments in the third to thirteenth embodiments.

[0098] In the fifteenth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 7 The group is selected from (C1-C4) alkyl, phenyl, and 4- to 6-membered monocyclic heterocyclic groups, wherein, where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 groups selected from R. Y The phenyl group is substituted with a group, and, where the valence allows, the phenyl group and the 4- to 6-membered monocyclic heterocyclic group are each optionally replaced by 1 to 3 groups selected from R. Z The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to fourteenth embodiments. Alternatively, as part of the fifteenth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. 7 The group is selected from (C1-C4)alkyl, phenyl, pyrrolidinyl, and aziridine, wherein, where the valence allows, the (C1-C4)alkyl group is optionally composed of 1 to 3 atoms selected from R Y The phenyl group is substituted, and, where the valence allows, the phenyl group, the pyrrolidinyl group, and the azacyclobutane group are each optionally replaced by 1 to 3 groups selected from R. Z The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to fourteenth embodiments.

[0099] In the sixteenth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof Z Selected from halogenated, -(C1-C4)alkylC(O)NR c R d Hydroxyl, phenyl, 4- to 6-membered heterocyclic groups, 5- to 6-membered monocyclic heteroaryl groups, -C(O)NR c R d and -C(O)R gWhere the valence permits, the phenyl group is optionally substituted with one to three groups selected from the group consisting of: halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, and halo(C1-C4)alkoxy, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to fifteenth examples. Alternatively, as part of the sixteenth example, R in the compound of Formula I, II, III, or IV or a pharmaceutically acceptable salt thereof Z Selected from halogenated, -(C1-C4)alkylC(O)NR c R d , hydroxyl, phenyl, tetrahydropyran, tetrahydrofuran, oxetane, pyridyl, pyrazolyl, pyridazinyl, -C(O)NR c R d and -C(O)R g The remaining variables are as described for Formula I or any of the embodiments in the third through fifteenth embodiments. Alternatively, as part of the sixteenth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. Z Selected from halogenated, -(C1-C4)alkylC(O)NR c R d , hydroxyl, phenyl, tetrahydropyran, tetrahydrofuran, oxetane, pyridinyl, pyrimidinyl, imidazole, triazolyl, pyrazolyl, pyridazinyl, -C(O)NR c R d and -C(O)R g The pyridyl, imidazolyl, and triazolyl groups are optionally substituted with one or two groups selected from halogenated and methyl groups, and the remaining variables are as described with respect to Formula I or any of the embodiments in the third to fifteenth examples.

[0100] In the seventeenth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof Y Selected from hydroxyl and 5- to 10-membered monocyclic or bicyclic heteroaryl groups, wherein, where the valence allows, the 5- to 10-membered monocyclic or bicyclic group is optionally surrounded by 1 to 3 groups selected from R. X The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to sixteenth embodiments. Alternatively, as part of the seventeenth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. Y The group is selected from hydroxyl, pyridyl, and pyrrolopyridyl, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to sixteenth examples.

[0101] In the eighteenth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereofc and R d Each is hydrogen, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to seventeenth embodiments.

[0102] In the nineteenth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof g It is -(C1-C4)alkyl, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to eighteenth examples.

[0103] In the twentieth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a 4- to 14-membered monocyclic or bicyclic heterocyclic group or a 5- to 12-membered monocyclic or bicyclic heteroaryl group, wherein, where the valence allows, each of the 4- to 14-membered monocyclic or bicyclic heterocyclic group or the 5- to 12-membered monocyclic or bicyclic heteroaryl group is optionally surrounded by 1 to 3 atoms selected from R Q The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to nineteenth embodiments. Alternatively, as part of the twentieth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. 6 and R 7 Together with the nitrogen atom to which it is attached, it forms aziridine, 2,5-diazaspiro[3.4]octyl, pyrrolidine, 2,6-diazaspiro[3.3]heptyl, piperazine, spiro[indoline-3,3'-pyrrolidine]yl, 6',7'-dihydrospiro[aziridine-3,5'-pyrrolo[1,2-a]imidazolyl]yl, each of which, where valence permits, is optionally bounded by 1 to 3 atoms selected from R Q The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to nineteenth embodiments. Alternatively, as part of the twentieth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. 6 and R 7Together with the nitrogen atom to which it is attached, it forms azircyclic butyl, 2,5-diazaspiro[3.4]octyl, pyrrolyl, 2,6-diazaspiro[3.3]heptyl, 2,6-diazabicyclo[3.2.0]heptyl, piperazine, spiro[indoline-3,3'-pyrrolidine]yl, 6',7'-dihydrospiro[azircyclic butane-3,5'-pyrrolo[1,2-a]imidazolyl]yl, and 3,4-dihydro-2H-benzo[b][1,4] Oxaazinyl, 3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazine, 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine, 2,3,4,5-tetrahydrobenzo[b][1,4]oxazine, 1,2,3,4-tetrahydroquinoxalinyl, 1-azaspiro[3.5]nonyl, 4-azaspiro[2.4]heptyl, wherein, where valence permits, each of them is optionally composed of 1 to 3 molecules selected from R Q The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to nineteenth embodiments.

[0104] In the twenty-first embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof Q Selected from halogenated, (C1-C4)alkyl, (C1-C4)alkoxy, cyano, phenyl, hydroxyl, 4- to 6-membered heterocyclic, 5- to 10-membered monocyclic or bicyclic heteroaryl, oxo, and -C(O)R g Where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 atoms selected from R M The phenyl group is substituted with a group thereof, and wherein, where the valence allows, the phenyl group, the 5- to 10-membered monocyclic or bicyclic heteroaryl group, and the 4- to 6-membered heterocyclic group are each optionally and independently replaced by 1 to 3 groups selected from R F The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to twentieth embodiments. Alternatively, as part of the twentieth embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt is... Q Selected from halogenated, (C1-C4)alkyl, -OR e Cyano, phenyl, hydroxyl, 4- to 6-membered heterocyclic groups, 5- to 10-membered monocyclic or bicyclic heteroaryl groups, oxo and -C(O)R g Where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 atoms selected from R M The phenyl group is substituted with a group thereof, and wherein, where the valence permits, the phenyl group, the 5- to 10-membered monocyclic or bicyclic heteroaryl group, and the 4- to 6-membered heterocyclic group are each optionally and independently substituted with 1 to 3 groups selected from R F The group is substituted, and R is wherein eIt is (C1-C4) alkyl or 5- to 6-membered heteroaryl, wherein the remaining variables are as described for Formula I or any of the embodiments in the third to thirteenth and twentieth examples. Alternatively, as part of the twenty-first example, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt may be used. Q Selected from halogenated, (C1-C4)alkyl, (C1-C4)alkoxy, cyano, phenyl, hydroxyl, morpholino, tetrahydropyrano, thiomorpholino, piperidinyl, oxacyclobutane, pyrazolyl, pyridinyl, tetrazolyl, imidazolyl, pyrazinyl, oxadiazolyl, triazolyl, pyrimidinyl, benzimidazolyl, 2,4,5,6-tetrahydrocyclopentadien[c]pyrazolyl, oxo, and -C(O)R g Where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 atoms selected from R M The groups are substituted, and wherein, where the valence allows, the morpholino, the tetrahydropyrano, the thiomorpholino, the piperidino, the oxacyclobutane, the pyrazolo, the pyridino, the tetrazolo, the imidazolyl, the pyrazino, the oxacyclobutane, the triazolo, the pyrimidino, the benzimidazolo, and the 2,4,5,6-tetrahydrocyclopentadien[c]pyrazolo are each optionally and independently replaced by 1 to 3 groups selected from R F The group substitutions, wherein the remaining variables are as described for any of the embodiments of Formula I or the third to thirteenth and twentieth embodiments. Alternatively, as part of the twenty-first embodiment, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt is... Q Selected from halogenated, (C1-C4)alkyl, -OR e Cyanoyl, phenyl, hydroxyl, morpholinyl, tetrahydropyranyl, thiomorpholinyl, piperidinyl, oxacyclobutane, pyrazolyl, pyridinyl, tetrazolyl, imidazolyl, pyrazinyl, isoxazolyl, oxazolyl, oxadiazolyl, triazolyl, pyrimidinyl, benzimidazolyl, 1H-pyrrolo[3,2-c]pyridine, 2,4,5,6-tetrahydrocyclopentadien[c]pyrazolyl, oxo and -C(O)R g Where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 atoms selected from R M The groups are substituted, and wherein, where the valence allows, the morpholino, the tetrahydropyrano, the thiomorpholino, the piperidino, the oxacyclobutane, the pyrazolo, the pyridino, the tetrazolo, the imidazolyl, the pyrazino, the isoxazolo, the oxazolo, the oxadiazolo, the triazolo, the pyrimidino, the benzimidazolo, the 1H-pyrrolo[3,2-c]pyridine, and the 2,4,5,6-tetrahydrocyclopentadien[c]pyrazolo are each optionally and independently replaced by 1 to 3 groups selected from R FThe group is substituted, wherein R is a group that is substituted for R. e It is (C1-C4) alkyl, pyridyl, pyrazinyl, pyrimidinyl, pyrazole, and wherein R is a derivative thereof. e The pyrazolyl group is optionally substituted with a (C1-C4) alkyl group, and the remaining variables are as described with respect to Formula I or any of the embodiments in the third to thirteenth and twentieth examples.

[0105] In the twenty-second embodiment, where the valence permits, R in a compound of formula I, II, III, or IV or a pharmaceutically acceptable salt thereof g The group is selected from (C1-C4) alkyl, 4- to 6-membered monocyclic heterocyclic groups, and 5- to 6-membered heteroaryl groups, wherein, where the valence allows, the (C1-C4) alkyl group is optionally surrounded by 1 to 3 groups selected from R J The groups are substituted, and where the valence allows, the 4- to 6-membered heterocyclic groups and the 5- to 6-membered heteroaryl groups are each independently and optionally substituted by 1 to 3 groups selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, benzyl, and hydroxyl, wherein the remaining variables are as described with respect to any of the embodiments of Formula I or the third to twenty-first embodiments.

[0106] In the twenty-third embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof J The phenyl group is used, and the remaining variables are as described for any of the embodiments of Formula I or the third to twenty-second embodiments.

[0107] In the twenty-fourth embodiment, where the valence permits, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof g The group is selected from (C1-C4)alkyl, morpholinyl, aziridine, tetrahydropyranyl, oxadiazine, pyrrolidinyl, and pyrazolyl, wherein, where the valence allows, the (C1-C4)alkyl group is optionally surrounded by 1 to 3 radicals selected from R. J The groups are substituted, and where the valence allows, the morpholino group, the aziridine group, the tetrahydropyrano group, the oxaziridine group, the pyrrolidinyl group, and the pyrazolyl group are each independently and optionally substituted by 1 to 3 groups selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, benzyl, and hydroxyl, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to twenty-third examples.

[0108] In the twenty-fifth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof FThe compounds are selected from cyano, (C1-C4)alkyl, hydroxy, and oxo groups, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to twenty-fourth examples. Alternatively, as part of the twenty-fifth example, R in the compounds of Formula I, II, III, or IV or their pharmaceutically acceptable salts is selected. F Selected from halogen, cyano, (C1-C4)alkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, hydroxy, -N[(C 1- [C4)alkyl]2, morpholino, piperazine, aziridine, pyrrolidinyl, and oxo, wherein the piperazine, pyrrolidinyl, and aziridine are each optionally substituted by one or two groups selected from: cyano, halogen, (C1-C4)alkyl, (C1-C4)alkoxy, and (C1-C4)alkyl(C1-C4)alkoxy, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to twenty-fourth examples.

[0109] In the twenty-sixth embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof M Selected from halogenated, (C1-C4)alkoxy, -S(O)2R f And -S(O) = NH(C1-C4)alkyl, wherein the remaining variables are as described for any of the embodiments of Formula I or the embodiments of the third to the twenty-fifth examples. Alternatively, as part of the twenty-sixth example, R in the compound of Formula I, II, III or IV or its pharmaceutically acceptable salt. M Selected from halogenated, hydroxylated, (C1-C4)alkoxy, -S(O)2R f -S(O) = NH(C1-C4)alkyl, pyridyl, pyrazolyl and phenyl optionally substituted with one or two halogens, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to twenty-fifth examples.

[0110] In the twenty-seventh embodiment, R in a compound of formula I, II, III or IV or a pharmaceutically acceptable salt thereof f The alkyl group is (C1-C4)alkyl, and wherein, where the valence permits, the (C1-C4)alkyl group is optionally substituted with 1 to 3 halogens, wherein the remaining variables are as described with respect to Formula I or any of the embodiments in the third to twenty-sixth examples.

[0111] Compounds having Formula I are further disclosed in the examples and are included in this disclosure. They include pharmaceutically acceptable salts as well as neutral forms.

[0112] 4. Uses, preparation and application

[0113] The compounds and compositions described herein are generally used to modulate the activity of STAT proteins, specifically STAT3 and / or STAT6. In some respects, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein inhibit the activity of STAT3 and / or STAT6.

[0114] In some respects, the compounds and pharmaceutical compositions described herein can be used for conditions that respond to regulation of STAT3 and / or STAT6. Therefore, this document provides a method for treating a subject for conditions that respond to regulation (e.g., inhibition) of STAT3 and / or STAT6, the method comprising administering to a subject in need a therapeutically effective amount of the compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof.

[0115] Also provided is the use of the compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof, for the preparation of a medicament for treating symptoms responsive to regulation (e.g., inhibition) of STAT3 and / or STAT6. Also provided are the compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof, for treating symptoms responsive to regulation (e.g., inhibition) of STAT3 and / or STAT6.

[0116] On the one hand, conditions that respond to regulation (e.g., inhibition) of STAT3 and / or STAT6 include, but are not limited to, cancer, neurodegenerative diseases, viral diseases, autoimmune diseases, inflammatory diseases, genetic diseases, hormone-related diseases, metabolic diseases, organ transplant-related conditions, immunodeficiency diseases, destructive bone diseases, proliferative diseases, infectious diseases, cell death-related conditions, thrombin-induced platelet aggregation, liver diseases, pathological immune conditions involving T cell activation, cardiovascular diseases, or CNS diseases.

[0117] On the other hand, conditions that respond to regulation (e.g., inhibition) of STAT3 and / or STAT6 include, but are not limited to, cancer (see, for example, Turkson and Jove, Oncogene 2000, 19:6613-6626), diabetes (see, for example, Gurzov et al., FEBS 2016, 283:3002), cardiovascular disease (see, for example, Grote et al., Vasc. Pharmacol. 2005, 43:2005), viral diseases (see, for example, Gao et al., J Hepatol. 2012, 57(2):430), autoimmune diseases such as lupus (see, for example, Goropevsek et al., Clin. Rev. Alleg. & Immun. 2017, 52(2):164), and rheumatoid arthritis. Inflammation (see, for example, Walker and Smith, Journal of Rheumatology, 2005, 32(9):1650), autoinflammatory syndromes (see, for example, Rauch et al., Jak-Stat, 2013, 2(l):e23820), atherosclerosis (see, for example, Ortiz-Munoz et al., Arteriosclerosis, Thrombosis and Vascular Biology, 2009, 29:525), psoriasis (see, for example, Andres et al., Experimental Dermatology, 2013, 22(5):323), allergic diseases (see, for example, Oh et al., European Respiratory Society, 2019, 19(115):46), inflammatory bowel disease (see, for example, Sugimoto, World Journal of Gastroenterology, 2019, 19(115):46), J Gastroenterol. 2008, 14(33):5110), inflammation (see, for example, Tamiya et al., Arteriosclerosis, Thrombosis and Vascular Biology 2011, 31:980), acute and chronic gout and gouty arthritis, neurological disorders (see, for example, Campbell, Brain Research Review 2005, 48(2):166), metabolic syndrome, immunodeficiency disorders such as AIDS and HIV (see, for example, O'Shea et al., The New England Journal of Medicine 2013, 368:161), destructive bone disorders (see, for example, Jatiani et al., Genes & Cancer ...(See, for example, Hodge et al., Blood, 2011, 1(10): 979), osteoarthritis, proliferative disorders, Waldenstrom's macroglobulinemia (see, for example, Hodge et al., Blood, 2014, 123(7): 1055), infectious diseases, cell death-related conditions, pathological immune conditions involving T cell activation, and CNS disorders.

[0118] Proliferative disorders include, but are not limited to, benign or malignant tumors, solid tumors, fluid-filled tumors, brain cancer, kidney cancer, liver cancer, adrenal cancer, bladder cancer, breast cancer, stomach cancer, gastric tumors, ovarian cancer, colon cancer, rectal cancer, prostate cancer, pancreatic cancer, lung cancer, vaginal cancer, cervical cancer, testicular cancer, urogenital cancer, esophageal cancer, laryngeal cancer, skin cancer, bone cancer or thyroid cancer, sarcoma, glioblastoma, neuroblastoma, multiple myeloma, gastrointestinal cancer (especially colon cancer or colorectal adenoma), neck and head tumors, epidermal hyperplasia, psoriasis, benign prostatic hyperplasia, tumor formation, epithelial characteristic tumor formation, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, lymphoma, Hodgkin's disease and non-Hodgkin's disease. and Non-Hodgkins), breast cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, IL-I driven disease, MyD88 driven disease, smoldering or blood malignancies of indolent multiple myeloma (including leukemia, diffuse large B-cell lymphoma (DLBCL), ABC DLBCL, chronic lymphocytic leukemia (CLL), chronic lymphocytic lymphoma, primary exudative lymphoma, Burkitt lymphoma / leukemia, acute lymphoblastic leukemia, B-cell prelymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenström's macroglobulinemia (WM), splenic marginal zone lymphoma, multiple myeloma, plasmacytoma, intravascular large B-cell lymphoma).

[0119] In some embodiments, the cancer to be treated is selected from glioma, breast cancer, prostate cancer, head and neck squamous cell carcinoma, melanoma of the skin, ovarian cancer, malignant peripheral nerve sheath tumor (MPNST), and pancreatic cancer. In other embodiments, the cancer to be treated is selected from the following cancers: glioma, breast cancer, prostate cancer, head and neck squamous cell carcinoma, melanoma of the skin, ovarian cancer, malignant peripheral nerve sheath tumor (MPNST), pancreatic cancer, small cell lung cancer (NSCLC) (including EGFR-mutant NSCLC), urothelial carcinoma, liver cancer, bile duct cancer, kidney cancer, colon cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, and hematologic malignancies, including lymphoma, leukemia, myeloma, myeloproliferative neoplasm, and myelodysplastic syndrome. In other embodiments, the cancer is selected from solid tumors (e.g., prostate cancer, kidney cancer, liver 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.), hematologic malignancies (e.g., lymphoma, leukemia such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), or multiple myeloma), and skin cancers (e.g., cutaneous T-cell lymphoma (CTCL) and cutaneous B-cell lymphoma). Example CTCLs include Sezary syndrome and mycosis fungoides.

[0120] The compounds, salts, and compositions described herein may also be used to treat inflammatory or obstructive airway diseases, resulting in, for example, reduction of tissue damage, airway inflammation, bronchial hyperresponsiveness, remodeling, or disease progression. Inflammatory or obstructive airway diseases include asthma of any type or cause, including intrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mild asthma, moderate asthma, severe asthma, bronchitis-related asthma, exercise-induced asthma, occupational asthma, and asthma induced by bacterial infection. Treatment of asthma should also be understood to include treatment of subjects, such as those under 4 or 5 years of age, who present with wheezing symptoms and are diagnosed or can be diagnosed as “wheezy infants,” an established patient category of primary medical concern, and now commonly identified as having initial or early-stage asthma.

[0121] The compounds, salts, and compositions described herein may also be used to treat heterologous immune diseases, including but not limited to graft-versus-host disease, transplantation, blood transfusion, allergic reactions, allergies (e.g., allergies to plant pollen, latex, drugs, food, insect venom, animal hair, animal dander, dust mites, or cockroach calyxes), type I hypersensitivity reactions, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

[0122] The compounds, salts, and compositions described herein may also be used to treat other inflammatory or obstructive airway diseases and conditions to which this invention applies, including acute lung injury (ALI), adult / acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease, airway or lung diseases (COPD, COAD, or COLD), including chronic bronchitis or associated dyspnea, emphysema, and exacerbations of airway hyperresponsiveness caused by other drug therapies (specifically other inhaled drug therapies). The compounds, salts, and compositions described herein may also be used to treat bronchitis, including but not limited to acute, peanut inhalational, catarrhal, gravid, chronic, or tuberculous bronchitis. The compounds, salts, and compositions described herein may also be used to treat any type or cause of pneumoconiosis (an inflammatory, common occupational lung disease, often accompanied by chronic or acute airway obstruction and caused by repeated inhalation of dust), including, for example, aluminosis, coal mine disease, asbestosis, stone lung, eyelash loss, iron pneumoconiosis, silicosis, smoke pneumoconiosis, and cotton dust disease.

[0123] The compounds, salts, and compositions described herein may also be used to treat inflammatory or allergic skin conditions such as psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, herpetic dermatitis, scleroderma, vitiligo, hypersensitivity vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, systemic lupus erythematosus, pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, acquired bullous epidermolysis bullosa, acne vulgaris, and other inflammatory or allergic skin conditions.

[0124] The compounds, salts, and compositions described herein may also be used to treat other diseases or conditions, such as those with inflammatory components, for example, eye diseases and conditions such as ocular allergies, conjunctivitis, dry keratoconjunctivitis, and vernal conjunctivitis; diseases affecting the nose, including allergic rhinitis; and inflammatory diseases involving autoimmune reactions or having autoimmune components or causes, including autoimmune hematologic disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell anemia, and idiopathic thrombocytopenic purpura); systemic lupus erythematosus, rheumatoid arthritis, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Steven-Johnson syndrome, idiopathic stomatitis, and autoimmune inflammatory bowel diseases (e.g., ulcerative colitis and Crohn's disease). Diseases, irritable bowel syndrome, celiac disease, periodontitis, hyaline membrane disease, nephropathy, glomerulonephropathy, alcoholic liver disease, multiple sclerosis, endocrine ophthalmopathy, Graves' disease, sarcoidosis, alveolitis, chronic hypersensitivity localized pneumonia, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), Sjogren's syndrome, keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitial pulmonary fibrosis, psoriatic arthritis, systemic juvenile idiopathic arthritis, cryopyrin-associated periodic syndrome, nephritis, vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis (with and without nephrotic syndrome, for example,This includes idiopathic nephrotic syndrome or pure mineral change nephropathy, chronic granulomatous disease, endometriosis, leptospirosis renaldisease, glaucoma, retinal disease, aging, headache, pain, complex regional pain syndrome, cardiomegaly, muscular atrophy, catabolism, obesity, fetal growth retardation, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, anhidrotic ectodermal dysplasia, Behcet's disease, pigmentary disorders, and Paget's disease. Diseases, pancreatitis, hereditary periodic fever syndrome, asthma (allergic and non-allergic, mild, moderate, severe, bronchitis and exercise-induced asthma), acute lung injury, acute respiratory distress syndrome, eosinophilia, hypersensitivity, allergic reaction, sinusitis, ocular allergy, silica-induced diseases, COPD (damage reduction, airway inflammation, bronchial hyperresponsiveness, remodeling or disease progression), lung diseases, 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. Diseases, 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-Schonlein purpura. (purpura), hepatitis, hidradenitis suppurativa, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, localized pneumonia, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendinitis, tonsillitis, ulcerative colitis, uveitis, vaginitis, vasculitis, or vulvitis.

[0125] In some embodiments, cardiovascular diseases treatable by the method according to the present invention include, but are not limited to, restenosis, cardiac hypertrophy, atherosclerosis, myocardial infarction, ischemic stroke, congestive heart failure, angina pectoris, post-angioplasty restenosis, post-angioplasty restenosis, aortocoronary shunt restenosis, aortocoronary shunt restenosis, stroke, transient ischemic attack, peripheral artery occlusive disease, pulmonary embolism, and deep vein thrombosis.

[0126] In some embodiments, neurodegenerative diseases treatable according to the method of the present invention include, but are not limited to, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, cerebral ischemia, and neurodegenerative diseases caused by traumatic injury, glutamate neurotoxicity, hypoxia, epilepsy, diabetes treatment, metabolic syndrome, obesity, organ transplantation, and graft-versus-host disease.

[0127] In some respects, the pharmaceutical compositions described herein are formulated for administration to patients in need of such compositions. The pharmaceutical compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implantable reservoir. As used herein, the term “parenterical” includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally, or intravenously. The sterile injectable form of the pharmaceutical compositions described herein can be an aqueous or oily suspension. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents according to techniques known in the art.

[0128] In some respects, the pharmaceutical composition is administered orally.

[0129] 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, weight, general health condition, sex, diet, timing of administration, excretion rate, drug combination, the judgment of the treating physician, and the severity of the specific disease being treated. The amount of the compounds described herein in the composition will also depend on the specific compounds in the pharmaceutical composition.

[0130] Example

[0131] Preparation of compounds

[0132] The compounds claimed herein were prepared according to the procedure outlined in the following scheme. The compound names were generated using software built into ChemDraw. The described chemical structure shall be considered the appropriate compound in respect of any discrepancy between the compound name and the structure it depicts.

[0133] Nucleosynthesis

[0134] Synthesis of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid and (3S,6S,7aR,8aS,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid

[0135]

[0136] Step 1: Preparation of methyl (2S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)pyrrolidine-2-carboxylate

[0137] Add Et3N (406 mL, 2.92 mol, 3.00 equivalents) and 2-chloro-1-methylpyrrolidine-2-carboxylate hydrochloride (200 g, 972 mmol, 1.00 equivalents) to a cooled (0 °C) solution of (2S)-5-allylpyrrolidine-2-carboxylate hydrochloride (200 g, 972 mmol, 1.00 equivalents) in CH2Cl2 (1.60 L) to a solution of (S)-2-((tert-butoxycarbonyl)amino)pentan-4-enoic acid (209 g, 972 mmol, 1.00 equivalents) (273 g, 1.07 mol, 1.10 equivalents). Heat the solution to 25 °C and stir for 1 hour. Pour the mixture into water (5.0 L) and extract with CH2Cl2 (2.00 L x 3). Wash the combined organic layers with brine (2.0 L), dry to Na2SO4, filter, and concentrate under reduced pressure. Six separate batches of the same size were performed in parallel and combined during processing. The resulting residues were 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 HNMR(400MHz, CDCl3)δ5.84–5.78(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).

[0138] Step 2: Preparation of (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate

[0139] At 25 °C, a first-generation Grubbs catalyst (44.9 g, 54.6 mmol, 0.10 equivalent) was added to a solution of (2S)-5-allyl-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)pyrrolidine-2-carboxylate (200 g, 546 mmol, 1.00 equivalent) in CH2Cl2 (2.00 L). The solution was then heated to 50 °C and stirred for 36 h. Six separate batches of the same scale were performed in parallel and combined during the process. The combined reaction mixtures were concentrated to give a residue. The residue was purified twice by column chromatography (petroleum ether:EtOAc = 100:1 to 0:1) to give a crude product. The crude product was milled with petroleum ether (2.00 L) for 12 h and filtered. The filter cake was dried under reduced pressure to give solid (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (510 g, 1.40 mol, 51.3% yield, 92.9% purity). LCMS (ESI) m / z = 361.2 [M+Na] + ; 1 H NMR (400MHz, CDCl3) δ5.82–5.80(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).

[0140] Step 3: Preparation of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate and (3S,6S,7aR,8aS,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate

[0141] A solution of KOH (144 g, 2.57 mol, 5.30 equivalents) in H2O (150 mL) was slowly added dropwise to a cooled (0 °C) solution of 1-methyl-1-nitrosourea (50.0 g, 485 mmol, 1.00 equivalents) in Et2O (1.50 L). The mixture was stirred at 0 °C for 30 minutes, at which point the organic layer turned yellow, indicating the completion of the reaction. The aqueous layer of the reaction mixture was separated, and the yellow Et2O layer was ready for use in the next step without further processing. [Et2O (1.50 L) containing diazomethane (20.0 g, 475 mmol, 98.0% yield) was obtained].

[0142] A solution of approximately 85% diazomethane (20.0 g, 476 mmol, 16.1 equivalents) in Et₂O (1.50 L) was carefully transferred to a solution of (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (10.0 g, 29.5 mmol, 1.00 equivalents) in THF (320 mL) (0 °C). A solution of approximately 50% Pd(OAc)₂ (1.99 g, 8.87 mmol, 0.30 equivalents) in THF (60.0 mL) was slowly added to the resulting mixture. The reaction mixture was stirred at 0 °C for 2 minutes, followed by the continuous addition of the remaining diazomethane ether solution and palladium solution. The reaction was stirred at 0°C for 20 minutes, then the ice-water bath was removed, and the reaction was warmed to 20°C. After stirring for 60 minutes, the solvent was removed by a nitrogen stream. The resulting dried reaction mixture was diluted with EtOAc and filtered through diatomaceous earth. The residue (total: 37.0 g) was purified by column chromatography: Phenomenex Luna C18 250 x 80 mm x 10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 45%–70%, for 21 minutes. The appropriate peaks corresponding to the products were collected. The pH of the corresponding samples was adjusted to approximately 8 with saturated NaHCO3 aqueous solution and concentrated separately. The corresponding aqueous layer was extracted with EtOAc (2 x 1.00 L), washed with brine (1.00 L), dried over Na2SO4, filtered, and concentrated to produce methyl (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (12.13) as a white solid. (33.5 mmol, 31.5% yield, 97.3% purity) and (3S,6S,7aR,8aS,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (9.12 g, 25.5 mmol, 24.0% yield, 98.6% purity), a white solid. Additionally, (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (7.60 g, 99.0% purity) was recovered as a yellow oil. (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylate: LCMS(ESI) m / z = 353.0 [M+H] + ;1 H NMR(400MHz, DMSO-d6)δ6.81(d,J=7.6Hz,0.8H),6.50(d,J=3.6Hz,0.2H),4.44-4.27(m,2H),4.08-3.99(m,1H),3.58(s,3H),2.1 9-2.07(m,4H),1.88-1.86(m,2H),1.73-1.59(m,2H),1.35-1.31(m,9H),1.08-1.05(m,1H),0.73-0.71(m,2H),0.02-0.03(m,1H).

[0143] (3S,6S,10aR,Z)-6-((tert-butoxycarbonyl)amino)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate: LCMS(ESI) m / z = 353.0 [M+H] + ; 1 H NMR(400MHz, DMSO-d6)δ6.75(d,J=7.6Hz,0.9H),6.22(d,J=2.0Hz,0.1H),4.33-4.25(m,2H),4.14-4.12(m,1H),3.60(s,3H),2.20-2.10 (m,1H),2.08-2.03(m,3H),1.85-1.84(m,1H),1.74-1.57(m,2H),1.38(s,9H),1.34-1.14(m,3H),0.75-0.73(m,1H),0.03-0.01(m,1H).

[0144] Step 4: Preparation of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylic acid

[0145] LiOH·H₂O (270 mg, 11.3 mmol, 2 equivalents) was added to a solution of methyl (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (2.00 g, 5.67 mmol, 1 equivalent) in a mixture of MeOH (1.0 mL), THF (1.0 mL), and water (1.0 mL). After stirring at ambient temperature for 2 hours, the reaction mixture turned dark white. The reaction mixture was carefully concentrated under reduced pressure to obtain a residue. The residue was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The pH of the aqueous phase was acidified to about 4 to 5 with 1 N HCl aqueous solution. The resulting acidic phase was extracted with EtOAc (2 x 50 mL), and the combined organic layers were concentrated under reduced pressure to obtain a residue. The isolated product (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (2.00 g) is a yellow oil and can be used without further purification or manipulation. LCMS (ESI) m / z = 339.0 [M+H] + .

[0146] Under the conditions described above for the preparation of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid, the intermediates shown in Table 1 were synthesized using appropriate starting materials and reagents.

[0147] Table 1.

[0148]

[0149] Synthesis of (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate and (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-8-methyl-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate

[0150]

[0151] Step 1: Preparation of (S)-2-((tert-butoxycarbonyl)amino)-5-oxo-7-(trimethylsilyl)hept-6-ynyl ester

[0152] Ethynyltrimethylsilane (1.86 kg, 18.9 mol, 2.62 L, 1.15 equivalents) was added to a cooled (0 °C) solution of isopropyl magnesium chloride (2.0 M, 9.04 L, 1.10 equivalents) and THF (4.00 L). The reaction mixture was stirred for 1 hour, followed by the slow addition of (S)-5-oxopyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester 2-methyl ester (4.00 kg, 16.4 mol, 1.00 equivalents) to a solution of THF (8.0 L) over 1.5 hours. After stirring for another 30 minutes, the reaction mixture was transferred to a stirred and cooled (0 °C) two-phase mixture of iPAc (5.00 L) and 20% NH4Cl aqueous solution (16.0 L). The two-phase mixture was separated, and the aqueous layer was extracted with iPAc (8.00 L). The combined organic layers were washed with 20% NH4Cl aqueous solution (8.00 L) and brine (8.00 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain methyl (S)-2-((tert-butoxycarbonyl)amino)-5-oxo-7-(trimethylsilyl)hept-6-acetylacetic acid (5.50 kg) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ5.11–5.09(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).

