6-oxodecahydropyrrolo [1, 2-a] [1, 5] diazacyclooctatetraene and 6-oxodecahydro-4h-pyrrolo [2, 1-d] [1, 5] thiazacyclooctatetraene derivatives as modulators of STAT3 and STAT6 for treatment of cancer and inflammatory conditions

Abstract

Provided are compounds of Formula (I): and pharmaceutically acceptable salts thereof, and compositions thereof, which are useful in the treatment of various conditions associated with STAT3 and / or STAT6, such as cancer and inflammatory conditions, etc. Preferred compounds are 6-oxodecahydropyrrolo [1, 2-a] [1, 5] diazacyclooctane and 6-oxodecahydro-4H-pyrrolo [2, 1-d] [1, 5] thiazacyclooctane derivatives, as well as methods for preparing such compounds. # imgabs0 #

Description

[0001] Related Applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 313,916, filed on February 25, 2022, and U.S. Provisional Application No. 63 / 337,428, filed on May 2, 2022, the entire contents of which are incorporated herein by reference. Background Art

[0003] The signal transducer and activator of transcription (STAT) protein family consists of transcription factors that play an important role 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 promoting tumor growth and immune evasion and is the only STAT family member whose genetic deletion results in embryonic lethality. In fact, it is estimated that more than 70% of human cancers have abnormally elevated STAT3 activity. Activated STAT3 can mediate key gene expression changes and molecular events that dysregulate cell growth and apoptosis, promote the development of angiogenesis, invasion, metastasis, and resistance to apoptosis, and inhibit host immune surveillance of tumors, thus making constitutively active STAT3 a key mediator of carcinogenesis and tumor progression.

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

[0006] Given their roles in the regulation of cellular processes, modulating the activity of one or more STAT proteins, specifically STAT3 and / or STAT6, is a key area of ​​research for the treatment of cancer, inflammatory conditions and other therapeutic needs. Summary of the invention

[0007] Provided herein are modulators of STAT3 and / or STAT6. Such modulators include modulators having structural formula I:

[0008]

[0009] and pharmaceutically acceptable salts thereof and compositions thereof, wherein R 1 , R 2 , R3 , R 4 , R 5 , R 6 , R 7 , x, q, t and p are as described herein.

[0010] In one aspect, the disclosed compounds of Formula I and pharmaceutically acceptable salts thereof inhibit STAT3 and / or STAT6 and are useful in a variety of therapeutic applications, such as the treatment of cancer and inflammatory conditions.

[0011] Also included are pharmaceutical compositions comprising the disclosed compounds of formula I and pharmaceutically acceptable salts of the compounds and methods for preparing the same.

[0012] Also included are methods of using the disclosed compounds, pharmaceutically acceptable salts, and compositions thereof to treat conditions responsive to modulation of STAT3 and / or STAT6. DETAILED DESCRIPTION

[0013] 1. General description of the compound

[0014] In a second embodiment, provided herein is a compound of formula I:

[0015]

[0016] q is 0 or 1, and t is 0, 1 or 2, provided that at least one of q or t is 1;

[0017] p is 1 or 2;

[0018] X is selected from S, SO 2 , -S(=O)=NH and NR 8 ;

[0019] R 1 is selected from 8- to 10-membered fused bicyclic heteroaryl, the 8- to 10-membered fused bicyclic heteroaryl being substituted with: -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T]; 8- to 10-membered fused bicyclic heterocyclic group, the 8- to 10-membered fused bicyclic heterocyclic group is 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 ]、-P(O)OR 1b OR 2b 、-

[0020] [P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; aryl, said aryl being 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ], wherein the aryl group may be further optionally substituted by 1 or 2 independently selected from cyano, (C 1 -C 4 ) substituted with alkoxy and halogenated groups; -(C 1 -C 4 )alkyl(aryl), wherein -(C 1 -C 4 )alkyl(aryl) wherein the aryl portion is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHRTy ][NH(AA)C(O)OR T ], -[P(O)[NH(AA)C(O)ORT][NH(AA)C(O)ORT] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; and -(C 2 -C 4 )alkenyl(aryl), wherein -(C 2 -C 4 )alkenyl(aryl) wherein the aryl portion is 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 ]、-P(O)OR 1b OR 2b 、-

[0021] [P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ];

[0022] R 1a and R 2a are each independently selected from hydrogen, cyano, (C 1 -C 4 ) alkyl, hydroxyl (C 1 -C 4 ) alkyl and fluorine; or R 1a and R 2a Together with the carbon to which it is attached, it forms an oxo group;

[0023] R 1b and R 2b are each independently selected from hydrogen, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 )alkyl, -[(C 1 -C 4 )alkyl]-OC(O)-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-C(O)O-[(C 1 -C 4)alkyl], -[(C 1 -C 4 )alkyl]-O-[(C 1 -C 20 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)-[halo(C 1 -C 4 )alkyl], [(C 1 -C 4 )alkyl]-OC(O)O-[5- to 7-membered heterocyclic group], [(C 1 -C 4 )alkyl]-OC(O)-[5- to 7-membered heterocyclic group], -[(C 1 -C 4 )alkyl]-OC(O)-[(C 1 -C 4 )alkyl]-OH, -[(C 1 -C 4 )alkyl]-OC(O)-[(C 1 -C 4 )alkyl]-O-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)O-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)O-[halo(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)O-[(C 1 -C 4 )alkyl]-OH, -[(C 1 -C 4 )alkyl]-OC(O)O-[(C 1 -C 4 )alkyl]-O-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-SC(O)-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-SC(O)-[halo(C 1 -C 4 )alkyl], -[(C1 -C 4 )alkyl]-SC(O)-[(C 1 -C 4 )alkyl]-OH, -[(C 1 -C 4 )alkyl]-SC(O)-[(C 1 -C 4 )alkyl]-O-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)NH(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)N[(C 1 -C 4 )alkyl] 2 and aryl, wherein the 5- to 6-membered heteroaryl and aryl are each optionally and independently substituted with 1 to 2 groups selected from halo, cyano and (C 1 -C 4 ) alkyl group, and wherein [(C 1 -C 4 )alkyl]-OC(O)O-[5- to 7-membered heterocyclic group] and [(C 1 -C 4 The 5- to 7-membered heterocyclic group in the [5- to 7-membered heterocyclic group] is optionally and independently substituted with 1 to 2 groups selected from C(O)OR h The group substitution;

[0024] R 2 Selected from hydrogen, halogen, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, hydroxyl (C 1 -C 4 ) alkyl, cyano and hydroxyl;

[0025] R 3 and R 4 Each independently selected from hydrogen, halo, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, hydroxyl (C1 -C 4 )alkyl, -(C 1 -C 4 ) alkylphenyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, -(C 1 -C 4 )alkyl(C 1 -C 4 ) alkoxy, hydroxy, cyano, -NR a R b , phenyl, (C 3 -C 6 )cycloalkyl, 5- to 6-membered heteroaryl and 4- to 6-membered heterocyclic group, wherein the phenyl, (C 3 -C 6 ) cycloalkyl, 5- to 6-membered heteroaryl and 4- to 6-membered heterocyclyl are each optionally substituted, where valence permits, with 1 to 3 selected from R S The group substitution;

[0026] R 5 and R 6 Each independently selected from hydrogen, phenyl and (C 1 -C 4 )alkyl;

[0027] R 7 Selected from (C 1 -C 4 )alkyl, phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic group and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein said (C 1 -C 4 ) alkyl is optionally substituted, when valence permits, with 1 to 3 selected from R Y The phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic group and 5- to 10-membered monocyclic or bicyclic heteroaryl group are each optionally substituted with 1 to 3 groups selected from R Z or

[0028] 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, where valence permits, by 1 to 3 selected from R Q The group substitution;

[0029] R 8 Selected from hydrogen, (C 1 -C 4 ) alkyl, halogenated (C1 -C 4 ) alkyl, (C 3 -C 6 )cycloalkyl, 5- to 7-membered heterocyclic group, -(C 1 -C 4 )[5- to 7-membered heterocyclic group], 5- to 10-membered heteroaryl, -(C 1 -C 4 )[5- to 10-membered heteroaryl], phenyl, -(C 1 -C 4 )alkylphenyl, -C(O)R Ha 、-C(O)OR Ha 、-C(O)NR Ha R Hb 、-C(O)OR Ha 、-SOR Ha R Hb and-SO 2 R Ha , wherein (C 3 -C 6 )cycloalkyl, 5- to 7-membered heterocyclyl, 5- to 10-membered heteroaryl, phenyl, (C 1 -C 4 ) the phenyl group on the alkylphenyl group, -(C 1 -C 4 )[5- to 7-membered heterocyclic group] and -(C 1 -C 4 )[5- to 6-membered heteroaryl], each of which is optionally substituted with 1 to 3 selected from R U The group substitution;

[0030] R Ha Selected from (C 1 -C 10 ) alkyl, (C 2 -C 10 ) alkenyl, (C 2 -C 10 ) alkynyl, phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, and 4- to 10-membered monocyclic or bicyclic heterocyclyl, wherein said (C 1 -C 10 ) alkyl, (C 2 -C 10 ) alkenyl, (C 2 -C 10 ) alkynyl groups are each optionally substituted, where valence permits, with 1 to 2 selected from R Owherein the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 10-membered monocyclic or bicyclic heterocyclic group are each optionally substituted by 1 to 3 groups selected from the following groups when valence permits: halo, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, oxo, cyano and optionally C 1 -C 4 an alkyl-substituted 4- to 6-membered heterocyclic group;

[0031] R O Selected from halogenated, (C 1 -C 4 ) alkoxy, OH, phenyl, NH 2 、-NH(C 1 -C 10 )alkyl, -N[(C 1 -C 10 )alkyl], (C 3-6 cycloalkyl), 4- to 10-membered monocyclic or fused bicyclic heterocyclic groups and 5- to 10-membered monocyclic or bicyclic heteroaryl groups, wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl groups and the 4- to 10-membered monocyclic or bicyclic heterocyclic groups are each optionally substituted, where valence permits, by 1 to 3 groups selected from the group consisting of halo, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, oxo and cyano;

[0032] R Hb is hydrogen or (C 1 -C 4 )alkyl;

[0033] AA is the residue of an α or β natural or unnatural amino acid;

[0034] R T and R Ty Each independently selected from (C 1 -C 4 )alkyl, benzyl and phenyl, wherein the phenyl is optionally substituted by 1 or 2 groups selected from halo, (C 1 -C 4 ) alkyl and halogen (C1 -C 4 ) alkyl radical substitution;

[0035] R Q and R U Each independently selected from halogenated, (C 2 -C 4 ) alkenyl, (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, cyano, phenyl, hydroxy, 4- to 9-membered monocyclic or bicyclic heterocyclic group, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C 3 -C 6 )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) 2 R f 、-S(O)=NH(C 1 -C 4 )alkyl, -S(O)NR e R f and -S(O) 2 NR e R f , wherein (C 2 -C 4 ) alkenyl and (C 1 -C 4 ) alkyl groups are each optionally and independently substituted with 1 to 3 groups selected from R M wherein the phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C 3 -C 6 ) cycloalkyl and 4- to 9-membered monocyclic or bicyclic heterocyclic groups are each optionally and independently substituted with 1 to 3 groups selected from R F The group substitution;