[0153] Step 2: Preparation of 2-methyl (2S,5R)-5-((trimethylsilyl)ethynyl)pyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester

[0154] A solution of (S)-2-((tert-butoxycarbonyl)amino)-5-oxo-7-(trimethylsilyl)hepta-6-acetylic acid methyl ester in iPAc (20.0 L) was added to a suspension of NaBH(OAc)3 (4.03 kg, 19.0 mol, 1.30 equivalent) in iPAc (5.00 L). The mixture was cooled in portions to (-10 °C), followed by the slow addition of TFA (7.18 kg, 62.9 mol, 4.66 L, 4.30 equivalent) over 1.5 hours. The mixture was warmed to 10 °C and stirred for another 2 hours. The reaction mixture was transferred to a cooled (0 °C) 25% K₂HPO₄ aqueous solution (40.0 L). The pH of the suspension was adjusted to pH 6–7 with a saturated NaHCO₃ aqueous solution. The two-phase mixture was separated, and the aqueous layer was extracted with iPAc (8.00 L). The combined organic layers were washed with 25% K₂HPO₄ solution (8.00 L) and brine (5.00 L), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 30 / 1 to 3 / 1) to produce 2-methyl (2S,5R)-5-((trimethylsilyl)ethynyl)pyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester, which was a yellow oil (2.27 kg, 6.97 mol, 42.4% yield). 1 HNMR (400MHz, DMSO-d6) δ4.49–4.32(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).

[0155] Step 3: Preparation of 2-methyl (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester

[0156] A solution of 1.0 M TBAF (1.84 L, 1.20 equivalent) in THF was added to a cooled (0 °C) solution of (2S,5R)-5-((trimethylsilyl)ethynyl)pyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester 2-methyl ester (500 g, 1.54 mol, 1.00 equivalent) in THF (2.00 L), and the mixture was then stirred for 1 hour. Four batches of the same scale were performed in parallel, and the reaction mixtures were combined and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 1 / 0 to 0 / 1) to give (920 g, 2.95 mol, 48.0% yield, 81.2% purity) of (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester 2-methyl ester as a yellow oil. 1HNMR (400MHz, CDCl3) δ4.62–4.50(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).

[0157] Step 4: Preparation of 2-methyl (2S,5R)-5-vinylpyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester

[0158] Under N2(g), Lindlar's catalyst (7.50 g, 1.82 mmol, 5.0% wt) and quinoline (163 g, 1.27 mol, 150 mL, 2.14 equivalents) were added to a suspension of (2S,5R)-5-ethynylpyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester 2-methyl ester (150 g, 592 mmol, 1.00 equivalents) in EtOAc (1.50 L). The suspension was degassed under vacuum and purged with H2(g) (3x). The mixture was stirred under H2(g) (50 psi) for 1 hour. Six batches of the same scale were performed in parallel, filtered, and the filtrates were combined during the treatment. The mixture was filtered, and the filtrates were washed, separated, and concentrated with 1N HCl aqueous solution (9.00 L). The combined residues were purified by silica gel column chromatography (petroleum ether / EtOAc = 1 / 0 to 0 / 1) to produce 2-methyl (2S,5R)-5-vinylpyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester, which was a yellow oil (745 g, 2.23 mol, 62.8% yield, 76.5% purity). 1 H NMR(400MHz,DMSO-d6)δ5.84–5.76(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).

[0159] Step 5: Preparation of (2S,5R)-5-vinylpyrrolidine-2-carboxylate methyl ester

[0160] A solution of 2-methyl (2S,5R)-5-vinylpyrrolidine-1,2-dicarboxylic acid 1-(tert-butyl) ester (245 g, 959 mmol, 1.00 equivalent) in EtOAc (1.25 L) was added to a solution of EtOAc (959 mL, 4.00 equivalent). Three batches of the same scale were performed in parallel, and the mixture was stirred at 25 °C for 2 hours. The reaction mixtures were combined and concentrated under reduced pressure to give the product (2S,5R)-5-vinylpyrrolidine-2-carboxylic acid methyl ester (520 g, HCl) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6)δ6.03–5.95(m,1H),5.44–5.33(m,2H),4.49–4.45(m,1H),4.1 1–4.05(m,1H),3.76(s,3H),2.31–2.28(m,1H),2.16–2.10(m,1H),1.81–1.76(m,1H).

[0161] Step 6: Preparation of methyl (2S,5R)-1-((S)-2-((tert-butoxycarbonyl)amino)pent-4-enoyl)-5-vinylpyrrolidine-2-carboxylate

[0162] Methyl (2S,5R)-5-vinylpyrrolidine-2-carboxylate (32.0 g, 167 mmol, 1.00 equivalent, HCl) and Et3N (69.7 mL, 0.501 mol, 3.00 equivalent) were added to a solution of (S)-2-((tert-butoxycarbonyl)amino)pent-4-enoic acid (35.9 g, 167 mmol, 1.00 equivalent, HCl) in CH2Cl2 (320 mL). The mixture was cooled to 0 °C and CMPI (98.8 g, 387 mmol, 1.10 equivalent) was added. The reaction mixture was then warmed to 25 °C and stirred for 3 hours. The reaction mixture was poured into water (500 mL) and extracted with CH2Cl2 (200 mL x 3). The combined organic layers were washed with saturated NH4Cl aqueous solution (300 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the 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)pent-4-enoyl)-5-vinylpyrrolidine-2-carboxylate (47.0 g) as a yellow oil. LCMS (ESI) m / z = 367.2 [M+H] + ; 1H NMR (400MHz, CDCl3) δ6.03-5.94(m,1H),5.77-5.66(m,1H),5.52(d,J=16.8Hz,1H),5.22(d,J=10.4Hz,1H),5.12-5.00(m,4H),4.81(br d,J=6.4Hz,1H),4.53-4.44(m,2H),3.73(s,3H),2.45-2.38(m,1H),2.32-2.27(m,1H),2.22-2.15(m,2H),2.02-1.83(m,3H),1.41(s,9H).

[0163] Under the reaction conditions described above, the intermediates shown in Table 2 were synthesized using methyl (2S,5R)-5-vinylpyrrolidine-2-carboxylate, a suitable N-protected amino acid, CMPI, triethylamine, and CH2Cl2. The intermediates were purified using standard methods.

[0164] Table 2.

[0165]

[0166] Step 7: Preparation of methyl (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate

[0167] A first-generation Grubb's catalyst (4.90 g, 5.96 mmol, 0.10 equivalent) was added to a solution of methyl (21.0 g, 59.6 mmol, 1.00 equivalent) in CH2Cl2 (2.10 L). The mixture was heated to 50 °C and stirred for 12 hours. Two batches of the same scale were performed in parallel. The reaction mixtures were then combined and concentrated to give the residue. The residue was purified by column chromatography (petroleum ether / EtOAc = 10 / 1 to 1 / 1) to give (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate as a gray solid (35.0 g, 94.8 mmol, 56.6% yield, 87.9% purity). LCMS (ESI) m / z = 225.0 [(M–Boc)+H] + ; 1HNMR (400MHz, CDCl3) δ5.78-5.71(m,2H),5.55(d,J=11.6Hz,1H),4.75-4.63(m,3H),3.72(s, 3H),2.64-2.58(m,1H),2.38-2.29(m,2H),2.11-2.05(m,2H),1.93-1.85(m,1H),1.44(s,9H).

[0168] The intermediates shown in Table 3 were synthesized using appropriate starting materials and reagents under the conditions described above for the preparation of (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate.

[0169] Table 3.

[0170]

[0171] Synthesis of (3S,6S,7aS,8aS,8bR)-6-((tert-butoxycarbonyl)amino)-7a-methyl-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylic acid and (3S,6S,7aR,8aR,8bR)-6-((tert-butoxycarbonyl)amino)-7a-methyl-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylic acid

[0172]

[0173] Step 1: Preparation of (3S,6S,7aS,8aS,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate and (3S,6S,7aR,8aR,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate

[0174] A solution of (3S,6S,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxo-2,3,5,6,7,9a-hexahydro-1H-pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate (1.00 g, 3.08 mmol, 1 equivalent) in DCM was added to a stirred and cooled (-40 °C) solution of 0.5 M diazomethane (1.29 g, 30.8 mmol, 1 equivalent) in MTBE (40 mL). DCM containing Pd(OAc)₂ (6.91 mg, 30.8 μmol, 0.01 equivalent) was added to the cooled mixture. The mixture was then warmed to -10 °C, and bubbling of N₂(g) gas was observed. The reaction mixture was aged at this temperature for 2 hours or until N₂(g) emission ceased. The reaction mixture was warmed to room temperature and filtered. The filtrate was concentrated to give a crude product as a mixture of diastereomers. The resulting mixture was purified by HPLC and separated to produce methyl (3S,6S,7aS,8aS,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate (100 mg, 295 μmol, 9.61%) and methyl (3S,6S,7aR,8aR,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylate (110 mg, 325 μmol, 10.5%).

[0175] (3S,6S,7aS,8aS,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylic acid methyl ester: LCMS(ESI) m / z = 239.0 [(M-Boc)+H] + .

[0176] (3S,6S,7aR,8aR,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylic acid methyl ester: LCMS(ESI) m / z = 239.0 [(M-Boc)+H] + .

[0177] Step 2: Preparation of (3S,6S,7aS,8aS,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylic acid and (3S,6S,7aR,8aR,8bR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydrocyclopropane[c]pyrrolo[1,2-a]azacycloheptatrien-3-carboxylic acid

[0178] The intermediates shown in Table 4 were synthesized using appropriate starting materials and LiOH·H2O under the hydrolysis conditions described in step 4 for the synthesis of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid and / or (3S,6S,7aR,8aS,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid.

[0179] Table 4.

[0180]

[0181] Synthesis of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-8,8-difluoro-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylic acid

[0182]

[0183] Step 1: Preparation of (3S,6S,10aR,Z)-6-amino-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraene-3-carboxylate

[0184] Under nitrogen atmosphere, trifluoroacetic acid (4.0 mL, 52 mmol, 17 equivalents) was added 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]azacyclooctatetraen-3-carboxylate (1.0 g, 2.95 mmol, 1 equivalent) in dichloromethane (15 mL). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated to dryness and directly purified by reversed-phase chromatography using a gradient of 5–100% MeCN / alkaline water (10 mM NH4HCO3, pH = 10) to produce methyl (3S,6S,10aR,Z)-6-amino-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate as a white solid (588 mg, 2.46 mmol, 83% yield). 1H NMR (400MHz, CD3OD) δ5.78-5.64(m,2H),4.45-4.41(m,1H),4.25-4.17(m,1H),4.11(dd,1H,J=9.7 ,6.0Hz),3.66(s,3H),2.81-2.67(m,2H),2.41-2.33(m,1H),2.22-2.05(m,3H),1.97-1.86(m,2H).

[0185] Step 2: Preparation of (3S,6S,10aR,Z)-6-(1,3-dioxoisoindoline-2-yl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid methyl ester

[0186] A solution of (3S,6S,10aR,Z)-6-amino-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (500 mg, 2.09 mmol, 1 equivalent) and phthalic anhydride (318 mg, 2.15 mmol, 1.03 equivalent) in toluene (20 mL) was heated under reflux for 24 hours. The solvent was then evaporated and the product was directly purified by reversed-phase chromatography using a gradient of 5–100% MeCN / water to produce (3S,6S,10aR,Z)-6-(1,3-dioxoisoindoline-2-yl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctate-3-carboxylate as a grayish-white solid (659 mg, 1.78 mmol, 85% yield). 1 H NMR (400MHz, CDCl3) δ7.86-7.80(m,2H),7.72-7.66(m,2H),6.18-6.09(m,1H) ,5.88-5.81(m,1H),5.31(dd,J=7.9,4.5Hz,1H),4.46-4.38(m,2H),3.70(s,3 H),3.13-3.03(m,1H),2.96-2.88(m,1H),2.63-2.54(m,1H),2.49-2.40(m,1H ),2.32-2.23(m,1H),2.22-2.12(m,1H),2.05-1.94(m,1H),1.92-1.84(m,1H).

[0187] Step 3: Preparation of (3S,6S,7aS,8aR,9aR)-6-(1,3-dioxoisoindoline-2-yl)-8,8-difluoro-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylate

[0188] A round-bottom flask equipped with a magnetic stirrer and a water condenser was filled with methyl (3S,6S,10aR,Z)-6-(1,3-dioxoisoindoline-2-yl)-5-oxo-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (150 mg, 0.407 mmol, 1 equivalent), sodium fluoride (5.12 mg, 0.122 mmol, 0.3 equivalent), and anhydrous toluene (2 mL). The mixture was heated to reflux for 30 minutes, and then, over 1 hour, a solution of 2,2-difluoro-2-(fluorosulfonyl)acetic acid trimethylsilane ester (159 μL, 0.814 mmol, 2 equivalent) in toluene (2 mL) was added via a syringe pump. The clear solution turned pale orange. The reaction was stirred under reflux for 1.5 hours, then toluene (2 mL) containing 2,2-difluoro-2-(fluorosulfonyl)acetic acid trimethylsilane ester (159 μL, 0.814 mmol, 2 equivalents) was added via syringe pump over 40 minutes, and the reaction was stirred under reflux for 1 hour. The reaction mixture was cooled to room temperature, diluted with saturated aqueous sodium bicarbonate solution and EtOAc, and stirred for 10 minutes. The layers were separated. The organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated to dryness to produce coarse orange foam (238 mg, 140%). Methyl 3-(3S,6S,7aS,8aR,9aR)-6-(1,3-dioxoisoindoline-2-yl)-8,8-difluoro-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (48 mg, 0.114 mmol, 28% yield) was purified by reversed-phase chromatography with elution of 5-100% MeCN / water (containing 0.1% formic acid). 1 H NMR (400MHz, CDCl3) δ7.86-7.81(m,2H),7.74-7.68(m,2H),5.25-5.20(m,1H),4.40(t ,J=8.1Hz,1H),4.28-4.20(m,1H),3.72(s,3H),2.74-2.61(m,2H),2.38-1.80(m,8H).

[0189] Step 4: Preparation of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-8,8-difluoro-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid

[0190] Lithium hydroxide monohydrate (43.2 mg, 1.03 mmol, 3 equivalents) was added to a solution of (3S,6S,7aS,8aR,9aR)-6-(1,3-dioxoisoindoline-2-yl)-8,8-difluoro-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (145 mg, 0.346 mmol, 1 equivalent) in water (2 mL) and tetrahydrofuran (6 mL). The reaction mixture was stirred at 20 °C for 1 h. The volatiles were removed under vacuum. The aqueous residue was diluted with 3M hydrochloric acid aqueous solution (5 mL) and the solution was heated to 100 °C for 20 h. The reaction mixture was concentrated to dryness. The residue was diluted with 1M NaOH aqueous solution (6 mL) and di-tert-butyl dicarbonate (377 mg, 1.73 mmol, 5 equivalents) was added. The reaction was stirred at 20°C for 20 hours. More di-tert-butyl dicarbonate (800 mg) was added, and the reaction was stirred at 20°C for 16 hours. Volatile substances were removed under vacuum. The resulting aqueous solution was acidified to pH 2 using 6N HCl and the product was extracted with EtOAc (2 x 15 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The product was purified by reversed-phase chromatography using a gradient of 5–100% MeCN / water (containing 0.1% formic acid) to yield (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-8,8-difluoro-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (84.0 mg, 0.224 mmol, 65%) as a white solid. 1 H NMR (400MHz, CDCl3) δ5.40-5.33(m,1H),4.72(d,J=7.4Hz,1H),4.64(q,J=8.5Hz,1H),4.28-4.18 (m,1H),2.43-2.26(m,2H),2.25-2.16(m,1H),2.11-1.90(m,5H),1.74-1.63(m,2H),1.43(s,9H).

[0191] Joint assembly

[0192] Prepare the following intermediates in Table 5 according to the scheme described in WO 2020205467.

[0193] Table 5.

[0194]

[0195] Representative procedures for the preparation of activated phosphonic esters

[0196] Synthesis of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0197]

[0198] Step 1: Preparation of 4-nitrobenzene 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0199] A mixture of 5-[(diethoxyphosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylic acid (L1) (10.0 g, 27.4 mmol), EDCI (7.85 g, 41.0 mmol), and DMAP (836 mg, 6.85 mmol) in CH2Cl2 (80 mL) was stirred at room temperature. After 15 minutes, 4-nitrophenol (4.75 g, 34.2 mmol) was added, and the resulting yellow mixture was stirred at room temperature for 18 hours. The reaction was quenched with water (30 mL), and the product was extracted with CH2Cl2 (10 mL x 2). The combined extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated under vacuum. The crude residue was purified by reversed-phase chromatography [C18 cartridge, eluting with a gradient of 5-100% acetonitrile / water], and the appropriate fractions were concentrated to give 4-nitrobenzene 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (7.80 g, 16.0 mmol, 59.0% yield) as a yellow solid. LCMS m / z = 486.2 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.32–8.37(m,3H),8.22(s,1H),8.01(d,J=9.1Hz,1H),7. 77(d,J=7.8Hz,1H),7.51–7.45(m,2H),4.14–4.32(m,4H),1.34(t,J=7.8Hz,6H).

[0200] Step 2: Preparation of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0201] N,O-bis(trimethylsilyl)trifluoroacetamide (12.1 mL, 46.0 mmol) and trimethyliodosilane (5.23 mL, 36.8 mmol) were added to a cooled (0 °C) solution of 4-nitrobenzene 4-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (4.47 g, 9.20 mmol) in CH2Cl2 (39 mL) as a solution in CH2Cl2 (10 mL). The reaction mixture was gradually warmed to ambient temperature. A mixture of 2:1 H2O / acetonitrile (containing 0.1% TFA) (50 mL) was added to the reaction mixture, and precipitation of the product was observed. The volatiles were removed under vacuum, and the crude residue was suspended in a mixture of acetonitrile / water solution (1:1 v / v, 100 mL). The suspension was filtered, the solid was washed with a 2:1 mixture of acetonitrile and aqueous solution, and the solid was dried under reduced pressure to produce (difluoro(2-(((4-nitrophenoxy)carbonyl)benzo[b]thiophene-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 mixture of acetonitrile and aqueous solution. The solid was dried under reduced pressure to produce another (difluoro(2-(((4-nitrophenoxy)carbonyl)benzo[b]thiophene-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]thiophene-5-yl)methyl)phosphonic acid (6.90 g, 16.0 mmol, 98.0% yield). 1 H NMR (400MHz, DMSO-d6) δ7.66–7.74(m,3H),8.27(d,J=8.3Hz,1H),8.30(s,1H),8.36–8.41(m,2H),8.66(s,1H).

[0202] Using a similar scheme to that outlined above for the synthesis of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid, and with suitable advanced intermediates as starting materials, the following intermediates in Table 6 were prepared.

[0203] Table 6.

[0204]

[0205] Synthesis of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0206]

[0207] Preparation of 5-methylbenzo[b]thiophene-2-carboxylic acid

[0208] 5-Methylbenzo[b]thiophene-2-carboxylic acid was prepared according to the procedure described in WO 2016100184 A1.

[0209] Step 1: Preparation of 5-methylbenzo[b]thiophene-2-carboxylic acid benzyl ester

[0210] Benzyl bromide (20.6 g, 121.0 mmol, 1.1 equivalent) was added to a solution of 5-methylbenzo[b]thiophene-2-carboxylic acid (21.2 g, 110.0 mmol, 1.0 equivalent) and K₂CO₃ (30.4 g, 220.0 mmol, 2.0 equivalent) in DMF (200 mL). 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 under vacuum to give benzyl 5-methylbenzo[b]thiophene-2-carboxylate (30.1 g, 106.0 mmol, 97% yield) as a yellow solid. 1 H NMR (400MHz, CDCl3) δ8.01 (s, 1H), 7.71 (t, J = 12.2Hz, 1H), 7.65 (s, 1H), 7.46 (d ,J=6.8Hz,2H),7.42–7.35(m,3H),7.29–7.26(m,1H),5.38(s,2H),2.47(s,3H).

[0211] Step 2: Preparation of benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate

[0212] Benzoyl peroxide (1.3 g, 5.31 mmol, 0.1 equivalent) was added to a solution of 5-methylbenzo[b]thiophene-2-carboxylate (15.0 g, 53.1 mmol, 1.0 equivalent) and NBS (10.3 g, 58.4 mmol, 1.1 equivalent) in CCl4 (30 mL). The reaction flask was subjected to three cycles of evacuation and backfilled 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 silica gel rapid column chromatography to give 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (6.80 g, 18.8 mmol, 36% yield) as a yellow solid.

[0213] Step 3: Preparation of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0214] A solution of 5-(bromomethyl)benzo[b]thiophene-2-carboxylate benzyl ester (10.3 g, 28.5 mmol, 1.0 equivalent) dissolved in triethyl phosphite (30.0 g, 180.0 mmol, 6.3 equivalents) was stirred at 100 °C for 5 hours. The reaction mixture was directly concentrated under reduced pressure, and the residue was purified by silica gel rapid column chromatography to give 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate benzyl ester (6.5 g, 15.5 mmol, 55% yield) as a colorless oil. LCMS (ESI) m / z = 419 [M+H] + ; 1 H NMR(400MHz, CDCl3)δ8.04(s,1H),7.80(d,J=8.3Hz,2H),7.49–7.33(m,6H),5 .39(s,2H),4.08–3.93(m,4H),3.26(d,J=21.5Hz,2H),1.24(t,J=7.1Hz,6H).

[0215] Step 4: Preparation of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0216] LiOH (1.10 g, 26.6 mmol, 2.0 equivalent) was added to a solution of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (5.6 g, 13.3 mmol, 1.0 equivalent) dissolved in a mixture of THF (80 mL) and H₂O (10 mL). The mixture was stirred at room temperature for 3 hours and then acidified with 1 N HCl aqueous solution (adjusted to pH ~3-4). The product precipitated from the solution during acidification. The resulting solid was filtered, the filter cake was washed with water (20 mL x 2), and the solid was dried under vacuum 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] + .

[0217] Step 5: Preparation of perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0218] Oxaloyl chloride (2.2 g, 17.7 mmol, 1.5 equivalence) was added to a cooled (0 °C) solution of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (3.9 g, 11.8 mmol, 1.0 equivalence) in CH₂Cl₂ (50 mL), followed by two drops of DMF. The mixture was stirred at 0 °C for 30 min, and then the reaction mixture was evaporated to dryness. The resulting solid was dissolved in CH₂Cl₂ (50 mL), followed by the addition of Et₃N (3.6 g, 35.4 mmol, 3.0 equivalence) and pentafluorophenol (2.6 g, 14.1 mmol, 1.2 equivalence). The resulting mixture was stirred at room temperature for another 2 h, and then poured into H₂O (30 mL). The biphasic solution was extracted with EtOAc (30 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (4.7 g, 9.5 mmol, 81% yield) as a white solid. LCMS (ESI) m / z = 419 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ8.29 (s, 1H), 7.88 (d, J = 9.1Hz, 2H), 7.51 (d, J = 8.4Hz, 1H), 4.14–3.94 (m, 4H), 3.29 (d, J = 21.5Hz, 2H), 1.26 (t, J = 7.0Hz, 6H).

[0219] Step 6: Preparation of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0220] Trimethylbromosilane (12 mL) was added to a solution of perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (4.7 g, 9.5 mmol, 1.0 equivalent) in CH2Cl2 (60 mL). The mixture was stirred at room temperature for 14 hours and then concentrated under reduced pressure. The residue was purified by C18 column chromatography to give ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (3.7 g, 8.4 mmol, 89% yield) as a white solid. LCMS (ESI) m / z = 439 [M+H] + .

[0221] 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

[0222]

[0223] Step 1: rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0224] A solution of 1 M NaHMDS (11.4 mL, 11.4 mmol, 2 equivalents) in THF was added to a cooled (-78 °C) solution of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (2.4 g, 5.73 mmol, 1 equivalent) in THF (75 mL) and 2-(benzenesulfonyl)-3-phenyloxazine propane (2.97 g, 11.4 mmol, 2 equivalents) in THF. A deep purple solution was observed after the addition of the base, which turned orange after the base addition was complete. The mixture was stirred for another 10 minutes, followed by the addition of saturated NH4Cl aqueous solution (50 mL). The mixture was warmed to ambient temperature, and EtOAc (75 mL) and water (25 mL) were added. After stirring for another 30 minutes, the phases were separated. The aqueous layer was extracted with EtOAc (125 mL x 2). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Another batch of the same size was prepared and combined for purification. The combined material (6.42 mmol, total 12.15 mmol) was purified by rapid chromatography (20%–100% = EtOAc:heptane) to give rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (3.69 g, 8.49 mmol, 70%) as a white, viscous solid. LCMS (ESI) m / z = 869.4 [2M+H] + ; 1 H NMR(400MHz, CDCl3)δ8.10(s,1H),8.04–8.00(m,1H),7.88(d,J=8.6Hz,1H),7.61(d,J=8.6Hz,1H),7.51–7.47(m,2H), 7.46–7.35(m,3H),5.42(s,2H),5.17(dd,J=10.4,4.5Hz,1H),4.18–3.95(m,4H),3.10–2.99(m,1H),1.33–1.20(m,6H).

[0225] Step 2: rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0226] Add (diethylamino)sulfur trifluoride (568 μL, 4.30 mmol, 1.2 equivalents) to a cooled (-78 °C) solution of rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylate (cc) (1.56 g, 3.59 mmol, 1 equivalent) in CH2Cl2 (30 mL) (under N2 (g)). Stir the reaction for 15 min, then add saturated aqueous NaHCO3 solution (50 mL). After warming to room temperature, extract the product with CH2Cl2 (50 mL x 3). Dry the combined organic layers with sodium sulfate, filter, and concentrate under reduced pressure. The crude residue was purified by C18 reversed-phase chromatography (eluting with 5-80% acetonitrile / water) to obtain rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (650 mg, 1.48 mmol, 41.6%) as a viscous, clear oil. LCMS (ESI) m / z = 873.2 [2M+H] + ; 1 H NMR (400MHz, CDCl3) δ8.12(s,1H),8.02–7.99(m,1H),7.92(d,J=8.7Hz,1H),7.61(d,J=8.7Hz,1H),7.51–7.5 1(m,2H),7.46–7.36(m,3H),5.82(dd,J=44.4,7.5Hz,1H),5.42(s,2H),4.21–4.02(m,4H),1.34–1.26(m,6H).

[0227] Step 3: Preparation of (R)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester and (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0228] rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate benzyl ester (650 mg, 1.48 mmol) was separated by chiral SFC (column: Lux i-amyl starch 3, 21.2 x 250 mm 5 μm column, 75 mL / min, 40% MeOH) to give (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate benzyl ester (304 mg, 0.70 mmol, 46.8% recovery, 99.9% ee) (peak 1) and (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate benzyl ester (317 mg, 0.73 mmol, 49% recovery, 99.9% ee) (peak 2) as a viscous, clear oil.

[0229] Note: The enantiomer that elutes fastest by SFC is arbitrarily designated as (R)-5-(fluoro(phosphono)methyl)benzo[b]thiophen-2-carboxylic acid, and the enantiomer that elutes slowest by SFC is designated as (S)-5-(fluoro(phosphono)methyl)benzo[b]thiophen-2-carboxylic acid.

[0230] HPLC method for analyzing enantiomer excess: Lux cellulose-3 150 mm 45% H2O + 0.05% TFA / 55% MeCN 1 mL / min 8 min.

[0231] Step 4: Preparation of (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0232] A mixture of 10% Pd / C (60 mg, 50% humidity) and (R)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (peak 1) (60 mg, 0.1374 mmol, 1 equivalent) in THF (5 mL) was degassed with H2(g) for 5 min. H2(g) was bubbled into the mixture for 5 min, and then the reaction was stirred at room temperature with H2(g) (1 atm). The reaction mixture was stirred until the consumption of the starting material was detected by LCMS. The reaction mixture was then sprayed with N2(g) for 15 min and subjected to… Filtering was performed using a pad filter. 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 viscous, clear oil. LCMS (ESI) m / z = 347.2 [M+H] + .

[0233] Using the procedure outlined above (in step 4), begin with (S)- or (R)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (peak 2) and prepare the following intermediates in Table 7 using appropriate reagents.

[0234] Table 7.

[0235]

[0236] Representative methods for synthesizing activated connectors

[0237] Method 1: Stepwise acid chloride method for synthesizing mixed joints

[0238] Representative procedure for the synthesis of 4-nitrobenzene 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid ester

[0239]

[0240] Step 1: Preparation of 4-nitrobenzene 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoyl)methyl)benzo[b]thiophene-2-carboxylic acid

[0241] Catalytic DMF (2 drops) was added to a cooled (0°C) heterogeneous solution of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (100 mg, 0.2329 mmol, 1 equivalent) in CH2Cl2 (4 mL), followed by dropwise addition of oxaloyl chloride (198 μL, 2.32 mmol, 10 equivalents). The homogeneous reaction mixture was warmed to room temperature and stirred for 2 h. The reaction was concentrated under vacuum and further dried under high vacuum for 30 min to give a yellow solid. The yellow solid was diluted in CH2Cl2 (4 mL) and cooled to -78°C. A solution of pyridine-3-ol (22.0 mg, 232 μmol, 1 equivalent) and triethylamine (64.7 μL, 465 μmol, 2 equivalents) in CH2Cl2 (1 mL) was slowly added [sonicated for 1 min to dissolve the solution]. The homogenized reaction mixture was stirred at -78°C for 2 minutes, then warmed to ambient temperature and stirred overnight. After 24 hours, the reaction mixture became heterogeneous, and was concentrated under reduced pressure. The crude product, 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoyl)methyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester, could be used directly in the next step without further purification or treatment. LCMS m / z = 507.2 [M+H] + .

[0242] Step 2: Preparation of 4-nitrobenzene 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0243] Two drops of DMF were added to a cooled (0°C) solution of 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid nitrobenzene ester (117 mg, 0.2329 mmol, 1 equivalent) in CH₂Cl₂ (5 mL), followed by dropwise addition of oxaloyl chloride (198 μL, 2.32 mmol, 10 equivalents). The reaction was warmed to room temperature and stirred for 1.5 h. LCMS analysis showed partial conversion to the desired activated intermediate. An additional oxaloyl chloride (198 μL, 2.32 mmol, 10 equivalents) was introduced into the reaction mixture, and the mixture was stirred for another 1 h. The reaction was concentrated under vacuum and further dried under high vacuum for 30 min to give a yellow solid. The yellow solid was diluted in CH₂Cl₂ (5 mL) and cooled to -78°C. A solution of 1-[(2-hydroxyethyl)sulfonyl]but-1-one (103 mg, 698 μmol, 3 equivalents) diluted in CH2Cl2 (1 mL) [previously dried by passing through anhydrous Na2SO4] was slowly added to the cooled solution, followed by the addition of triethylamine (134 μL, 967 μmol, 2 equivalents). After stirring for 2 minutes, the resulting mixture was warmed to ambient temperature and stirred overnight. Add... The mixture was carefully concentrated under vacuum. The crude residue was purified by rapid chromatography (gradient elution 0-60% EtOAc / heptane) to give 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoyl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester (22.0 mg, 0.03455 mmol, 14.9% yield) as a clear oil. LCMS m / z = 637.2 [M+H] + ; 1 HNMR: (400MHz, DMSO-d6) δ 8.50–8.44 (m, 2H), 8.33–8.38 (m, 3H), 8.25 (s, 1H), 8.04 (d, J = 8.0Hz, 1H), 7.79 (d, J = 8.6Hz, 1H), 7.57–7.53 (m, 1H), 7.50–7.45 (m, 2H), 7.31–7.26 (m, 1H), 4.37–4.24 (m, 2H), 3.20–3.08 (m, 2H), 2.52 (t, J = 7.6Hz, 2H), 1.67 (sextet, J = 7.3Hz, 2H), 0.94 (t, J = 7.6Hz, 3H).

[0244] Using a similar scheme described above for the synthesis of 4-nitrobenzene 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid, and with suitable advanced intermediates as starting materials, the following intermediates in Table 8 were prepared.

[0245] Table 8.

[0246]

[0247]

[0248]

[0249] Method 2: One-pot acid chloride method for synthesizing activated linkers

[0250] Representative procedure for the synthesis of perfluorophenyl 5-((bis(4-((3-methylbutyryl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0251]

[0252] Oxaloyl chloride (266 mg, 2.10 mmol, 5.0 equivalent) was added dropwise to a cooled (0 °C) solution of 3 (200 mg, 0.42 mmol, 1.0 equivalent) in dry CH2Cl2 (15 mL) and catalytic DMF (3.2 μL, 42.1 μmol, 0.1 equivalent). The reaction mixture was warmed to 40 °C. After stirring for 2 hours, the reaction mixture was concentrated under vacuum and dried (to remove excess oxaloyl chloride). The resulting solid was redissolved in anhydrous CH2Cl2 (5 mL) and cooled to 0 °C. Add 239 mg of 3-methylthiobutyrate-S-(4-hydroxybutyl) ester (3.0 equivalence), DMAP (5.14 mg, 42.1 μmol, 0.1 equivalence), and N,N-diisopropylethylamine (217 mg, 1.68 mmol, 4.0 equivalence) to a solution of anhydrous CH2Cl2 (10 mL). Warm the reaction mixture to room temperature and stir for an additional 18 hours. Quench the reaction by adding 10 mL of H2O and extract with CH2Cl2 (3 x 10 mL). Combine the organic layers, wash with 20 mL of brine, dry to anhydrous Na2SO4, and concentrate under vacuum. The residue was purified by rapid column chromatography to yield perfluorophenyl 5-((bis(4-((3-methylbutyryl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (15.0 mg, 18.3 μmol, 4.4% yield). LCMS (ESI) m / z = 819 [M+H] + .

[0253] Using a similar scheme described above for the synthesis of 5-((bis(4-((3-methylbutyryl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester, and with suitable advanced intermediates as starting materials, the following intermediates in Table 9 were prepared.