[0036] R Y Selected from halogenated, (C 1 -C 4 ) alkoxy, halogenated (C1 -C 4 )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) 2 R f 、-S(O)NR e R f 、-S(O)=NH(C 1 -C 4 )alkyl, -S(O) 2 NR e R f , hydroxy, phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl are each optionally substituted with 1 to 3 selected from R where valence permits. X The group substitution;

[0037] R M and R J Each independently selected from halogenated, (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 )alkoxy, cyano, -C(O)R g 、-C(O)OR e 、-NHC(O)R e 、-C(O)NR c R d 、-NR a R b 、-S(O)R e R f 、-S(O) 2 R f 、-S(O)NR e R f 、-S(O)=NH(C 1 -C 4 )alkyl, -S(O) 2 NR e R f, hydroxy, phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl are each optionally substituted with 1 to 3 selected from R where valence permits. X The group substitution;

[0038] R F , R S , R X and R Z Each independently selected from halo, cyano, (C 1 -C 4 ) alkyl, (C 3 -C 6 Cycloalkyl), halo (C 1 -C 4 )alkyl, -(C 1 -C 4 )alkyl(C 1 -C 4 ) alkoxy, hydroxyl (C 1 -C 4 )alkyl, -(C 1 -C 4 ) alkylphenyl, -(C 1 -C 4 ) alkylheteroaryl, (C 2 -C 4 )alkenyl, halo (C 2 -C 4 ) alkenyl, (C 2 -C 4 ) alkynyl, halo (C 2 -C 4 ) alkynyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 )alkoxy, -OR e , oxo, imino, phenyl, 4- to 6-membered heterocyclic group, -S(O)R e R f 、-S(O) 2 R f 、-S(O)=NH(C 1 -C 4 )alkyl, -S(O)NR e R f and -S(O) 2 NR e R f 、-C(O)OR e 、-NR c C(O)R e 、-(C 1 -C4 alkyl)C(O)R g 、-C(O)R g 、-(C 1 -C 4 Alkyl)C(O)NR c R d 、-C(O)NR c R d 、-NO 2 and-NR a R b , wherein (C 1 -C 4 )alkyl is optionally substituted with cyano, wherein the phenyl, the 4-membered to 6-membered heterocyclic group and -(C 1 -C 4 The phenyl group of the alkylphenyl group may be optionally and independently substituted with 1 to 3 groups selected from the group consisting of halo, cyano, oxo, (C 1 -C 10 ) alkyl, (C 2 -C 10 ) alkenyl, (C 2 -C 10 ) alkynyl, halo (C 1 -C 10 ) alkyl, (C 1 -C 10 ) alkoxy and halogen (C 1 -C 10 ) alkoxy, wherein said (C 1 -C 10 ) alkyl, (C 2 -C 10 ) alkenyl and (C 2 -C 10 ) alkynyl is each optionally substituted, where valence permits, by 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 with a 5- to 7-membered heterocyclyl optionally substituted with 1 to 2 oxo; and

[0039] R a , R b , R c , R d , R e , R f , R g and R h Where valence permits, each independently selected from hydrogen, (C 1 -C 4 ) alkyl, (C 2-C 4 ) alkynyl, -(C 1 -C 4 ) alkylphenyl, phenyl, (C 3 -C 6 )cycloalkyl, 4-membered to 6-membered heterocyclyl and 5-membered to 6-membered heteroaryl, wherein said (C 1 -C 4 ) alkyl is optionally substituted, when valence permits, with 1 to 3 selected from R J The phenyl group, (C 3 -C 6 )cycloalkyl, 4- to 6-membered heterocyclyl and 5- to 6-membered heteroaryl are each independently optionally substituted by 1 to 3 groups selected from the following groups when valence permits: halo, cyano, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, hydroxy, phenyl and benzyl.

[0040] 2. definition

[0041] When used conjunctively to describe a chemical group that may have multiple points of attachment, the hyphen (-) indicates the point of attachment of the group to the variable defining it. For example, -NR c C(O)R e This means that the point of attachment of this group occurs on the nitrogen atom.

[0042] The terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br) and iodine (iodo, -I).

[0043] Unless otherwise stated, the term "alkyl" when used alone or as part of a larger moiety such as "haloalkyl", etc., means a saturated straight or branched chain monovalent hydrocarbon radical.

[0044] The term "haloalkyl" embraces monohaloalkyl, polyhaloalkyl and perhaloalkyl wherein the halogen is independently selected from fluorine, chlorine, bromine and iodine.

[0045] "Alkoxy" means an alkyl group represented by -O-alkyl attached through an oxygen linking atom. For example, "(C 1 -C 4 The term "alkoxy" includes methoxy, ethoxy, propoxy and butoxy.

[0046] "Haloalkoxy" is a haloalkyl group attached to another moiety through an oxygen atom, for example, -OCHF 2 or -OCF 3 .

[0047] The term "oxo" means the group =0.

[0048] The term "imino" refers to the group =NH.

[0049] Unless otherwise indicated, 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 may be quaternized. The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring", "heteroaryl group" or "heteroaromatic". The heteroaryl group can be monocyclic or bicyclic. Monocyclic heteroaryl includes, for example, thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, etc. Bicyclic heteroaryl includes a group in which a monocyclic heteroaryl ring is fused to one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, benzoxazolyl, benzoxadiazolyl, indazolyl, benzimidazolyl, benzothiazolyl, benzothiophenyl, quinolyl, quinazolinyl, quinoxalinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrrolopyridinyl, thienopyridinyl, thienopyrimidinyl, indolizinyl, purinyl, cinnolinyl, naphthyridinyl, and pteridinyl. It should be understood that, when specified, optional substituents on heteroaryl can be present in any substitutable position and include, for example, the position of attachment of the heteroaryl (where valence permits).

[0050] Unless otherwise indicated, the term "heterocyclyl" means a 4- to 12-membered saturated or partially unsaturated heterocyclic ring containing 1 to 4 heteroatoms independently selected from N, O and S. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclic moiety" and "heterocyclic radical" are used interchangeably herein. The heterocyclyl ring can be attached to its side group at any heteroatom or carbon atom that forms a stable structure. The heterocyclyl group can be a monocyclic or bicyclic ring (e.g., a bridged, fused or spirocyclic bicyclic ring). Examples of monocyclic saturated or partially unsaturated heterocyclic groups include, but are not limited to, azetidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, dihydrooxadiazolyl, and dihydroisoxazolyl. Bicyclic heterocyclyls include, for example, an unsaturated heterocyclyl fused to another unsaturated heterocyclyl, cycloalkyl, aryl or heteroaryl ring, such as benzoxolyl, dihydrobenzodioxinyl, dihydroxybenzofuranyl, dihydroxybenzothiophenyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl, 6-thia-2,7-diazaspiro[3.4]octanyl, 2,6-diazaspiro[3.3]heptanyl, spiro[indoline-3,3'-pyrrolidinyl]-yl, thiochromanyl, etc. It is also understood that, when specified, optional substituents on the heterocyclyl may be present at any substitutable position and include, for example, the position of attachment of the heterocyclyl where valence permits.

[0051] The term "spirocycle" refers to two rings that share one ring atom (eg, carbon).

[0052] The term "fused" refers to two rings that share two adjacent ring atoms with each other.

[0053] The term "bridged" refers to two rings that share three adjacent ring atoms with each other.

[0054] The term "aryl" refers to an aromatic carbocyclic monocyclic or bifused ring system containing 6 to 10 carbon atoms. Examples include phenyl, indanyl, tetrahydronaphthalene, and naphthyl. In one aspect, aryl is phenyl or naphthyl.

[0055] The term "cycloalkyl", when used alone or as part of a larger molecule, refers to a saturated cyclic aliphatic monocyclic or bicyclic ring system as described herein, unless otherwise specified, having 3 to 10 carbon ring atoms. Monocyclic cycloalkyls 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 cycloalkyl or alicyclic groups can be present at any substitutable position and include, for example, the position of attachment of the cycloalkyl.

[0056] The "residue of an amino acid" is what remains after a bond is formed between a reactive group in another compound (e.g., an amino group) and a carboxylic acid in an amino acid, after a bond is formed between a reactive group in another compound (e.g., a carboxylic acid) and an amino group in an amino acid, or both. As a result of the bond formation, the carboxylic acid in the amino acid no longer has an OH group, but instead a bond is formed between the carbonyl group and the reactive group in the compound; the amino group has only one hydrogen atom, but instead a bond is formed between the reactive group in the other compound and the nitrogen of the amino group; or both. For example, an "α amino acid residue" can be structurally depicted as NH 2 CR'RC(O)-, -NHCR'RC(O)OH or -NHCR'RC(O)-; and the "β-amino acid residue" can be structurally described as NH 2 CR'RCH 2 -C(O)-、-NHCR'RCH 2 -C(O)OH or -NHCR'RCH 2 -C(O)-, where R' is H or C 1 -C 6 Alkyl, and R is H or C 1 -C 6 Alkyl, the C 1 -C 6 The alkyl group is optionally substituted with 1 to 3 groups selected from the group consisting of halo, (C 1 -C 3 ) alkoxy, OH, NH 2 、-NH(C 1 -C 4 alkyl), -N[(C 1 -C 4 alkyl)] 2 SH, S(C 1 -C 4 alkyl), imino, COOH, -COO(C 1 -C 4 Alkyl), -CO(C 1 -C 4 alkyl), -CONH(C1 -C 4 alkyl)phenyl, phenyl and 5- to 10-membered heteroaryl, wherein the C 1 -C 6 The alkyl group may also be optionally interrupted by sulfur or nitrogen heteroatoms, and wherein the phenyl group may be optionally substituted by 1 to 3 groups selected from the group consisting of OH, cyano, (C 1 -C 4 Alkyl) and halo (C 1 -C 4 alkyl); or R together with the nitrogen atom from an α or β amino acid residue forms a 4- to 6-membered heterocyclic group. For naturally occurring α amino acids (i.e., amino acids occurring in nature), R' is H, and R is selected from hydrogen, methyl, isopropyl, -CH 2 CH(CH 3 ) 2 、-(CH 2 ) 2 SCH 3 、-CH(CH 3 )(CH 2 CH 3 ), CH 2 OH, -CH(OH)(CH 3 ), CH 2 SH, -CH 2 C(O)NH 2 、-(CH 2 ) 2 C(O)NH 2 , benzyl, p-hydroxybenzyl, -CH 2 (indolyl), -(CH 2 ) 4 NH 2 、-(CH 2 ) 3 NHC (=NH 2 )NH 2 、-CH 2 (imidazolyl), -(CH 2 )COOH and -(CH 2 ) 2 COOH; or R together with the nitrogen atom of the α or β amino acid residue forms a pyrrolidinyl ring.