[0254] Table 9.

[0255]

[0256]

[0257] Method 3: One-pot silver salt method for synthesizing activated junctions

[0258] Representative procedure for the synthesis of perfluorophenyl 5-((bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0259]

[0260] Step 1: Preparation of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)silver phosphonate (I)

[0261] Add Amberlite to a solution of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (300 mg, 684 μmol, 1.0 equivalent) in a mixture of deionized H₂O (4 mL) and THF (2 mL). Resin (Na) + (Form) (1.5 g). The resulting mixture was stirred at room temperature for 1 hour, and then the suspension was filtered. A solution of AgNO3 (463 mg, 2.73 mmol, 4.0 equivalent) in deionized H2O (2 mL) was added to the filtrate, and the resulting mixture was stirred at room temperature for another 1 hour. The formation of a white precipitate was observed, and the solid was collected by filtration. The filter cake was then washed with cold H2O (3 x 2 mL), and the solid was dried under reduced pressure to produce silver ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonate (I) as a dry powder. The silver salt can be used without further purification.

[0262] Step 2: Preparation of 5-((bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0263] A suspension of silver (I) 2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonate was suspended in anhydrous toluene (10 mL), and methyl iodide 2-methylpropionate (500 mg, 2.05 mmol, 3.0 equivalent) was added dropwise. After addition, the resulting mixture was stirred at room temperature for an additional 12 hours. The reaction progress was monitored by LCMS, and unreacted silver salt was recovered by filtration upon completion. The filtrate solution was concentrated under vacuum, and the resulting residue was purified by reversed-phase chromatography [C18 column gradient elution water / acetonitrile = 90% to 1%] to give perfluorophenyl 5-((bis((((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (165 mg, 246 μmol, 36% yield) as a white solid. LCMS (ESI) m / z = 671 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ8.28 (s, 1H), 7.96-7.80 (m, 2H), 7.48 (d, J = 8.5Hz, 1H), 5.70-5.5 3(m,4H), 4.90(dt,J=12.6,6.2Hz,2H), 3.42(d,J=22.2Hz,2H), 1.31(d,J=6.2Hz,12H).

[0264] Using a similar scheme described above for the synthesis of 5-((bis(4-((3-methylbutyryl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester, and with suitable advanced intermediates as starting materials, the following intermediates in Table 10 were prepared.

[0265] Table 10.

[0266]

[0267]

[0268]

[0269] (R)- or (S)-5-((bis((neovaleryloxy)methoxy)phosphoyl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0270]

[0271] Step 1: Preparation of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid

[0272] Add BSTFA (746 μL, 2.81 mmol) to a cooled (0 °C) solution of (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (peak 1) (205 mg, 0.4697 mmol) in CH2Cl2 (8.0 mL), followed by the addition of a solution of 1.0 M trimethyliodosilane (1.87 mL, 1.87 mmol) in CH2Cl2. After stirring for 1 hour, add a mixture of acetonitrile (0.66 mL), water (0.33 mL), and 0.1% TFA. Remove the solvent under reduced pressure at 0 °C. The crude residue was purified by reversed-phase chromatography (C18 cartridge, eluted with 5-40% acetonitrile / water) to give (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid (163 mg, 0.4285 mmol) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ8.33(s,1H),8.11–8.06(m,2H),7.58(d,J=8.6Hz,1H),7 .51–7.47(m,2H),7.45–7.33(m,3H),5.84(dd,J=44.6,8.3Hz,1H),5.40(s,2H).

[0273] Step 2: Preparation of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid

[0274] [The following reactions were carried out in a foil-covered container and in the absence of ambient light]: A solution of sodium hydroxide aqueous solution (34.2 mg, 857 μmol, 2 equivalents) in water was added to a suspension of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid (163 mg, 0.4285 mmol, 1 equivalent) in water (5 mL). Silver nitrate (I) (181 mg, 1.07 mmol, 2.5 equivalents) was added to the yellow solution, and the resulting off-white suspension was stirred at room temperature for 1.5 hours. 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 hours). The resulting dark yellow powder was suspended in toluene (10 mL), and methyl iodide 2,2-dimethylpropionate (191 μL, 1.28 mmol, 3 equivalents) was added. After stirring for 20 hours, the reaction mixture was stirred at room temperature for another 20 hours. The reaction mixture was filtered and washed with toluene. The filtrate was concentrated under reduced pressure. The crude residue was purified (C18 cartridge, eluted with 5-100% acetonitrile / water) to give (R)- or (S)-((((2-(((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionate) (122 mg, 0.2004 mmol, 46.9%) as a clear oil. 1 H NMR (400MHz, CDCl3) δ8.11(s,1H),8.01–7.99(m,1H),7.92(d,J=8.3Hz,1H),7.59(d,J=8.3Hz,1H),7.52–7 .47(m,2H),7.46–7.36(m,3H),8.87(dd,J=44.3,7.5Hz,1H),5.72–5.62(m,4H),5.43(s,2H),1.21(s,18H).

[0275] Step 3: Preparation of (R)- or (S)-5-((bis((neovaleryloxy)methoxy)phosphoyl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0276] Under N2(g), 10% Pd / C (50% humidity, 120 mg, 0.1127 mmol, 0.56 equivalents) was added to a solution of (R)-((((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionate) (122 mg, 0.200 mmol, 1 equivalent) in THF (10 mL). The suspension was bubbled with H2(g) for 5 minutes. The reaction mixture was stirred at room temperature under H2(g) (1 atm). After stirring for 22 hours, the reaction mixture was purged with N2(g) and subjected to… Filtration. The filter pad was washed with THF and concentrated under reduced pressure to give (R)-5-((bis((neovaleryloxy)methoxy)phosphoyl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid (0.200 mmol, 99.9%) as a viscous, clear oil. LC-MS (ESI) m / z [M+H] + =519.1.

[0277] Using a similar scheme described above for the synthesis of (R)- or (S)-5-((bis((neovaleroxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid, and utilizing suitable advanced intermediates as starting materials, the following intermediates in Table 11 were prepared. The absolute configuration of the starting material, (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid was not determined.

[0278] Table 11.

[0279]

[0280]

[0281] Method 4: Stepwise silver salt method for synthesizing hybrid joints

[0282] Representative procedure for 5-(((2-(butyrylthio)ethoxy)(2-(neovalerylthio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0283]

[0284] Step 1: Preparation of 5-(difluoro(hydroxy(2-(neovalerylthio)ethoxy)phospho)methyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0285] Using the method described in step 1, method 3 for synthesizing silver (2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonate (I), silver (2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid is synthesized from (difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid.

[0286] Silver 2,2-dimethylthiopropionic acid-S-(2-iodoethyl) phosphonate (I) (647 mg, 940 μmol, 1.0 equivalent) was added dropwise to a suspension of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonate (I) in anhydrous toluene (10 mL) (310 mg, 1.14 mmol, 1.2 equivalent). After the addition of alcohol was complete, the resulting mixture was stirred at room temperature for an additional 12 hours. The heterogeneous mixture was filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified by reversed-phase chromatography to give perfluorophenyl 5-(difluoro(hydroxy(2-(neovalerylthio)ethoxy)phosphoyl)methyl)benzo[b]thiophene-2-carboxylic acid (240 mg, 388 μmol, 41% yield). LCMS (ESI) m / z = 617 [MH] - .

[0287] Step 2: Preparation of 5-(((2-(butyrylthio)ethoxy)(2-(neovalerylthio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0288] 5-(((2-(butyrylthio)ethoxy)(2-(neovalerylthio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester was synthesized using a similar scheme outlined above. Starting with 5-(difluoro(hydroxy(2-(neopentylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester (190 mg, 307 μmol, 1.0 equivalent), AgNO3 (207 mg, 1.22 mmol, 4.0 equivalent), and thiobutyrate-S-(2-iodoethyl) ester (94.9 mg, 368 μmol, 1.2 equivalent), a white solid 5-(((2-(butyrylthio)ethoxy)(2-(neopentylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester (55.0 mg, 73.4 μmol, 24%) is produced. LCMS(ESI) m / z = 749 [M+H] + .

[0289] Using a similar scheme described above for the synthesis of 5-(((2-(butyrylthio)ethoxy)(2-(neovalerylthio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester, and using suitable advanced intermediates as starting materials, the following intermediates in Table 12 were prepared.

[0290] Table 12.

[0291]

[0292] Preparation of 5-(((((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0293]

[0294] Step 1: Preparation of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0295] 3-bromoprop-1-ene (440 mg, 3.6 mmol, 1.2 equivalence) was added to a suspension of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (1.0 g, 3.0 mmol, 1.0 equivalence) and K₂CO₃ (839 mg, 6.1 mmol, 2.0 equivalence) in DMF (20 mL). The mixture was stirred at room temperature for 14 hours and then poured into water (30 mL). The mixture was extracted with EtOAc (25 mL x 3). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to give allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (0.980 g, 2.7 mmol, 88% yield) as a pale yellow solid. LCMS (ESI) m / z = 369 [M+H] + ; 1 H NMR(400MHz, CDCl3)δ8.04(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,1 H), 5.36–5.28 (m, 1H), 4.87–4.83 (m, 2H), 4.08–3.97 (m, 4H), 3.27 (d, J = 21.4Hz, 2H), 1.25 (t, J = 7.1Hz, 6H).

[0296] Step 2: Preparation of ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0297] Trimethylbromosilane (3 mL) was added to a solution of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (980 mg, 2.7 mmol, 1.0 equivalent) in CH2Cl2 (15 mL). The mixture was stirred at room temperature for 14 hours and then concentrated under reduced pressure. The resulting residue was ground with H2O (5 mL) and the resulting precipitate was filtered. The filter cake was washed with H2O (5 mL x 2) and dried under reduced pressure to give ((2-((allyloxy)carbonyl)benzo[b]thiophene-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(400MHz, DMSO-d6)δ8.20(s,1H),7.96(d,J=8.4Hz,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=21.2Hz,2H).

[0298] Step 3: Preparation of 5-(((((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0299] Oxaloyl chloride (3.40 g, 26.8 mmol, 1 equivalent) was added to a cooled (0 °C) solution of ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (2.80 g, 8.96 mmol, 1 equivalent) and catalytic DMF (1 drop) in dry CH2Cl2 (50 mL) under a constant flow of N2 (g). After the gas stopped bubbling, the mixture was heated at 40 °C. After 2 hours, the mixture was cooled to room temperature and concentrated under vacuum to give a yellow solid. The solid was then diluted with CH2Cl2 (50 mL) and cooled to 0 °C. Phenol (0.843 g, 8.96 mmol, 1 equivalent) and Et3N (4.53 g, 44.8 mmol, 5 equivalent) were added to the cooled solution. After the addition was complete, the mixture was warmed to room temperature and stirred for 1 hour, followed by the introduction of (2S)-2-aminopropionic acid propionate (1.75 g, 13.4 mmol, 1.5 equivalents). After stirring for another 2 hours, the mixture was concentrated to dryness. The residue was purified by C18 column chromatography (eluting with 50%–80% acetonitrile / water) to give 5-(((((S)-1-isopropoxy-1-oxopropion-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (2.23 g, 4.44 mmol, 49.6% yield) as a white solid. LCMS (ESI) m / z = 502.0 [M+H] + ; 1 H NMR(400MHz, CDCl3)δ8.05(d,J=6.9Hz,1H),7.91–7.80(m,2H),7.53–7.43(m,1H),7.2 9(d,J=8.1Hz,2H),7.18–7.09(m,3H),6.05(ddd,J=16.1,10.9,5.6Hz,1H),5.48–5.40 (m,1H),5.32(dd,J=10.4,1.2Hz,1H),4.98–4.87(m,1H),4.85(d,J=5.7Hz,2H),4.04– 3.85(m,1H),3.44(dd,J=20.7,14.1Hz,2H),3.12(t,J=10.9Hz,1H),1.21–1.10(m,9H).

[0300] Step 4: Preparation of 5-(((((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0301] A solution of 5-((((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (90 mg, 0.1794 mmol, 1 equivalent), pyrrolidine (12.7 mg, 179 μmol, 1 equivalent), and Pd(PPh3)4 (10.3 mg, 8.97 μmol, 0.05 equivalent) in CH2Cl2 (5 mL) was stirred under N2 (g). After 2 hours, the reaction was concentrated under vacuum. The residue was purified by C18 column elution (30%–70% acetonitrile / water) to yield 5-(((((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid as a white solid (64.0 mg, 0.1386 mmol, 77.4% yield). LCMS (ESI) m / z = 462.1 [M+H] + .

[0302] The following intermediates in Table 13 were prepared using the methods described above for the synthesis of 5-(((((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid, with appropriate starting materials and modifications.

[0303] Table 13.

[0304]

[0305] Synthesis scheme:

[0306] Synthesis of 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid

[0307]

[0308] Step 1: Preparation of 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0309] At -78 °C, a solution of sodium bis(trimethylsilylamide) (1 M in THF) (1.43 mL, 1.43 mmol, 3.0 equivalents) was added dropwise to a solution of 5-[(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylate (200 mg, 0.4779 mmol, 1 equivalent) in tetrahydrofuran (10 mL). The reaction was stirred at -78 °C for 5 min, followed by a single addition of 1H-benzotriazole-1-methanol (142 mg, 0.955 mmol, 2 equivalents). The reaction was stirred at -78 °C for 2 h. The reaction mixture was quenched at -78 °C with saturated ammonium chloride aqueous solution (10 mL), and then the ice bath was removed. The product was extracted with EtOAc (3 x 30 mL). The combined extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 50 g C 18 Purified on a elution chamber (eluted with a gradient of 5-80% MeCN / water (containing 0.1% formic acid)) to give benzyl 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylate (24 mg, 0.05351 mmol, 11.2%) as a clear, viscous oil. LCMS: m / z = 449.2 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.27(s,1H),8.02(d,J=8.5Hz,1H),7.99(s,1H),5.40(s,2H),4.87(t,J=5.5Hz,1 H), 4.07-3.94 (m, 3H), 3.93-3.71 (m, 3H), 3.52-3.41 (m, 1H), 1.21 (t, J = 7.3Hz, 3H), 1.03 (t, J = 7.3Hz, 3H).

[0310] Step 2: Preparation of 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid

[0311] A mixture of 10% palladium / carbon (50% wet) (80 mg, 0.03758 mmol, 0.163 equivalents) and benzyl 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylate, and benzyl 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylate (38 mg, 0.08473 mmol, 1 equivalent) in tetrahydrofuran (8 mL) was degassed with nitrogen for 5 minutes. Hydrogen was bubbled for 5 minutes, and then the reaction was stirred at room temperature under hydrogen (1 atm) for 20 hours. The reaction mixture was filtered through diatomaceous earth and eluted with MeOH. The filtrate was concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 50 g C 18The solution was purified on a syringe (eluted with 5-80% MeCN / water (containing 0.1% formic acid)), then concentrated under reduced pressure and freeze-dried to give 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid (8.00 mg, 0.02232 mmol, 26.4%) as a white solid. LCMS: m / z = 359.0 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ8.02-7.92(m,2H),7.90(s,1H),7.48-7.42(m,1H),4.92-4.79(m ,1H),4.08-3.69(m,6H),3.52-3.39(m,1H),1.21(t,J=7.2Hz,3H),1.03(t,J=7.3Hz,3H).

[0312] Synthesis of 5-(1-(diethoxyphosphoryl)ethyl)benzo[b]thiophene-2-carboxylic acid and 5-(2-(diethoxyphosphoryl)prop-2-yl)benzo[b]thiophene-2-carboxylic acid

[0313]

[0314] Step 1: Preparation of rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester and 5-[2-(diethoxyphosphoryl)propyl-2-yl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0315] A solution of bis(trimethylsilylamide) sodium ... 18Purification was performed on a elution chamber (eluted with a gradient of 5-80% MeCN / water (containing 0.1% formic acid)) to give rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (89.0 mg, 0.1993 mmol, 41.9%) as a clear oil and 5-[2-(diethoxyphosphoryl)propyl-2-yl]-1-benzothiophene-2-carboxylic acid benzyl ester (22.0 mg, 0.5087 mmol, 10.6%) as a clear oil.

[0316] rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester LCMS: m / z = 433.0 [M+H] + ; 1 H NMR(400MHz, DMSO-d6)δ8.08(s,1H),7.88-7.85(m,1H),7.83(d,J=8.7Hz,1H),7.52-7.47(m,3H),7.45-7.36(m,3H),5.42(s,2H),4.12-4.02 (m,2H),4.00-3.91(m,1H),3.89-3.78(m,1H),3.38-3.25(m,1H),1.66(dt,J=18.3,7.6Hz,3H),1.30(t,J=7.3Hz,3H),1.16(t,J=7.3Hz,3H).

[0317] 5-[2-(diethoxyphosphoryl)prop-2-yl]-1-benzothiophene-2-carboxylic acid benzyl ester: LCMS: m / z = 447.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.01(s,1H),7.93-7.90(m,1J),7.74(d,J=8.6Hz,1H),7.69-7.63(m,1H),7.39(d,J= 8.6Hz,1H),7.36-7.25(m,3H),5.32(s,2H),3.92-3.73(m,4H),1.61(d,J=16.8Hz,6H),1.11(t,J=7.2Hz,6H).

[0318] Step 2: Preparation of rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid

[0319] A mixture of 10% Pd / C (50% wet) (80 mg, 0.03758 mmol, 0.163 equivalents) and 5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (100 mg, 0.2312 mmol, 1 equivalent) in tetrahydrofuran (8 mL) was degassed with nitrogen for 5 minutes. Hydrogen was bubbled for 5 minutes, and then the reaction was stirred at room temperature under hydrogen (1 atm). The reaction mixture was filtered through diatomaceous earth and eluted with THF. The filtrate was concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 50 g C 18 Purified on a elution tube (eluted with a gradient of 5-80% MeCN / water (containing 0.1% formic acid)) to give rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid (51 mg, 0.1489 mmol, 65%) as a white solid. LCMS: m / z = 343.0 [M+H] + .

[0320] Step 3: Preparation of 5-[2-(diethoxyphosphoryl)prop-2-yl]-1-benzothiophene-2-carboxylic acid

[0321] A mixture of 10% palladium / carbon (50% wet) (5 mg, 0.002349 mmol, equivalent) and 5-[2-(diethoxyphosphoryl)propyl-2-yl]-1-benzothiophene-2-carboxylic acid benzyl ester (33 mg, 0.07390 mmol, 1 equivalent) in tetrahydrofuran (2 mL) was degassed with nitrogen for 5 minutes. The mixture was bubbled with hydrogen for 5 minutes, and then stirred at room temperature under hydrogen (1 atm). The reaction was filtered through a syringe filter and eluted with MeOH. The filtrate was concentrated under reduced pressure. The crude residue was diluted in tetrahydrofuran (2 mL). 10% palladium / carbon (50% wet) (20 mg, 0.009396 mmol, 0.13 equivalent) was added under nitrogen bubbling, and then bubbled with hydrogen for 5 minutes. The reaction was stirred overnight at room temperature under hydrogen (1 atm). The reaction was filtered through a syringe filter and eluted with MeOH. The filtrate was concentrated under reduced pressure. The crude residue was diluted in tetrahydrofuran (2 mL), and then 10% palladium / carbon (50% wet) (20 mg, 0.009396 mmol, 0.13 equivalents) was added under bubbling nitrogen, followed by bubbling with hydrogen for 5 minutes. The reaction was stirred at room temperature under hydrogen (1 atm) for 40 hours. The reaction was filtered through a syringe filter and eluted with MeOH. The filtrate was concentrated under reduced pressure to give the crude product 5-[2-(diethoxyphosphoryl)propyl-2-yl]-1-benzothiophene-2-carboxylic acid (26.3 mg, 0.0738 mmol, 99%) as a clear oil. LCMS: m / z = 357.2 [M+H] + .

[0322] Synthesis of 5-((bis((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0323]

[0324] Step 1: Preparation of 2-methoxyethyl chloromethyl carbonate

[0325] A solution of 2-methoxyethyl-1-ol (1 g, 13.1 mmol, 1 equivalent) and chloroacetyl chloride (1.68 g, 13.1 mmol, 1 equivalent) in diethyl ether (30 mL) was cooled to 0 °C under nitrogen atmosphere. Pyridine (1.04 mL, 13.1 mmol, 1.0 equivalent) was added dropwise, and the reaction was then stirred at 0 °C for 15 min, followed by stirring at room temperature for 16 h. The white suspension was filtered and washed with diethyl ether (30 mL). The filtrate was washed with 1 N HCl (20 mL) and water (2 x 20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 2-methoxyethyl chloromethyl carbonate (1.63 g, 9.66 mmol, 74.0% yield) as a clear liquid. 1 H NMR (400MHz, CDCl3) δ5.76 (s, 2H), 4.42-4.37 (m, 2H), 3.68-3.64 (m, 2H), 3.42 (s, 3H).

[0326] Step 2: Preparation of 5-((bis((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0327] Water (2 mL) containing sodium hydroxide (59.9 mg, 1.50 mmol, 2 equivalents) was added dropwise to a stirred solution of ((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (300 mg, 0.7531 mmol, 1 equivalent) in water (15 mL). When the mixture reached pH ~8, silver nitrate (382 mg, 2.25 mmol, 3 equivalents) was added in a single batch. After 2 hours at room temperature, the gray suspension was cooled to 0 °C. The precipitate was collected by filtration, washed with water, absorbed in MeCN, dried under reduced pressure, and then dried under high vacuum for 2 hours. The solid was suspended in dry toluene (10 mL), and methyl chlorocarbonate 2-methoxyethyl ester (379 mg, 2.25 mmol, 3 equivalents) was added. The mixture was stirred at room temperature for 18 hours, then heated to 50 °C for 5 days. The mixture was adsorbed onto silica gel and concentrated under reduced pressure. The crude residue was purified by rapid chromatography on a 24 g silica gel cartridge (eluted with 0-90% EtOAc / heptane) to give 5-((bis(((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (290 mg, 0.4377 mmol, 58.2% yield) as a clear oil. LCMS: m / z = 663.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.17(s,1H),8.16-8.14(m,1H),7.98(d,J=8.4Hz,1H),7.69(d,J=8.4Hz,1H),7.51-7.48(m,2H),7.46-7.36(m ,3H),5.75(dd,J=12.0,5.5,Hz,2H),5.69(dd,J=12.0,5.5Hz,2H),5.43(s,2H),4.36-4.31(m,4H),3.64-3.60(m,4H),3.39(s,6H).

[0328] Step 3: Preparation of 5-((bis((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0329] 10% palladium / carbon (50% wet) (145 mg, 0.1353 mmol, equivalent) was added to a mixture of 5-((bis((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (145 mg, 0.2188 mmol, 1 equivalent) in anhydrous tetrahydrofuran (15 mL). Hydrogen was bubbled into the suspension for 2 minutes, and the reaction mixture was then stirred under hydrogen (1 atm) for 20 hours. Nitrogen was bubbled, and the reaction was filtered through diatomaceous earth, washed with 2-MeTHF, and then concentrated under reduced pressure to give the crude product 5-((bis((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (144 mg, 0.2188 mmol, 99% yield) as a gray solid. LCMS: m / z = 573.2 [M+H] + .

[0330] The following connectors in Table 14 are prepared using appropriate starting materials according to the described procedure for the synthesis of 5-((bis((((2-methoxyethoxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid.

[0331] Table 14.

[0332]

[0333] Synthesis of 5-((bis(4-ethoxy-4-oxobutoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0334]

[0335] Step 1: Preparation of diethyl 4,4'-((((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphoryl)bis(oxy))diethyl dibutyrate

[0336] Add dropwise 2 mL of H₂O containing sodium hydroxide (59.9 mg, 1.50 mmol) to a stirred solution of ((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (300 mg, 0.7531 mmol, 1 equivalent) in 15 mL of H₂O. When the mixture reaches pH ~8, add silver nitrate (382 mg, 2.25 mmol). After 2 hours at room temperature, cool the gray suspension to 0 °C. Collect the precipitate by filtration, wash with water, absorb in MeCN, concentrate under reduced pressure, and then dry under high vacuum. Suspend the solid in dry toluene (10 mL) and add ethyl 4-bromobutyrate (438 mg, 2.25 mmol, 3 equivalents). Stir the mixture at 50 °C for 18 hours. Adsorb the mixture onto silica gel and concentrate under reduced pressure. The crude residue was purified by rapid chromatography on a 24 g silica gel cartridge (eluted with 0-90% EtOAc / heptane) to give 4,4'-((((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphoryl)bis(oxy))diethyl dibutyrate (320 mg, 0.5106 mmol, 67.9% yield) as a clear oil. 1 HNMR (400MHz, CDCl3) δ8.18-8.13(m,2H),7.97(d,J=8.6Hz,1H),7.70(d,J=8.5Hz,1H),7.53-7.35(m,5H). 5.43(s,2H),4.28-4.09(m,8H),2.44-2.36(m,4H),2.04-1.92(m,4H),1.32-1.22(m,6H).

[0337] Step 2: Preparation of 5-((bis(4-ethoxy-4-oxobutoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0338] 10% palladium / carbon (50% wet) (100 mg, 0.04698 mmol, 0.3 equivalents) was added to a mixture of 4,4'-((((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphoryl)bis(oxy))diethyl dibutyrate (100 mg, 0.1595 mmol, 1 equivalent) in anhydrous tetrahydrofuran (10 mL). Hydrogen was bubbled into the suspension for 2 minutes, and the reaction mixture was then stirred at 1 atm·H₂O for 16 hours. Nitrogen was bubbled into the reaction mixture, and the mixture was then filtered through diatomaceous earth (washed with 2-MeTHF) and concentrated under reduced pressure to give 5-((bis(4-ethoxy-4-oxobutoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (85.3 mg, 0.1595 mmol, 100% yield) as a clear oil. LCMS: m / z = 559.2 [M + Na] + .

[0339] Synthesis of 5-(cyano(ethoxy(hydroxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0340]

[0341] Step 1: Preparation of ethyl 5-(cyano(diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0342] Sodium hydride (60% in mineral oil) (50.3 mg, 1.26 mmol, 2.1 equivalents) was added to a solution of diethyl cyanomethylphosphonate (127 mg, 722 μmol, 1.2 equivalents) in anhydrous 1,2-dimethoxyethane (5 mL) under nitrogen atmosphere. The reaction was stirred at room temperature for 10 min, and then tetrakis(triphenylphosphine)palladium (34.7 mg, 30.1 μmol, 0.05 equivalents) and ethyl 5-iodo-1-benzothiophene-2-carboxylate (200 mg, 602 μmol, 1 equivalent) were added. The reaction mixture was stirred at 85 °C for 16 h. The reaction mixture was filtered through diatomaceous earth (eluted with DCM), and the filtrate was concentrated under reduced pressure. The crude residue was analyzed by reversed-phase chromatography at 50 g C 18 Purified on a cartridge (eluted with 5-80% MeCN / water). The combined fractions were concentrated to give ethyl 5-(cyano(diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate as an orange solid (50.0 mg, 131 μmol, 21.8% yield). LCMS: m / z = 382.2 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ8.33 (s, 1H), 8.18-8.06 (m, 2H), 7.54 (d, J = 8.6Hz, 1H), 5.67-5.56 (m, 1H ), 4.37 (q, J = 7.1Hz, 2H), 4.11-3.97 (m, 4H), 1.34 (t, J = 7.1Hz, 3H), 1.20 (dt, J = 9.8, 7.1Hz, 6H).

[0343] Step 2: Preparation of 5-(cyano(ethoxy(hydroxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0344] Hydrochloric acid (6 mL, 3 N) was added to a solution of ethyl 5-(cyano(diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (80 mg, 209 μmol, 1 equivalent) in acetonitrile (4 mL). The reaction mixture was stirred at 70 °C for 7 hours. More hydrochloric acid (1.5 mL, 3 N) was added, and the reaction mixture was stirred at 70 °C for another 22 hours. Hydrochloric acid (1.5 mL, 3 N) was added, and the reaction mixture was stirred at 80 °C for another 20 hours. The reaction mixture was concentrated under reduced pressure. The crude product was analyzed by reversed-phase chromatography using 50 g C 18 The solution was directly purified by elution with a MeCN / water gradient (5% in 3CV, then 5% to 100% in 18CV) to give 5-(cyano(ethoxy(hydroxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (52.0 mg, 159 μmol, 76.5% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.19 (s, 1H), 8.12-7.98 (m, 2H), 7.52 (d, J = 8.6Hz, 1H), 5.23-5.12 (m, 1H), 3.97 (quin, J = 7.3Hz, 2H), 1.22-1.13 (m, 3H).

[0345] Synthesis of rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid

[0346]

[0347] Step 1: Preparation of rac-5-[(diethoxyphosphoryl)(hydroxy)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0348] A solution of 5-[(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (2.4 g, 5.73 mmol, 1 equivalent) in tetrahydrofuran (75 mL) was cooled to -78 °C, and then 2-(benzenesulfonyl)-3-phenyloxazine propane (2.97 g, 11.4 mmol, 2 equivalents) was added dropwise, followed by the dropwise addition of sodium bis(trimethylsilylamide) solution (1.0 M in THF) (11.4 mL, 11.4 mmol, 2 equivalents) (internal T... o = -75℃ to -69℃). A deep purple solution was observed after the addition of alkali, which rapidly turned orange. The mixture was stirred at -78℃ for 10 minutes. The reaction mixture was quenched at -78℃ with a saturated aqueous solution of NH4Cl (50 mL), and then the dry ice bath was removed. EtOAc (75 mL) and water (25 mL) were added, and the mixture was stirred for 30 minutes (until the internal temperature reached 15℃). The phases were separated, and the aqueous layer was then back-extracted with EtOAc (125 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, adsorbed onto silica gel, and concentrated under reduced pressure. The crude material was combined with another batch (6.42 mmol, total 12.15 mmol) and purified by rapid chromatography on a 330 g silica gel cartridge (eluted with 20–100% EtOAc / heptane) to give rac-5-[(diethoxyphosphoryl)(hydroxy)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (3.69 g, 8.49 mmol, 70% yield) as a white, viscous solid. LCMS: m / z = 869.4 (2M+H) + ; 1 H NMR(400MHz, CDCl3)δ8.10(s,1H),8.04-8.00(m,1H),7.88(d,J=8.6Hz,1H),7.61(d,J=8.6Hz,1H),7.51-7.47(m,2H), 7.46-7.35(m,3H),5.42(s,2H),5.17(dd,J=10.4,4.5Hz,1H),4.18-3.95(m,4H),3.10-2.99(m,1H),1.33-1.20(m,6H).

[0349] Step 2: Preparation of rac-5-[(diethoxyphosphoryl)(fluoro)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0350] (Diethylamino)sulfur trifluoride (568 μL, 4.30 mmol, 1.2 equivalent) was added to a solution of rac-5-[(diethoxyphosphoryl)(hydroxy)methyl]-1-benzothiophene-2-carboxylate (1.56 g, 3.59 mmol, 1 equivalent) in methylene chloride (30 mL) under nitrogen at -78 °C. The reaction was stirred at -78 °C for 15 min. The reaction was quenched by adding saturated aqueous sodium bicarbonate solution (50 mL) and the product was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 275 g C. 18 Purified on a flask (eluted with 5-80% MeCN / water) to give rac-5-[(diethoxyphosphoryl)(fluoro)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (650 mg, 1.48 mmol, 41.6% yield) as a viscous, clear oil. LCMS: m / z = 873.2 (2M+H) + ; 1 H NMR (400MHz, CDCl3) δ8.12(s,1H),8.02-7.99(m,1H),7.92(d,J=8.7Hz,1H),7.61(d,J=8.7Hz,1H),7.51-7.5 1(m,2H),7.46-7.36(m,3H),5.82(dd,J=44.4,7.5Hz,1H),5.42(s,2H),4.21-4.02(m,4H),1.34-1.26(m,6H).

[0351] Step 3: Preparation of rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0352] At -78 °C, benzyl bis(trimethylsilylamide)carboxylate (100 mg, 0.2291 mmol, 1 equivalent) and iodomethane (42.7 μL, 687 μmol, 3 equivalents) in tetrahydrofuran (4 mL) solution (458 μL, 458 μmol, 2 equivalents) was added dropwise. The mixture was stirred at -78 °C for 5 min. The reaction was quenched with 1 M HCl (10 mL), warmed to room temperature, and extracted with DCM (2 x 25 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was subjected to reversed-phase chromatography at 50 g C. 18Purified on a syringe (eluted with 5-80% MeCN / water (containing 0.1% formic acid)). Fractions were combined, concentrated under reduced pressure, and then lyophilized to give rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (67.0 mg, 0.1487 mmol, 65.0% yield) as a pale yellow oil. LCMS: m / z = 451.2 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ8.13(s,1H),8.05-8.02(m,1H),7.90(d,J=8.7Hz,1H),7.65(d,J=8.7Hz,1H),7.52-7.47(m,2H),7.46-7.36(m,3H),5.42 (s,2H),4.27-4.16(m,2H),4.05-3.94(m,1H),3.90-3.79(m,1H),2.03(dd,J=25.0,14.0Hz,3H),1.37(t,J=7.1Hz,3H),1.17(t,J=7.1Hz,3H).

[0353] Step 4: Preparation of rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid

[0354] A mixture of 10% palladium / carbon (50% wet) (99 mg, 0.04651 mmol, 0.21 equivalents) and benzyl 5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylate, and benzyl 5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylate (99 mg, 0.2197 mmol, 1 equivalent) in tetrahydrofuran (6 mL) was degassed with nitrogen for 5 min. Hydrogen was bubbled for 5 min, and the reaction was then stirred at room temperature under hydrogen (1 atm) for 20 h. The reaction mixture was filtered through a syringe filter and washed with THF. The filtrate was concentrated under reduced pressure to give crude rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid (78.9 mg, 0.2197 mmol, 100% yield) as a clear oil. LCMS: m / z = 361.2 [M+H] + .

[0355] Synthesis of rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid

[0356]

[0357] Step 1: Preparation of 7-bromonaphthalene-2-carboxylic acid

[0358] A solution of 2,7-dibromonaphthalene (2 g, 6.99 mmol, 1 equivalent) in tetrahydrofuran (24 mL) was cooled to -78 °C under nitrogen atmosphere. A solution of n-butyllithium (1.6 M in hexane) (4.58 mL, 7.33 mmol, 1.05 equivalent) was added dropwise. The reaction was stirred at -78 °C for 15 min, then CO2 was bubbled into the reaction mixture. The ice bath was removed, and the reaction was stirred under CO2 bubbling for 1 h. The reaction was quenched by adding 1 N HCl (50 mL, pH 2), and the product was extracted with EtOAc (3 x 75 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was ground in heptane, and the resulting solid was filtered and dried to give 7-bromonaphth-2-carboxylic acid as a white solid (1.27 g, 5.05 mmol, 72.5% yield). 1 H NMR (400MHz, CDCl3) δ13.21(s,1H),8.61(s,1H),8.44(s,1H),8.08-7.96(m,3H),7.81-7.76(m,1H).

[0359] Step 2: Preparation of tert-butyl 7-bromonaphthalene-2-carboxylate

[0360] N,N-dimethylformamide dibutyl acetal (3.80 mL, 15.9 mmol, 1 equivalent) was added to a stirred suspension of 7-bromonaphthalene-2-carboxylic acid (1.00 g, 3.98 mmol, 1 equivalent) in toluene (9 mL) over 15 minutes at 90 °C. The reaction was stirred at 90 °C for 2 hours, followed by dropwise addition of an additional amount of N,N-dimethylformamide dibutyl acetal (0.9 mL, 3.38 mmol, 1 equivalent), and the reaction was stirred at 90 °C for 30 minutes. The reaction was cooled to room temperature and stirred overnight. The reaction was adsorbed onto silica gel and concentrated under reduced pressure. The crude residue was purified by rapid chromatography on a 40 g silica gel cartridge (eluted with a gradient of 0–10% EtOAc / heptane) to give tert-butyl 7-bromonaphthalene-2-carboxylic acid as a white solid (1.11 g, 3.61 mmol, 90.9% yield). 1 H NMR (400MHz, CDCl3) δ8.46 (s, 1H), 8.14 (s, 1H), 8.06 (dd, J = 8.6, 1.6Hz, 1H), 7.86 ( d, J=8.6Hz, 1H), 7.77 (d, J=8.6, Hz, 1H), 7.66 (dd, J=8.6, 1.6Hz, 1H), 1.67 (s, 9H).

[0361] Step 3: Preparation of tert-butyl 7-methylnaphthalene-2-carboxylate

[0362] A mixture of tert-butyl 7-bromonaphthyl-2-carboxylate (1.11 g, 3.61 mmol, 1 equivalent), trimethylcycloboroxane (906 mg, 7.22 mmol, 2 equivalents), tetrakis(triphenylphosphine)palladium (834 mg, 722 μmol, 0.2 equivalents), and potassium carbonate (1.99 g, 14.4 mmol, 4 equivalents) in 1,4-dioxane (20 mL) was degassed for 5 min, and then sealed and heated to 110 °C for 20 h. The reaction mixture was diluted with EtOAc (20 mL) and adsorbed onto silica gel. The crude residue was purified by rapid chromatography by elution with 0–10% EtOAc / heptane to give tert-butyl 7-methylnaphthyl-2-carboxylate (808 mg, 3.33 mmol, 92.4% yield) as a white crystalline solid. 1 H NMR (400MHz, CDCl3) δ8.45(s,1H),7.95(d,J=8.7,Hz,1H),7.80(d,J=8.5,Hz,1H),7 .77(d,J=8.5Hz,1H),7.71(s,1H),7.40(d,J=8.7Hz,1H),2.53(s,3H),1.65(s,9H).

[0363] Step 4: Preparation of tert-butyl 7-(bromomethyl)naphthalene-2-carboxylate

[0364] N-bromosuccinimide (621 mg, 3.49 mmol) and benzoyl peroxide (32.2 mg, 133 μmol) were added to a solution of tert-butyl 7-methylnaphthalene-2-carboxylate (808 mg, 3.33 mmol) in anhydrous carbon tetrachloride (25 mL) under a nitrogen atmosphere. The reaction mixture was heated under reflux and stirred at this temperature for 20 hours. The precipitate was filtered off, washed with carbon tetrachloride (10 mL), and the filtrate was adsorbed onto silica gel and concentrated under reduced pressure. The crude material was purified by rapid chromatography on an 80 g silica gel cartridge (eluted with 0–10% EtOAc / heptane) to give tert-butyl 7-(bromomethyl)naphthalene-2-carboxylate (720 mg, 2.24 mmol, 67.9% yield) as a white solid containing residual starting material (about 25%). 1 H NMR (400MHz, CDCl3) δ8.50 (s, 1H), 8.04 (dd, J = 8.6, 1.6Hz, 1H), 7.93 (s, 1H), 7.85(t,J=8.8Hz,2H), 7.60(dd,J=8.6,1.6Hz,1H), 4.67(s,2H), 1.64(s,9H).

[0365] Step 5: Preparation of tert-butyl 7-[(diethoxyphosphoryl)methyl]naphthalene-2-carboxylate

[0366] 7-(bromomethyl)naphthyl-2-carboxylic acid tert-butyl ester (720 mg, 2.24 mmol) was suspended in triethyl phosphite (2 mL, 11.6 mmol), and the reaction mixture was heated under reflux for 1.5 hours (becoming a solution at 110 °C). The reaction was cooled to room temperature and analyzed by reversed-phase chromatography at 50 g C. 18 7-[(diethoxyphosphoryl)methyl]naphthalene-2-carboxylic acid tert-butyl ester (580 mg, 1.53 mmol, 68.4% yield) was purified to a viscous, pale yellow oil using a gradient of 5–80% MeCN / water. LCMS: m / z = 379.3 [M+H] + ; 1 H NMR(400MHz, CDCl3)δ8.52(s,1H),8.08-8.00(m,1H),7.89-7.83(m,3H),7.58-7.54( m, 1H), 4.11-3.97 (m, 4H), 3.36 (d, J = 21.6Hz, 2H), 1.67 (s, 9H), 1.26 (t, J = 7.0Hz, 6H).

[0367] Step 6: Preparation of rac-7-[(diethoxyphosphoryl)(hydroxy)methyl]naphthalene-2-carboxylic acid tert-butyl ester

[0368] At -78°C, sodium bis(trimethylsilylamide) sodium solution (1M in THF) (792 μL, 792 μmol, 1.5 equivalents) was added dropwise to a solution of tert-butyl 7-[(diethoxyphosphoryl)methyl]naphthalene-2-carboxylate (200 mg, 0.5285 mmol, 1 equivalent) in tetrahydrofuran (5 mL). The mixture was stirred for 2 min, and then 2-(benzenesulfonyl)-3-phenyloxazine propane (274 mg, 1.05 mmol, 2.0 equivalents) was added in portions. The deep red solution was stirred at -78°C for 20 min. The reaction mixture was quenched at -78°C with saturated ammonium chloride aqueous solution (50 mL), and then the ice bath was removed. EtOAc (25 mL) and water (25 mL) were added, and the mixture was stirred for 30 min. The phases were separated, and the aqueous layer was then back-extracted with EtOAc (2 x 25 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by rapid chromatography on a 24 g silica gel cartridge (eluted with 20–100% EtOAc / heptane) to give rac-7-[(diethoxyphosphoryl)(hydroxy)methyl]naphthalene-2-carboxylic acid tert-butyl ester (140 mg, 0.3549 mmol, 67.3% yield) as a white, viscous solid. LCMS: m / z = 394.6 [M+H] + ; 1H NMR (400MHz, CDCl3) δ8.57(s,1H),8.08(s,1H),8.05(d,J=8.6,Hz,1H),7.90(d,J=8.3Hz,1H),7.87(d,J=8.3Hz,1H),7.72(d, J=8.3Hz,1H),5.24(dd,J=11.1,4.7Hz,1H),4.18–3.99(m,4H),3.31(dd,J=11.1,4.7Hz,1H),1.66(s,9H),1.32-1.22(m,6H).

[0369] Step 7: Preparation of rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid tert-butyl ester

[0370] Under nitrogen atmosphere at -78°C, a solution of (diethylamino)sulfur trifluoride (56.1 μL, 425 μmol, 1.2 equivalents) was added dropwise to a solution of tert-butyl 7-[(diethoxyphosphoryl)(hydroxy)methyl]naphthalene-2-carboxylate (140 mg, 0.3549 mmol, 1 equivalent) in methylene chloride (5 mL). The reaction was stirred at -78°C for 20 min. The reaction was quenched by adding saturated sodium bicarbonate aqueous solution (50 mL) and extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 50 g C 18 Purified on a elution chamber (using a gradient of 5-80% MeCN / water) to give rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid tert-butyl ester as a clear oil (80.0 mg, 0.2018 mmol, 57.1% yield). LCMS: m / z = 397.4 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ8.59(s,1H),8.11-8.04(m,2H),7.94(d,J=8.3,Hz,1H),7.90(d,J=8.3Hz ,1H),7.72(d,J=8.6,1H),5.90(dd,J=44.6,8.1Hz,1H),4.24-4.02(m,4H),1.35-1.25(m,6H).

[0371] Step 8: Preparation of rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid

[0372] Trifluoroacetic acid (1 mL) was added to a solution of tert-butyl rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid (80 mg, 0.2018 mmol) in methylene chloride (4 mL). The resulting yellow solution was stirred at room temperature for 2.5 hours. The reaction was concentrated under reduced pressure to give crude rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid (68.3 mg, 0.2018 mmol) as a clear oil. LCMS: m / z = 341.2 [M+H] + .

[0373] Synthesis of 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid

[0374]

[0375] Step 1: Preparation of (2S)-pyrrolidine-1,2-dicarboxylic acid 1-chloromethyl ester and 2-methyl ester

[0376] Chloroacetyl chloride (777 mg, 6.03 mmol) was added to a solution of (2S)-pyrrolidine-2-carboxylate methyl hydrochloride (1.00 g, 6.03 mmol) and N,N-diisopropylethylamine (818 mg, 6.33 mmol, 1.05 equivalence) in dichloromethane (15 mL) at 0 °C under a nitrogen atmosphere. After stirring for 0.5 hours, the reaction mixture was washed successively with 1N hydrochloric acid aqueous solution (50 mL), water (50 mL), and brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The product was purified by rapid chromatography using a 25 g silica gel cartridge (eluted with a gradient of EtOAc / heptane (0 to 50%)). The combined fractions were concentrated under reduced pressure to yield (2S)-pyrrolidine-1,2-dicarboxylate 1-chloromethyl ester 2-methyl ester (701 mg, 3.16 mmol, 52.7% yield). 1 HNMR(400MHz, CDCl3) δ5.88(d,J=6.0Hz,0.5H),5.76-5.70(m,1H),5.65(d,J=6.2Hz,0.5H),4.39(ddd,J =18.8,8.5,3.6Hz,1H),3.74(d,J=3.4Hz,3H),3.68-3.45(m,2H),2.32-2.16(m,1H),2.10-1.87(m,3H).

[0377] Step 2: Preparation of (2S)-pyrrolidine-1,2-dicarboxylic acid 1-{[({2-[(benzyloxy)carbonyl]-1-benzothiophene-5-yl}difluoromethyl)({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]oxy}methyl 2-methyl ester

[0378] Water (2 mL) containing sodium hydroxide (59.9 mg, 1.50 mmol) was added dropwise to a stirred solution of ({2-[(benzyloxy)carbonyl]-1-benzothiophene-5-yl}difluoromethyl)phosphonic acid (300 mg, 753 μmol, 1 equivalent) in water (15 mL), followed by the addition of silver nitrate (382 mg, 2.25 mmol). After incubation at room temperature for 2 hours, the suspension was cooled to 0 °C. The precipitate was collected by filtration, washed with water, co-evaporated with toluene (2x), and dried under high vacuum. The powder was suspended in dry toluene (10 mL) and (2S)-pyrrolidine-1,2-dicarboxylic acid 1-chloromethyl ester 2-methyl ester (582 mg, 2.63 mmol, 3.5 equivalents) was added. The mixture was stirred at room temperature for 18 hours. The crude product was directly purified by normal-phase chromatography using a 12 g silica gel cartridge, eluting with a gradient of EtOAc / heptane (0 to 100% in 18CV) to produce (2S)-pyrrolidine-1,2-dicarboxylic acid 1-{[({2-[(benzyloxy)carbonyl]-1-benzothiophene-5-yl}difluoromethyl)({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]oxy}methyl 2-methyl ester (90.0 mg, 117 μmol, 15.5% yield). 1 H NMR (400MHz, CDCl3) δ8.20-8.12(m,2H),7.97-7.91(m,1H),7.69-7.64(m,1H),7.48-7.33(m,5H),5.79-5.66(m,2H),5.66-5. 52(m,2H),5.41-5.38(m,2H),4.40-4.31(m,2H),3.75-3.65(m,6H),3.63–3.40(m,4H),2.25–2.11(m,2H),2.04–1.81(m,6H).

[0379] Step 3: Preparation of 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid

[0380] Palladium / carbon (10% loaded, 50% wet supported) (62.1 mg, 58.5 μmol, 0.5 equivalents) was added to a mixture of (2S)-pyrrolidine-1,2-dicarboxylic acid 1-{[({2-[(benzyloxy)carbonyl]-1-benzothiophene-5-yl}difluoromethyl)({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]oxy}methyl 2-methyl ester (90 mg, 117 μmol, 1 equivalent) in anhydrous tetrahydrofuran (10 mL). Hydrogen was bubbled into the suspension for 2 minutes, and then the reaction mixture was stirred at 1 atm (balloon) hydrogen for 16 hours. The reaction mixture was filtered on a diatomaceous earth mat and washed with Me-THF. The filtrate was concentrated under vacuum to give crude product 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid (95.0 mg, 140 μmol, 119% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ8.18-8.13(m,1H),8.13-8.07(m,1H),7.97-7.92(m,1H),7.69-7.63(m,1H),5.82-5. 58(m,4H),4.43-4.34(m,2H),3.77-3.67(m,6H),3.64-3.42(m,4H),2.27-2.15(m,2H),2.09-1.82(m,6H).

[0381] According to the described procedure for the synthesis of 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid, the following connectors in Table 15 are prepared using appropriate starting materials.

[0382] Table 15.

[0383]

[0384] Synthesis of 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid:

[0385]

[0386] Step 1: Preparation of 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0387] At 0 °C, oxalyl chloride (814.4 mg, 6.4 mmol, 10 equivalents) was added dropwise to a solution of ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (200 mg, 0.64 mmol, 1.0 equivalents) in dry DCM (6 mL) and DMF (1 drop). The reaction was heated to 40 °C and then stirred for another 1–2 hours. The reaction was monitored by removing a small amount of crude sample and quenching it with MeOH to ensure that bis-Cl phosphoryl chloride 5-((dichlorophosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (mono-methoxy phosphonic acid ester observed by LCMS). After completion, excess oxalyl chloride and solvent were removed under reduced pressure, and the residue was redissolved in anhydrous DCM (5 mL). Anhydrous DCM (2 mL) containing BnNH2 (64.8 mg, 0.64 mmol, 1.0 equivalent) and Et3N (194.32 mg, 1.92 mmol, 3.0 equivalent) was then added to the solution at -40 °C. The reaction was monitored by removing a small amount of crude sample and quenching with MeOH to ensure that the majority of the product was 5-(((benzylamino)chlorophosphoyl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (mono-methoxy phosphonate ester observed by LCMS). Anhydrous DCM (2 mL) containing isopropyl-L-alanine (107.2 mg, 0.64 mmol, 1.0 equivalent) was then added to the solution at -40 °C. The reaction was warmed to room temperature and stirred for another 2 hours. After completion, the reaction was quenched by adding H2O (10 mL) and extracted with DCM (10 mL x 3). The organic layers were combined and washed with brine (20 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to yield 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (100 mg, 194.33 μmol, 30% yield). LCMS (ESI): m / z = 515.2 [M+H] + .

[0388] Step 2: Preparation of 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0389] A solution of 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (100 mg, 194 μmol, 1 equivalent), Pd(PPh3)4 (22.47 mg, 19.4 μmol, 0.1 equivalent), and pyrrolidine (13.8 mg, 194 μmol, 1 equivalent) in DCM (3 mL) was stirred at room temperature for 1 hour. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to yield 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (70 mg, 147.5 μmol, 76% yield). LCMS (ESI): m / z = 475.1 [M+H] + .

[0390] Synthesis of 5-((bis(((propoxycarbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester

[0391]

[0392] Step 1: Preparation of methyl chloropropyl carbonate

[0393] A solution of propan-1-ol (1.24 mL, 16.5 mmol, 1 equivalent) and chloroacetyl chloride (1.46 mL, 16.5 mmol, 1 equivalent) in diethyl ether (50 mL) was cooled to 0 °C under nitrogen atmosphere. Pyridine (1.32 mL, 16.5 mmol, 1.0 equivalent) was added dropwise, and the reaction mixture was stirred at 0 °C for 15 min, followed by stirring at room temperature for 3 h. The white suspension was filtered and washed with diethyl ether (20 mL). The filtrate was washed with 1 N HCl (20 mL) and water (2 x 15 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give methyl chloropropyl carbonate (1.26 g, 8.25 mmol, 50.1% yield) as a clear oil. 1 H NMR (400MHz, CDCl3) δ5.74 (s, 2H), 4.25-4.17 (m, 2H), 1.74 (sxt, J = 7.1Hz, 2H), 1.02-0.94 (m, 3H).

[0394] Step 2: Preparation of 5-((bis(((((1-methoxy-2-methylprop-2-yl)oxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester

[0395] H₂O (1 mL) containing sodium hydroxide (37.1 mg, 930 μmol, 2 equivalents) was added dropwise to a stirred suspension of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (200 mg, 465 μmol) in water (4 mL, 1 equivalent). When the mixture reached pH ~8, the product began to precipitate, and silver nitrate (236 mg, 1.39 mmol, 3 equivalents) was added in a single step. After 2 hours at 0 °C, the yellow precipitate was collected by filtration, washed with water, and dried under vacuum. The powder was suspended in dry toluene (10 mL), and methyl chloropropyl carbonate (282 mg, 1.85 mmol, 4 equivalents) was added. The mixture was stirred at room temperature for 18 hours. The crude product was directly purified by normal-phase chromatography using a 24 g silica gel cartridge, eluting with a gradient of EtOAc / heptane (0 to 60% in 18CV) to give a white semi-solid 5-((bis(((((1-methoxy-2-methylprop-2-yl)oxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester (10 mg, 15.1 μmol, 3.25% yield): LCMS: m / z = 662.2 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ8.64(s,1H),8.43-8.27(m,4H),7.72-7.65(m,3H), 5.75-5.65(m,4H),4.12-4.02(m,4H),1.67-1.55(m,4H),0.90-0.84(m,6H).

[0396] Synthesis of 5-((bis(((((1-methoxy-2-methylprop-2-yl)oxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester

[0397]

[0398] Step 1: Preparation of 1-methoxy-2-methylpropyl-2-yl methyl chloride carbonate

[0399] A solution of 1-methoxy-2-methylprop-2-ol (2 g, 19.2 mmol, 1 equivalent) and chloroacetyl chloride (1.7 mL, 19.2 mmol, 1 equivalent) in diethyl ether (60 mL) was cooled to 0 °C under nitrogen atmosphere. Pyridine (1.53 mL, 19.2 mmol, 1.0 equivalent) was added dropwise, and the reaction mixture was stirred at 0 °C for 15 min, followed by stirring at room temperature for 3 h. The white suspension was filtered and washed with diethyl ether (20 mL). The filtrate was washed with 1N HCl (30 mL) and water (2 x 30 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give methyl chloride 1-methoxy-2-methylprop-2-yl ester as a clear oil (860 mg, 4.37 mmol, 22.8% yield). 1 H NMR (400MHz, CDCl3) δ7.20-7.17(m,2H),5.04-5.00(m,2H),4.90(s,3H),3.03-2.99(m,6H).

[0400] Step 2: Preparation of 5-((bis(((((1-methoxy-2-methylprop-2-yl)oxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester

[0401] H₂O (1 mL) containing sodium hydroxide (37.1 mg, 930 μmol, 2 equivalents) was added dropwise to a stirred suspension of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (200 mg, 465 μmol) in water (5 mL, 1 equivalent). When the mixture reached pH ~8, the product began to precipitate, and silver nitrate (236 mg, 1.39 mmol, 3 equivalents) was added in a single step. After 2 hours at 0 °C, the white precipitate was collected by filtration, washed with water, and dried under vacuum. The powder was suspended in dry toluene (10 mL), and methyl chloride 1-methoxy-2-methylpropyl-2-yl ester (318 mg, 1.62 mmol, 3.5 equivalents) was added. The mixture was stirred at 50 °C for 18 hours. The crude mixture was purified directly by normal-phase chromatography using a 24 g silica gel cartridge, eluting with a gradient of EtOAc / heptane (0 to 60% in 18CV) to give 5-((bis(((((1-methoxy-2-methylprop-2-yl)oxy)carbonyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester (51.0 mg, 68.0 μmol, 14.6% yield) as a white semi-solid. 1H NMR (400MHz, CDCl3) δ8.38-8.33(m,3H),8.24(s,1H),8.03(d,J=8.8Hz,1H),7.76(d,J=8.8Hz, 1H),7.51-7.47(m,2H),5.74-5.62(m,4H),3.53-3.50(m,4H),3.41-3.38(m,6H),1.51(s,12H).

[0402] Synthesis of 5-((bis(((dipropylcarbamoyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester

[0403]

[0404] Step 1: Preparation of dipropylcarbamate chloromethyl ester

[0405] Dipropylamine (5.85 g, 57.9 mmol, 2.5 equivalents) was added to a solution of chloroacetyl chloride (2.06 mL, 23.2 mmol, 1 equivalent) in hexane (30 mL) under nitrogen atmosphere at 0 °C and stirred for 30 min at the same temperature. The reaction mixture was then diluted with EtOAc (50 mL). The phases were separated, and the organic layer was washed successively with 1 N hydrochloric acid (50 mL), water, and brine. The organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The product was purified by rapid chromatography using an 80 g silica gel cartridge (eluted with a gradient of 0-10% EtOAc / heptane). The pure, combined fractions were dried under reduced pressure to give dipropylcarbamate chloromethyl dipropylcarbamate (2.89 g, 14.9 mmol, 64.3% yield) as a colorless oil. LCMS: m / z = 194.2 [M+H] + ; 1 HNMR (400MHz, CDCl3) δ5.75 (s, 2H), 3.21-3.11 (m, 4H), 1.59-1.47 (m, 4H), 0.87-0.82 (m, 6H).

[0406] Step 2: Preparation of 5-((bis(((dipropylcarbamoyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester

[0407] H₂O (1 mL) containing sodium hydroxide (37.1 mg, 930 μmol, 2 equivalents) was added dropwise to a stirred suspension of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (200 mg, 465 μmol) in H₂O (5 mL, 1 equivalent). When the mixture reached pH ~8, the product began to precipitate, and silver nitrate (236 mg, 1.39 mmol, 3 equivalents) was added in a single step. After 2 hours at 0 °C, the yellow precipitate was collected by filtration, washed with water, and dried under vacuum. The powder was suspended in dry toluene (10 mL), and dipropylcarbamate (313 mg, 1.62 mmol, 3.5 equivalents) was added. The mixture was stirred at room temperature for 18 hours. The crude product was directly purified by normal-phase chromatography using a 12 g silica gel cartridge, eluting with a gradient of EtOAc / heptane (0 to 60% in 18CV) to give 5-((bis(((dipropylcarbamoyl)oxy)methoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester (110 mg, 147 μmol, 31.8% yield) as a clear oil. 1 H NMR(400MHz,DMSO-d6)δ8.65-8.56(m,1H),8.41-8.21(m,4H),7.76-7.59(m,3H ),5.77-5.60(m,4H),3.17-2.99(m,8H),1.53-1.38(m,8H),0.85-0.75(m,12H).

[0408] Synthesis of 4-nitrobenzene 5-((bis(1-((propoxycarbonyl)oxy)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0409]

[0410] Step 1: Preparation of 1-chloroethylpropyl carbonate

[0411] A solution of propan-1-ol (2 g, 33.2 mmol, 1 equivalent) and 1-chloroethyl chloroformate (4.74 g, 33.2 mmol, 1 equivalent) in diethyl ether (100 mL) was cooled to 0 °C under nitrogen atmosphere. Pyridine (2.66 mL, 33.2 mmol, 1.0 equivalent) was added dropwise, and the reaction mixture was stirred at 0 °C for 15 min, followed by stirring at room temperature for 3 h. The white suspension was filtered and washed with diethyl ether (20 mL). The filtrate was washed with 1 N HCl (20 mL) and water (2 x 15 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 1-chloroethyl propyl carbonate (4.62 g, 27.7 mmol, 83.5% yield) as a clear oil. 1H NMR (400MHz, CDCl3) δ6.44 (q, J=5.8Hz, 1H), 4.24-4.10 (m, 2H), 1.84 (d, J=5.9Hz, 3H), 1.78-1.66 (m, 2H), 1.01-0.94 (m, 3H).

[0412] Step 2: Preparation of 4-nitrobenzene 5-((bis(1-((propoxycarbonyl)oxy)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0413] Add dropwise 1 mL of H₂O containing sodium hydroxide (37.1 mg, 930 μmol, 2 equivalents) to a stirred suspension of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (200 mg, 465 μmol) in H₂O (4 mL, 1 equivalent). When the mixture becomes clear (pH ~8), silver nitrate (236 mg, 1.39 mmol, 2 equivalents) is added in one step. After 2 hours at 0 °C, the yellow precipitate is collected by filtration and dried under vacuum. The powder is suspended in dry toluene (10 mL) and 1-chloroethylpropyl carbonate (269 mg, 1.62 mmol, 3.5 equivalents) is added. The mixture is stirred at room temperature for 18 hours. The crude product was directly purified by normal-phase chromatography using a 12 g silica gel cartridge, eluting with a gradient of EtOAc / heptane (0 to 60% in 18CV) to give 5-((bis(1-(((propoxycarbonyl)oxy)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester (113 mg, 163 μmol, 35.3% yield) as a clear oil. LCMS: m / z = 712.2 (M+Na) + .

[0414] Synthesis of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methylacrylic acid

[0415]

[0416] Step 1: Preparation of tert-butyl 2-(diethoxyphosphoryl)propionate

[0417] A mixture of tert-butyl 2-bromopropionate (3 g, 14.3 mmol, 1 equivalent) and diethyl 2-(2-yl-1-ylphosphite) (2.82 g, 17.1 mmol, 1.2 equivalent) was stirred at 110 °C for 16 hours. Afterward, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to yield tert-butyl 2-(diethoxyphosphoryl)propionate (3.0 g, 11.3 mmol, 79% yield) as a white solid. LCMS (ESI): m / z = 267 [M+H] + .

[0418] Step 2: Preparation of (E)-3-(3-iodophenyl)-2-methylacrylate tert-butyl ester

[0419] n-BuLi (1.72 mL, 4.30 mmol, 1.0 equivalent) was added to a solution of tert-butyl 2-(diethoxyphosphoryl)propionate (1.14 g, 4.30 mmol, 1 equivalent) in THF (10 mL) at -78 °C under nitrogen atmosphere, and the resulting mixture was stirred at this temperature for 0.5 h. Then, a solution of 3-iodobenzaldehyde (1.0 g, 4.30 mmol, 1 equivalent) in THF (5 mL) was added, and the reaction mixture was warmed to room temperature and stirred for 14 h. After completion, the reaction mixture was quenched by adding H₂O (20 mL) and then extracted with EtOAc (15 mL x 3). The organic layers were combined and washed with brine (15 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to yield (E)-3-(3-iodophenyl)-2-methacrylate tert-butyl ester (739.6 mg, 2.15 mmol, 50% yield) as a white solid. LC-MS (ESI) m / z = 345 [M+H] + .

[0420] Step 3: Preparation of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methylacrylate tert-butyl ester

[0421] A solution of diethyl (bromodifluoromethyl)phosphonate (1.08 g, 4.06 mmol, 2 equivalents) and zinc (265 mg, 4.06 mmol, 2 equivalents) in DMAc (7 mL) was stirred at 60 °C for 1 hour under a nitrogen atmosphere. Then, CuBr (582 mg, 4.06 mmol, 2 equivalents) was added, and the mixture was stirred at 60 °C for another hour. Next, (E)-3-(3-iodophenyl)-2-methylacrylate tert-butyl ester (700 mg, 2.03 mmol, 1 equivalent) was added, and the mixture was stirred at 60 °C for another 12 hours. After completion, the suspension was strained... The mixture was filtered through a filter pad, and the filter cake was concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to give (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylate tert-butyl ester (180 mg, 445 μmol, 22% yield) as a yellow oil. LCMS (ESI): m / z = 405 [M+H] + .

[0422] Step 4: Preparation of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylic acid

[0423] A solution of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylate tert-butyl ester (180 mg, 445 μmol, 1 equivalent) in a mixture of DCM (8 mL) and TFA (4 mL) was stirred at room temperature for 1 hour. After completion, the reaction mixture was concentrated under reduced pressure to give (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylic acid (154 mg, quantitative) as a white solid, which could be used directly in the next step without further purification. LCMS (ESI): m / z = 349 [M+H] + .

[0424] Synthesis of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid

[0425]

[0426] Step 1: Preparation of tert-butyl (E)-3-(3-iodophenyl)but-2-enoic acid

[0427] n-BuLi (1.72 mL, 4.06 mmol, 1.0 equivalent) was added to 20 mL of THF containing tert-butyl 2-(diethoxyphosphoryl)acetate (1.02 g, 4.06 mmol, 1 equivalent) at -78 °C, and the mixture was stirred at -78 °C for 0.5 h. Then, a solution of 1-(3-iodophenyl)ethyl-1-one (1 g, 4.06 mmol, 1 equivalent) in 5 mL of THF was added dropwise to the reaction mixture. After addition, the reaction mixture was warmed to room temperature and stirred for 18 h. Upon completion, the reaction mixture was quenched by adding 20 mL of H₂O and then extracted with EtOAc (20 mL x 3). The organic layers were combined and washed with brine (15 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to produce tert-butyl (E)-3-(3-iodophenyl)but-2-enoate in oil (800 mg, 2.32 mmol, 58% yield). 1H NMR (400MHz, CDCl3) δ7.79(t,J=1.6Hz,1H),7.66(d,J=7.9Hz,1H),7.40(d,J=7.9Hz, 1H), 7.09 (t, J = 7.9Hz, 1H), 6.01 (d, J = 1.2Hz, 1H), 2.49 (d, J = 1.2Hz, 3H), 1.52 (s, 9H).

[0428] Step 2: Preparation of tert-butyl (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid

[0429] A solution of diethyl (bromodifluoromethyl)phosphonate (1.23 g, 4.64 mmol, 2 equivalents) in DMAc (2 mL) was slowly added to a stirred suspension of Zn (301 mg, 4.64 mmol, 2 equivalents) in dry DMAc (2 mL) under a nitrogen atmosphere. The reaction mixture was stirred at 45 °C for 2 h, and then CuBr (665 mg, 4.64 mmol, 2 equivalents) was added, and the resulting mixture was stirred at room temperature for 45 min. A suspension of (E)-3-(3-iodophenyl)but-2-enoic acid tert-butyl ester (800 mg, 2.32 mmol, 1 equivalent) in DMAc (3 mL) was added to the reaction mixture. The mixture was stirred at 45 °C under a nitrogen atmosphere for 24 h. The reaction mixture was then partitioned between water and diethyl ether. The mixture was passed through diatomaceous earth and extracted with diethyl ether. The organic extract was washed with brine and dried over anhydrous Na₂SO₄. The solvent was removed under vacuum, and the residue was purified by silica rapid column chromatography to give (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid tert-butyl ester (350 mg, 0.87 mmol, 37% yield) in oil. 1 H NMR(400MHz, CDCl3) δ7.67(d,J=13.2Hz,1H),7.63-7.54(m,2H),7.45(t,J=7.7Hz,1H),6.07( d,J=1.2Hz,1H),4.27-4.11(m,4H),2.54(d,J=1.0Hz,3H),1.52(s,9H),1.32(t,J=7.1Hz,6H).

[0430] Step 3: Preparation of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid

[0431] TFA (6.13 g, 53.8 mmol, 62.2 equivalents) was added to a solution of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid tert-butyl ester (350 mg, 0.87 mmol, 1 equivalent) in DCM (4 mL) at room temperature, and the resulting mixture was stirred at room temperature for 18 hours. After completion, the reaction mixture was concentrated under reduced pressure to produce (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid in oil (250 mg, 0.72 mmol, 83% yield), which could be used in the next step without further purification. LC-MS (ESI) m / z = 347.2 [MH] - .