[0057] Unnatural amino acids known in the art include α-alkyl amino acids (e.g., α-methyl), α-alkylalkoxy amino acids (e.g., α-CH 2 OCH 3 ), N-methyl amino acids, homologous amino acids, etc.

[0058] Compounds with one or more chiral centers can exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in spatial arrangement. Stereoisomers include all diastereomeric forms, enantiomeric forms and epimeric forms, as well as racemates and mixtures thereof. "Geometric isomers" refer to isomers with different orientations of substituents and carbon-carbon double bonds, cycloalkyl rings or bridged bicyclic systems. The atoms on each side of the carbon-carbon double bond (except H) can be in E (substituents on the opposite side of the carbon-carbon double bond) or Z (substituents toward the same side) configuration. "Cis" refers to substituents toward the same side of the ring, while "trans" refers to substituents toward the opposite side of the ring.

[0059] When the stereochemical configuration at a chiral center in a compound having one or more chiral centers is depicted by its chemical name (e.g., where the configuration is represented by "R" or "S" in the chemical name) or structure (e.g., the configuration is represented by a "wedge-shaped" bond), the enrichment of the indicated configuration relative to the opposite configuration is greater than 50%, 60%, 70%, 80%, 90%, 99%, or 99.9%. The "enrichment of a specified configuration relative to the opposite configuration" is a molar percentage and is determined by dividing the number of compounds having a specified stereochemical configuration at a chiral center by the total number of all compounds in the mixture having the same or opposite stereochemical configuration.

[0060] When a geometric isomer is depicted 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%. The "enrichment of a specified isomer relative to the opposite isomer" is a molar percentage and is determined by dividing the number of compounds having the specified geometric configuration by the total number of all compounds in the mixture having the same or opposite geometric configuration.

[0061] When a disclosed compound is named or depicted by structure without indicating stereochemistry, it is understood that the name or structure encompasses one possible stereoisomer or geometric isomer without other stereoisomers, or mixtures of stereoisomers or geometric isomers encompassed.

[0062] In some cases, compounds were isolated and tested as a 1:1 mixture of diastereomers. In such cases, the relative stereochemistry is indicated by "rel-" in the compound name, with a flat bond replacing a wedge-shaped bond. For example, ((2-(((5S,8S,10aR)-8-((rel-trans)-3-cyano-4-phenylpyrrolidine-1-carboxyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid, has the following structure: means trans with respect to the substituents on the pyrrolidine ring and encompasses a mixture of both diastereoisomers

[0063]

[0064] The terms "subject" and "patient" are used interchangeably and refer to mammals in need of treatment, e.g., companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, the subject is a human in need of treatment.

[0065] The terms "inhibit," "inhibition," or "inhibiting" encompass reducing the baseline activity of a biological activity or process.

[0066] As used herein, the term "treatment (treatment, treat and treating)" refers to reversing, alleviating, delaying the onset of a disease or disorder as described herein or one or more of its symptoms or inhibiting its progress. In some aspects, treatment can be administered after one or more symptoms occur, i.e., therapeutic treatment. In other aspects, treatment can be administered in the absence of symptoms. For example, treatment can be administered to susceptible individuals before the onset of symptoms (e.g., according to symptom history and / or according to exposure to a specific organism or other susceptible factors), i.e., preventive treatment. Treatment can also be continued after the symptoms have subsided, for example, to delay its recurrence.

[0067] The term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated with it. Pharmaceutically acceptable carriers, adjuvants or vehicles 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), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin).

[0068] For use in medicine, the salts of the compounds described herein refer to non-toxic "pharmaceutically acceptable salts". Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid and sulfuric acid) and salts of organic acids (such as acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid and p-toluenesulfonic acid). The compounds taught by the present 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 with quaternary ammonium groups also contain counter anions such as chloride, bromide, iodide, acetate, perchlorate, etc. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates, and salts containing amino acids such as glutamic acid.

[0069] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound described herein sufficient to achieve the desired therapeutic effect (eg, treating a condition described herein) under the conditions of administration, such as a dosage between 0.01-100 mg / kg body weight / day.

[0070] 3. Compound

[0071] In a first embodiment, a compound of formula I is provided:

[0072]

[0073] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above.

[0074] In a second embodiment, R in the compound of Formula I or a pharmaceutically acceptable salt thereof 1 is selected from 8- to 10-membered fused bicyclic heteroaryl, the 8- to 10-membered fused bicyclic heteroaryl being substituted with: -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-

[0075] [P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b][NH(AA)C(O)OR T ]; 8- to 10-membered fused bicyclic heterocyclic group, the 8- to 10-membered fused bicyclic heterocyclic group is 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 ]、-P(O)OR 1b OR 2b 、-

[0076] [P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; aryl, said aryl being 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ];-(C 1 -C 4 )alkyl(aryl), wherein -(C 1 -C 4 )alkyl(aryl) wherein the aryl portion is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR1b ][NH(AA)C(O)OR T ]; and -(C 2 -C 4 )alkenyl(aryl), wherein -(C 2 -C 4 )alkenyl(aryl) wherein the aryl portion is 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 ]、-P(O)OR 1b OR 2b 、-

[0077] [P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ];

[0078] R 8 Selected from hydrogen, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 3 -C 6 )cycloalkyl, 5- to 7-membered heterocyclic group, -(C 1 -C 4 )[5- to 7-membered heterocyclic group], 5- to 6-membered heteroaryl, -(C 1 -C 4 )[5- to 6-membered heteroaryl], phenyl, -(C 1 -C 4 )alkylphenyl, -C(O)R Ha 、-C(O)OR Ha 、-C(O)NR Ha R Hb 、-C(O)OR Ha 、-SOR Ha R Hb and-SO 2 R Ha , wherein (C 3 -C 6 )cycloalkyl, 5- to 7-membered heterocyclyl, 5- to 6-membered heteroaryl, phenyl, (C 1 -C 4 ) the phenyl group on the alkylphenyl group, -(C1 -C 4 )[5- to 7-membered heterocyclic group] and -(C 1 -C 4 )[5- to 6-membered heteroaryl], each of which is optionally substituted by 1 to 3 selected from R U The group substitution;

[0079] R Ha Selected from (C 1 -C 10 ) alkyl, (C 2 -C 10 ) alkenyl, (C 2 -C 10 ) alkynyl, phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, and 4- to 10-membered monocyclic or bicyclic heterocyclyl, wherein said (C 1 -C 10 ) alkyl, (C 2 -C 10 ) alkenyl, (C 2 -C 10 ) alkynyl groups are each optionally substituted, where valence permits, with 1 to 2 selected from R O wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 10-membered monocyclic or bicyclic heterocyclic group are each optionally substituted by 1 to 3 groups selected from the following groups when valence permits: halo, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 ) alkoxy, oxo and cyano; and

[0080] R O Selected from halogenated, (C 1 -C 4 ) alkoxy, OH, phenyl, NH 2 、-NH(C 1 -C 10 )alkyl, -N[(C 1 -C 10 )alkyl], a 4- to 10-membered monocyclic or fused bicyclic heterocyclic group, and a 5- to 10-membered monocyclic or bicyclic heteroaryl group, wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl group and the 4- to 10-membered monocyclic or bicyclic heterocyclic group are each optionally substituted, as valence permits, by 1 to 3 groups selected from the group consisting of halo, (C 1 -C 4) alkyl, halogenated (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 )alkoxy, oxo and cyano, with the remainder of the variables being as described above for Formula I.

[0081] In a third embodiment, the compound of formula I has structural formula II:

[0082]

[0083] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I or the Second Embodiment.

[0084] In a fourth embodiment, the compound of Formula I or II, or a pharmaceutically acceptable salt thereof, wherein p is 1, wherein the remaining variables are as described above for Formula I or the second embodiment.

[0085] In a fifth embodiment, R in the compound of Formula I or II or a pharmaceutically acceptable salt thereof 2 is hydrogen, wherein the remainder of the variables are as described above for Formula I or any of the second or fourth embodiments.

[0086] In a sixth embodiment, R in the compound of Formula I or II or a pharmaceutically acceptable salt thereof 3 is hydrogen, wherein the remainder of the variables are as described above for Formula I or any of the second, fourth or fifth embodiments.

[0087] In a seventh embodiment, R in the compound of Formula I or II or a pharmaceutically acceptable salt thereof 4 is hydrogen, wherein the remainder of the variables are as described above for Formula I or any one of the second and fourth through sixth embodiments.

[0088] In an eighth embodiment, q in the compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is 0, and t is 2, wherein the remaining variables are as described above for Formula I or any one of the second and fourth to seventh embodiments. Alternatively, as part of the eighth embodiment, q in the compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is 1, and t is 1, wherein the remaining variables are as described above for Formula I or any one of the second and fourth to seventh embodiments. In another alternative, as part of the eighth embodiment, q in the compound of Formula I or II, or a pharmaceutically acceptable salt thereof, is 0, and t is 1, wherein the remaining variables are as described above for Formula I or any one of the second and fourth to seventh embodiments.

[0089] In a ninth embodiment, the compound of formula I has structural formula III, IV, V, VI, VII or VIII:

[0090]

[0091] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I or the Second Embodiment.

[0092] In the tenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof 5 is hydrogen, wherein the remainder of the variables are as described above for Formula I or any one of the second and fourth through eighth embodiments.

[0093] In an eleventh embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof 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 substituted with, wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth and tenth embodiments. Alternatively, as part of the eleventh embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII, or a pharmaceutically acceptable salt thereof, is 1 is selected from benzothienyl, indolyl, naphthyl, thienopyridinyl, benzothiazolyl, quinolyl, isoquinolyl, 4,5,6,7-tetrahydropyrrolopyridinyl, 5,6,7,8-tetrahydroimidazopyrazinyl, each of which is replaced by -CR 1a R 2a P(O)OR 1b OR 2b substituted with the remaining variables as described above for Formula I or any one of the second, fourth to eighth and tenth embodiments. In another alternative, as part of the eleventh embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII, or a pharmaceutically acceptable salt thereof, is 1 Selected from

[0094]

[0095] wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth and tenth embodiments. In another alternative, as part of the eleventh embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof is 1 yes wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth embodiments.

[0096] In a twelfth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof 1a is hydrogen and R 2a is fluorine, or R 1a is fluorine and R 2a is fluoro, wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, tenth and eleventh embodiments. Alternatively, as part of the twelfth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII, or a pharmaceutically acceptable salt thereof, is 1a is fluorine and R 2a is fluoro, wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, tenth and eleventh embodiments.