[0432] Synthesis of perfluorophenyl 7-((bis(2-(butyrylthio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoic acid and perfluorophenyl 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid

[0433]

[0434] Oxaloyl chloride (266 mg, 2.13 mmol, 1.0 equivalent) was added dropwise to a solution of (difluoro(7-((perfluorophenoxy)carbonyl)naphth-2-yl)methyl)phosphonic acid (1 g, 2.10 mmol, 1 equivalent) in dry DCM (18 mL) at 25 °C, followed by the addition of DMF (155 mg, 2.13 mmol, 1.0 equivalent). The reaction mixture was warmed to 40 °C and then stirred under reflux for an additional 1.2 h. The reaction was monitored by removing a small amount of crude sample and quenching it with MeOH to ensure complete formation of bis-Cl phosphoryl chloride (observed by LCMS as bis-methoxy phosphonate). After completion, excess oxaloyl chloride and solvent were removed under reduced pressure. The residue was redissolved in anhydrous DCM (14 mL) at 0 °C and then added to a mixture of thiobutyric acid-S-(2-hydroxyethyl) ester (641 mg, 4.33 mmol, 2.2 equivalents) and triethylamine (198 mg, 1.97 mmol, 1.0 equivalents) in anhydrous DCM (15 mL). The reaction was warmed to room temperature and stirred for an additional 12 hours. The reaction progress was monitored by LCMS, and upon completion, the reaction was quenched by adding H2O (10 mL) and concentrated under reduced pressure. The residue was purified by C18 column chromatography to yield two products: perfluorophenyl 7-((bis(2-(butyrylthio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoic acid (680 mg, 1.13 mmol, 58% yield) as a white solid. LCMS (ESI): m / z = 729 [M+H] +And a white solid, 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid perfluorophenyl ester (65.0 mg, 89.2 μmol). LCMS (ESI): m / z = 599 [M+H] + .

[0435] Synthesis of perfluorophenyl 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid

[0436]

[0437] Amberlite was added to a solution of perfluorophenyl 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid (300 mg, 501 μmol, 1 equivalent) in a mixture of deionized H₂O (8 mL) and THF (2 mL). Resin (Na) + (Formula) (233 mg, 751 μmol, 1.5 equivalents). The resulting mixture was stirred at room temperature for 1 hour, and then excess resin was removed by filtration. Then, deionized H2O (2 mL) containing AgNO3 (127 mg, 751 μmol, 1.5 equivalents) was added to the resulting solution. After addition, the resulting mixture was stirred at room temperature for another 1 hour. During this time, silver salt formed a white precipitate, which was collected by filtration. The filter cake was washed with cold H2O (2 mL x 3), and the silver salt was further dried under reduced pressure to obtain a dry powder that was sufficiently pure for the next step without requiring further purification.

[0438] The separated mono-Ag salt was suspended in ACN (8 mL), and methyl isopropyl iodide carbonate (244 mg, 1.00 mmol, 2 equivalents) was added dropwise. After addition, the resulting mixture was stirred at 40 °C for an additional 12 hours. The reaction progress was monitored by LCMS, and after completion, the unreacted silver salt was recovered by filtration, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative TLC to give 7-(((2-(butyrylthio)ethoxy)(((isopropoxycarbonyl)oxy)methoxy)phospho)difluoromethyl)-2-naphthoic acid perfluorophenyl ester (30 mg, 41.9 μmol, 8%) as a colorless oil. LCMS (ESI) m / z = 737 [M + Na] + .

[0439] Synthesis of 5-((bis(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0440]

[0441] Step 1: Preparation of diisopropyl 2,2'-((((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphoryl)bis(aminoxylidene))(2S,2'S)-diisopropyl propionate

[0442] Oxaloyl chloride (609 mg, 4.80 mmol, 5.0 equivalent) was added dropwise to a solution of ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (300 mg, 960 μmol, 1.0 equivalent) in dry DCM (10 mL) and DMF (catalytic amount) at 0 °C. The reaction mixture was warmed to 40 °C and then stirred for another 1–2 hours. The reaction was monitored by removing a small amount of crude sample and quenching it with MeOH to ensure that bis-Cl phosphoryl chloride (bis-methoxy phosphonate ester observed by LCMS) had been completely formed. After completion, excess oxaloyl chloride and solvent were removed under reduced pressure, and the residue was redissolved in anhydrous DCM (5 mL). This solution was then added at 0 °C to a mixture of L-alanine isopropyl ester (502 mg, 3.83 mmol, 4.0 equivalence) and N,N-diisopropylethylamine (620 mg, 4.80 mmol, 5.0 equivalence) in anhydrous DCM (10 mL). The reaction was warmed to room temperature and stirred for an additional 18 hours. Progress was monitored by LCMS. After completion, the reaction was quenched by adding H2O (10 mL) and extracted with DCM (10 mL x 3). The organic layers were combined and washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to yield 2,2'-((((2-(((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphoryl)bis(aminoxylidene))(2S,2'S)-diisopropyl propionate (70.0 mg, 129 μmol, 14% yield). LCMS(ESI): m / z = 539.2 [M+H] + .

[0443] Step 2: Preparation of 5-((bis(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0444] Pyrrolidine (9.17 mg, 129 μmol, 1.0 equivalent) and Pd(PPh3)4 (14.9 mg, 12.9 μmol, 0.1 equivalent) were added to a solution of 2,2'-((((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphoryl)bis(aminoxylidene))(2S,2'S)-diisopropyl propionate (70 mg, 129 μmol, 1.0 equivalent) in DCM (1 mL) under N2 conditions, and the resulting mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was cooled in an ice bath and then carefully neutralized with HCl (1 M aqueous solution) until the pH was adjusted to pH 4-6. The resulting mixture was extracted with DCM (10 mL x 3), and the combined organic layers were washed with brine (10 mL x 2), dried over anhydrous Na2SO4, and then concentrated under reduced pressure. The residue was strained through... Purified by C18 column chromatography to obtain 5-((bis(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (70.0 mg, 140 μmol, 108%, 60% purity) as a colorless oil. LCMS (ESI): m / z = 499.2 [M+H] + .

[0445] Synthesis of 5-(difluoro(((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0446]

[0447] Step 1: Preparation of 5-(difluoro(((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0448] At 0 °C, two drops of DMF (catalytic amount) were added to a solution of ((2-((benzyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (300 mg, 753 μmol, 1 equivalent) in methylene chloride (8 mL), followed by dropwise addition of oxaloyl chloride (192 μL, 2.25 mmol, 3 equivalents). The reaction was warmed to room temperature and stirred for 2 hours. The reaction was initially insoluble, but became a clear solution after the addition of oxaloyl chloride and warming to room temperature. The reaction was concentrated under reduced pressure. The latter was then diluted in methylene chloride (8 mL) and the solution was cooled to -78 °C. A solution of phenol (56.6 mg, 602 μmol, 0.8 equivalents) and triethylamine (155 μL, 1.12 mmol, 1.5 equivalents) in DCM (1 mL) was slowly added to the yellow solution over 5 minutes. The reaction mixture was stirred at -78°C for 15 minutes, then warmed to room temperature and stirred for 2 hours. The reaction mixture was cooled to -78°C. A solution of 2-aminoacetic acid propion-2-yl ester (88.2 mg, 753 μmol, 1 equivalent) and triethylamine (155 μL, 1.12 mmol, 1.5 equivalent) in DCM (1 mL) was slowly added to the yellow solution over 5 minutes. The reaction mixture was stirred at -78°C for 15 minutes, then warmed to room temperature and stirred for 18 hours. Water (2-3 drops) was added and the reaction was concentrated under reduced pressure. The crude residue was analyzed by reversed-phase chromatography at 50 g C. 18 Purified on a elution vessel (eluted with 5-100% MeCN / water (containing 0.1% formic acid)) to give 5-(difluoro(((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (90.0 mg, 156 μmol, 20.8% yield) as a yellow oil. LCMS: m / z = 574.2 [M+H] + .

[0449] Step 2: Preparation of 5-(difluoro(((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0450] Under nitrogen atmosphere, 10% palladium / carbon (50% wet) (166 mg, 78.0 μmol, 0.5 equivalent) was added to a solution of 5-(difluoro((((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (90 mg, 156 μmol, 1 equivalent) in anhydrous tetrahydrofuran (10 mL). Hydrogen was bubbled for 5 minutes, and the reaction mixture was stirred at room temperature under hydrogen (1 atm) for 18 hours. Nitrogen was bubbled into the mixture, which was then filtered through diatomaceous earth (washed with 2-MeTHF) and concentrated under reduced pressure. The isolated product, 5-(difluoro((((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid, was ready for use without further purification. LCMS: m / z = 484.2 [M+H] + .

[0451] According to the described procedure for the synthesis of 5-(difluoro(((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid, the following connectors in Table 16 are prepared using appropriate starting materials.

[0452] Table 16.

[0453]

[0454]

[0455] Synthesis of 4-nitrobenzene 5-(difluoro((2-isopropoxy-2-oxoethoxy)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0456]

[0457] Step 1: Preparation of 4-nitrobenzene 5-(difluoro(hydroxy(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0458] At 0 °C, 2 drops of DMF (catalytic amount) were added to a solution of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (1 g, 2.32 mmol, 1 equivalent) in methylene chloride (6 mL), followed by dropwise addition of oxaloyl chloride (1.18 mL, 13.9 mmol, 6 equivalents). The reaction was warmed to room temperature and stirred for 2 hours. The reaction was initially insoluble (a white solid floated on the surface), but became a clear solution after the addition of oxaloyl chloride and warming to room temperature. The reaction was concentrated under reduced pressure and completely dried under high vacuum for 30 minutes to give a yellow solid. The latter was then diluted in methylene chloride (10 mL) and cooled to -78 °C. A solution of phenol (218 mg, 2.32 mmol, 1 equivalent) and triethylamine (646 μL, 4.64 mmol, 2 equivalent) in DCM (1 mL, dried on Na₂SO₄) was slowly added to the yellow solution over 5 minutes. The reaction mixture was stirred at -78 °C for 15 minutes, then warmed to room temperature and stirred for 2 hours. Water (1 mL) was added and the reaction mixture was concentrated under reduced pressure. The crude residue was analyzed by reversed-phase chromatography at 50 g C. 18 Purified on a cartridge (eluted with a gradient of 5-100% MeCN / water (containing 0.1% formic acid)) to give 4-nitrobenzene 5-(difluoro(hydroxy(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (848 mg, 1.67 mmol, 72.4%) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ9.04(br.s.,1H),8.63(s,1H),8.37(d,J=9.0Hz,2H),8.29(s,1H),8.19(d,J=8.8Hz,1 H),7.78(d,J=8.8Hz,1H),7.71(d,J=9.0Hz,2H),7.27-7.18(m,2H),7.10(d,J=8.3Hz,2H),7.02-6.95(m,1H).

[0459] Step 2: Preparation of 4-nitrobenzene 5-(difluoro((2-isopropoxy-2-oxoethoxy)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0460] Two drops of DMF (catalytic amount) were added to a solution of 4-nitrobenzene 5-(difluoro(hydroxy(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (100 mg, 197 μmol, 1 equivalent) in methylene chloride (5 mL) at 0 °C, followed by dropwise addition of oxaloyl chloride (168 μL, 1.97 mmol, 10 equivalents). The reaction mixture was warmed to room temperature and stirred for 18 h. The reaction mixture was concentrated under reduced pressure and dried completely under high vacuum. The latter was then diluted in methylene chloride (5 mL) and cooled to 0 °C. A solution of propyl-2-hydroxyacetate (30.2 mg, 256 μmol, 1.3 equivalents) and triethylamine (82.3 μL, 591 μmol, 3 equivalents) in DCM (1 mL) (dried on Na2SO4) was slowly added to the orange solution. The reaction mixture was stirred at 0 °C for 5 min, then warmed to room temperature and stirred for 5 h. The reaction mixture was concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on a 50 g C18 cylinder (eluted with 5-100% MeCN / alkaline water (NH4HCO3 10 mM, pH = 10)) to give 5-(difluoro((2-isopropoxy-2-oxoethoxy)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester (65.0 mg, 107 μmol, 54.6%) as a yellow oil. LCMS: m / z = 606.2 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.66(s,1H),8.46(s,1H),8.41-8.35(m,3H),7.83(d,J=8.3Hz,1H),7.72-7.67(m,2 H),7.44-7.38(m,2H),7.28-7.20(m,3H),5.02-4.94(m,1H),4.91-4.73(m,2H),1.18(dd,J=6.4,2.4Hz,6H).

[0461] The following connectors in Table 17 are prepared using appropriate starting materials according to the described procedure for 5-(difluoro((2-isopropoxy-2-oxoethoxy)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid 4-nitrobenzene ester.

[0462] Table 17.

[0463]

[0464] Synthesis of 5-(((((S)-1-(benzyloxy)-1-oxopropyl-2-yl)amino)(2-(((3-methylbutyryl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0465]

[0466] Step 1: Preparation of 5-(((((S)-1-(benzyloxy)-1-oxopropyl-2-yl)amino)(2-(((3-methylbutyryl)thio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0467] At 0 °C, 3 drops of DMF were added to a solution of ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (200 mg, 0.5742 mmol, 1 equivalent) in methylene chloride (10 mL), followed by dropwise addition of oxaloyl chloride (147 μL, 1.72 mmol, 3 equivalents). The reaction was warmed to room temperature and stirred for 3 hours. The reaction was concentrated under reduced pressure and dried under high vacuum for 30 minutes to give a grayish-white solid. The crude solid was diluted in methylene chloride (10 mL) and cooled to 0 °C. A solution of 1-[(2-hydroxyethyl)thioalkyl]-3-methylbut-1-one (93.1 mg, 574 μmol, 1 equivalent) in DCM (2 mL, dried on Na2SO4) was added dropwise, followed by a solution of triethylamine (299 μL, 2.87 mmol, 5 equivalent) in DCM (2 mL, dried on Na2SO4) over 1 minute. The reaction mixture was stirred at 0 °C for 1 minute, then warmed to room temperature and stirred for 1 hour. The reaction was cooled to 0 °C, and then (2R)-2-aminopropionate benzyl ester 4-toluene-1-sulfonate (302 mg, 861 μmol, 1.5 equivalent) was added in a single batch. The reaction was warmed to room temperature and stirred for 20 hours. The reaction was concentrated under reduced pressure and then diluted with DMSO / MeCN / water (3 mL). The crude residue was analyzed by reversed-phase chromatography at 100 g C 18 Purified on a flask (eluted with 5-100% MeCN / water) to give 5-(((((S)-1-(benzyloxy)-1-oxopropyl-2-yl)amino)(2-(((3-methylbutyryl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (180 mg, 0.2753 mmol, 48.0% yield) as a viscous, slightly brown oil. 1H NMR (400MHz, CDCl3) δ8.16-8.12(m,1H),7.94(d,J=8.5Hz,0.5H),7.78(d,J=8.5Hz,0.5H),7.7 0-7.64(m,1H),7.43-7.31(m,6H),6.12-6.01(m,1H),5.47(d,J=16.8Hz,1H),5.35(d,J=10.4H z,1H),5.25-5.11(m,2H),4.90-4.86(m,2H),4.24-4.00(m,2H),3.82-3.69(m,1H),3.24-2.92 (m,2H),2.45-2.40(m,2H),2.18-2.10(m,1H),1.44(dd,J=16.0,7.1Hz,3H),0.98-0.94(m,6H).

[0468] Step 2: Preparation of 5-(((((S)-1-(benzyloxy)-1-oxopropyl-2-yl)amino)(2-(((3-methylbutyryl)thio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0469] Under nitrogen atmosphere, morpholine (39.5 μL, 458 μmol, 5 equivalents) and tetra(triphenylphosphine)palladium (10.5 mg, 9.17 μmol, 0.10 equivalents) were added to a stirred solution of 5-((((S)-1-(benzyloxy)-1-oxopropyl-2-yl)amino)(2-(((3-methylbutyryl)thio)ethoxy)phospho)difluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (60 mg, 0.09178 mmol, 1 equivalent) in tetrahydrofuran (3 mL). The reaction mixture was stirred at room temperature for 1.5 h. The reaction was injected directly onto a column. The product was analyzed by reversed-phase chromatography at 50 g C 18 The solution was purified on a syringe (eluted with 5-80% MeCN / water (containing 0.1% formic acid)) and then freeze-dried to give 5-((((S)-1-(benzyloxy)-1-oxopropyl-2-yl)amino)(2-(((3-methylbutyryl)thio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid as a grayish-white solid (40.0 mg, 0.06518 mmol, 71.1% yield). LCMS: m / z = 611.9 [M+H] + ; 1H NMR(400MHz, CDCl3) δ8.10(d,J=16.8Hz,1H),7.86-7.78(m,2H),7.61-7.54(m,1H),7.45-7.35(m,5H),5.25(d,J=13.0Hz,2H),4.60 -4.45(m,1H),4.40-4.14(m,3H),3.36-3.14(m,2H),2.48(t,J=7.1Hz,3H),2.25-2.13(m,1H),1.58-1.51(m,3H),1.02-0.96(m,6H).

[0470] Composition of building blocks

[0471] Synthesis of 6-phenyl-4-azaspiro[2,4]heptane

[0472]

[0473] Step 1: Preparation of methyl 3-cyano-2-phenylpropionate

[0474] A solution of 2M LDA (20 mL, 40.0 mmol, 1.2 equivalence) in THF was slowly added to a cooled (-78°C) solution of methyl 2-phenylacetate (5.0 g, 33.3 mmol, 1.0 equivalence) in THF. After 1 hour, 2-bromoacetonitrile (4.2 g, 35.0 mmol, 1.1 equivalence) was slowly added dropwise, and the reaction was further aged at -78°C for another hour. A saturated aqueous solution of NH4Cl (5 mL) was added to the mixture, and the mixture was warmed to room temperature. The mixture was treated with EtOAc (20 mL x 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 silica gel rapid column chromatography to give methyl 3-cyano-2-phenylpropionate (5.0 g, 26.5 mmol, 79% yield) as a white solid. LCMS(ESI)m / z=190.1[M+H] + .

[0475] Step 2: Preparation of 6-phenyl-4-azaspiro[2.4]hepta-5-one

[0476] A 3M EtMgBr solution (20 mL, 59.6 mmol, 2.25 equivalents) was slowly added to a cooled (0 °C) solution of methyl 3-cyano-2-phenylpropionate (5.0 g, 26.5 mmol, 1.0 equivalent) and Ti(OiPr)4 (9.0 g, 31.7 mmol, 1.2 equivalent) in THF (100 mL) while maintaining an internal temperature between -5 °C and 0 °C. After the addition of EtMgBr was complete, the mixture was stirred at 0 °C for an additional 1 hour. The reaction mixture was quenched by adding 2N HCl aqueous solution (60 mL). The resulting acidic solution of EtOAc was extracted (50 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by rapid column chromatography to yield 6-phenyl-4-azaspiro[2,4]hepta-5-one (2.1 g, 11.2 mmol, 42% yield) as a white solid. LCMS (ESI) m / z = 188 [M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.39-7.24(m,6H),3.86(dd,J=9.4,7.6Hz,1H),2.51(dd,J=12.9,9.4Hz ,1H),2.29(dd,J=12.9,7.6Hz,1H),0.85-0.88(m,1H),0.87-0.81(m,1H),0.76-0.64(m,2H).

[0477] Step 3: Preparation of 6-phenyl-4-azaspiro[2,4]heptane

[0478] NaBH4 (2.1 g, 56.0 mmol, 5.0 equivalent) was added aliquots to a cooled (10 °C) solution of 6-phenyl-4-azaspiro[2.4]hepta-5-one (2.1 g, 11.2 mmol, 1.0 equivalent) in dry THF (50 mL). BF3·Et2O (6.7 mL, 56.0 mmol, 5.0 equivalent) was added dropwise to the mixture. The reaction mixture was heated at 60 °C for 16 hours. The mixture was cooled to ambient temperature and slowly introduced into a 2N HCl aqueous solution (30 mL). The acidic mixture was extracted with EtOAc (30 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was strained... C18 column purification was performed to produce 6-phenyl-4-azaspiro[2,4]heptane as a colorless oil (1.5 g, 8.7 mmol, 78% yield). LCMS (ESI) m / z = 174 [M+H] + ; 1H NMR(400MHz, CDCl3)8.59(s,1H),δ7.38-7.27(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).

[0479] Synthesis of pyrrolidine building blocks:

[0480] Synthesis of rel-(trans)-4-cyclohexylpyrrolidine-3-carboxynitrile

[0481]

[0482] Step 1: Preparation of (E)-3-cyclohexylacrylonitrile

[0483] At room temperature, t-BuOK (998 mg, 8.90 mmol, 2.0 equivalence) and diethyl (cyanomethyl)phosphonate (788 mg, 4.45 mmol, 1.0 equivalence) were added to a solution of cyclohexaneformaldehyde (500 mg, 4.45 mmol, 1.0 equivalence) in THF (15 mL). The solution was stirred at room temperature for 1 hour. After completion, the reaction mixture was quenched by adding H2O (10 mL) and then extracted with EtOAc (10 mL x 3). The organic layers were combined and washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to produce (E)-3-cyclohexylacrylonitrile (300 mg, 2.21 mmol, 50% yield) as a colorless oil. LC-MS (ESI) m / z = 136 [M+H] + .

[0484] Step 2: Preparation of rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carboxynitrile

[0485] N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methylamine (524 mg, 2.21 mmol, 1.0 equivalent) and TFA (25.1 mg, 221 μmol, 0.1 equivalent) were added to a solution of (E)-3-cyclohexylacrylonitrile (300 mg, 2.21 mmol, 1.0 equivalent) in CH2Cl2 (5 mL). The reaction mixture was stirred at room temperature for 12 hours. After stirring, the reaction mixture was diluted with CH2Cl2 (10 mL) and washed with saturated NaHCO3 aqueous solution (5 mL). The organic layer was separated and concentrated under reduced pressure to yield the crude product rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carboxynitrile (700 mg) as a colorless oil. LC-MS (ESI) m / z = 269 [M+H]+ .

[0486] Step 3: Preparation of rel-(trans)-4-cyclohexylpyrrolidine-3-carboxynitrile

[0487] Add 1-chloroethyl chloroformate (3.71 g, 26.0 mmol, 10.0 equivalent) to a solution of rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carboxynitrile (700 mg) in dry 1,2-dichloroethane (15 mL). Stir the resulting mixture at 70 °C for 12 h, then concentrate under reduced pressure. Dissolve the crude product in MeOH (5 mL) and stir at 70 °C for 1 h. After completion, cool the reaction solution to room temperature and concentrate under reduced pressure. Purify the residue by silica gel rapid column chromatography to yield rel-(trans)-4-cyclohexylpyrrolidine-3-carboxynitrile (211 mg, 1.18 mmol, 46% yield) as a colorless oil. LC-MS (ESI) m / z = 179 [M+H] + .

[0488] The following intermediates in Table 18 were prepared using appropriate starting materials and modifications according to the representative procedure described for rel-(trans)-4-cyclohexylpyrrolidine-3-carboxynitrile (steps 1 to 3). The compounds were prepared as racemates having trans stereochemistry relative to the C3 and C4 stereocenters within the pyrrolidine ring.

[0489] Table 18.

[0490]

[0491] The tert-butyl rel-(trans)-3-cyano-4-phenylpyrrolidine-1-carboxylate and (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-carboxylate. The racemic mixture of the trans isomers was purified under SFC conditions, and the absolute stereochemistry is arbitrarily specified as shown in the figure.

[0492]

[0493] Preparative separation methods:

[0494] Instrumentation: Waters Thar 80 preparative SFC; Column: ChiralPak C-IG, 100 × 4.6 mm ID, 5 μm; Mobile phase: A represents CO2, and B represents methanol (0.05% diethylamine); Gradient: 10% to 40% B over 8 minutes; Flow rate: 2.5 mL / min; Back pressure: 100 bar; Column temperature: 40 °C; Wavelength: 210 nm; Cycle time: 2 minutes

[0495] The following intermediates in Table 19 are prepared using appropriate starting materials and modifications according to the described procedure for rel-(trans)-4-cyclohexylpyrrolidine-3-carboxynitrile.

[0496] Table 19.

[0497]

[0498] Synthesis of 2-(4-amino-[1,1'-biphenyl]-3-yl)acetamide

[0499]

[0500] Step 1: Preparation of 2-(5-bromo-2-nitrophenyl)acetyl chloride

[0501] A solution of 2-(5-bromo-2-nitrophenyl)acetic acid (0.30 g, 1.2 mmol, 1 equivalent) in thionyl chloride (2 mL) was stirred at 80 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to give 2-(5-bromo-2-nitrophenyl)acetyl chloride (0.30 g, crude product) as a white solid.

[0502] Step 2: Preparation of 2-(5-bromo-2-nitrophenyl)acetamide

[0503] A solution of 2-(5-bromo-2-nitrophenyl)acetyl chloride (0.30 g, 1.1 mmol, 1 equivalent) in tetrahydrofuran (3 mL) and NH3·H2O (4 mL) was stirred at 25 °C for 30 min. The reaction mixture was concentrated under reduced pressure to give 2-(5-bromo-2-nitrophenyl)acetamide (0.30 g, crude product) as a white solid. LCMS: m / z [M+H] + =258.9.

[0504] Step 3: Preparation of 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide

[0505] Sodium carbonate (0.12 g, 1.2 mmol, 3 equivalents), phenylboronic acid (52 mg, 0.42 mmol, 1.1 equivalents), and iron(2+)bis(cyclopentan-2,4-diyn-1-yldiphenyl-λ4-phosphine)palladium dichloride (14 mg, 0.019 mmol, 0.05 equivalents) were added to a solution of 2-(5-bromo-2-nitrophenyl)acetamide (0.10 g, 0.39 mmol, 1 equivalent) in dioxane (1 mL) and water (0.2 mL). The mixture was stirred at 100 °C for 12 hours. The mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL × 2). The combined organic layers were washed with saturated brine (20 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography to give 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (60 mg, 60% yield) as a white solid. LCMS: m / z (M+Na) + =279.0. 1 H NMR (400MHz, CD3OD) δ8.17 (d, J=8.4Hz, 1H), 7.79-7.69 (m, 4H), 7.52-7.40 (m, 3H), 4.06 (s, 2H)

[0506] Step 4: Preparation of 2-(4-amino-[1,1'-biphenyl]-3-yl)acetamide

[0507] Ammonium chloride (0.038 g, 0.70 mmol, 3 equivalents) and iron (0.13 g, 2.4 mmol, 10 equivalents) were added to a solution of 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (60 mg, 0.23 mmol, 1 equivalent) in ethanol (0.8 mL) and water (0.2 mL). The mixture was stirred at 25 °C for 4 hours. The reaction mixture was filtered and the filtrate was concentrated to give 2-(4-amino-[1,1'-biphenyl]-3-yl)acetamide (0.043 g, crude product) as a white solid. LCMS: m / z [M+H] + =227.0. 1 H NMR (400MHz, DMSO-d6) δ7.53(d,J=7.6Hz,3H),7.41-7.35(m,3H),7.30-7.20(m,2H),6.98(s,1H),6.73(d,J=8.4Hz,1H),5.26(s,2H),3.32(s,2H).

[0508] Synthesis of 1-((3S,4R)-3-amino-4-fluoropyrrolidone-1-yl)ethyl-1-one

[0509]

[0510] Step 1: Preparation of (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester

[0511] Benzyl chloroformate (416 mg, 2.44 mmol, 2.0 equivalent) was added to a solution of (3S,4R)-3-amino-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester (250 mg, 1.22 mmol, 1.0 equivalent) and sodium bicarbonate (512 mg, 6.10 mmol, 5.0 equivalent) in THF (10 mL) and water (2 mL) at room temperature. After addition, the reaction mixture was stirred overnight at room temperature. After completion, the reaction mixture was poured into water (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to produce a yellow oil of (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester (2,400 mg, 1.18 mmol, 97%), which could be used directly in the next step without further purification. LCMS (ESI): m / z = 361.1 [M + Na] + .

[0512] Step 2: Preparation of ((3S,4R)-4-fluoropyrrolidine-3-yl)carbamate benzyl ester

[0513] A solution of (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester (200 mg, 0.59 mmol, 1.0 equivalent) in DCM (1 mL) and TFA (0.5 mL) was stirred overnight at 25 °C under N2. After completion, the reaction mixture was concentrated under reduced pressure to give a crude product ((3S,4R)-4-fluoropyrrolidine-3-yl)carbamate (140 mg, quantitative) as a white solid, which could be used directly in the next step without further purification. LCMS (ESI): m / z = 239.3 [M+H] + .

[0514] Step 3: Preparation of ((3S,4R)-1-acetyl-4-fluoropyrrolidine-3-yl)carbamate benzyl ester

[0515] Acetyl chloride (46.0 mg, 0.59 mmol, 1.0 equivalent) was added to a solution of ((3S,4R)-4-fluoropyrrolidone-3-yl)carbamate (140 mg, 0.59 mmol, 1.0 equivalent) and TEA (179 mg, 1.77 mmol, 3.0 equivalent) in dry DCM (6 mL) at 0 °C. After addition, the resulting mixture was warmed to 20 °C and stirred under N2 for 2 hours. After completion, the reaction mixture was decanted into water (10 mL) and extracted with DCM (10 mL x 3). The organic layers were combined and washed with brine (10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel rapid column chromatography to yield ((3S,4R)-1-acetyl-4-fluoropyrrolidone-3-yl)carbamate (611 mg, 1.29 mmol, 62%) as a white solid. LCMS(ESI): m / z = 281.1[M+H] + .

[0516] Step 4: Preparation of 1-((3S,4R)-3-amino-4-fluoropyrrolidone-1-yl)ethyl-1-one

[0517] Pd / C (50 mg) was added to a solution of ((3S,4R)-1-acetyl-4-fluoropyrrolidine-3-yl)carbamate (250 mg, 1.22 mmol, 1.0 equivalent) in EtOH (10 mL) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The resulting mixture was stirred overnight at room temperature. After completion, the suspension was passed through... The mixture was filtered through a filter pad, and the filter cake was washed with EtOH (20 mL). The combined filtrates were concentrated to dryness to give 1-((3S,4R)-3-amino-4-fluoropyrrolidone-1-yl)ethyl-1-one (375 mg, 2.10 mmol, 87%) as a colorless oil. LCMS (ESI): m / z = 147.1 [M+H] + .

[0518] Synthesis of (R)-azacyclobutane-3-yl(2-methylmorpholino)methyl ketone

[0519]

[0520] Step 1: Preparation of (R)-3-(2-methylmorpholine-4-carbonyl)azacyclobutane-1-carboxylic acid tert-butyl ester

[0521] To a solution of 1-[(tert-butoxy)carbonyl]azacyclobutane-3-carboxylic acid (1,100 mg, 496 μmol, 1.0 equivalent), HATU (226 mg, 595 μmol, 1.2 equivalent), and triethylamine (100 mg, 992 μmol, 2.0 equivalent) in DMF (2 mL), (2R)-2-methylmorpholine (55.1 mg, 545 μmol, 1.1 equivalent) was added, and the resulting mixture was stirred overnight at 20 °C under N2. After completion, the reaction mixture was filtered and concentrated under reduced pressure. The residue was strained... Purified by C18 column chromatography to obtain (R)-3-(2-methylmorpholino-4-carbonyl)azacyclobutane-1-carboxylic acid tert-butyl ester (2,140 mg, 492 μmol, 99%) as a white solid. LCMS (ESI): m / z = 229.2 [(M-56)+H] + .

[0522] Step 2: Preparation of (R)-azacyclobutane-3-yl(2-methylmorpholino) methyl ketone

[0523] A solution of (R)-3-(2-methylmorpholino-4-carbonyl)azacyclobutane-1-carboxylic acid tert-butyl ester (2, 50 mg, 175 μmol, 1.0 equivalent) in DCM (6 mL) and TFA (2 mL) was stirred overnight at 25 °C under N2. After completion, the reaction mixture was concentrated to give crude (R)-azacyclobutane-3-yl(2-methylmorpholino) methyl ketone (3, 40.0 mg, quantitative) as a white solid, which could be used directly in the next step without further purification. LCMS (ESI): m / z = 185.3 [M+H] + .

[0524] The following building blocks were prepared according to the above scheme:

[0525] The following intermediates in Table 20 are prepared using appropriate starting materials and modifications according to the described procedure for (R)-azacyclobutane-3-yl(2-methylmorpholino) methyl ketone.

[0526] Table 20.

[0527]

[0528] Synthesis of (R)-morpholino(pyrrolidone-3-yl)methyl ketone

[0529]

[0530] Step 1: Preparation of (R)-3-(morpholino-4-carbonyl)pyrrolidine-1-carboxylic acid benzyl ester

[0531] Morpholine (58 mg, 0.66 mmol) and DIEA (0.23 g, 1.8 mmol) were added to a solution of (R)-1-((benzyloxy)carbonyl)pyrrolidine-3-carboxylic acid (0.15 g, 0.60 mmol) and HATU (0.34 g, 0.90 mmol) in DMF (1 mL), and the mixture was stirred at 15 °C for 1 h to obtain a brown solution. The solution was quenched with water (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with saturated brine (10 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give benzyl (R)-3-(morpholine-4-carbonyl)pyrrolidine-1-carboxylic acid (0.10 g, 52% yield) as a brown oil. 1 H NMR (400MHz, CDCl3) δ7.41-7.28 (m, 5H), 5.14 (s, 2H), 3.68 (d, J = 4.4Hz, 9H), 3.53-3.40(m,3H),3.24-3.16(m,1H),2.34-2.13(m,1H),1.49-1.42(m,1H).