[0097] In the thirteenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof 1b and R 2b are each independently selected from hydrogen, (C 1 -C 4 )alkyl, -[(C 1 -C 4 )alkyl]-OC(O)-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-OC(O)O-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-SC(O)-[(C 1 -C 4 )alkyl], -[(C 1 -C 4 )alkyl]-SC(O)-[halo(C 1 -C 4 )alkyl], -[(C 1 -C 4)alkyl]-OC(O)NR 2c R 2d ]、[(C 1 -C 4 )alkyl]-OC(O)O-[5- to 7-membered heterocyclic group] and phenyl, wherein [(C 1 -C 4 )alkyl]-OC(O)O-[5- to 7-membered heterocyclic group], the 5- to 7-membered heterocyclic group is optionally substituted by C(O)OR 2c Substituted, wherein said (C 1 -C) 4 Any of the alkyl groups is optionally replaced by 1 or 2 (C 1 -C 4 ) alkyl, wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twelfth embodiments. Alternatively, as part of the thirteenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII, or a pharmaceutically acceptable salt thereof, is 1b and R 2b Each is -[(C 1 -C 4 )alkyl]-OC(O)-[(C 1 -C 4 )alkyl], wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twelfth embodiments. In another alternative, as part of the thirteenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII, or a pharmaceutically acceptable salt thereof, is 1b and R 2b and R is hydrogen, with the remainder of the variables being as described above for Formula I or any one of the second, fourth through eighth, and tenth through twelfth embodiments.

[0098] In the fourteenth embodiment, -CR in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof 1a R 2a P(O)OR 1b OR 2b Selected from

[0099]

[0100] wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to thirteenth embodiments. Alternatively, as part of the fourteenth embodiment, -CR in the compound of Formula I, II, III, IV, V, VI, VII, or VIII, or a pharmaceutically acceptable salt thereof, is 1a R 2a P(O)OR 1b OR 2b Selected from wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to thirteenth embodiments.

[0101] In the fifteenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof 7 Selected from (C 1 -C 4 )alkyl, phenyl, 4- to 6-membered monocyclic heterocyclic group, 9- or 10-membered fused bicyclic heterocyclic group, 5- or 6-membered monocyclic heteroaryl and 9- or 10-membered fused bicyclic heteroaryl, wherein said (C 1 -C 4 ) alkyl is optionally substituted, when valence permits, with 1 to 3 selected from R Y The phenyl, 5-membered or 6-membered monocyclic heterocyclic group, 9-membered or 10-membered fused bicyclic heterocyclic group, 5-membered or 6-membered monocyclic heteroaryl group and 9-membered or 10-membered fused bicyclic heteroaryl group are each optionally substituted with 1 to 3 groups selected from R Z wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to fourteenth embodiments. Alternatively, as part of the fifteenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII, or VIII, or a pharmaceutically acceptable salt thereof, is 7 Selected from (C 1 -C 4 )alkyl, phenyl, pyridyl, pyrimidinyl, naphthyl, cinnolinyl, quinoxalinyl, quinazolinyl, quinolyl, isoquinolyl, chromanyl, pyrazolyl, indazolyl, benzisoxazolyl, imidazo[1,2-a]pyridinyl, pyrrolidinyl, cyclopentyl, cyclohexyl, azetidinyl, piperidinyl and dihydropyridinyl, and wherein said (C 1 -C 4 ) alkyl is optionally substituted, when valence permits, with 1 to 3 selected from R Ysubstituted with a group, and the phenyl, pyridyl, pyrimidinyl, naphthyl, cinnolinyl, quinoxalinyl, quinazolinyl, quinolyl, isoquinolyl, chromanyl, pyrazolyl, indazolyl, benzisoxazolyl, imidazo[1,2-a]pyridinyl, pyrrolidinyl, cyclopentyl, cyclohexyl, azetidinyl, piperidinyl and dihydropyridinyl are each optionally substituted with 1 to 3 selected from R where valence permits Z wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to fourteenth embodiments.

[0102] In the sixteenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof Y Selected from cyano, (C 1 -C 4 )alkyl, -NR a R b and hydroxy, wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth through eighth, and tenth through fifteenth embodiments.

[0103] In the seventeenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof Z Selected from halo, cyano, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, hydroxyl (C 1 -C 4 )alkyl, -(C 1 -C 4 ) alkylphenyl, -(C 1 -C 4 ) alkylheteroaryl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 )alkoxy, -OR e , oxo, phenyl, 4- to 6-membered heterocyclic group, -S(O) 2 R f 、-(C 1 -C 4 alkyl)C(O)R g 、-C(O)R g 、-(C 1 -C 4 Alkyl)C(O)NR c R d 、-C(O)NR c R d 、-NO2 and-NR a R b , wherein (C 1 -C 4 )alkyl is optionally substituted with cyano, and wherein the phenyl and the 4- to 6-membered heterocyclyl are each optionally and independently substituted with 1 to 3 groups selected from cyano and oxo, where valence permits, and wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to sixteenth embodiments. Alternatively, as part of the seventeenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII, or VIII, or a pharmaceutically acceptable salt thereof, Z Select halo, cyano, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 ) alkyl, hydroxyl (C 1 -C 4 )alkyl, -(C 1 -C 4 ) alkylphenyl, -(C 1 -C 4 ) alkylheteroaryl, (C 1 -C 4 ) alkoxy, halogenated (C 1 -C 4 )alkoxy, -OR e , oxo, -S(O) 2 R f 、-(C 1 -C 4 alkyl)C(O)R g 、-C(O)R g 、-(C 1 -C 4 Alkyl)C(O)NR c R d 、-C(O)NR c R d 、-NO 2 、-NR a R b , phenyl and dihydropyridyl, wherein said (C 1 -C 4 )alkyl is optionally substituted with cyano, and wherein the phenyl and the dihydropyridinyl are each optionally and independently substituted, as valence permits, with 1 to 3 groups selected from cyano and oxo, and wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to sixteenth embodiments.

[0104] In the eighteenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII 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 6-membered monocyclic heterocyclic group, a 7- to 13-membered spiro bicyclic heterocyclic group, or a 9- to 10-membered fused bicyclic heterocyclic group, each of which is optionally substituted, where valence permits, by 1 to 3 selected from R Q wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to seventeenth embodiments. Alternatively, as part of the eighteenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII, or VIII, or a pharmaceutically acceptable salt thereof, is 6 and R 7 Together with the nitrogen atom to which it is attached, it forms an azetidinyl, spiro[indoline-3,3'-pyrrolidinyl], pyrrolidinyl, 3,4-dihydrobenzoxazinyl, 1,2,3,4-tetrahydroquinolinyl, octahydrothiopyranopyrrolyl, octahydro-1H-thieno[3',4':3,4]cyclobutane[1,2-c]pyrrolyl, 2,6-diazabicyclo[3.2.0]heptanyl, piperazinyl, morpholinyl, 1,4,5,6-tetrahydropyrrolopyrrolyl, oxazolyl, 4-azaspiro[2.4]heptyl, 2-oxa-5-azabicyclo[2.2.1]heptyl, 7-oxa-4-azaspiro[2.5]octanyl, 8-oxa-3-azabicyclo[3.2.1]octanyl, 4,6-diazaspiro[2.4]heptyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl or piperidinyl, each of which is optionally substituted, where valence permits, by 1 to 3 selected from R Q wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to seventeenth embodiments. In another alternative, as part of the eighteenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof is 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a pyrrolidinyl and an azetidinyl group, each of which is optionally substituted, where valence permits, with 1 to 3 selected from R Q wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to seventeenth embodiments.

[0105] In the nineteenth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof Q Selected from halogenated, (C 1-C 4 ) alkyl, halogenated (C 1 -C 4 )alkyl, cyano, phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic group, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C 3 -C 6 )cycloalkyl, oxo, -OR e 、-C(O)R g 、-C(O)NR c R d 、-C(O)OR e 、-NR c C(O)R e , wherein (C 1 -C 4 ) alkyl groups are each optionally and independently substituted with 1 to 3 groups selected from R M wherein the phenyl, 4- to 6-membered heterocyclic group, 5- to 10-membered monocyclic or bicyclic heteroaryl and (C 3 -C 6 ) cycloalkyl is optionally and independently substituted with 1 to 3 selected from R F wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to eighteenth embodiments. Alternatively, as part of the nineteenth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII, or VIII, or a pharmaceutically acceptable salt thereof, is Q Selected from cyano, (C 1 -C 4 ) alkyl, halogenated (C 1 -C 4 )alkyl, oxo, -C(O)R g 、-OR e 、-C(O)OR e 、-C(O)NR c R d 、-NR c C(O)R e , cyclobutyl, cyclopropyl, cyclohexyl, phenyl, pyridyl, pyrimidinyl, dihydropyridyl, pyrazolyl, piperidinyl, tetrahydroimidazopyridinyl and isothiazolidinyl, wherein said (C 1 -C 4 ) alkyl is optionally and independently substituted with 1 to 3 selected from R M wherein the cyclobutyl, cyclopropyl, cyclohexyl, phenyl, pyridyl, pyrimidinyl, dihydropyridyl, pyrazolyl, piperidinyl, tetrahydroimidazopyridinyl and isothiazolidinyl are each optionally and independently substituted with 1 to 3 selected from R where valence permitsF wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to eighteenth embodiments.

[0106] In the twentieth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof M phenyl and 5- to 6-membered monocyclic heteroaryl, wherein the phenyl and 5- to 6-membered monocyclic heteroaryl and the phenyl are each optionally substituted by 1 to 3 selected from R X wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to nineteenth embodiments. Alternatively, as part of the twentieth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII, or VIII, or a pharmaceutically acceptable salt thereof, is M is selected from phenyl and pyridyl, wherein the phenyl and pyridyl are each optionally substituted, where valence permits, with 1 to 3 selected from R X wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to nineteenth embodiments.

[0107] In the twenty-first embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof X Selected from halogenated and (C 1 -C 4 )alkoxy, wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth through eighth, and tenth through twentieth embodiments.

[0108] In the twenty-second embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof F Selected from halo, cyano, (C 1 -C 4 ) alkoxy, oxo and -NR c C(O)R e , wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twenty-first embodiments.

[0109] In the twenty-third embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof c and R d where valence permits, are each independently selected from hydrogen and (C 1 -C 4) alkyl, and wherein said (C 1 -C 4 ) alkyl is optionally substituted, when valence permits, with 1 to 3 selected from R J wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twenty-second embodiments.

[0110] In the twenty-fourth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof e are independently selected from hydrogen, phenyl, 5- to 6-membered heteroaryl and (C 1 -C 4 ) alkyl, and wherein said (C 1 -C 4 ) alkyl is optionally substituted, when valence permits, with 1 to 3 selected from R J wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twenty-third embodiments.

[0111] In the twenty-fifth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof J is independently selected from phenyl and 5- to 6-membered heteroaryl, and wherein the 5- to 6-membered heteroaryl is optionally replaced by (C 3 -C 6 ) cycloalkyl, wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twenty-fourth embodiments. Alternatively, as part of the twenty-fifth embodiment, R in the compound of Formula I, II, III, IV, V, VI, VII or VIII, or a pharmaceutically acceptable salt thereof, is J is phenyl, wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth through eighth, and tenth through twenty-fourth embodiments.

[0112] In the twenty-sixth embodiment, R in the compound of formula I, II, III, IV, V, VI, VII or VIII or a pharmaceutically acceptable salt thereof g is independently selected from piperidinyl, morpholinyl, and piperazinyl, wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twenty-fifth embodiments.