[0532] Step 2: Preparation of (R)-morpholino(pyrrolidine-3-yl)methyl ketone

[0533] A solution of (R)-3-(morpholino-4-carbonyl)pyrrolidine-1-carboxylate (0.10 g, 0.31 mmol) in MeOH (5 mL) was mixed with wet Pd / C (0.10 g) and stirred at 15 °C for 2 hours under H2 (15 psi) to give a black suspension. The reaction mixture was filtered and concentrated under vacuum to give (R)-morpholino(pyrrolidine-3-yl) methyl ketone (60 mg, crude product) as a colorless oil. LCMS: (ESI) m / z [M+H] + =185.3

[0534] The following intermediates in Table 21 are prepared using appropriate starting materials and modifications according to the described procedure for (R)-morpholino(pyrrolidine-3-yl)methyl ketone.

[0535] Table 21.

[0536]

[0537] Synthesis of (S)-1-(tetrahydrofuran-3-yl)azacyclobutane-3-amine

[0538]

[0539] Step 1: Preparation of (R)-tetrahydrofuran-3-yl mesylate

[0540] Methanesulfonyl chloride (3.4 g, 29 mmol, 1.3 equivalent) was added dropwise to a solution of (R)-tetrahydrofuran-3-ol (2.0 g, 23 mmol, 1.0 equivalent) and TEA (6.9 g, 69 mmol, 3.0 equivalent) in DCM (20 mL) at 0 °C. The mixture was stirred at 25 °C for 2 hours to obtain a yellow mixture. The mixture was quenched with H2O (50 mL) and extracted with DCM (50 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated to give (R)-tetrahydrofuran-3-yl methanesulfonic acid (3.5 g, crude product) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ5.28-5.20(m,1H),3.99-3.75(m,4H),2.97(s,3H),2.23-2.11(m,2H)

[0541] Step 2: Preparation of (S)-(1-(tetrahydrofuran-3-yl)azacyclobutane-3-yl)tert-butyl carbamate

[0542] K₂CO₃ (5.6 g, 41 mmol, 4.5 equivalence) was added to a solution of (R)-tetrahydrofuran-3-yl methanesulfonic acid (1.5 g, 9.0 mmol, 1.0 equivalence) and tert-butyl carbamate aziridine-3-yl ester (4.7 g, 27 mmol, 3.0 equivalence) in MeCN (15 mL). The mixture was stirred at 80 °C for 12 hours to obtain a pale yellow suspension. The mixture was filtered and concentrated to obtain a residue. The residue was purified by column chromatography to obtain (S)-(1-(tetrahydrofuran-3-yl)aziridine-3-yl)carbamate tert-butyl ester (0.6 g, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =243.0

[0543] Step 3: Preparation of (S)-1-(tetrahydrofuran-3-yl)azacyclobutane-3-amine

[0544] A solution of (S)-(1-(tetrahydrofuran-3-yl)azacyclobutane-3-yl)carbamate tert-butyl ester (0.10 mg, 0.41 mmol, 1 equivalent) in DCM (1.5 mL) and TFA (0.50 mL) was stirred at 25 °C for 2 hours to give a pink solution. The reaction mixture was concentrated under reduced pressure to give (S)-1-(tetrahydrofuran-3-yl)azacyclobutane-3-amine (0.10 mg, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =143.3

[0545] The following intermediates in Table 22 are prepared using appropriate starting materials and modifications according to the described procedure for (S)-1-(tetrahydrofuran-3-yl)azacyclobutane-3-amine.

[0546] Table 22.

[0547]

[0548] Synthesis of (3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-amine

[0549]

[0550] Step 1: Preparation of ((3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-yl)tert-butyl carbamate

[0551] To a solution of ((3S,4R)-4-fluoropyrrolidone-3-yl)carbamate tert-butyl ester (0.1 g, 0.49 mmol, 1.0 equivalent) in dioxane (2.5 mL), 3-bromopyridine (93 mg, 0.59 mmol, 1.2 equivalent), Xantphos (25 mg, 44 μmol, 0.1 equivalent), and cesium carbonate (0.25 g, 0.78 mmol, 1.6 equivalent) were added. The reaction mixture was degassed and purged three times with N2, followed by the addition of Pd2(dba)3 (0.13 g, 14 μmol, 0.3 equivalent). The resulting reaction mixture was stirred at 100 °C for 72 hours to obtain a black suspension. The reaction mixture was filtered, and the filtrate was concentrated to obtain a residue. The residue was diluted with water (50 mL) and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by rapid silica gel chromatography to give tert-butyl ((3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-yl)carbamate (0.1 g, 73% yield) as a yellow oil. LCMS: (ESI) m / z [M+H] + =282.1

[0552] Step 2: Preparation of (3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-amine

[0553] A solution of ((3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-yl)carbamate tert-butyl ester (0.1 g, 0.36 mmol, 1.0 equivalent) in DCM (0.3 mL) and trifluoroacetic acid (0.1 mL) was stirred at 20 °C for 1 hour to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give (3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-amine (0.1 g, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =182.1

[0554] The following intermediates in Table 23 are prepared using appropriate starting materials and modifications according to the described procedure for (3S,4R)-4-fluoro-1-(pyridin-3-yl)pyrrolidine-3-amine.

[0555] Table 23.

[0556]

[0557] Preparation of 1-((3S,4S)-3-amino-4-fluoropyrrolidone-1-yl)ethyl-1-one

[0558]

[0559] Step 1: Preparation of ((3S,4S)-1-acetyl-4-fluoropyrrolidine-3-yl)tert-butyl carbamate

[0560] TEA (0.1 g, 0.98 mmol, 2 equivalents) was added to a solution of ((3S,4S)-4-fluoropyrrolidone-3-yl)carbamate tert-butyl ester (0.1 g, 0.49 mmol, 1 equivalent) in DCM (0.5 mL). The mixture was stirred at 0 °C for 5 min, and then Ac₂O (0.05 g, 0.49 mmol, 1 equivalent) was added to a solution of DCM (0.5 mL). The mixture was stirred at 25 °C for 30 min. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL × 2). The combined organic layers were washed with saturated brine (50 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 10:1 to 3:1) to give tert-butyl ((3S,4S)-1-acetyl-4-fluoropyrrolidine-3-yl)carbamate (60 mg, 49% yield) as a yellow oil. LCMS: (ESI) m / z [M+H] + =247.1

[0561] Step 2: Preparation of 1-((3S,4S)-3-amino-4-fluoropyrrolidone-1-yl)acetone

[0562] A solution of ((3S,4S)-1-acetyl-4-fluoropyrrolidone-3-yl)carbamate tert-butyl ester (60 mg, 0.24 mmol, 1 equivalent) in TFA (0.3 mL) and DCM (0.9 mL) was stirred at 25 °C for 30 min to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give 1-((3S,4S)-3-amino-4-fluoropyrrolidone-1-yl)acetone (60 mg, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =147.2

[0563] The following intermediates in Table 24 are prepared using appropriate starting materials and modifications according to the described procedure for 1-((3S,4S)-3-amino-4-fluoropyrrolidine-1-yl)ethyl-1-one.

[0564] Table 24.

[0565]

[0566] Synthesis of 2-phenyl-1-(2,6-diazaspiro[3.3]hept-2-yl)ethyl-1-one

[0567]

[0568] Step 1: Preparation of tert-butyl 6-(2-phenylacetyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0569] HATU (0.83 g, 2.2 mmol, 1.5 equivalent) and DIEA (0.38 g, 3.0 mmol, 2 equivalent) were added to a solution of 2-phenylacetic acid (0.20 g, 1.5 mmol, 1 equivalent) in DMF (1 mL), followed by a solution of 2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (0.35 g, 1.8 mmol, 1.2 equivalent) in DMF (1 mL) and DIEA (0.57 g, 4.5 mmol, 3 equivalent). The mixture was stirred at 25 °C for 2 hours to obtain a yellow solution. The mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL × 2). The combined organic layers were washed with saturated brine (10 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM:MeOH = 100:1 to 10:1) to give tert-butyl 6-(2-phenylacetyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (0.100 g, 23% yield) as a yellow oil. LCMS: (ESI) m / z [M+H] + =317.0

[0570] Step 2: Preparation of 2-phenyl-1-(2,6-diazaspiro[3.3]hept-2-yl)acetone

[0571] To a solution of 0.1 g (0.32 mmol, 1 equivalent) of tert-butyl 6-(2-phenylacetyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate in DCM (3 mL) and TFA (1 mL), the mixture was stirred at 25 °C for 2 hours to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give 0.100 g (crude product) of 2-phenyl-1-(2,6-diazaspiro[3.3]hept-2-yl)acetone (3-methylazacyclobutane-3-yl)(piperidin-1-yl) methyl ketone as a yellow oil: 1 H NMR (400MHz, CDCl3) δ4.07-3.96(m,2H),3.67(s,2H),3.24-3.13(m,2H),3.09(s,2H),2.99(s,2H),2.91(s,2H),1.74(s,3H).

[0572] According to the described procedure for 2-phenyl-1-(2,6-diazaspiro[3.3]hept-2-yl)ethyl-1-one, the following intermediates in Table 25 are prepared using appropriate starting materials and modifications.

[0573] Table 25.

[0574]

[0575]

[0576] Synthesis of 4-(3-fluorozacricyclobutane-3-yl)benzylnitrile

[0577]

[0578] Step 1: Preparation of tert-butyl 3-(4-cyanophenyl)-3-fluorozacriane-1-carboxylate

[0579] To a solution of tert-butyl 3-(4-bromophenyl)-3-fluorozacricyclobutane-1-carboxylate (0.50 g, 1.5 mmol, 1.0 equivalent) in DMF (10 mL), K4Fe(CN)6 (0.16 g, 0.38 mmol, 0.25 equivalent), sodium carbonate (0.16 mg, 1.51 mmol, 1.0 equivalent), and Pd(dppf)Cl2 (55 mg, 75.5 μmol, 0.05 equivalent) were added. The mixture was stirred at 25 °C for 12 hours to obtain a black mixture. The reaction mixture was filtered, and the filtrate was diluted with water (50 mL) and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by rapid silica gel chromatography to give tert-butyl 3-(4-cyanophenyl)-3-fluorozacriane-1-carboxylate (0.35 g, 83% yield) as a white solid. LCMS: (ESI) m / z [M+H] + =277.0

[0580] Step 2: Preparation of 4-(3-fluorozacriane-3-yl)benzylnitrile

[0581] A solution of tert-butyl 3-(4-cyanophenyl)-3-fluorozacricyclobutane-1-carboxylate (0.25 mg, 0.90 mmol, 1.0 equivalent) in DCM (1.5 mL) and TFA (0.50 mL) was stirred at 25 °C for 30 min to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give 0.16 g of crude 4-(3-fluorozacricyclobutane-3-yl)benzyl nitrile as a yellow oil. LCMS: (ESI) m / z [M+H] + =177.3

[0582] Synthesis of 2-(4-amino-[1,1'-biphenyl]-3-yl)propionamide

[0583]

[0584] Step 1: Preparation of 2-(5-bromo-2-nitrophenyl)acetyl chloride

[0585] A solution of 2-(5-bromo-2-nitrophenyl)acetic acid (1.0 g, 3.8 mmol, 1 equivalent) in SOCl2 was stirred at 80 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to give 2-(5-bromo-2-nitrophenyl)acetyl chloride (1 g, crude product) as a yellow oil. LCMS: (ESI) m / z (M+MeOH) + =273.9.

[0586] Step 2: Preparation of 2-(5-bromo-2-nitrophenyl)acetamide

[0587] NH3·H2O (0.63 g, 17 mmol, 5 equivalents) was added dropwise to a solution of 2-(5-bromo-2-nitrophenyl)acetyl chloride (1.0 g, 3.6 mmol, 1 equivalent) in THF (20 mL) at 0 °C, and the reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give 2-(5-bromo-2-nitrophenyl)acetamide (0.8 g, 86% yield) as a white solid. LCMS: (ESI) m / z [M+H] + =258.9.

[0588] Step 3: Preparation of 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide

[0589] To a solution of 2-(5-bromo-2-nitrophenyl)acetamide (0.8 g, 3.1 mmol, 1 equivalent) in dioxane (10 mL) and H₂O (1 mL), Na₂CO₃ (0.65 g, 6.2 mmol, 2 equivalents) and phenylboronic acid (0.45 g, 3.7 mmol, 1.2 equivalents) were added, followed by the addition of Pd(dppf)Cl₂ (0.22 g, 0.31 mmol, 0.1 equivalents) under N₂. The mixture was stirred at 100 °C for 12 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (0.5 g, 63% yield) as a white solid. LCMS: (ESI) m / z [M+H] + =257.0. Step 4: Preparation of 2-(4-nitro-[1,1'-biphenyl]-3-yl)acrylamide

[0590] Cesium carbonate (0.13 g, 0.38 mmol, 2 equivalents) and methyl iodine (41 mg, 0.29 mmol, 1.5 equivalents) were added to a solution of 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (50 mg, 0.19 mmol, 1 equivalent) in dimethylformamide (10 mL). The mixture was stirred at 80 °C for 2 hours under microwave irradiation. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give 2-(4-nitro-[1,1'-biphenyl]-3-yl)propionamide (34 mg, 64% yield) as a white solid. LCMS: (ESI) m / z [M+H] + =271.2.

[0591] Step 5: Preparation of 2-(4-amino-[1,1'-biphenyl]-3-yl)propionamide

[0592] Ammonium chloride (18 mg, 0.33 mmol, 3 equivalents) and iron (61 mg, 1.1 mmol, 10 equivalents) were added to a solution of 2-(4-nitro-[1,1'-biphenyl]-3-yl)propionamide (30 mg, 0.11 mmol, 1 equivalent) in ethanol (0.4 mL) and water (0.1 mL). The mixture was stirred at 25 °C for 4 hours. The reaction mixture was filtered and the filtrate was concentrated to give 2-(4-amino-[1,1'-biphenyl]-3-yl)propionamide (30 mg, crude product) as a white solid. LCMS: (ESI) m / z [M+H] + =241.1

[0593] Synthesis of (R)-azacyclobutane-3-yl(3-methoxypyrrolidine-1-yl)methyl ketone

[0594]

[0595] Step 1: Preparation of (R)-3-(3-hydroxypyrrolidine-1-carbonyl)azacyclobutane-1-carboxylic acid benzyl ester

[0596] HATU (1.2 g, 3.2 mmol, 1.5 equivalent), DIEA (0.82 g, 6.3 mmol, 3 equivalent), and (R)-pyrrolidine-3-ol (0.18 g, 2.1 mmol, 1 equivalent) were added to a solution of 1-((benzyloxy)carbonyl)azacyclobutane-3-carboxylic acid (0.5 g, 2.1 mmol, 1 equivalent) in DMF (5 mL). The mixture was stirred at 25 °C for 2 hours to obtain a yellow solution. The mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 2). The combined organic layers were washed with saturated brine (20 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by rapid silica gel chromatography to give (R)-3-(3-hydroxypyrrolidine-1-carbonyl)azacyclobutane-1-carboxylate benzyl ester (0.55 g, 85% yield) as a yellow oil. 1 HNMR (400MHz, CDCl3) δ7.44-7.30(m,5H),5.16-5.02(m,2H),4.59-4.49(m,1H),4.37-4.06(m,5H),3.74-3.16(m,6H).

[0597] Step 2: Preparation of (R)-3-(3-methoxypyrrolidine-1-carbonyl)azacyclobutane-1-carboxylic acid benzyl ester

[0598] A solution of (R)-3-(3-hydroxypyrrolidine-1-carbonyl)azacyclobutane-1-carboxylate (50 mg, 0.16 mmol, 1 equivalent) in THF (1 mL) was added with NaH (13 mg, 0.33 mmol, 2 equivalents) and stirred under N2 for 10 min, followed by the addition of methyl iodine (47 mg, 0.33 mmol, 2 equivalents). The mixture was stirred at 25 °C for 2 h to obtain a white suspension. The reaction mixture was quenched with H2O (1 mL) and then concentrated under reduced pressure to obtain a residue. The residue was purified by rapid silica gel chromatography to give (R)-3-(3-methoxypyrrolidine-1-carbonyl)azacyclobutane-1-carboxylate (22 mg, 42% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ7.44-7.30(m,5H),5.11(s,2H),4.17-8.15(m,2H),4.07-3.92(m ,1H),3.79-3.38(m,6H),3.34(d,J=3.2Hz,3H),2.22-1.79(m,2H),1.77-1.41(m,1H).

[0599] Step 3: Preparation of (R)-azacyclobutane-3-yl(3-methoxypyrrolidine-1-yl)methyl ketone

[0600] Pd / C (10 mg) was added to a solution of (R)-3-(3-methoxypyrrolidine-1-carbonyl)azacyclobutane-1-carboxylic acid benzyl ester (50 mg, 0.16 mmol, 1 equivalent) in MeOH (2.5 mL) under N2. The mixture was stirred at 25 °C for 16 hours under H2 (15 psi) to give a black suspension. The reaction mixture was filtered and the filtrate was concentrated to give (R)-azacyclobutane-3-yl(3-methoxypyrrolidine-1-yl) methyl ketone (40 mg, crude product) as a yellow oil.

[0601] Synthesis of (R)-1-(1H-pyrazol-4-yl)pyrrolidine-3-amine

[0602]

[0603] Step 1: Preparation of 4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole

[0604] A solution of 4-bromo-1H-pyrazole (2.0 g, 14 mmol, 1 equivalent) in THF (30 mL) was cooled to 0 °C, and sodium hydride (1.1 g, 27 mmol, 1.5 equivalent) was added at 0 °C under N2. The mixture was stirred at 0 °C for 30 min, and then SEMCl (3.4 g, 20 mmol, 1.5 equivalent) was added dropwise at 0 °C. The mixture was stirred at 25 °C under N2 for 12 h to obtain a yellow solution. The reaction mixture was quenched at 0 °C with H2O (20 mL), diluted with water (50 mL), and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 1:0 to 95:5) to give 4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (1.2 g, 31% yield) as a white solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 7.93 (s, 1H), 7.54 (s, 1H), 5.40 (s, 2H), 3.62–3.50 (m, 2H), 0.91–0.83 (m, 2H), 0.05–0.09 (m, 9H).

[0605] Step 2: Preparation of (R)-(1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-yl)pyrrolidine-3-yl)tert-butyl carbamate

[0606] A solution of 4-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (0.50 g, 1.8 mmol, 1 equivalent), tert-butyl carbamate(R)-pyrrolidine-3-yl ester (0.67 g, 3.6 mmol, 2 equivalents), and potassium tert-butoxide (0.40 g, 3.6 mmol, 2 equivalents) in o-xylene (10 mL) was degassed and purged three times with N2. Then, Pd2(dba)3 (0.16 g, 0.18 mmol, 0.1 equivalents) and t-BuDavephos (0.12 g, 0.36 mmol, 0.2 equivalents) were added to the mixture. The mixture was stirred at 90 °C for 12 hours to obtain a black solution. The mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with saturated brine (50 mL x 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 1:0 to 6:1) to give (R)-(1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)pyrrolidine-3-yl)carbamate tert-butyl ester (60 mg, 8.7% yield) as a white solid. 1 H NMR (400MHz, methanol-d4) δ7.26-7.13(m,2H),5.38-5.25(m,2H),4.24-4.14(m,1H),3.57-3.48(m,2H),3.21(m,1H),3.05(m,1H) ),2.93–2.89(m,1H),2.33-2.22(m,1H),1.90-1.79(m,1H),1.48-1.41(m,9H),1.36-1.27(m,1H),0.92-0.82(m,2H),0.03 -0.06(m,9H).

[0607] Step 3: Preparation of (R)-1-(1H-pyrazol-4-yl)pyrrolidine-3-amine

[0608] A solution of (R)-(1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-yl)pyrrolidine-3-yl)carbamate (30 mg, 78 μmol, 1 equivalent) in DCM (2 mL) and TFA (2 mL) was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to give (R)-1-(1H-pyrazole-4-yl)pyrrolidine-3-amine (30 mg, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =153.0

[0609] Synthesis of (R)-3-aminopyrrolidine-1-carboxamide

[0610]

[0611] Step 1: Preparation of (R)-(1-carbamoylpyrrolidone-3-yl)tert-butyl carbamate

[0612] Urea (1.3 g, 16 mmol, 3.0 equivalent) was added to a solution of tert-butyl carbamate (R)-pyrrolidine-3-yl ester (1.0 g, 5.4 mmol, 1.0 equivalent) in 1,4-dioxane (20 mL). The mixture was stirred at 140 °C for 12 h to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was washed with DCM and filtered. The filtrate was concentrated to give a residue. The residue was purified by rapid silica gel chromatography to give (R)-(1-carbamoylpyrrolidine-3-yl) tert-butyl carbamate (0.800 g, 66% yield) as a white solid. 1 HNMR (400MHz, CDCl3) δ5.06-4.06 (m, 4H), 3.62 (dd, J = 6.0, 10.4Hz, 1H), 3.48 (s, 2H), 3. 25(dd,J=3.6,10.0Hz,1H),2.28-2.08(m,1H),1.90(dd,J=6.4,12.0Hz,1H),1.45(s,9H)

[0613] Step 2: Preparation of (R)-3-aminopyrrolidine-1-carboxamide

[0614] A solution of (R)-(1-carbamoylpyrrolidone-3-yl)carbamate tert-butyl ester (0.8 g, 3.5 mmol, 1 equivalent) in DCM (10 mL) and trifluoroacetic acid (3 mL) was stirred at 20 °C for 1 hour to give a white solution. The reaction mixture was concentrated under reduced pressure to give (R)-3-aminopyrrolidone-1-carboxamide (1 g, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =130.1.

[0615] Synthesis of 1-(pyridin-2-yl)azacyclobutane-3-amine

[0616]

[0617] Step 1: Preparation of (1-(pyridin-2-yl)azacyclobutane-3-yl)tert-butyl carbamate

[0618] K₂CO₃ (1.3 g, 9.5 mmol, 3 equivalents) was added to a solution of 2-bromopyridine (0.50 g, 3.2 mmol, 1 equivalent) and tert-butyl carbamate aziridine-3-yl ester (0.54 g, 3.2 mmol, 1 equivalent) in DMF (5 mL). The mixture was stirred at 80 °C for 16 h to obtain a grayish-white suspension. The reaction mixture was quenched with water (30 mL) and extracted with EtOAc 90 mL (30 mL x 3). The combined organic layers were washed with saturated brine (30 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: YMC Triart C18 250×50mm×7um; mobile phase: [water(FA)-ACN]; B%: 7%-37%, 10 min), followed by lyophilization to give tert-butyl 1-(pyridin-2-yl)azacyclobutane-3-yl)carbamate as a white solid (0.12 g, 15% yield). LCMS: (ESI) m / z [M+H] + =250.1

[0619] Step 2: Preparation of 1-(pyridin-2-yl)azacyclobutane-3-amine

[0620] A solution of (1-(pyridin-2-yl)azacyclobutane-3-yl)carbamate tert-butyl ester (0.10 g, 0.40 mmol, 1 equivalent) in DCM (0.9 mL) and TFA (0.3 mL) was stirred at 25 °C for 1 hour to give a pale yellow solution. The reaction mixture was concentrated under reduced pressure to give 1-(pyridin-2-yl)azacyclobutane-3-amine (0.10 g, crude product) as a yellow oil. LCMS: (ESI) m / z [M+H] + =150.2

[0621] SFC method:

[0622]

[0623] Following separation by chiral SFC (column name: cellulose 4, 15% methanol (0.1% DEA), flow rate: 4 mL / min, injection volume: 14 μL, outlet pressure: 100 bar), (3R,4'R)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carboxynitrile (the fastest eluting enantiomer) and (3S,4'S)-2-oxospiro[indoline-3,3'-pyrrolidine]-4'-carboxynitrile (the slowest eluting enantiomer) were separated. The relative stereochemistry of the oxospiro[indoline-3,3'-pyrrolidine] system is shown in the figure, but the absolute stereochemistry is unknown.

[0624] Representative procedures of the general scheme

[0625] Representative procedure: General scheme 1 (phosphonic acid):

[0626] Synthesis of difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(((R)-2-methylmorpholine-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (Example 1)

[0627]

[0628] Step 1: Preparation of tert-butyl carbamate ((3S,6S,7aS,8aR,9aR)-3-(3-((R)-2-methylmorpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamate

[0629] To a solution of (3S,6S,7aS,8aR,9aR)-6-((tert-butoxycarbonyl)amino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (50 mg, 147 μmol, 1.0 equivalent), HATU (66.8 mg, 176 μmol, 1.2 equivalent), and triethylamine (44.6 mg, 441 μmol, 3.0 equivalent) in DMF (2 mL), (R)-azacyclobutane-3-yl(2-methylmorpholino) ketone (29.6 mg, 161 μmol, 1.1 equivalent) was added, and the mixture was stirred overnight at 20 °C under N2. After completion, the reaction mixture was filtered and concentrated under reduced pressure. The residue was strained... Purified by C18 column chromatography to obtain ((3S,6S,7aS,8aR,9aR)-3-(3-((R)-2-methylmorpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamate tert-butyl (70.0 mg, 138 μmol, 94%) as a white solid. LCMS (ESI): m / z = 505.4 [M+H] + .

[0630] Step 2: Preparation of (5S,8S,10aR)-3-acetyl-5-amino-8-(6-phenyl-4-azaspiro[2,4]heptane-4-carbonyl)octahydropyrrolo[1,2-a][1,5]diazacyclic octatetraen-6(1H)-one

[0631] A solution of ((3S,6S,7aS,8aR,9aR)-3-(3-((R)-2-methylmorpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamate (80 mg, 158 μmol, 1.0 equivalent) in DCM (6 mL) and TFA (2 mL) was stirred overnight at 25 °C under N2. After completion, the reaction mixture was concentrated to give a crude product (5S,8S,10aR)-3-acetyl-5-amino-8-(6-phenyl-4-azaspiro[2,4]heptane-4-carbonyl)octahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-6(1H)-one (63.0 mg, quantified) as a white solid, which could be used directly in the next step without further purification. LCMS (ESI): m / z = 405.4 [M+H] + .

[0632] Step 3: Preparation of (difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(((R)-2-methylmorpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0633] A solution of (5S,8S,10aR)-3-acetyl-5-amino-8-(6-phenyl-4-azaspiro[2,4]heptane-4-carbonyl)octahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-6(1H)-one (7,60 mg, 148 μmol, 1.0 equivalent), triethylamine (44.8 mg, 443 μmol, 3.0 equivalent), DMAP (9.03 mg, 74.0 μmol, 0.5 equivalent), and (difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (83.9 mg, 177 μmol, 1.2 equivalent) in DMF (2 mL) was stirred overnight at 30 °C under N2. After completion, the reaction mixture was filtered and concentrated under reduced pressure. The residue was strained... Purified by C18 column chromatography to obtain a white solid (difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(((R)-2-methylmorpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (80.0 mg, 115 μmol, 78%). LCMS (ESI): m / z = 695.3 [M+H] + ;1 H NMR (400MHz, DMSO-d6) δ9.00-8.84(m,1H),8.34(s,1H),8.13(d,J=8.5Hz,1H),8.08(s,1H),7.58(d,J=8.6Hz, 1H),5.00-4.86(m,1H),4.68-4.46(m,1H),4.40-4.25(m,2H),4.22-4.09(m,2H),4.05-3.85(m,2H),3.81-3.6 3(m,2H),3.48-3.27(m,3H),3.11-2.98(m,1H),2.78-2.62(m,1H),2.39-2.28(m,1H),2.18-1.96(m,4H),1.91 -1.75(m,2H),1.70-1.55(m,1H),1.42-1.27(m,1H),1.12-1.01(m,3H),0.84-0.66(m,2H),0.03-0.04(m,1H).

[0634] Representative procedure: General scheme 2: (phosphonate or amide)

[0635] Synthesis of ((difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)glycine isopropyl ester (Example 2)

[0636]

[0637] HATU (63.1 mg, 166 μmol, 1.67 equivalent) was added to a solution of (3S,6S,7aS,8aR,9aR)-6-amino-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)decahydro-5H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-5-one (50 mg, 99.1 μmol, 1 equivalent), DIPEA (99.2 mg, 768 μmol, 7.75 equivalent), and 5-(difluoro(((2-isopropoxy-2-oxoethyl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (73.9 mg, 153 μmol, 1.54 equivalent) in DMF (3 mL). The reaction was stirred at room temperature for 1 hour. The reaction mixture was directly purified by reversed-phase chromatography on a 50 g C18 cartridge (eluted with 5-100% MeCN / water) to give a white solid ((difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)glycine isopropyl ester (88.2 mg, 103 μmol, 104%). LCMS: (ESI) m / z = 856.1 [M+H] + ; 1 HNMR(400MHz,CD3OD)δ8.23-8.12(m,2H),8.07-7.98(m,1H),7.75-7.69(m,1H),7.36-7.29(m,2H),7.23-7.15(m,3 H),5.07-4.92(m,2H),4.73-4.70(m,1H),4.54-4.37(m,2H),4.33-4.16(m,2H),4.11-4.02(m,1H),3.82-3.71(m,1H ),3.71-3.44(m,8H),3.38-3.34(m,1H),3.30-3.20(m,1H),2.50-2.38(m,1H),2.35-2.06(m,4H),2.04-1.92(m,2H) ,1.78-1.67(m,1H),1.45-1.36(m,1H),1.20-1.16(m,6H),0.98-0.89(m,1H),0.88-0.79(m,1H),0.10-0.02(m,1H).

[0638] Representative procedure: General scheme 3 (phosphonic acid):

[0639] Synthesis of ((2-(((3S,6S,7aS,8aR,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (Example 3)

[0640]

[0641] Step 1: Preparation of (3S,6S,7aR,8aS,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxymethyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylate

[0642] HATU (45 mg, 1.2 mmol, 1.5 equivalent) was added to a solution of 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (29 mg, 0.79 mmol, 1 equivalent) in DMF (1 mL), followed by a solution of (3S,6S,7aR,8aS,9aR)-6-amino-5-oxodecahydro-1H-cyclopropion[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylic acid methyl ester (20 mg, 0.79 mmol, 1.2 equivalent) in DMF (1 mL) and DIEA (41 mg, 3.2 mmol, 4 equivalent). The mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with saturated brine (10 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / EtOAc = 1:1 to 15:85) to give (3S,6S,7aR,8aS,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylate (15 mg, 41% yield) as a yellow solid. LCMS: m / z [M+H] + =599.4.

[0643] Step 2: Preparation of (3S,6S,7aR,8aS,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxymethyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylic acid

[0644] Methyl (3S,6S,7aR,8aS,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxymethyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylate (15 mg, 0.25 mmol, 1 equivalent) was added to a solution of ACN (0.9 mL) and 2N HCl (0.3 mL), and the mixture was stirred at 70 °C for 5 hours. The reaction mixture was filtered, and the filtration residue was purified by preparative HPLC (column: Phenomenex Luna C18150 × 25 mm × 10 μm, mobile phase: water (0.1% TFA)-ACN; B%: 28%-58%, 10 min), followed by lyophilization to give (3S,6S,7aR,8aS,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (4 mg, 43% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ8.11(s,1H),7.95(m,2H),7.85(d,J=5.6Hz,1H),7.66(d,J=8.4Hz ,1H),4.96(d,J=5.2Hz,1H),4.56-4.45(m,1H),4.32-4.10(m,5H),2.39-2.27(m,2H),2 .24-2.12(m,2H),2.10-2.00(m,1H),1.99-1.88(m,1H),1.82-1.72(m,1H),1.72-1.64( m,2H),1.37-1.29(m,6H),1.27-1.18(m,1H),0.87-0.75(m,1H),0.10(d,J=4.8Hz,1H).

[0645] Step 3: Preparation of ((2-(((3S,6S,7aR,8aS,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonate diethyl ester

[0646] HATU (3.8 mg, 10 μmol, 1.5 equivalent) was added to a solution of (3S,6S,7aR,8aS,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (4 mg, 6.8 μmol, 1 equivalent) in DMF (0.3 mL), followed by a solution of [1,1'-biphenyl]-4-amine (1.4 mg, 8.2 μmol, 1.2 equivalent) in DMF (0.3 mL) and DIEA (4.0 mg, 0.33 mmol, 4 equivalent). The mixture was stirred at 25 °C for 1 hour. The mixture was directly purified by column chromatography (SiO2, petroleum ether / EtOAc = 10:1 to 3:1) to obtain a yellow oil of ((2-(((3S,6S,7aR,8aS,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonate diethyl ester (12 mg, crude product). LCMS: m / z [M+H] + =736.2

[0647] Step 4: Preparation of ((2-(((3S,6S,7aR,8aS,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid

[0648] At 0 °C, BSTFA (25 mg, 0.96 mmol, 6 equivalents) was added to a solution of ((2-(((3S,6S,7aR,8aS,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonate diethyl ester (12 mg, 0.16 mmol, 1 equivalent) in DCM (0.5 mL), and DCM containing TMSI (13 mg, 0.64 mmol, 4 equivalents) was added dropwise. The mixture was stirred at 0 °C for 15 minutes. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150×25mm×10um, mobile phase: water (0.1% TFA)-ACN; B%: 40%-70%, 10 min), followed by lyophilization to give a white solid of ((2-(((3S,6S,7aR,8aS,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (1.22 mg, 9.45% yield). LCMS: (ESI) m / z [M+H] + =680; 1 H NMR (400MHz, CD3OD) δ8.18-8.08(m,2H),7.98(d,J=8.4Hz,1H),7.69(d,J=8.4Hz,1H),7.66-7.61(m, 2H),7.61-7.54(m,4H),7.41(t,J=7.6Hz,2H),7.33-7.26(m,1H),5.00(d,J=4.4Hz,1H),4.55(t,J=8. 4Hz,1H),4.37(t,J=7.6Hz,1H),2.42-2.31(m,2H),2.27-2.14(m,2H),2.12-2.04(m,1H),2.03-1.96( m,2H),1.89-1.76(m,1H),1.54-1.41(m,1H),1.38-1.23(m,3H),0.92-0.84(m,1H),0.29-0.22(m,1H)

[0649] Specific analogue synthesis

[0650] Synthesis of (((difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphoryl)bis(oxy))bis(methylene)divalerate (Example 4)

[0651]

[0652] Step 1: Preparation of methyl valerate

[0653] A solution of NaHCO3 (1.63 g, 19.5 mmol, 2.0 equivalent) in water (10 mL) was added dropwise to a solution of valeric acid (1.1 g, 9.79 mmol, 1.0 equivalent), tetrabutylammonium hydrogen sulfate (332 mg, 979 μmol, 0.1 equivalent), and methyl thiochloride (1.76 g, 10.7 mmol, 1.1 equivalent) in DCM (10 mL). The reaction mixture was then heated to room temperature and stirred overnight under N2. After completion, the reaction mixture was extracted with dichloromethane (20 mL x 3). The organic layers were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give methyl valerate (1.40 g, 9.29 mmol, 95% yield) as a clear oil. 1 HNMR (400MHz, CDCl3) δ5.71 (s, 2H), 2.39 (t, J = 7.5Hz, 2H), 1.69-1.60 (m, 2H), 1.41-1.33 (m, 2H), 0.93 (t, J = 7.4Hz, 3H).