[0113] In the twenty-seventh embodiment, R in the compound of formula I, II, III, VI or VIII or a pharmaceutically acceptable salt thereof 8 is hydrogen, -C(O)R Ha 、-C(O)ORHa , (C 1 -C 4 )alkyl, -SO 2 R Ha 、-C(O)NR Ha R Hb , halogenated (C 1 -C 4 ) alkyl, phenyl, -(C 1 -C 4 )[4- to 10-membered monocyclic or bicyclic heterocyclic group], -(C 1 -C 4 )[5- to 10-membered monocyclic or bicyclic heteroaryl], -(C 3 -C 6 )cycloalkyl or 4- to 10-membered monocyclic or bicyclic heterocyclic group, wherein the -(C 1 -C 4 The phenyl group on the alkylphenyl group is optionally substituted by 1 or 2 groups independently selected from -(C 1-3 alkoxy) and optionally -(C 1 -C 4 ) alkyl-substituted 5- to 6-membered heteroaryl group, and wherein said (C 1 -C 4 ) alkyl is optionally substituted with 1 to 3 phenyl groups; R Ha Yes (C 1 -C 4 )alkyl, -(C 1 -C 4 )[5- to 6-membered heteroaryl], -(C 1 -C 4 )[phenyl], -(C 1 -C 4 )[(C 3 -C 6 )cycloalkyl], phenyl, 4- to 10-membered monocyclic or bicyclic heterocyclic group, wherein the phenyl and 4- to 10-membered monocyclic or bicyclic heterocyclic group are each optionally substituted by 1 to 3 groups independently selected from the following: halo, (C 1 -C 4 ) alkyl, (C 1 -C 4 ) alkoxy and optionally C 1 -C 4 alkyl-substituted 4- to 6-membered monocyclic heterocyclic group; and R Hb is H, wherein the remaining variables are as described above for Formula I or any one of the second, fourth to eighth, and tenth to twenty-sixth embodiments. Alternatively, as part of the twenty-seventh embodiment, R in the compound of Formula I, II, III, VI or VIII, or a pharmaceutically acceptable salt thereof, is 8Yes -H, -CH 2 CH 3 、-CH 2 CH(CH 3 ) 2 、-CH 2 CF 2 、-CH 2 CF 3 , cyclobutyl, -CH 2 -quinuclidine, -CH 2 -imidazo[1,5-a]pyridinyl, -CH 2 -pyrimidinyl, -CH 2 -phenyl, phenyl, pyrimidinyl, pyridinyl, isoquinolyl, -C(O)R Ha 、-C(O)OR Ha 、-SO 2 R Ha or -C(O)NR Ha R Hb , or R 8 Represented by the following structure: And R Ha It is methyl, ethyl, propyl, isopropyl, -CH 2 -Quinolinyl, -CH 2 -phenyl, -CH 2 CH 2 -phenyl, -CH(CH 3 )-phenyl, -CH(CH 3 )CH 2 -phenyl, -CH 2 CH(CH 3 )-phenyl, -CH 2 CH 2 -pyrimidinyl, -CH 2 C(CH 3 ) 2 -cyclohexyl, quinolyl, indazolyl, benzisoxazolyl, imidazo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, 5,6,7,8-tetrahydroimidazo[1,5-a]pyridinyl, 4,5,6,7-tetrahydrobenzo[d]isoxazolyl or 6,7-dihydro-4H-pyrano[3,4-d]isoxazolyl, and wherein the quinolyl, indazolyl, benzisoxazolyl isoxazolyl, imidazo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, 5,6,7,8-tetrahydroimidazo[1,5-a]pyridinyl, 4,5,6,7-tetrahydrobenzo[d]isoxazolyl, 6,7-dihydro-4H-pyrano[3,4-d]isoxazolyl are each optionally substituted with 1 to 3 groups independently selected from: -Cl, -CH 3 、-CH 2 CH3 、-OCH 3 or N-methylpiperazinyl, wherein the remainder of the variables are as described above for Formula I or any one of the second, fourth through eighth, and tenth through twenty-sixth embodiments.

[0114] Compounds having Formula I are further disclosed in the Examples and are included in the present disclosure, including pharmaceutically acceptable salts and neutral forms thereof.

[0115] 4. Use, preparation and application

[0116] The compounds and compositions described herein are generally useful for regulating the activity of STAT proteins, specifically STAT3 and / or STAT6. In some aspects, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein inhibit STAT3 and / or STAT6 activity.

[0117] In some aspects, the compounds and pharmaceutical compositions described herein can be used for conditions that respond to the regulation of STAT3 and / or STAT6. Therefore, provided herein are methods for treating conditions that respond to the regulation (e.g., inhibition) of STAT3 and / or STAT6 in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or a pharmaceutically acceptable salt thereof.

[0118] Also provided are the uses of compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for treating a condition responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6. Also provided are compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof, for treating a condition responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6.

[0119] In one aspect, conditions responsive to modulation (e.g., inhibition) of STAT3 and / or STAT6 include, but are not limited to, cancer, neurodegenerative disorders, viral diseases, autoimmune diseases, inflammatory disorders, genetic disorders, hormone-related diseases, metabolic disorders, conditions associated with organ transplantation, immunodeficiency disorders, destructive bone disorders, proliferative disorders, infectious diseases, conditions associated with cell death, thrombin-induced platelet aggregation, liver disease, pathological immune conditions involving T cell activation, cardiovascular disorders, or CNS disorders.

[0120] On the other hand, conditions that respond to modulation (e.g., inhibition) of STAT3 and / or STAT6 include, but are not limited to, cancer (see, e.g., Turkson and Jove, Oncogene 2000, 19:6613-6626); diabetes (see, e.g., Gurzov et al., FEBS 2016, 283:3002); cardiovascular disease (see, e.g., Grote et al., Vasc. Pharmacol. 2005, 43:2005); viral disease (see, e.g., Gao et al., Journal of Hepatology (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 (see, for example, Walker & Smith, (J. Rheumat.) 2005, 32(9):1650); autoinflammatory syndromes (see, for example, Rauch et al., Jak-Stat 2013, 2(l):e23820); atherosclerosis (see, e.g., Ortiz-Munoz et al., Arterio., Thrombo., Vase. Bio. 2009, 29:525); psoriasis (see, e.g., Andres et al., Exp. Derm. 2013, 22(5):323); allergic disorders (see, e.g., Oh et al., Eur. Respir. Rev. 2019, 19(115):46); inflammatory bowel disease (see, e.g., Sugimoto, World J Gastroenterol. Gastroenterol. 2008, 14(33):5110); inflammation (see, e.g., Tamiya et al., Arteriosclerosis Thrombosis and Vascular Biology 2011, 31:980); acute and chronic gout and gouty arthritis; neurological disorders (see, e.g., Campbell, Brain Res. Rev. 2005, 48(2):166); metabolic syndrome; immunodeficiency disorders (such as AIDS and HIV) (see, e.g., O'Shea et al., N. Engl. J. Med. 2013, 368:161); destructive bone disorders (see, e.g., Jatiani et al., Genes & Can.)》2011,1(10):979); osteoarthritis; proliferative disorders; Waldenstrom's Macroglobulinemia (see, e.g., Hodge et al., Blood 2014,123(7):1055); infectious diseases; conditions associated with cell death; pathological immune conditions and CNS disorders involving T cell activation. .

[0121] Proliferative disorders include, but are not limited to, benign or malignant tumors, solid tumors, liquid tumors, carcinomas, sarcomas, glioblastomas, neuroblastomas, multiple myeloma, gastrointestinal cancer, especially colon cancer or colorectal adenomas, neck and head tumors, epidermal hyperproliferation, psoriasis, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate cancer, prostate hyperplasia, neoplasia, epithelial neoplasia, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, lymphoma, Hodgkin and non-Hodgkin carcinoma, breast cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, IL-1 driven disorder, MyD88 driven disorder, smoldering indolent multiple myeloma, or hematologic malignancies (including leukemia, diffuse large B-cell lymphoma (DLBCL), ABC DLBCL, chronic lymphocytic leukemia (CLL), chronic lymphocytic lymphoma, primary effusion lymphoma, Burkitt lymphoma / leukemia, acute lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenstrom's macroglobulinemia (WM), splenic marginal zone lymphoma, multiple myeloma, plasmacytoma, intravascular large B-cell lymphoma).

[0122] In some embodiments, the cancer to be treated is selected from glioma, breast cancer, prostate cancer, squamous cell carcinoma of the head and neck, melanoma of the skin, ovarian cancer, malignant peripheral nerve sheath tumor (MPNST), and pancreatic cancer. In other embodiments, the cancer to be treated is a cancer selected from the following: glioma, breast cancer, prostate cancer, squamous cell carcinoma of the head and neck, melanoma of the skin, ovarian cancer, malignant peripheral nerve sheath tumor (MPNST), pancreatic cancer, non-small cell lung cancer (NSCLC) (including EGFR mutant NSCLC), urothelial carcinoma, lung cancer, bile duct cancer, kidney cancer, colon cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors and hematological 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.), hematological cancers (e.g., lymphoma, leukemia, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) or multiple myeloma), and skin cancers, such as cutaneous T-cell lymphoma (CTCL) and cutaneous B-cell lymphoma. Example CTCLs include Sezary syndrome and mycosis fungoides.

[0123] The compounds, salts and compositions described herein can also be used to treat inflammatory or obstructive airway diseases, such as reducing tissue damage, airway inflammation, bronchial hyperresponsiveness, remodeling or disease progression. Inflammatory or obstructive airway diseases include asthma of any type or origin, including intrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mild asthma, moderate asthma, severe asthma, bronchial asthma, exercise-induced asthma, occupational asthma and asthma induced after bacterial infection. The treatment of asthma should also be understood to cover the treatment of subjects who are, for example, less than 4 or 5 years old, show symptoms of wheezing, and are diagnosed or can be diagnosed as "wheezing babies", which is a recognized major medical problem patient category and is now generally identified as an initial or early asthma patient.

[0124] The compounds, salts and compositions described herein may also be used to treat heteroimmune diseases, including but not limited to graft-versus-host disease, transplantation, transfusion, anaphylaxis, allergies (e.g., to plant pollens, latex, drugs, foods, pesticides, animal hair, animal dander, dust mites, or cockroach calyx), type I hypersensitivity reactions, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

[0125] The compounds, salts and compositions described herein can also be used to treat other inflammatory or obstructive airway diseases and conditions to which the present invention is applicable, and include acute lung injury (ALI), adult / acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease, airway or lung disease (COPD, COAD or COLD), including chronic bronchitis or associated dyspnea, emphysema, and other drug therapies, specifically other inhaled drug therapies causing increased airway hyperresponsiveness. The compounds, salts and compositions described herein can also be used to treat bronchitis, including but not limited to acute, arachidonic, catarrhal, croup, chronic or tuberculous bronchitis. The compounds, salts and compositions described herein can also be used to treat bronchitis, including but not limited to pneumoconiosis of any type or origin (inflammatory (usually occupational) lung disease, often accompanied by airway obstruction, whether chronic or acute, and caused by repeated inhalation of dust), including, for example, aluminum pneumoconiosis, coal lung disease, asbestosis, stone end lung, eyelash loss, siderosis, silicosis, fume and cotton lung.