[0654] Step 2: Preparation of iodomethyl valerate

[0655] A solution of methyl valerate (400 mg, 2.65 mmol, 1.0 equivalent) and NaI (794 mg, 5.30 mmol, 2.0 equivalent) in MeCN (10 mL) was stirred overnight at 40 °C under N2. After completion, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel rapid chromatography to give iodomethyl valerate (3, 100 mg, 413 μmol, 16%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ5.91 (s, 2H), 2.34 (t, J = 7.5Hz, 2H), 1.68-1.58 (m, 2H), 1.41-1.31 (m, 2H), 0.92 (t, J = 7.3Hz, 3H).

[0656] Step 3: Preparation of (((difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphoryl)bis(oxy))bis(methylene)divalerate

[0657] A solution of NaOH (7.03 mg, 176 μmol, 2.0 equivalent) in water (2 mL) was added dropwise to a stirred suspension of (difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (60 mg, 88.1 μmol, 1.0 equivalent) in H2O (6 mL). When the mixture became clear (pH ~ 9), AgNO3 (32.7 mg, 193 μmol, 2.2 equivalent) was added. After stirring at 0 °C for 2 hours, the gray precipitate was collected by filtration and dried under vacuum. The powder was suspended in dry toluene (1 mL) and methyl iodide valerate (3, 63.9 g, 264 μmol, 3.0 equivalent) was added. The mixture was stirred at room temperature for 24 hours. After filtration, the solvent was removed under vacuum. The crude residue was purified directly by silica gel rapid chromatography to give (((difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphoryl)bis(oxy))bis(methylene)divalerate (ST-214-2, 2.00 mg, 2.20 μmol, 3% yield). LCMS(ESI): m / z = 909.3 [M+H] + ; 1H NMR(400MHz, DMSO-d6)δ8.99(dd,J=22.8,7.5Hz,1H),8.39-8.00(m,3H),7.55(d,J=8.6Hz,1H),5.86-5.36(m, 4H),4.99-4.89(m,1H),4.68-4.50(m,1H),4.37-4.24(m,2H),4.20-4.11(m,1H),4.05-3.85(m,2H),3.75-3.6 4(m,1H),3.56-3.50(m,4H),3.48-3.44(m,2H),3.28-3.23(m,2H),2.34-2.24(m,4H),2.17-1.94(m,4H),1.90 -1.75(m,2H),1.68-1.57(m,1H),1.54-1.40(m,4H),1.36-1.19(m,6H),0.89-0.67(m,8H),0.01--0.02(m,1H).

[0658] ((R)-((2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)-L-alanine isopropyl ester and ((S)- Preparation of ((2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)-L-alanine isopropyl ester (Examples 5 and 6)

[0659]

[0660] Preparative separation method: Instrument: Waters Thale 80 preparative SFC; Column: ChiralPak IB, 250 × 21.2 mm ID, 5 μm; Mobile phase: A represents CO2, and B represents MeOH + 0.1% NH3H2O; Gradient: B 35%; Flow rate: 40 mL / min; Back pressure: 100 bar; Column temperature: 35 °C; Wavelength: 220 nm; Cycle time: 4 min.

[0661] (((2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)-L-alanine isopropyl ester (60 mg, 71.9 μmol, 1.0 equivalent) was purified by preparative SFC to produce:

[0662] Peak 1: A white solid ((R)-((2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)-L-alanine isopropyl ester or ((S)-((2-(((3S,6S,7aS,8aR,9aR)- 3-(3-(morpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)-L-alanine isopropyl ester (20.0 mg, 24.0 μmol, 99.9% ee, 33% yield) Peak 1 data (Example 5), p-chirality arbitrarily specified LCMS (ESI): m / z = 834.4 [M+H] + ; 1H NMR (400MHz, CDCl3) δ7.89-7.73(m,3H),7.49-7.42(m,1H),7.37-7.27(m,3H),7.16-7.08(m,3H),5.10-5. 01(m,1H),4.99-4.73(m,2H),4.58-4.50(m,1H),4.48-4.39(m,1H),4.35-4.26(m,1H),4.22-4.05(m,2H), 4.00-3.91(m,1H),3.74-3.53(m,7H),3.45(d,J=20.8Hz,2H),3.33-3.23(m,2H),3.18-3.08(m,1H),2.37- 2.24(m,3H),2.16-1.97(m,4H),1.67-1.61(m,1H),1.50-1.38(m,1H),1.21-1.16(m,6H),1.12(d,J=7.1Hz, 3H), 1.01-0.91(m,1H), 0.86-0.77(m,1H), 0.06-0.00(m,1H) and peak 2: white solid ((S)-((2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy (Phosphoyl)-L-alanine isopropyl ester or ((R)-((2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoyl)-L-alanine isopropyl ester (30.0 mg, 36.0 μmol, 99.7% ee, 50% yield). LCMS(ESI): m / z = 834.4 [M+H] + Peak 2 data (Example 6), P-chirality arbitrarily specified: 1H NMR (400MHz, CDCl3) δ7.86-7.69(m,3H),7.55-7.33(m,2H),7.32-7.27(m,2H),7.19-7.06(m,3H),5.11-5 .00(m,1H),4.95-4.72(m,2H),4.58-4.48(m,1H),4.46-4.37(m,1H),4.35-4.26(m,1H),4.22-4.05(m,2H) ,3.98-3.87(m,1H),3.74-3.51(m,7H),3.46-3.20(m,5H),2.37-2.23(m,3H),2.15-1.96(m,4H),1.72-1. 62(m,1H),1.50-1.39(m,1H),1.21-1.08(m,9H),1.00-0.89(m,1H),0.86-0.76(m,1H),0.04-0.00(m,1H).

[0663] Synthesis of difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((R)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (Example 7)

[0664]

[0665] Step 1: Preparation of tert-butyl 4-((3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylamino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carbonyl)piperazine-1-carboxylate

[0666] To a solution of (3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (0.4 g, 0.68 mmol, 1.0 equivalent) in DCM (1 mL), piperazine-1-carboxylic acid tert-butyl ester (0.13 g, 0.68 mmol, 1.0 equivalent), T3P (0.46 g, 1.0 mmol, 1.5 equivalent, 50% purity) and DIEA (0.13 g, 1.0 mmol, 1.5 equivalent) were added. The mixture was stirred at 20 °C for 16 hours to give a yellow turbid solution. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by rapid silica gel chromatography to give 4-((3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylammono)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carbonyl)piperazine-1-carboxylic acid tert-butyl ester (0.43 g, 83% yield) as a white solid. LCMS: (ESI) m / z [M-Boc] + =653.2

[0667] Step 2: Preparation of diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(piperazin-1-carbonyl)decahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate

[0668] TFA (1.0 mL) was added to a solution of 4-((3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carbonyl)piperazine-1-carboxylic acid tert-butyl ester (0.43 g, 0.56 mmol, 1.0 equivalent) in DCM (3.0 mL). The resulting mixture was stirred at 25 °C for 1 hour to obtain a yellow solution. The reaction mixture was concentrated under reduced pressure to give diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(piperazin-1-carbonyl)decahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate (0.43 g, crude product).

[0669] Step 3: Preparation of diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((R)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate

[0670] DIEA (30 mg, 0.23 mmol, 3.0 equivalent) was added to a solution of diethyl phosphonate (2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(piperazin-1-carbonyl)decahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate (50 mg, 77 μmol, 1.0 equivalent) in DMF (1.0 mL), and the reaction was stirred at 20 °C for 30 min to obtain a colorless solution. DIEA (30 mg, 0.23 mmol, 3.0 equivalence) was added to a separate solution of (3R)-oxacyclopentane-3-carboxylic acid (8.9 mg, 77 μmol, 1.0 equivalence) in DMF (1.0 mL), and HATU (43 mg, 0.1 mmol, 1.5 equivalence) was added. The mixture was stirred at 20 °C for 30 min to obtain a black solution. The two mixtures were combined and stirred at 20 °C for 1 h to obtain a solution. The mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL × 2). The combined organic layers were washed with saturated brine (10 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give a yellow oil of diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((R)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate (50 mg, crude product). LCMS: (ESI) m / z [M+H] + =751.3

[0671] Step 4: Preparation of (difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((R)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0672] BSTFA (0.1 g, 0.4 mmol, 6.0 equivalence) was added to a solution of diethyl phosphonate (50 mg, 0.07 mmol, 1.0 equivalence) in DCM (1.0 mL) at 0 °C, followed by dropwise addition of trimethyliodosilane (54 mg, 0.27 mmol, 4.0 equivalence) and stirring at 0 °C for 0.5 h to give a yellow mixture. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (neutral; column: Waters Xbridge 150×25mm 10µm, mobile phase: water (10mMNH4HCO3)-ACN; B%: 5%-35%, 10 min), followed by lyophilization to give a white solid of (difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((R)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (1.2 mg, 2.6% yield). LCMS: (ESI) m / z [M+H] + =695.2; 1 H NMR (400MHz, CD3OD) δ8.17(s,1H),8.11(s,1H),7.93(d,J=8.4Hz,1H),7.73(d,J=8.4Hz,1H),5.09-5 .01(m,1H),5.00-4.95(m,1H),4.31(d,J=6.4Hz,1H),3.95(s,1H),3.91-3.65(m,8H),3.64-3.51(m, 2H),3.50-3.40(m,2H),2.53-2.42(m,1H),2.33(dd,J=2.0,14.4Hz,1H),2.19(s,5H),2.05-1.87(m, 2H),1.85-1.68(m,1H),1.45(s,1H),1.02(d,J=8.8Hz,1H),0.91-0.79(m,1H),0.08(d,J=5.2Hz,1H)

[0673] According to the procedure described above for (difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((R)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid, the following (difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((S)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid was prepared using appropriate starting materials (Example 8):

[0674]

[0675] (difluoro(2-(((3S,6S,7aS,8aR,9aR)-5-oxo-3-(4-(((S)-tetrahydrofuran-3-carbonyl)piperazine-1-carbonyl)decahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid: 1 HNMR(400MHz,CD3OD)δ8.27-8.07(m,2H),7.93(d,J=8.4Hz,1H),7.73(d,J =8.8Hz,1H),5.13-4.94(m,2H),4.36-4.19(m,1H),4.04-3.38(m,13H),2.5 6-2.42(m,1H),2.38-2.29(m,1H),2.24-1.88(m,7H),1.78-1.75(m,1H),1 .51-1.36(m,1H),1.08-0.97(m,1H),1.11-0.87(m,1H),0.12-0.08(m,1H).

[0676] Synthesis of ((2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-1-acetyl-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (Example 9)

[0677]

[0678] Step 1: Preparation of (3S,4S)-3-((3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxyamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxyamido)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester

[0679] A solution of (3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (0.1 g, 0.17 mmol, 1 equivalent) was added to DCM (5 mL), CMPI (44 mg, 0.34 mmol, 2 equivalents), and TEA (52 mg, 0.51 mmol, 3 equivalents). The mixture was stirred at 20 °C for 1 hour to obtain a yellow solution. The solution was quenched with water (10 mL) and extracted with EtOAc (10 mL × 2). The combined organic layers were washed with saturated brine (10 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by rapid silica gel chromatography to give (3S,4S)-3-((3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylamino)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylamino)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester (90 mg, 79% yield) as a yellow solid. LCMS: (ESI) m / z [M+H] + =769.2

[0680] Step 2: Preparation of diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate

[0681] A solution of (3S,4S)-3-((3S,6S,7aS,8aR,9aR)-6-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamid)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxamid)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester (90 mg, 0.12 mmol, 1 equivalent) in DCM (3 mL) and TFA (0.6 mL) was stirred at 20 °C for 1 hour to obtain a yellow solution. The mixture was concentrated under vacuum to give diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate (90 mg, crude product). LCMS: (ESI) m / z [M+H] + =669.2

[0682] Step 3: Preparation of ((2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-1-acetyl-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonate diethyl ester

[0683] A solution of diethyl difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonate (90 mg, 0.12 mmol, 1 equivalent) in DCM (5 mL) and triethylamine (24 mg, 0.24 mmol, 2 equivalents) was stirred at 20 °C for 10 min. Then, Ac₂O (15 mg, 0.14 mmol, 1.2 equivalents) was added dropwise at 0 °C, and the mixture was stirred at 20 °C for 2 h to obtain a yellow solution. The mixture was concentrated under vacuum to obtain a yellow solid. The solid was purified by rapid silica gel chromatography to give a yellow solid ((2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-1-acetyl-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonate diethyl ester (47 mg, 55% yield).1 H NMR (400MHz, CDCl3) δ8.10 (s, 1H), 7.98-7.89 (m, 2H), 7.71 (d, J = 3.2Hz, 1H), 7.63 (d, J = 8.4Hz, 1H) ,5.18-5.06(m,1H),4.72-4.57(m,1H),4.32-4.14(m,6H),3.81(d,J=5.6Hz,1H),3.72(q,J=7.2Hz, 3H),3.08-3.06(m,1H),3.14(dd,J=4.4,7.2Hz,1H),2.41-2.18(m,4H),2.09-2.05(m,2H),1.95(s, 3H),1.61(dd,J=3.6,7.6Hz,1H),1.34-1.30(m,6H),0.86-0.71(m,2H),0.07(dd,J=4.0,7.6Hz,1H)

[0684] Step 4: Preparation of ((2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-1-acetyl-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid

[0685] BSTFA (89 mg, 0.34 mmol, 4 equivalents) and trimethyliodosilane (69 mg, 0.34 mmol, 4 equivalents) were added dropwise to a solution of ((2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-1-acetyl-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropyl[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonate diethyl ester (47 mg, 86 μmol, 1 equivalent) in DCM (3 mL). The reaction mixture was stirred at 0 °C for 1 hour to give a yellow suspension. The mixture was concentrated under vacuum to give a brown solid. The solid was purified by preparative HPLC (column: Phenomenex Luna C18 100×30mm×5um, conditions: water (10mMNH4HCO3)-ACN, flow rate (mL / min): 25 min), followed by lyophilization to obtain a yellow solid of ((2-(((3S,6S,7aS,8aR,9aR)-3-(((3S,4S)-1-acetyl-4-hydroxypyrrolidine-3-yl)carbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (11 mg, 20% yield). LCMS: (ESI) m / z [M+H] + =655.1; 1 H NMR (400MHz, CD3OD) δ8.19(s,1H),8.13(s,1H),7.93(d,J=8.4Hz,1H),7.74(d,J=8.4Hz,1H),5.03( t,J=9.2Hz,1H),4.61-4.50(m,1H),4.31-4.25(m,1H),4.19-4.05(m,2H),3.89-3.53(m,2H),3.49- 3.36(m,2H),2.54-2.40(m,1H),2.34-2.25(m,1H),2.23-2.09(m,3H),2.05(d,J=10.4Hz,4H),2.01 -1.92(m,1H),1.81-1.62(m,1H),1.25-0.99(m,1H),0.93-0.81(m,2H),0.10(dd,J=3.2,5.4Hz,1H)

[0686] Phosphonic acid data sheet

[0687] Phosphonic acid was prepared according to general scheme 1.

[0688] Based on the representative procedure described above for the synthesis of (difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(((R)-2-methylmorpholino-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (Example 1), the following compounds in Table 26 were prepared using appropriate starting materials and modifications.

[0689] Table 26.

[0690]

[0691]

[0692]

[0693]

[0694]

[0695]

[0696]

[0697]

[0698]

[0699]

[0700]

[0701]

[0702]

[0703]

[0704]

[0705]

[0706]

[0707]

[0708]

[0709]

[0710]

[0711]

[0712]

[0713]

[0714]

[0715]

[0716]

[0717]

[0718]

[0719]

[0720]

[0721]

[0722] Prepare according to general scheme 2: Phosphonic acid

[0723] Based on the representative procedure described above for the synthesis of ((2-(((3S,6S,7aS,8aR,9aR)-3-([1,1'-biphenyl]-4-ylcarbamoyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)difluoromethyl)phosphonic acid (Example 3), the following compounds in Table 27 were prepared using appropriate starting materials and modifications.

[0724] Table 27.

[0725]

[0726]

[0727]

[0728]

[0729]

[0730]

[0731]

[0732]

[0733]

[0734]

[0735]

[0736]

[0737]

[0738]

[0739]

[0740]

[0741] Data sheet for phosphonates and amides:

[0742] General Option 1: Phosphate or Amide

[0743] Based on the representative procedure described above for the synthesis of ((difluoro(2-(((3S,6S,7aS,8aR,9aR)-3-(3-(morpholin-4-carbonyl)azacyclobutane-1-carbonyl)-5-oxodecahydro-1H-cyclopropane[d]pyrrolo[1,2-a]azacyclooctatetraen-6-yl)carbamoyl)benzo[b]thiophene-5-yl)methyl)(phenoxy)phosphoryl)glycine isopropyl ester (Example 2), the following compounds in Table 28 were prepared using appropriate starting materials and modifications.

[0744] Table 28.

[0745]

[0746]

[0747]

[0748]

[0749]

[0750]

[0751]

[0752]

[0753]

[0754]

[0755]

[0756]

[0757]

[0758]

[0759]

[0760]

[0761]

[0762]

[0763]

[0764]

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771]

[0772]

[0773]

[0774]

[0775]

[0776]

[0777]

[0778]

[0779]

[0780]

[0781]

[0782]

[0783]

[0784]

[0785]

[0786]

[0787]

[0788]

[0789]

[0790]

[0791] Synthesis of (3S,6S,8aR,9aS,9bR)-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid

[0792]

[0793] Step 1: (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-methyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid

[0794]

[0795] Lithium hydroxide monohydrate (356 mg, 8.50 mmol, 2.0 equivalent) was added to a solution of (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-methyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (1.5 g, 4.25 mmol, 1 equivalent) in tetrahydrofuran (6 mL) and water (2 mL). The reaction was stirred at room temperature for 16 hours. The reaction was then partially concentrated under reduced pressure to remove THF. The reaction mixture was diluted with water (20 mL) and EtOAc (50 mL). The phases were separated and the aqueous phase was acidified to pH 1–2 with 1 N HCl aqueous solution. The product was then extracted with DCM (3 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure to give (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-methyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (1.43 g, 100%) as a white solid. LCMS (ESI) m / z = 339.2 (M+H) + .

[0796] Step 2: (3S,6S,8aR,9aR,9bR)-9,9-dibromo-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid and (3S,6S,8aS,9aS,9bR)-9,9-dibromo-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid

[0797]

[0798] Under a nitrogen atmosphere, bromoform (5.22 mL, 59.9 mmol, 14 equivalents) and benzyltriethylammonium chloride (309 mg, 1.36 mmol, 0.32 equivalents) were added to a solution of (3S,6S,10aR)-6-((tert-butoxycarbonyl)amino)-9-methyl-5-oxo-1,2,3,5,6,7,8,10a-octahydropyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (1.45 g, 4.28 mmol, 1 equivalent) in methylene chloride (15 mL). A solution of sodium hydroxide (3.76 g, 94.1 mmol, 22 equivalents) in water (7.52 mL) was added. The reaction was heated under reflux for 72 hours. The reaction was then concentrated under reduced pressure to remove DCM, water, and most of the excess bromoform. The resulting dark brown residue was dissolved in water (50 mL) and acidified to pH 1-2 with 3N HCl aqueous solution. The aqueous layer was extracted with DCM (3 x 50 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 275 g C. 18 Purification was performed using a syringe (eluted with 5-60% MeCN / water (containing 0.1% formic acid)). The first eluent, (3S,6S,8aS,9aS,9bR)-9,9-dibromo-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (479 mg, 21.9%), was separated as a white solid. LCMS (ESI) m / z = 408.9 (M-Boc+H) + ; 1 H NMR(400MHz, CDCl3) δ5.87(d,J=6.8Hz,1H),4.54(t,J=8.8Hz,1H),4.30-4.22(m,1H),3 .96-3.88(m,1H),2.52-1.96(m,8H),1.69(d,J=10.8Hz,1H),1.44(s,9H),1.36(s,3H).

[0799] The second elution product (3S,6S,8aR,9aR,9bR)-9,9-dibromo-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (191 mg, 8.8%) was separated as a white solid. LCMS (ESI) m / z = 408.9 (M-Boc+H) + ; 1H NMR (400MHz, CDCl3) δ5.91(s,1H),5.36(d,J=8.8Hz,1H),4.78-4.70(m,1H),4.66(t,J=7.6Hz,1H),3.39-3.28(m,2H),2.89(dt,J=13.9 ,7.2Hz,2H),2.48-2.38(m,1H),2.34-2.24(m,1H),2.05-1.95(m,1H),1.87(s,3H),1.84-1.75(m,1H),1.61-1.52(m,1H),1.44(s,9H).

[0800] Step 3: (3S,6S,8aR,9aS,9bR)-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraene-3-carboxylic acid

[0801]

[0802] In a tube under a nitrogen atmosphere, (3S,6S,8aS,9aS,9bR)-9,9-dibromo-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (600 mg, 1.17 mmol, 1 equivalent) was dissolved in 10 mL of 2-propanol. Then, potassium hydroxide (393 mg, 7.02 mmol, 6 equivalents) and 10% palladium / carbon (50% wet) (400 mg, 188 μmol, 0.16 equivalents) were added. The tube was filled with hydrogen (40 psi) and sealed. The reaction mixture was heated to 70 °C for 20 hours. The reaction mixture was then cooled to room temperature, diluted with MeOH, and filtered through a diatomaceous earth mat. The filtrate was concentrated under reduced pressure. The crude residue was analyzed by reversed-phase chromatography at 50 g C. 18 Purified on a syringe (eluted with 5-60% MeCN / water (containing 0.1% formic acid)) to give (3S,6S,8aR,9aS,9bR)-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (120 mg, 29.1%) as a white solid after lyophilization. LCMS (ESI) m / z = 353.2 (M+H) + ; 1HNMR (400MHz, CDCl3) δ5.87(d,J=6.8Hz,1H),4.56(t,J=8.6Hz,1H),4.24(t,J=7.9Hz,1H),3.63(dd,J=11.0,6.4Hz,1H),2.68-2.51(m,1H),2.38- 2.20(m,2H),2.14-1.87(m,4H),1.53(dd,J=15.3,10.6Hz,1H),1.44(s,9 H), 1.04-0.93 (m, 4H), 0.54 (dd, J = 8.6, 4.9Hz, 1H), 0.33 (t, J = 5.0Hz, 1H).

[0803]

[0804] (3S,6S,8aS,9aR,9bR)-6-((tert-butoxycarbonyl)amino)-8a-methyl-5-oxodecahydro-1H-cyclopropane[c]pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid was synthesized under the same conditions as described above. LCMS (ESI) m / z = 353.2 (M+H) + .

[0805] synthesis

[0806] [(2-{[(1R,3S,5R,7S,10S)-3-methyl-8-oxo-10-[3-(pyridin-3-yl)azacyclobutane-1-carbonyl]-9-azatricyclic]

[0807] [7.3.0.0 3 [5]dodecane-7-yl]carbamoyl}-1-benzothiophene-5-yl)methyl]phosphonic acid

[0808]

[0809] Step 1: (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methyl-5-oxo-1H,2H,3H,5H,6H,7H,10H,10aH-pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid

[0810]

[0811] Lithium hydroxide monohydrate (1.04 g, 24.8 mmol, 2.5 equivalents) was added to a solution of (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methyl-5-oxo-1H,2H,3H,5H,6H,7H,10H,10aH-pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylate (3.5 g, 9.93 mmol, 1 equivalent) in tetrahydrofuran (24 mL) and water (8 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was partially concentrated under reduced pressure (to remove THF). The crude mixture was diluted with water (40 mL) and EtOAc (60 mL). The phases were separated, and the aqueous phase was acidified with 1N HCl (pH = 1–2), and the product was then extracted with DCM (3 x 60 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 150g C 18 Purified on a syringe (eluted with 5-60% MeCN / water (containing 0.1% formic acid)) to give (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methyl-5-oxo-1H,2H,3H,5H,6H,7H,10H,10aH-pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (2.85 g, 84.8%) as a white solid. LCMS (ESI): m / z = 339.1 (M+H) + ; 1 H NMR(400MHz,DMSO-d6)δ12.32(br.s.,1H),6.75(d,J=7.3Hz,1H),5.48-5.39(m,1H),4.47(q,J=7.4Hz,1H) ,4.19(d,J=7.1Hz,2H),2.77-2.52(m,2H),2.26-2.03(m,4H),1.83(br.s.,2H),1.74(s,3H),1.36(s,9H).

[0812] Step 2: (1R,7S,10S)-4,4-dibromo-7-{[(tert-butoxy)carbonyl]amino}-3-methyl-8-oxo-9-azatricyclic [7.3.0.0] 3 [5] Dodecane-10-carboxylic acid

[0813]

[0814] Under a nitrogen atmosphere, tribromomethane (3.84 mL, 44.3 mmol, 10 equivalents) and benzyltriethylammonium chloride (201 mg, 886 μmol, 0.2 equivalents) were added to a solution of (3S,6S,10aR)-6-{[(tert-butoxy)carbonyl]amino}-9-methyl-5-oxo-1H,2H,3H,5H,6H,7H,10H,10aH-pyrrolo[1,2-a]azacyclooctatetraen-3-carboxylic acid (1.5 g, 4.43 mmol, 1 equivalent) in methylene chloride (50 mL). A solution of sodium hydroxide (3.89 g, 97.4 mmol, 22 equivalents) in water (7.8 mL) was added. The reaction mixture was heated to 40 °C for 24 hours. The reaction mixture was concentrated under reduced pressure to remove DCM, water, and most of the bromoform. The dark brown residue was dissolved in water (30 mL) and slowly acidified to pH 2 with 3N HCl aqueous solution while stirring. The acidic aqueous layer was washed with DCM (3 x 30 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was subjected to reversed-phase chromatography at 150 g C. 18 Purified on a cartridge (eluted with 5-60% MeCN (containing 0.1% formic acid / water)) to give (1R,7S,10S)-4,4-dibromo-7-{[(tert-butoxy)carbonyl]amino}-3-methyl-8-oxo-9-azatricyclo[7.3.0.0] as a beige solid. 3 [5] Dodecane-10-carboxylic acid (1.42 g, 62.8%). 1 H NMR(400MHz,DMSO-d6)δ12.45(br.s.,1H),6.70(d,J=6.8Hz,1H),4.55-4.03(m,3H),2.43-2.08(m,4 H),2.06-1.97(m,1H),1.96-1.67(m,4H),1.55-1.50(m,1H),1.48-1.44(m,1H),1.42-1.24(m,10H).

[0815] Step 3: (1R,7S,10S)-4,4-dibromo-7-{[(tert-butoxy)carbonyl]amino}-3-methyl-8-oxo-9-azatricyclic [7.3.0.0] 3 [5] Dodecane-10-carboxylic acid

[0816]

[0817] In a pressure vessel, (1R,7S,10S)-4,4-dibromo-7-{[(tert-butoxy)carbonyl]amino}-3-methyl-8-oxo-9-azatricyclo[7.3.0.0] 3[5] Dodecane-10-carboxylic acid (1.4 g, 2.74 mmol, 1 equivalent) was dissolved in 2-propanol (40 mL), and then potassium hydroxide (920 mg, 16.4 mmol, 6 equivalent) and 10% palladium / carbon (50% wet) (1.44 g, 0.68 mmol, 0.25 equivalent) were added under a nitrogen atmosphere. The reaction mixture was heated to 70 °C at 40 psi for 21 hours. The reaction mixture was cooled to room temperature, diluted with MeOH, and filtered through a diatomaceous earth mat. The filtrate was concentrated under reduced pressure. The crude formate was diluted with a minimal amount of water. The crude product was analyzed by reversed-phase chromatography at 50 g C 18 Purification was performed on a tube (eluted with 5-60% MeCN / water (containing 0.1% formic acid / water)). The tube containing the pure product was concentrated under reduced pressure to give the desired product (460 mg) as a white solid (a mixture of the two isomers). The product was subjected to chiral SFC separation (SFC conditions: Lux amylose-121.2 x 250 mm 5 μm column, 10 mg / inj, concentration 1.67 mg / mL, column T = 40 °C, flow rate: 25 mL / min, 10% MeOH, circulation time: 4.6 min) to give the white solid (1R,3S,5R,7S,10S)-7-{[(tert-butoxy)carbonyl]amino}-3-methyl-8-oxo-9-azatricyclo[7.3.0.0] 3 [5] Dodecane-10-carboxylic acid (180 mg, 18.6%, fastest eluting isomer). LCMS (ESI): m / z = 353.2 (M+H) + . 1 ¹H NMR (400 MHz, benzene-d6) δ 6.30–6.18 (m, 1H), 4.52–4.32 (m, 2H), 3.60–3.47 (m, 1H), 2.25–2.20 (m, 1H), 1.77–1.72 (m, 1H), 1.66–1.53 (m, 3H), 1.46 (s, 9H), 1.42–1.25 (m, 4H), 1.17–1.06 (m, 1H), 0.81 (s, 3H), 0.33–0.30 (m, 1H), 0.04–0.10 (m, 1H). (1R,3R,5S,7S,10S)-7-{[(tert-butoxy)carbonyl]amino}-3-methyl-8-oxo-9-azatricyclo[7.3.0.0] 3 [5] Dodecane-10-carboxylic acid (256 mg, 26.5%, slowest eluting isomer) was separated as a white solid. LCMS (ESI): m / z = 353.2 (M+H) + ; 1H NMR (400MHz, benzene-d6) δ5.33-5.31(m,1H),4.52-4.46(m,1H),4.11-4.09(m,1H),3.30-3.26(m,1H),2.34-2.31(m,1H),2.02-1.86(m,1H),1.62 -1.45(m,3H),1.44(s,9H),1.36-1.09(m,2H),1.12-0.90(m,2H),0.75 (s,3H),0.46-0.32(m,1H),0.17-0.14(m,1H),-0.42(t,J=4.4Hz,1H).

[0818] Joint assembly:

[0819] ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid and ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)

[0820] Synthesis of fluoromethylphosphonic acid:

[0821]

[0822] 5-Methylbenzo[b]thiophene-2-carboxylate was prepared according to the procedure described in WO 2016 / 100184.

[0823] Step 1: Ethyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate

[0824] AIBN (544 g, 3.31 mol, 0.10 equivalent) and NBS (6.19 kg, 34.8 mol, 1.05 equivalent) were added to a solution of ethyl 5-methylbenzo[b]thiophene-2-carboxylate (1, 7.3 kg, 33.1 mol, 1.0 equivalent) in CHCl3 (58 L) with stirring at 20 °C. The mixture was heated from 30 °C to 50 °C over 4 hours, and then heated to 60 °C with stirring for 12 hours. After completion, the reaction mixture was cooled to 10 °C and 15% Na2SO3 (20 L) was added. The organic layer was washed with H2O (20 L * 2), dried over Na2SO4, and concentrated under reduced pressure at 45 °C to produce ethyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (2, 9.50 kg, 23.5 mol, 70.9% yield, 74.0% purity) as a yellow solid. LCMS(ESI): m / z = 298.9 [M+H] + ; 1H NMR (400MHz, CDCl3) δ8.034(s,1H),7.89–7.84(m,2H),7.50(d,J=9.6Hz,1H),4.64(s,2H),4.45-4.40(m,2H),1.45–1.41(m,3H).

[0825] Step 2: Ethyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0826] Triethyl phosphite (5.8 kg, 34.9 mol, 1.1 equivalent) was added to a solution of ethyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (2, 9.50 kg, 31.8 mol, 1.0 equivalent) in DMF (28.5 L) with stirring at 20 °C. The mixture was heated to 100 °C and stirred for 5 hours. After completion, the reaction mixture was cooled to 15 °C, poured into H2O (50.0 L), and extracted with EtOAc (20 L * 2). The combined organic matter was washed with H2O (20 L * 2) and brine (10 L), dried over Na2SO4, and concentrated under reduced pressure at 45 °C to obtain the residue. The crude residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 100 / 1 to 1 / 1, petroleum ether / ethyl acetate = 0 / 1) to produce ethyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate as a yellow solid (3, 5.24 kg, 14 mol, 44.4% yield). LCMS (ESI): m / z = 356.9 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.02 (s, 1H), 7.82–7.80 (m, 2H), 7.41 (d, J = 2.0Hz, 1H), 4.44–4.41 ( m,2H),4.05-4.01(m,4H),3.27(d,J=21.6Hz,2H),1.46–1.42(m,3H),1.27–1.23(m,6H).