[0126] The compounds, salts and compositions described herein can also be used to treat inflammatory or allergic conditions of the skin, such as psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, scleroderma, vitiligo, hypersensitivity vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, systemic lupus erythematosus, pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus, epidermolysis bullosa acquisita, acne vulgaris and other inflammatory or allergic conditions of the skin.

[0127] The compounds, salts and compositions described herein can also be used to treat other diseases or conditions, such as diseases or conditions with an inflammatory component, for example, to treat eye diseases and conditions, such as ocular allergies, conjunctivitis, keratoconjunctivitis sicca and vernal conjunctivitis, diseases affecting the nose, including allergic rhinitis, and inflammatory diseases involving an autoimmune reaction or having an autoimmune component or etiology, including autoimmune blood diseases (e.g., hemolytic anemia, aplastic anemia, pure red cell anemia and idiopathic thrombocytopenia), systemic lupus erythematosus, rheumatoid arthritis, polychondritis, scleroderma, Wegener granulamatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease), disease), irritable bowel syndrome, celiac disease, periodontitis, hyaline membrane disease, kidney disease, glomerular disease, alcoholic liver disease, multiple sclerosis, endocrine eye disease, Graves' disease, sarcoidosis, alveolitis, chronic hypersensitivity pneumonitis, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), Sjögren's syndrome, keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitial pulmonary fibrosis, psoriatic arthritis, systemic juvenile idiopathic arthritis, cryptopyrin-associated periodic syndromes, nephritis, vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis (with and without nephrotic syndrome,Examples include idiopathic nephrotic syndrome or minimal change disease), chronic granulomatous disease, endometriosis, leptospirosis, glaucoma, retinal disease, aging, headache, pain, complex regional pain syndrome, cardiac hypertrophy, muscle atrophy, catabolic disorders, obesity, fetal growth retardation, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, anhidrotic ectodermal dysplasia, Behcet's disease, incontinence pigmenti, Paget's disease disease), pancreatitis, hereditary periodic fever syndromes, asthma (allergic and nonallergic, mild, moderate, severe, bronchial and exercise-induced), acute lung injury, acute respiratory distress syndrome, eosinophilia, hypersensitivity, anaphylaxis, sinusitis, ocular allergies, silica-induced disease, COPD (damage reduction, airway inflammation, bronchial hyperresponsiveness, remodeling or disease progression), lung disease, cystic fibrosis, acid-induced lung injury, pulmonary hypertension, polyneuropathy, cataracts, muscle inflammation associated with systemic sclerosis, inclusion body myositis, myasthenia gravis, thyroiditis, Addison's disease disease), lichen planus, type 1 or type 2 diabetes, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic graft rejection, colitis, conjunctivitis, Crohn's disease, cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, encephalomyelitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis, Henoch-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 pneumonitis, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendinitis, tonsillitis, ulcerative colitis, uveitis, vaginitis, vasculitis, or vulvitis.

[0128] In some embodiments, cardiovascular diseases that can be treated according to the methods of the present invention include, but are not limited to, restenosis, cardiac hypertrophy, atherosclerosis, myocardial infarction, ischemic stroke, congestive heart failure, angina pectoris, reocclusion after angioplasty, restenosis after angioplasty, reocclusion after aortocoronary bypass, restenosis after aortocoronary bypass, stroke, transient ischemia, peripheral arterial occlusive disorder, pulmonary embolism, and deep vein thrombosis.

[0129] In some embodiments, neurodegenerative diseases that can be treated according to the methods 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 trauma, glutamate neurotoxicity, hypoxia, epilepsy, diabetes treatment, metabolic syndrome, obesity, organ transplantation, and graft-versus-host disease.

[0130] In some aspects, pharmaceutical compositions described herein are formulated to be used for patients who need such compositions. 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 "parenteral" includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In certain embodiments, the compositions are administered orally, intraperitoneally or intravenously. The sterile injectable form of pharmaceutical compositions described herein can be aqueous or oily suspensions. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents according to technology known in the art.

[0131] In some aspects, the pharmaceutical composition is administered orally.

[0132] The specific dosage and treatment regimen for any particular patient will depend on various factors including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination and the judgment of the treating physician and the severity of the specific disease being treated. The amount of the compound described herein in the composition will also depend on the specific compound in the pharmaceutical composition.

[0133] example

[0134] Preparation of compounds

[0135] The compounds claimed herein were prepared according to the procedures outlined in the following schemes. The compound names were generated using software built into ChemDraw. If the compound name was inconsistent with its depicted structure, the depicted chemical structure was used as the appropriate compound.

[0136] Nuclear synthesis

[0137] Synthesis of Methyl (5S,8S,10aR)-5-((tert-Butyloxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,4]diazacyclooctatetraene-8-carboxylate

[0138]

[0139] Step 1: Preparation of methyl (2S,5R)-1-((S)-4-(((benzyloxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)butanoyl)-5-vinylpyrrolidine-2-carboxylate

[0140] To a solution of methyl (2S,5R)-5-vinylpyrrolidine-2-carboxylate hydrochloride (12.1 g, 63.1 mmol, 1.00 equiv) in DCM (302 mL) were added BOPCl (16.1 g, 63.1 mmol, 1.00 equiv), NaHCO 3 The mixture was stirred for 2 hours at 40 ℃ for 1 h. ... 1 H NMR (400 MHz, CDCl 3 )δ7.43-7.28(m,5H),5.99-5.83(m,1H),5.43(br d,J=17.0Hz,1H),5.24-5.12(m,2H),5.12-5.03(m,2H),4.77-4.61(m,1H),4.58-4.43(m,2H),3.76-3.69(m,3H),3.56-3.41(m,1 H), 3.01 (tdd, J = 6.0, 9.4, 14.2Hz, 1H), 2.27-2.13 (m, 2H), 2.02-1.91 (m, 2H), 1.88-1.80 (m, 1H), 1.68-1.55 (m, 2H), 1.42 (s, 9H).

[0141] Step 2: Preparation of methyl (2S,5R)-1-((S)-4-(((benzyloxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)butanoyl)-5-formylpyrrolidine-2-carboxylate

[0142] Ozone (5 psi) was bubbled into a cooled (-40 °C) solution of methyl (2S,5R)-1-((S)-4-(((benzyloxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)butanoyl)-5-vinylpyrrolidine-2-carboxylate (18.3 g, 37.4 mmol, 1.00 equiv) in DCM (183 mL). After 1 h, the reaction mixture was sparged with N 2 (g) to remove excess ozone, followed by addition of Et 3 N (14.1mL, 101mmol, 2.71 equivalents). The reaction mixture was warmed to room temperature and stirred for 1 hour. The mixture was concentrated in vacuo, and the resulting residue was purified by column chromatography (gradient elution, petroleum ether / EtOAc=0 / 1 to 1 / 0) to give (2S, 5R)-1-((S)-4-(((benzyloxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)butanoyl)-5-formylpyrrolidine-2-carboxylic acid methyl ester (13.7g, 18.6mmol, two-step yield 49.7%, purity 66.6%) as a light yellow syrup. LCMS (ESI) m / z=492.2[M+H] +

[0143] Step 3: Preparation of methyl (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,4]diazacyclooctatetraene-8-carboxylate

[0144] In N 2 To a solution of methyl (2S,5R)-1-((S)-4-(((benzyloxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)butanoyl)-5-formylpyrrolidine-2-carboxylate (13.7 g, 18.6 mmol, 66.6% purity, 1.00 equiv) in i-PrOH (137 mL) was added Pd / C (6.30 g, 10.0% purity). The suspension was subjected to three evacuation cycles and treated with H 2 (g) (50 Psi) was purged. The mixture was heated to 40°C. After 12 hours, N 2(g) The mixture is purged. The suspension is filtered and the filtrate is concentrated under reduced pressure. The residue is purified by preparative HPLC (column: Kromasil Eternity XT 250x 80mmx 10um; mobile phase: [water (ammonia hydroxide v / v)-acetonitrile]; B%: 10%-40%, 20 minutes) and lyophilized to give (5S, 8S, 10aR)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,4]diazacyclooctatetraene-8-carboxylic acid methyl ester (3.39 g, 9.38 mmol, yield 50.5%, purity 94.4%) as a white solid. LCMS (ESI) m / z=342.1[M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ5.51(br d,J=8.0Hz,1H),4.80-4.72(m,1H),4.53(t,J=8.6Hz,1H),4.30-4.21(m,1H),3.76(s,3H),3.16-3.08(m,1H) ,2.99-2.84(m,3H),2.34-2.28(m,1H),2.20-2.16(m,2H),2.06-1.89(m,2H),1.77-1.59(m,2H),1.42(s,9H).

[0145] Synthesis of (5R,8S,10aR)-5-((tert-Butyloxycarbonyl)amino)-6-oxooctahydro-4H-pyrrolo[2,1-d][1,5]thiazoline-8-carboxylic acid

[0146]

[0147] Step 1: Preparation of (S)-1-benzyl-5-oxopyrrolidine-2-carboxylic acid tert-butyl ester

[0148] To a cooled (0°C) suspension of NaH (32.4 g, 810 mmol, 60% purity, 1.20 equiv) in THF (250 mL) was added dropwise a solution of (S)-tert-butyl 5-oxopyrrolidine-2-carboxylate (125 g, 675 mmol, 1.00 equiv) in THF (1.00 L). The mixture was warmed to 25°C and stirred for 1 hour. The mixture was then subsequently cooled to 0°C and benzyl bromide (88.2 mL, 742 mmol, 1.10 equiv) was subsequently added dropwise. After complete addition of benzyl bromide, the reaction mixture was warmed to 25°C and stirred for 2 hours. Four batches of equal scale were prepared and subsequently combined for post-processing. The reaction mixture was poured into a cooled saturated NH 4The biphasic mixture was extracted with EtOAc (500 mL x 3) and the organic phases were combined. The combined organic layers were washed with brine (800 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a crude product. The crude product was purified by column chromatography (gradient elution: petroleum ether / EtOAc=1 / 0 to 50 / 1) and concentrated under reduced pressure to give (S)-1-benzyl-5-oxopyrrolidine-2-carboxylic acid tert-butyl ester (397 g, 1.41 mol, yield 52.1%, purity 98.0%) as a yellow solid. LCMS (ESI) m / z=275.9 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ7.34-7.28(m,3H),7.23-7.21(m,2H),5.06(d,J=14.8Hz,1H),3.96(d,J=14.8Hz,1H),3.85-3.8 2(m,1H),2.56-2.44(m,1H),2.43-2.41(m,1H),2.24-2.19(m,1H),2.06-2.04(m,1H),1.44(s,9H).