[0827] Step 3: ethyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate

[0828] Three batches were carried out in parallel.

[0829] Under N2 and at -70°C, LiHMDS (1M, 702mL, 1.00 equivalent) was added dropwise to a solution of ethyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (3,250g, 702mmol, 1.00 equivalent) and N-(benzenesulfonyl)-N-fluorobenzenesulfonamide (221g, 702mmol, 1.00 equivalent) in THF (2.50L). The mixture was stirred at -70°C for 3 hours. After completion, the reaction mixture was slowly poured into a saturated aqueous solution of NH4Cl (5.00L) at 0°C, and the mixture was stirred at 0°C for 0.5 hours. The three batches were then combined for processing. The mixture was extracted with ethyl acetate (5.00L*3). The organic layers were combined, washed with brine (5.00L), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was subjected to column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0-3 / 1, R) f Purified with petroleum ether / ethyl acetate (0.30, petroleum ether / ethyl acetate = 1 / 1) to produce ethyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (4,357 g, 928 mmol, 44.1% yield) as a yellow oil. LCMS (ESI): m / z = 375.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.07(s,1H),8.00(s,1H),7.90(d,J=8.4Hz,1H),7.59(d,J= 8.8Hz,1H),5.88-5.75(m,1H),4.45-4.40(m,2H),4.15-4.05(m,4H),1.45-1.41(m 3H),1.31-1.28(m,6H).

[0830] Step 4: 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0831] Under N2 at 10–20 °C, H2O (760 mL) and LiOH·H2O (56.5 g, 1.35 mol, 2.00 equivalent) were added to a solution of ethyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (4,252 g, 673 mmol, 1.00 equivalent) in MeOH (1.80 L). The mixture was stirred at 10–20 °C for 1 hour. TLC (petroleum ether / ethyl acetate = 1 / 1) showed that compound 4 (R) was consumed. f =0.30) and formed the desired spot (R f=0.10). The reaction mixture was quenched with H2O (2.50 L) and the pH was adjusted to 3-4 with HCl (aqueous solution, 1 M). The mixture was extracted with dichloromethane (2.50 L * 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure to give 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid as a white solid (5,222 g, 626 mmol, 93.0% yield).

[0832] Step 5: 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0833] Two batches are carried out in parallel.

[0834] At 10–20 °C, K₂CO₃ (142 g, 1.03 mol, 2.00 equivalent) and allyl bromide (68.4 g, 565 mmol, 1.10 equivalent) were added to a solution of 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid (5, 178 g, 514 mmol, 1.00 equivalent) in DMF (1.78 L). The mixture was stirred at 10–20 °C for 12 hours. TLC (petroleum ether / ethyl acetate = 0 / 1) showed that compound 5 (R) was consumed. f =0.60) and new spots (R) were formed. f =0.70). The reaction mixture was diluted with H2O (6.00 L) and extracted with ethyl acetate (2.00 L * 3). The organic layers were combined. The mixture was washed with brine (2.00 L) and NH4Cl (2.00 L), dried over anhydrous Na2SO4 and concentrated under reduced pressure to give 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (6,388 g, 985 mmol, 95.8% yield) as a yellow oil.

[0835] Step 6: ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid

[0836] At 0 °C, TMSBr (411 g, 2.71 mol, 20.0 equivalent) was added dropwise to a solution of 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester (7, 50 g, 136 mmol, 1.0 equivalent) in DCM (5 L). After addition, the reaction mixture was warmed to room temperature and stirred for an additional 12 hours. The reaction progress was monitored by LCMS. After completion, the reaction mixture was concentrated under reduced pressure and water was added. The resulting mixture was filtered and the filter cake was washed with water (2 L) and then dried under vacuum to produce ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)methyl)phosphonic acid (40.3 g, 129 mmol, 95%) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.20(s,1H),7.96(d,J=8.4Hz,1H),7.87(s,1H),7.43(d,J=8.4Hz,1H),6. 10-5.98(m,1H),5.46-5.37(m,1H),5.33-5.24(m,1H),4.86-4.77(m,2H),3.08(d,J=21.2Hz,2H). LCMS(ESI)m / z=313.1[M+H] + .

[0837] Step 7: ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid

[0838] At 0 °C, TMSBr (41.1 g, 270 mmol, 20.0 equivalent) was added dropwise to a solution of 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (9, 5.2 g, 13.5 mmol, 1.0 equivalent) in DCM (500 mL). After addition, the reaction mixture was warmed to room temperature and stirred for an additional 12 hours. The reaction progress was monitored by LCMS. After completion, the reaction mixture was concentrated under reduced pressure and water was added. The resulting mixture was filtered and the filter cake was washed with water (200 mL) and then dried under vacuum to produce ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid (3.1 g, 9.37 mmol, 69%) as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ8.32 (s, 1H), 8.14-8.05 (m, 2H), 7.59 (d, J = 8.4Hz, 1H), 6.12-5.99 (m ,1H),5.84(dd,J=44.3,8.2Hz,1H),5.49-5.40(m,1H),5.35-5.27(m,1H),4.88-4.81(m,2H). LCMS(ESI):m / z=329.1[MH] - .

[0839] Chiral separation of 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0840] Rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (617 g, 1.62 mol, 1.00 equivalent) was purified by SFC to obtain (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (peak 1) and (R)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (peak 2).

[0841] Preparative SFC Method: Instrument: Waters 350 preparative SFC. Column: REGIS(S,S)WHELK-O1, 250×50mm ID, 10μm. Mobile Phase: A represents CO2, and B represents MEOH (neutralized). Gradient: B 30%. Flow Rate: 220 g / min. Back Pressure: 100 bar. Column Temperature: 35℃. Wavelength: 220 nm. Cycle Time: 3.3 min.

[0842] Analytical SFC method: Column: Kromasil (S,S)WHELK-O1, 50 × 4.6 mm ID, 3.5 μm. Mobile phase: A represents CO2, and B represents MEOH (0.05% DEA). Gradient: B 5 to 40%. Flow rate: 3 mL / min. Back pressure: 100 bar. Column temperature: 35 °C. Wavelength: 220 nm.

[0843] (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester was obtained as a yellow oil (peak 1, 267 g, 685 mmol, 39.7% yield, >99% ee, RT = 1.36 min). LCMS (ESI): m / z = 387.1 [M+H] + .

[0844] (R)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester was obtained as a yellow oil (peak 2, 270 g, 676 mmol, 39.2% yield, >99% ee, RT = 1.55 min). LCMS (ESI): m / z = 387.1 [M+H] + .

[0845] The absolute stereochemical configuration is specified by comparing experimental vibrational circular dichroism (VCD) spectra with theoretical VCD spectra obtained from DFT calculations.

[0846] The method for preparing ((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid described in step 7 above was used, and the following intermediates in Table 29 were prepared using appropriate starting materials and modifications.

[0847] Table 29

[0848]

[0849] Synthesis of perfluorophenyl 5-((S)-fluoro((R)-(((S)-1-oxo-1-propoxypropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid perfluorophenyl ester

[0850]

[0851] Step 1: Preparation of (S)-((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid

[0852] Trimethyliodosilane (21 g, 0.10 mol, 2 equivalents) was added to a solution of (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (20 g, 52 mmol, 1 equivalent) in methylene chloride (500 mL). The mixture was stirred at 0 °C for 1 h to give a brown solution. The reaction mixture was concentrated under reduced pressure to give a residue. The reaction residue was purified by preparative HPLC (TFA) to lyophilization to give (S)-((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid (12 g, 36 mmol, 70% yield) as a brown solid. LCMS (ESI) m / z = 330.9

[0853] Step 2: Preparation of (S)-5-((dichlorophosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0854] Under a nitrogen atmosphere at 0°C, dimethylformamide (0.24 g, 3.3 mmol, 0.1 equivalent) was added to a solution of (S)-((2-((allyloxy)carbonyl)benzo[b]thiophene-5-yl)fluoromethyl)phosphonic acid (11 g, 33 mmol, 1 equivalent) in methylene chloride (200 mL), followed by dropwise addition of oxaloyl chloride (13 g, 0.10 mol, 3 equivalent). The mixture was stirred at 0°C for 30 minutes and then heated to 40°C for 1 hour to obtain a brown solution. The reaction mixture was concentrated under reduced pressure to give (S)-5-((dichlorophosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid allyl ester (11 g, crude product) as a yellow solid. LCMS (ESI) m / z = 358.9.

[0855] Step 3: Preparation of 5-((1S)-fluoro((((S)-1-oxo-1-propoxypropyl-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid allyl ester

[0856] At 0 °C, a solution of (S)-5-((dichlorophosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (11 g, 30 mmol, 1 equivalent) in methylene chloride (150 mL) was added to a solution of methylene chloride (20 mL) containing phenol (2.2 g, 24 mmol, 0.8 equivalent) over 10 minutes. The...

Claims

1. A compound having structural formula I: or a pharmaceutically acceptable salt thereof, wherein: 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; R 1 Selected from the following: -CR 1a R 2a P(O)OR 1b OR 2b 、 -CR 1a R 2a P(O)[OR 1b [NH(AA)C(O)OR T 、 -CR 1a R 2a P(O)[NHR Ty [NH(AA)C(O)OR T 、 -P(O)OR 1b OR 2b 、 -[P(O)[NHR Ty [NH(AA)C(O)OR T 、 -CR 1a R 2a P(O)[NH(AA)C(O)OR T [NH(AA)C(O)OR T or -P(O)[OR 1b [NH(AA)C(O)OR T substituted 8 - to 10 - membered fused bicyclic heteroaryl; -CR 1a R 2a P(O)OR 1b OR 2b 、 -CR 1a R 2a P(O)[OR 1b [NH(AA)C(O)OR T 、 -CR 1a R 2a P(O)[NHR Ty [NH(AA)C(O)OR T 、 -P(O)OR 1b OR 2b 、 -[P(O)[NHR Ty [NH(AA)C(O)OR T 、 -CR 1a R 2a P(O)[NH(AA)C(O)OR T [NH(AA) C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ] substituted 8- to 10-membered fused bicyclic heterocyclic group; 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T ]][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ] substituted aryl; -(C1-C4)alkyl(aryl), wherein the aryl portion of -(C1-C4)alkyl(aryl) is replaced by -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -CR 1a R 2a P(O)[NH(AA)C(O)OR T ]] [NH(AA)C(O)OR T ]、-P(O)[OR 1b ][NH(AA)C(O)OR T ] substituted; and -(C2-C4)alkenyl(aryl), wherein the aryl portion of -(C2-C4)alkenyl(aryl) is replaced by -CR 1a R 2a P(O)OR 1b OR 2b ,–CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T ]][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]replace; R 1a and R 2a are each independently selected from hydrogen, cyano, (C1-C4)alkyl, hydroxy(C1-C4)alkyl and fluorine; or R 1a and R 2a Together with the carbon to which it is attached, it forms an oxo group; R 1b and R 2b Each is independently selected from 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-C4) alkyl] 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)alkylphenyl]-C(O)O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)-[NH(AA)C(O)OR T ], -[(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, -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl]-O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)NH(C1-C4)alkyl], -[(C1-C4)alkyl] wherein, when valence permits, the 5- to 6-membered heteroaryl and the aryl are each optionally and independently substituted with 1 to 2 groups selected from halo, cyano and (C1-C4) alkyl, and wherein, when valence permits, the 5- to 7-membered heterocyclyl of [(C1-C4) alkyl]-OC(O)O-[5- to 7-membered heterocyclyl] and [(C1-C4) alkyl]-OC(O)-[5- to 7-membered heterocyclyl] are each optionally and independently substituted with 1 to 2 groups selected from C(O)OR h The group substitution; R 2 is selected from hydrogen, halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, cyano and hydroxy; R 3 and R 4 are each independently selected from hydrogen, halo and (C1-C4)alkyl; R 5 and R 6 are each independently selected from hydrogen, phenyl and (C1-C4)alkyl; R 7 is selected from (C1-C4)alkyl, phenyl, 4 to 9-membered monocyclic or bicyclic heterocyclic group and 5 to 10-membered monocyclic or bicyclic heteroaryl, wherein when valence permits, the (C1-C4)alkyl is optionally substituted by 1 to 3 selected from R Y and when valence permits, the phenyl, the 4- to 9-membered monocyclic or bicyclic heterocyclic group and the 5- to 10-membered monocyclic or bicyclic heteroaryl are each optionally substituted by 1 to 3 groups selected from R Z or R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a 4- to 14-membered monocyclic or bicyclic heterocyclic group or a 5- to 12-membered monocyclic or bicyclic heteroaryl group, each of which is optionally substituted with 1 to 3 members selected from R Q The group substitution; R 8 is hydrogen or (C1-C4) alkyl; AA is the residue of an α or β natural or unnatural amino acid; R T and R Ty Each is independently selected from (C1-C4)alkyl, (C1-C4)alkyl-C(O)O(C1-C4)alkyl, benzyl and phenyl, wherein the phenyl is optionally substituted with 1 or 2 groups selected from halo, (C1-C4)alkyl and halo(C1-C4)alkyl; R Q is selected from halo, (C2-C4) alkenyl, (C1-C4) alkyl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, cyano, phenyl, hydroxy, 4-9 membered monocyclic or bicyclic heterocyclic group, 5-10 membered monocyclic or bicyclic heteroaryl, (C3-C6) cycloalkyl, oxo, imino, -OR e 、-C(O)R g 、-C(O)OR e 、-NR c C(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)2NR e R f , wherein the (C2-C4)alkenyl and the (C1-C4)alkyl are each optionally and independently substituted with 1 to 3 selected from R M wherein, when valence permits, the phenyl, the 5- to 10-membered monocyclic or bicyclic heteroaryl, the (C3-C6) cycloalkyl and the 4- to 9-membered monocyclic or bicyclic heterocyclyl are each optionally and independently substituted with 1 to 3 groups selected from R F The group substitution; R Y is selected from 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 when valence permits, the phenyl, the 4 to 6 membered heterocyclyl and the 5 to 10 membered monocyclic or bicyclic heteroaryl are each optionally substituted by 1 to 3 selected from R X The group substitution; R M and R J Each is independently selected from halo, (C1-C4) alkyl, (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)=NR e (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 when valence permits, the phenyl, the 4 to 6 membered heterocyclyl and the 5 to 10 membered monocyclic or bicyclic heteroaryl are each optionally substituted by 1 to 3 selected from R X The group substitution; R F , R X and R Z Each is independently selected from halo, cyano, (C1-C4) alkyl, cyano (C1-C4) alkyl, (C3-C6) cycloalkyl, halo (C1-C4) alkyl, -(C1-C4) alkyl C(O)NR c R d , -(C1-C4)alkyl(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, -(C1-C4)alkylphenyl, -(C1-C4)alkylheteroaryl, (C2-C4)alkenyl, halo(C2-C4)alkenyl, (C2-C4)alkynyl, halo(C2-C4)alkynyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, -OR e , oxo, imino, phenyl, 4- to 6-membered heterocyclic group, 5- to 6-membered monocyclic heteroaryl, -S(O)R e R f 、-S(O)2R f 、-S(O)=NH(C1-C4)alkyl、-S(O)NR e R f 、-S(O)2NR e R f 、-C(O)OR e 、-NR c C(O)R e 、-(C1-C4 alkyl)C(O)R g 、-C(O)R g 、-C(O)NR c R d , NO2 and -NR a R b , wherein the phenyl group, the 4- to 6-membered heterocyclyl group and the phenyl group of the -(C1-C4)alkylphenyl group are each optionally and independently substituted with 1 to 3 groups selected from the following groups: halo, cyano, oxo, (C1-C4)alkylphenyl, 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 ) alkynyl, halo(C1-C 10 )alkyl, (C1-C 10 )alkoxy, -(C1-C4)alkyl(C1-C4)alkoxy and halo(C1-C 10 ) alkoxy, wherein, when valence permits, the (C1-C 10 ) alkyl, the (C2-C 10 )alkenyl and said (C2-C 10 ) alkynyl is each optionally substituted with a 5- to 10-membered monocyclic or bicyclic heteroaryl, or a 4- to 10-membered monocyclic or bicyclic heterocyclyl, each of which is optionally substituted with oxo or a 5- to 7-membered heterocyclyl optionally substituted with 1 to 2 oxo; and When valence permits, R a , R b , R c , R d , R e , R f , R g and R h are independently selected from hydrogen, (C1-C4) alkyl, (C 2- C4) alkynyl, -(C1-C4) alkylphenyl, phenyl, (C3-C6) cycloalkyl, 4- to 6-membered heterocyclyl, and 5- to 6-membered heteroaryl, wherein the (C1-C4) alkyl is optionally substituted with 1 to 3 members selected from R J and when valence permits, the phenyl, the (C3-C6)cycloalkyl, the 4- to 6-membered heterocyclyl and the 5- to 6-membered heteroaryl are each independently and optionally substituted by 1 to 3 groups selected from the following: halo, cyano, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxy, phenyl and benzyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R F , R X and R Z Each is independently selected from halo, cyano, (C1-C4) alkyl, cyano (C1-C4) alkyl, (C3-C6) cycloalkyl, halo (C1-C4) alkyl, -(C1-C4) alkyl C(O)NR c R d , -(C1-C4)alkyl(C1-C4)alkoxy, hydroxy(C1-C4)alkyl, -(C1-C4)alkylphenyl, -(C1-C4)alkylheteroaryl, (C2-C4)alkenyl, halo(C2-C4)alkenyl, (C2-C4)alkynyl, halo(C2-C4)alkynyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, -OR e , oxo, imino, phenyl, 4- to 6-membered heterocyclic group, 5- to 6-membered monocyclic heteroaryl, -S(O)R e R f 、-S(O)2R f 、-S(O)=NH(C1-C4)alkyl、-S(O)NR e R f 、-S(O)2NR e R f 、-C(O)OR e 、-NR c C(O)R e 、-(C1-C4 alkyl)C(O)R g 、-C(O)R g 、-C(O)NR c R d , NO2 and -NR a R b , wherein the phenyl group, the 4- to 6-membered heterocyclyl group and the phenyl group of the -(C1-C4)alkylphenyl group are each optionally and independently substituted with 1 to 3 groups selected from the following groups: halo, cyano, oxo, (C1-C4)alkylphenyl, 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 ) alkynyl, halo(C1-C 10 )alkyl, (C1-C 10 ) alkoxy and halogen (C1-C 10 ) alkoxy, wherein, when valence permits, the (C1-C 10 ) alkyl, the (C2-C 10 )alkenyl and said (C2-C 10 )alkynyl is each optionally substituted with a 5- to 10-membered monocyclic or bicyclic heteroaryl or a 4- to 10-membered monocyclic or bicyclic heterocyclyl, each of which is optionally substituted with oxo or a 5- to 7-membered heterocyclyl optionally substituted with 1 to 2 oxo.

3. The compound according to claim 1 or 2, wherein the compound has structural formula II: or a pharmaceutically acceptable salt thereof. 4 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein q is 1. 5 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein t is 1. 6 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein p is 1.

7. A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R 2 For hydrogen.

8. The compound according to any one of claims 1 to 7, wherein the compound has structural formula III or IV: or a pharmaceutically acceptable salt thereof.

9. A compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, wherein R 5 For hydrogen.

10. The compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, wherein R 3 and R 4 are each independently selected from hydrogen and halo.

11. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, wherein R 3 and R 4 Each is hydrogen.

12. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, wherein R 3 and R 4 Each is fluorine.

13. A compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from 8- to 10-membered fused bicyclic heteroaryl and aryl, each of which is replaced by -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T ]][NH(AA)C(O)OR T ] or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]replace.

14. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from benzothienyl and naphthyl, each of which is replaced by -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[NHR Ty ][NH(AA)C(O)OR T ]、-CR 1a R 2a P(O)[NH(AA)C(O)OR T ]][NH(AA)C(O)OR T ] or -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]replace.

15. A compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, wherein R 1 Selected from 16. A compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, wherein R 1 Selected from 17. A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, wherein R 1 for 18. A compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, wherein R 1a and R 2a are each independently selected from hydrogen and fluorine.

19. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 1a and R 2a Each is hydrogen.

20. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 1a is hydrogen and R 2a For fluorine.

21. A compound according to any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, wherein R 1a and R 2a Each is fluorine.

22. A compound according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b each independently selected from 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)alkylphenyl]-C(O)O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)-[NH(AA)C(O)OR T ], -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl]-OH, -[(C1-C4)alkyl]-OC(O)O-[5- to 7-membered heterocyclyl], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl]-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)-[(C1-C4)alkyl]-OH and phenyl.

23. A compound according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b Each is independently selected from hydrogen, [(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl]-O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl] and -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl].

24. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b Each is -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl].

25. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b For hydrogen.

26. A compound according to any one of claims 1 to 17 and 22, or a pharmaceutically acceptable salt thereof, wherein -CR 1a R 2a P(O)OR 1b OR 2b Selected from 27. A compound according to any one of claims 1 to 17, 22 and 26, or a pharmaceutically acceptable salt thereof, wherein -CR 1a R 2a P(O)OR 1b OR 2b for 28. A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, wherein -(AA)C(O)OR T -C(R')(R)C(O)R T or -C(R')(R)CH2C(O)R T , wherein R' is hydrogen or methyl, and R is selected from hydrogen, methyl, ethyl, -CH2CH(CH3)2, -CH2OCH3, benzyl and -CH2CH2-phenyl.

29. A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, wherein -(AA)C(O)OR T -C(R')(R)C(O)R T or -C(R')(R)CH2C(O)R T , wherein R' is hydrogen, and R is selected from hydrogen, methyl, -CH2CH(CH3)2, benzyl and -CH2CH2-phenyl.

30. A compound according to any one of claims 1 to 16, 28 and 29, or a pharmaceutically acceptable salt thereof, wherein R T Selected from (C1-C4)alkyl, (C1-C4)alkyl-C(O)OC 1-4 Alkyl and benzyl.

31. A compound according to any one of claims 1 to 16 and 28 to 30, or a pharmaceutically acceptable salt thereof, wherein -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]Selected from 32. A compound according to any one of claims 1 to 16 and 28 to 30, or a pharmaceutically acceptable salt thereof, wherein -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]Selected from 33. A compound according to any one of claims 1 to 32, or a pharmaceutically acceptable salt thereof, wherein R 6 For hydrogen.

34. A compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, wherein R 7 is selected from (C1-C4) alkyl, phenyl and 4 to 6 membered monocyclic heterocyclic groups, wherein when valence permits, the (C1-C4) alkyl is optionally substituted by 1 to 3 selected from R Y and when valence permits, the phenyl group and the 4- to 6-membered monocyclic heterocyclic group are each optionally substituted by 1 to 3 groups selected from R Z The group is substituted.

35. A compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, wherein R 7 is selected from (C1-C4)alkyl, phenyl, pyrrolidinyl and azetidinyl, wherein when valence permits, the (C1-C4)alkyl is optionally substituted by 1 to 3 selected from R Y and when valence permits, the phenyl group, the pyrrolidinyl group and the azetidinyl group are each optionally substituted with 1 to 3 groups selected from R Z The group is substituted.

36. A compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, wherein R Z Selected from halo, -(C1-C4)alkylC(O)NR c R d , hydroxyl, phenyl, 4- to 6-membered heterocyclic group, 5- to 6-membered monocyclic heteroaryl, -C(O)NR c R d and -C(O)R g , wherein the phenyl is optionally substituted, when valence permits, with 1 to 3 groups selected from the group consisting of halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy and halo(C1-C4)alkoxy.

37. A compound according to any one of claims 1 to 36, or a pharmaceutically acceptable salt thereof, wherein R Z Selected from halo, -(C1-C4)alkylC(O)NR c R d , hydroxyl, phenyl, tetrahydropyran, tetrahydrofuran, oxetanyl, pyridyl, pyrimidyl, imidazolyl, triazolyl, pyrazolyl, pyridazinyl, -C(O)NR c R d and -C(O)R g , wherein the pyridyl, the imidazolyl and the triazolyl are optionally substituted by one or two groups selected from halo and methyl.

38. A compound according to any one of claims 1 to 36, or a pharmaceutically acceptable salt thereof, wherein R Z Selected from halo, -(C1-C4)alkylC(O)NR c R d , hydroxyl, phenyl, tetrahydropyran, tetrahydrofuran, oxetanyl, pyridyl, pyrazolyl, pyridazinyl, -C(O)NR c R d and -C(O)R g .

39. A compound according to any one of claims 1 to 38, or a pharmaceutically acceptable salt thereof, wherein R Y is selected from hydroxy and 5 to 10 membered monocyclic or bicyclic heteroaryl, wherein when valence permits, the 5 to 10 membered monocyclic or bicyclic ring is optionally substituted by 1 to 3 selected from R X The group is substituted.

40. A compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt thereof, wherein R Y is selected from hydroxy, pyridyl and pyrrolopyridyl.

41. A compound according to any one of claims 1 to 40, or a pharmaceutically acceptable salt thereof, wherein R c and R d Each is hydrogen.

42. A compound according to any one of claims 1 to 41 or a pharmaceutically acceptable salt thereof, wherein R g It is -(C1-C4)alkyl.

43. A compound according to any one of claims 1 to 32, or a pharmaceutically acceptable salt thereof, wherein R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a 4- to 14-membered monocyclic or bicyclic heterocyclic group or a 5- to 12-membered monocyclic or bicyclic heteroaryl group, each of which is optionally substituted with 1 to 3 members selected from R Q The group is substituted.

44. A compound according to any one of claims 1 to 32 and 43, or a pharmaceutically acceptable salt thereof, wherein R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms azetidinyl, 2,5-diazaspiro[3.4]octanyl, pyrrolidinyl, 2,6-diazaspiro[3.3]heptyl, 2,6-diazabicyclo[3.2.0]heptyl, piperazinyl, spiro[indoline-3,3'-pyrrolidinyl]yl, 6',7'-dihydrospiro[azetidine-3,5'-pyrrolo[1,2-a]imidazolyl]yl, 3,4-dihydro-2H-benzo[b][1,4] oxazinyl, 3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazine, 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine, 2,3,4,5-tetrahydrobenzo[b][1,4]oxazepine, 1,2,3,4-tetrahydroquinoxalinyl, 1-azaspiro[3.5]nonanyl, 4-azaspiro[2.4]heptanyl, each of which is optionally substituted, when valence permits, by 1 to 3 selected from R Q The group is substituted.

45. A compound according to any one of claims 1 to 32 and 43, or a pharmaceutically acceptable salt thereof, wherein R 6 and R 7 together with the nitrogen atom to which it is attached, form azetidinyl, 2,5-diazaspiro[3.4]octanyl, pyrrolidinyl, 2,6-diazaspiro[3.3]heptanyl, piperazinyl, spiro[indoline-3,3'-pyrrolidinyl]yl, 6',7'-dihydrospiro[azetidine-3,5'-pyrrolo[1,2-a]imidazolyl]yl, each of which is optionally substituted, as valence permits, by 1 to 3 selected from R Q The group is substituted.

46. ​​A compound according to any one of claims 1 to 32 and 43 to 45, or a pharmaceutically acceptable salt thereof, wherein R Q Selected from halo, (C1-C4) alkyl, -OR e , cyano, phenyl, hydroxy, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, oxo, and -C(O)R g , wherein the (C1-C4)alkyl group is optionally substituted with 1 to 3 selected from R M wherein the phenyl, the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 6-membered heterocyclic group are each optionally and independently substituted with 1 to 3 groups selected from R F substituted with a group, and wherein R e is a (C1-C4) alkyl group or a 5- to 6-membered heteroaryl group.

47. A compound according to any one of claims 1 to 32 and 43 to 45, or a pharmaceutically acceptable salt thereof, wherein R Q is selected from halo, (C1-C4) alkyl, (C1-C4) alkoxy, cyano, phenyl, hydroxy, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, oxo, and -C(O)R g , wherein the (C1-C4)alkyl group is optionally substituted with 1 to 3 selected from R M wherein the phenyl, the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 6-membered heterocyclic group are each optionally and independently substituted with 1 to 3 groups selected from R F The group is substituted.

48. A compound according to any one of claims 1 to 32 and 43 to 47, or a pharmaceutically acceptable salt thereof, wherein R Q Selected from halo, (C1-C4) alkyl, -OR e , cyano, phenyl, hydroxy, morpholinyl, tetrahydropyranyl, thiomorpholinyl, piperidinyl, oxetanyl, pyrazolyl, pyridinyl, tetrazolyl, imidazolyl, pyrazinyl, isoxazolyl, oxazolyl, oxadiazolyl, triazolyl, pyrimidinyl, benzimidazolyl, 1H-pyrrolo[3,2-c]pyridine, 2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl, oxo, and -C(O)R g , wherein the (C1-C4)alkyl group is optionally substituted with 1 to 3 selected from R M wherein the morpholinyl, tetrahydropyranyl, thiomorpholinyl, piperidinyl, oxetanyl, pyrazolyl, pyridinyl, tetrazolyl, imidazolyl, pyrazinyl, isoxazolyl, oxazolyl, oxadiazolyl, triazolyl, pyrimidinyl, benzimidazolyl, 1H-pyrrolo[3,2-c]pyridine and 2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl are each optionally and independently substituted with 1 to 3 groups selected from R F substituted by a group, wherein R e is (C1-C4)alkyl, pyridyl, pyrazinyl, pyrimidinyl, pyrazole, and wherein R e The pyrazolyl represented is optionally substituted by (C1-C4)alkyl.

49. A compound according to any one of claims 1 to 32 and 43 to 47, or a pharmaceutically acceptable salt thereof, wherein R Q is selected from the group consisting of halo, (C1-C4)alkyl, (C1-C4)alkoxy, cyano, phenyl, hydroxy, morpholinyl, tetrahydropyranyl, thiomorpholinyl, piperidinyl, oxetanyl, pyrazolyl, pyridinyl, tetrazolyl, imidazolyl, pyrazinyl, oxadiazolyl, triazolyl, pyrimidinyl, benzimidazolyl, 2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl, oxo, and -C(O)R g , wherein the (C1-C4)alkyl group is optionally substituted with 1 to 3 selected from R M wherein the morpholinyl, tetrahydropyranyl, thiomorpholinyl, piperidinyl, oxetanyl, pyrazolyl, pyridinyl, tetrazolyl, imidazolyl, pyrazinyl, oxetanyl, triazolyl, pyrimidinyl, benzimidazolyl and 2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl are each optionally and independently substituted with 1 to 3 groups selected from R F The group is substituted.

50. A compound according to any one of claims 1 to 49, or a pharmaceutically acceptable salt thereof, wherein, when valency permits, R g is selected from (C1-C4)alkyl, 4- to 6-membered heterocyclyl and 5- to 6-membered heteroaryl, wherein the (C1-C4)alkyl is optionally substituted with 1 to 3 members selected from R J The 4- to 6-membered heterocyclyl and the 5- to 6-membered heteroaryl are each independently and optionally substituted by 1 to 3 groups selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, benzyl and hydroxyl, when valence permits.

51. A compound according to any one of claims 1 to 50, or a pharmaceutically acceptable salt thereof, wherein R J It is phenyl.

52. A compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, wherein, when valence permits, R g is selected from (C1-C4)alkyl, morpholinyl, azetidinyl, tetrahydropyranyl, oxetanyl, pyrrolidinyl and pyrazolyl, wherein when valence permits, the (C1-C4)alkyl is optionally substituted by 1 to 3 selected from R J , and when valence permits, the morpholinyl, azetidinyl, tetrahydropyranyl, oxetanyl, pyrrolidinyl and pyrazolyl are each independently and optionally substituted with 1 to 3 groups selected from the following groups: (C1-C4) alkyl, (C1-C4) alkoxy, benzyl and hydroxy.

53. A compound according to any one of claims 1 to 52, or a pharmaceutically acceptable salt thereof, wherein R F is selected from halo, cyano, (C1-C4) alkyl, (C1-C4) haloalkyl, (C1-C4) alkoxy, hydroxy, -N[(C 1- C4)alkyl] 2, morpholinyl, piperazinyl, azetidinyl, pyrrolidinyl and oxo, wherein the piperazinyl, the pyrrolidinyl and the azetidinyl are each optionally substituted with 1 or 2 groups selected from the group consisting of cyano, halo, (C1-C4)alkyl, (C1-C4)alkoxy and (C1-C4)alkyl(C1-C4)alkoxy.

54. A compound according to any one of claims 1 to 52, or a pharmaceutically acceptable salt thereof, wherein R F Selected from cyano, (C1-C4)alkyl, hydroxy and oxo.

55. A compound according to any one of claims 1 to 54, or a pharmaceutically acceptable salt thereof, wherein R M Selected from halo, hydroxy, (C1-C4) alkoxy, -S(O)2R f , -S(O)=NH(C1-C4)alkyl, pyridyl, pyrazolyl and phenyl optionally substituted by 1 or 2 halogen groups.

56. A compound according to any one of claims 1 to 54, or a pharmaceutically acceptable salt thereof, wherein R M Selected from halo, (C1-C4) alkoxy, -S(O)2R f and -S(O)=NH(C1-C4)alkyl.

57. A compound according to any one of claims 1 to 55, or a pharmaceutically acceptable salt thereof, wherein R f is (C1-C4)alkyl, and wherein the (C1-C4)alkyl is optionally substituted with 1 to 3 halo groups when valence permits.

58. The compound according to claim 1, wherein the compound is selected from any one of Compounds 1 to 435 or a pharmaceutically acceptable salt thereof.

59. A pharmaceutically acceptable composition comprising a compound according to any one of claims 1 to 58 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

60. A method of treating a condition responsive to modulation of STAT3 or STAT6 in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 58, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition according to claim 59.

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