[0149] Step 2: Preparation of (S)-1-benzyl-5-thiopyridin-2-carboxylic acid tert-butyl ester

[0150] Lawesson's Reagent (131 g, 324 mmol, 0.50 equiv) was added to a solution of (S)-1-benzyl-5-oxopyrrolidine-2-carboxylic acid tert-butyl ester (179 g, 648 mmol, 1.00 equiv) in DCM (1.79 L) at 25 ° C. Two batches of equal scale preparation were carried out and then combined for post-processing. After stirring for 4 hours, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (gradient elution: petroleum ether / EtOAc=1 / 0 to 50 / 1) to give (S)-1-benzyl-5-sulfylidenepyrrolidine-2-carboxylic acid tert-butyl ester (324 g, 1.11 mol, yield 85.5%, purity 99.7%) as a yellow solid. LCMS (ESI) m / z=292.1 [M+H] + ; 1 H NMR (400 MHz, CDCl 3)δ7.35-7.30(m,5H),5.81(d,J=14.4Hz,1H),4.28(d,J=14.8Hz,1H),4.17-4.1 4(m,1H),3.16-3.09(m,2H),2.27-2.23(m,1H),2.22-2.10(m,1H),1.45(s,9H).

[0151] Step 3: Preparation of tert-butyl (S,Z)-1-benzyl-5-(2-methoxy-2-oxoethylidene)pyrrolidine-2-carboxylate

[0152] A solution of (S)-tert-butyl 1-benzyl-5-thiopyridinium-2-carboxylate (58.0 g, 199 mmol, 1.00 equiv) and methyl bromoacetate (37.7 g, 247 mmol, 23.3 mL, 1.24 equiv) in acetonitrile (160 mL) was stirred at 25°C. After stirring for 40 hours, the reaction mixture was diluted with dichloromethane (1.20 L) and cooled to 0°C. After another 10 minutes, PPh 3 (78.3 g, 298 mmol, 1.50 equiv) and Et 3 N (83.1 mL, 597 mmol, 3.00 equiv). The reaction mixture was allowed to warm to ambient temperature and stirred for 16 h. Six batches of equal size were prepared and subsequently combined for workup. The mixture was poured into 1N NaH 2 PO 4 The biphasic mixture was extracted with dichloromethane (800 mL x 3). The combined organic layers were washed with brine (800 mL) and purified by Na 2 SO 4 Dry, filter and concentrate to obtain a residue. The residue was purified by column chromatography (gradient elution: petroleum ether / EtOAc=80 / 1 to 15 / 1) to obtain (S,Z)-1-benzyl-5-(2-methoxy-2-oxoethylidene)pyrrolidine-2-carboxylic acid tert-butyl ester (255 g, 706 mmol, yield 59.2%, purity 91.8%) as a yellow oil. LCMS (ESI) m / z=332.1 [M+H] + ; 1 H NMR (400 MHz, CDCl 3)δ7.35-7.29(m,3H),7.20-7.18(m,2H),4.76(s,1H),4.54(d,J=15.6Hz,1H),4.20(d,J=15.6Hz,1H),3.98-3.9 5(m,1H),3.75(s,3H),3.62-3.40(m,1H),3.11-3.09(m,1H),2.25-2.20(m,1H),2.10-2.05(m,1H),1.44(s,9H).

[0153] Step 4: Preparation of tert-butyl (2S,5R)-1-benzyl-5-(2-methoxy-2-oxoethyl)pyrrolidine-2-carboxylate

[0154] In N 2 To a suspension of tert-butyl (S,Z)-1-benzyl-5-(2-methoxy-2-oxoethylidene)pyrrolidine-2-carboxylate (125 g, 377 mmol, 1.00 equiv) in EtOAc (1.25 L) was added 10% Pd / C (125 g). The suspension was subjected to three evacuation cycles and treated with H 2 (g) (50 psi) was purged. The mixture was heated at 25 °C under H 2 After stirring for 12 hours, the mixture was emptied three times and stirred with N 2 (g) Purge and The filtrate was transferred to another container and filtered under N 2 (g) Pt / C (125 g, 5% purity) was added under air flow. The suspension was evacuated three times and heated with H 2 (g) (50 psi) was purged. The reaction mixture was heated to 60°C and heated under H 2 The product was stirred for 16 hours at 50 psi. Two batches of preparations of equal scale were carried out and then merged for post-processing. The combined reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (gradient elution: petroleum ether / EtOAc=80 / 1 to 30 / 1) to obtain (2S, 5R)-1-benzyl-5-(2-methoxy-2-oxoethyl)pyrrolidine-2-carboxylic acid tert-butyl ester (119 g, 358 mmol, 47.5% yield, 100% purity) as a yellow oil. LCMS (ESI) m / z=334.1 [M+H] + ; 1 H NMR (400 MHz, CDCl 3)δ7.33-7.30(m,4H),7.24-7.22(m,1H),3.89-3.78(m,2H),3.63(s,3H),3.29-3.25(m,2H),2.60-2. 55(m,1H),2.37-2.31(m,1H),2.05-2.00(m,2H),1.99-1.88(m,1H),1.68-1.66(m,1H),1.45(s,9H).

[0155] Step 5: Preparation of tert-butyl (2S,5R)-1-benzyl-5-(2-hydroxyethyl)pyrrolidine-2-carboxylate

[0156] In N 2 (g) To a cooled (0 °C) solution of tert-butyl (2S,5R)-1-benzyl-5-(2-methoxy-2-oxoethyl)pyrrolidine-2-carboxylate (58.5 g, 175 mmol, 1.00 equiv) in THF (585 mL) was added LiBH 4 (2.00 M, 175 mL, 2.00 equiv.). The reaction mixture was warmed to 25 °C and stirred for 16 h. Two batches of equal size were prepared and subsequently combined for workup. The combined reaction mixture was poured into 1N K 2 CO 3 The combined organic layers were washed with brine (1.00 L) and purified by Na 2 SO 4 Dry, filter and concentrate to obtain a residue. The residue was purified by column chromatography (gradient elution: petroleum ether / EtOAc=80 / 1 to 10 / 1) to obtain tert-butyl (2S,5R)-1-benzyl-5-(2-hydroxyethyl)pyrrolidine-2-carboxylate (60.0 g, 196 mmol) as a yellow oil. LCMS (ESI) m / z=306.1 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ7.33-7.28(m,4H),7.26-7.24(m,1H),4.09-4.00(m,2H),3.76-3.73(m,1H),3.58-3.55(m,1H),3.30 -3.28(m,1H),3.27-3.21(m,1H),2.08-2.04(m,2H),1.91-1.87(m,3H),1.55-1.54(m,1H),1.50(s,9H).

[0157] Step 6: Preparation of tert-butyl (2S,5R)-1-benzyl-5-(2-((methylsulfonyl)oxy)ethyl)pyrrolidine-2-carboxylate

[0158] To a cooled (-10 °C) solution of tert-butyl (2S,5R)-1-benzyl-5-(2-hydroxyethyl)pyrrolidine-2-carboxylate (29.5 g, 96.6 mmol, 1.00 equiv) in DCM (295 mL) was added methanesulfonyl chloride (11.7 mL, 151 mmol, 1.57 equiv), DMAP (590 mg, 4.83 mmol, 0.05 equiv) and Et 3 N (40.3 mL, 290 mmol, 3.00 equiv). The mixture was warmed to 25 ° C and stirred for 1 hour. Two batches of equal scale were prepared and then combined for post-processing. The reaction mixtures were combined and diluted with dichloromethane (800 mL). The resulting mixture was washed with 0.5N HCl (1.00 L) and washed with saturated NaHCO 3 The mixture was washed with aqueous solution (1.00 L), washed with brine (1.00 L), and then with Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give tert-butyl (2S,5R)-1-benzyl-5-(2-((methylsulfonyl)oxy)ethyl)pyrrolidine-2-carboxylate (72.0 g, crude) as a yellow oil. LCMS (ESI) m / z=384.1 [M+H] +

[0159] Step 7: Preparation of S-(2-((2R,5S)-1-benzyl-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-N-(tert-butoxycarbonyl)-L-cysteine

[0160] In N 2 To a cooled (0°C) solution of tert-butyl (2S,5R)-1-benzyl-5-(2-((methylsulfonyl)oxy)ethyl)pyrrolidine-2-carboxylate (20.8 g, 93.9 mmol, 1.00 equiv) in DMF (360 mL) was added NaH (7.51 g, 187 mmol, 60% purity, 2.00 equiv) in portions under (g). The mixture was stirred at 0°C for 1 h, followed by addition of a solution of (tert-butoxycarbonyl)-L-cysteine ​​(36.0 g, 93.9 mmol, 1.00 equiv) in DMF (360 mL). The mixture was warmed to 25°C and stirred for 1 hour. Two batches of equal scale were prepared and subsequently combined for post-processing. The reaction mixtures were combined and the pH of the mixture was adjusted with 1N aqueous HCl (until pH was about 2). The acidic mixture was extracted with EtOAc (2.00 L x 2). The combined organic layers were washed with brine (2.00 L) and purified by Na2 SO 4 The mixture was dried, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (gradient elution; petroleum ether / EtOAc=20 / 1 to 0 / 1) to give S-(2-((2R,5S)-1-benzyl-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-N-(tert-butoxycarbonyl)-L-cysteine ​​(92.0 g, crude) as a yellow oil. LCMS (ESI) m / z=509.2 [M+H] +

[0161] Step 8: Preparation of N-(tert-butoxycarbonyl)-S-(2-((2R,5S)-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-L-cysteine

[0162] In N 2 To a solution of S-(2-((2R,5S)-1-benzyl-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-N-(tert-butoxycarbonyl)-L-cysteine ​​(46.0 g, 90.4 mmol, 1.00 equiv) in MeOH (460 mL) was added Raney-Ni (46.0 g). The suspension was subjected to three evacuation cycles and treated with H 2 (g) Purging. The mixture was heated at 60 °C under H 2 After stirring for 16 hours, the mixture was heated with N 2 (g) Purge and filter. The filtrate is transferred to a reaction vessel and heated under N 2 (g) Raney-Ni (92.0 g) was added to the filtrate. The suspension was subjected to three evacuation cycles and H 2 (g) Purging. The mixture was heated to 80°C and heated at 80°C under H 2 (g) (50 Psi) and stirred for 40 hours. LCMS analysis showed that about 54.0% of S-(2-((2R,5S)-1-benzyl-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-N-(tert-butoxycarbonyl)-L-cysteine ​​(Rt = 0.582 minutes) remained, and the expected MS was detected (Rt = 0.487 minutes). The mixture was filtered and heated to N 2 (g) was added to the filtrate. The suspension was degassed under vacuum and heated to 400 °C. 2 The mixture was heated to 80°C and heated at 80°C under H 2(50Psi) and stirred for 40 hours. LCMS analysis showed that S-(2-((2R,5S)-1-benzyl-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-N-(tert-butoxycarbonyl)-L-cysteine ​​(Rt=0.584 minutes) remained 39.0%, and the expected MS (Rt=0.492 minutes) was detected. Two batches of equal scale preparation were carried out and then combined for post-processing. The reaction mixture was filtered, and the filtrate was combined and concentrated to give a residue to give N-(tert-butoxycarbonyl)-S-(2-((2R,5S)-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-L-cysteine ​​(59.0 g, crude) as a yellow solid. LCMS (ESI) m / z=419.2[M+H] + .

[0163] Step 9: Preparation of tert-butyl (5R,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxooctahydro-4H-pyrrolo[2,1-d][1,5]thiazoline-8-carboxylate

[0164] PyBOP (54.1 g, 104 mmol, 3.00 equiv) and DIPEA (24.9 g, 192 mmol, 33.5 mL, 5.55 equiv) were added to a cooled (0 °C) solution of N-(tert-butoxycarbonyl)-S-(2-((2R,5S)-5-(tert-butoxycarbonyl)pyrrolidin-2-yl)ethyl)-L-cysteine ​​(14.5 g, 34.6 mmol, 1.00 equiv) in DCM (1.50 L). The mixture was then warmed to 25 °C and stirred for 3 hours. Four batches of equal scale were prepared and then combined for workup. The reaction mixture was poured into 0.5N aqueous HCl (4.50 L) and extracted with dichloromethane (2.00 L x 2). The combined organic layers were washed with saturated NaHCO 3 The aqueous layer was extracted with DCM (2.00 L x 2). The combined organic layers were washed with brine (1.00 L) and purified by Na 2 SO 4 The residue was purified by column chromatography (gradient elution: petroleum ether / EtOAc=1 / 0 to 20 / 1) and concentrated under reduced pressure to give (5R, 8S, 10aR)-5-((tert-butoxycarbonyl)amino)-6-oxooctahydro-4H-pyrrolo[2,1-d][1,5]thiazacyclooctatetraene-8-carboxylic acid tert-butyl ester (12.6 g, 31.1 mmol, yield 22.4%, purity 98.8%) as a white solid. LCMS (ESI) m / z=401.1 [M+H] + ; 1H NMR (400 MHz, CDCl 3 )δ5.98-5.96(m,1H),4.69-4.68(m,1H),4.66-4.58(m,1H),4.40-4.35(m,1H),2.94-2.90(m,2H) ,2.76-2.74(m,2H),2.73-2.74(m,1H),1.99-1.96(m,3H),1.71-1.67(m,2H),1.48-1.42(m,18H).

[0165] Step 10: Preparation of (5R,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxooctahydro-4H-pyrrolo[2,1-d][1,5]thiazoline-8-carboxylic acid

[0166] To a cooled (0 °C) solution of (5R,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxooctahydro-4H-pyrrolo[2,1-d][1,5]thiazazecin-8-carboxylic acid tert-butyl ester (6.30 g, 15.7 mmol, 1.00 equiv) in DCM (64.0 mL) was added triisopropylsilane (323 uL, 1.57 mmol, 0.10 equiv) followed by a solution of TFA (143 mL, 1.93 mol, 123 equiv) in DCM (64.0 mL). After stirring for 1 hour, the reaction mixture was concentrated to give an oil. The resulting amino acid (oil) was diluted in acetonitrile (176 mL) and H 2 O (8.80 mL), followed by the addition of Et 3 N (17.7 mL, 127 mmol, 8.10 equiv) and Boc 2 O (10.3 g, 47.2 mmol, 10.8 mL, 3.00 equiv). The mixture was warmed to 25 ° C for 2 hours. Two batches of equal scale were prepared and then combined for post-processing. The reaction mixtures were combined and concentrated under reduced pressure. The resulting residue was washed with H 2 The mixture was diluted with 1% HCl (80.0 mL) and extracted with EtOAc (60.0 mL x 2). The aqueous layer was adjusted to pH about 2 with 1N HCl aqueous solution and extracted with EtOAc (50.0 mL x 3). The combined organic layers were washed with brine (40.0 mL x 2) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to obtain a residue. The residue was purified by HPLC. 2O (20.0 mL) and acetonitrile (6.00 mL) and lyophilized to give (5R, 8S, 10aR) -5- ((tert-butoxycarbonyl) amino) -6-oxooctane-4H-pyrrolo [2, 1-d] [1, 5] thiazoline-8-carboxylic acid (5.50 g, 15.4 mmol, yield 48.9%, purity 96.4%) as a white solid. LCMS (ESI) m / z = 367.1 [M + Na] + ; 1 H NMR (400 MHz, CDCl 3 )δ6.10(brs,1H),5.87-5.85(m,1H),4.73-4.70(m,2H),4.68-4.58(m,1H),2.98-2.94(m,2H),2.89-2.74(m,1H ),2.72-2.71(m,1H),2.54-2.38(m,2H),2.05-2.02(m,1H),1.89-1.86(m,2H),1.75-1.48(m,1H),1.44(s,9H).

[0167] (5S,8S,10aR)-3-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid:

[0168]

[0169] This is commercially available: CAS 2497590-06-2

[0170] Synthesis of linkers

[0171] Synthesis procedure of linker:

[0172] The following intermediates in Table 1 were prepared according to the protocol described in WO 2020 / 205467.

[0173] Table 1

[0174]

[0175] Synthesis of (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylic acid

[0176]

[0177] (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylic acid was prepared according to the protocol described in US2004 / 0225146.

[0178] Representative preparation steps of activated ester phosphonic acids

[0179] Synthesis of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0180]

[0181] Step 1: Preparation of 4-nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0182] 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) were dissolved in CH 2 Cl 2 The mixture in 4% paraformaldehyde (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 washed with CH 2 Cl 2 The product was purified by reverse phase chromatography [C18 cartridge, eluting with a gradient of 5-100% acetonitrile / water] and the appropriate fractions were concentrated to give 4-nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (7.80 g, 16.0 mmol, 59.0% yield) as a yellow solid. LCMS m / z=486.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )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).

[0183] Step 2: Preparation of (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0184] 4-Nitrophenyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (4.47 g, 9.20 mmol) was added to CH 2 Cl 2To a cooled (0°C) solution of 1% iodine (39 mL) was added N,O-bis(trimethylsilyl)trifluoroacetamide (12.1 mL, 46.0 mmol) and iodotrimethylsilane (5.23 mL, 36.8 mmol) in CH 2 Cl 2 The reaction mixture was allowed to gradually warm to ambient temperature. 2:1 H 2 In 40% ethyl acetate, 1:10 mol / l 4- nitropropene was added to 4- nitropropene (1:1 v / v, 100 mL) and 0.05% ethyl acetate (50 mL). The mixture of O / acetonitrile (containing 0.1% TFA) (50 mL), and the precipitation of product was observed. Volatiles were removed in a vacuum, and the crude residue was suspended in a mixture of acetonitrile / aqueous solution (1:1 v / v, 100 mL). The suspension was filtered, and the mixture of the acetonitrile / aqueous solution of solid 2:1 was washed, and the solid was dried under reduced pressure to obtain [difluoro ({ 2- [(4- nitrophenoxy) carbonyl] -1- benzothiophene -5- bases}) methyl] phosphonic acid (6.5 g, 94%) in a beige solid. The filtrate was concentrated to 50% of the solvent volume, and the resulting suspension was filtered, and washed with 1:2 acetonitrile / aqueous solution. The solid was dried under reduced pressure to obtain [difluoro ({ 2- [(4- nitrophenoxy) carbonyl] -1- benzothiophene -5- bases}) methyl] phosphonic acid (0.4 g) in a beige solid. Both products were lyophilized to give (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (6.90 g, 16.0 mmol, 98.0% yield). 1 H NMR (400 MHz, DMSO-d 6 )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).

[0185] The following intermediates in Table 2 were prepared using a similar protocol outlined above for the synthesis of [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid and utilizing the appropriate higher intermediates as starting materials.

[0186] Table 2

[0187]

[0188] Synthesis of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0189]

[0190] Preparation of 5-methylbenzo[b]thiophene-2-carboxylic acid

[0191] 5-Methylbenzo[b]thiophene-2-carboxylic acid was prepared according to the procedure described in WO 2016 / 100184 A1.

[0192] Step 1: Preparation of benzyl 5-methylbenzo[b]thiophene-2-carboxylate

[0193] 5-Methylbenzo[b]thiophene-2-carboxylic acid (21.2 g, 110.0 mmol, 1.0 equiv) and K 2 CO 3 To a solution of 5-methylbenzo[b]thiophene-2-carboxylic acid benzyl ester (30.1 g, 106.0 mmol, 97% yield) in DMF (200 mL) was added benzyl bromide (20.6 g, 121.0 mmol, 1.1 eq). The mixture was stirred at room temperature for 14 h. The reaction mixture was poured into ice water (400 mL) and stirred for 5 min. The resulting solid was filtered and the filter cake was washed with water (50 mL) and dried in vacuo to give benzyl 5-methylbenzo[b]thiophene-2-carboxylate (30.1 g, 106.0 mmol, 97% yield) as a yellow solid. 1 H NMR (400 MHz, CDCl 3 )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).

[0194] Step 2: Preparation of benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate

[0195] Benzyl 5-methylbenzo[b]thiophene-2-carboxylate (15.0 g, 53.1 mmol, 1.0 eq.) and NBS (10.3 g, 58.4 mmol, 1.1 eq.) were dissolved in CCl. 4 To the solution in 1% ethanol (30 mL), benzoyl peroxide (1.3 g, 5.31 mmol, 0.1 eq) was added. The reaction flask was evacuated three times and the mixture was heated with N 2 (g) Backfill. The mixture is heated at a constant N 2 (g) The mixture was stirred at 80°C for 16 hours under an atmosphere of 4% paraformaldehyde (pH 7.0). The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography to give benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (6.80 g, 18.8 mmol, 36% yield) as a yellow solid.

[0196] Step 3: Preparation of benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0197] A solution of benzyl 5-(bromomethyl)benzo[b]thiophene-2-carboxylate (10.3 g, 28.5 mmol, 1.0 equiv) dissolved in triethyl phosphite (30.0 g, 180.0 mmol, 6.3 equiv) 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 flash column chromatography to give benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (6.5 g, 15.5 mmol, 55% yield) as a colorless oil. LCMS (ESI) m / z=419 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )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).

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

[0199] Benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (5.6 g, 13.3 mmol, 1.0 equiv) was dissolved in THF (80 mL) and H 2 LiOH (1.10 g, 26.6 mmol, 2.0 equiv) was added to the solution in the mixture in 0 (10 mL). The mixture was stirred at room temperature for 3 hours and then acidified with 1N HCl aqueous solution (adjusted to a pH of about 3-4). After acidification, the product was precipitated from the solution. 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] + .

[0200] Step 5: Preparation of perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0201] 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (3.9 g, 11.8 mmol, 1.0 equiv) was added to CH 2 Cl 2To a cooled (0 °C) solution of 1% paraformaldehyde (50 mL) was added oxalyl chloride (2.2 g, 17.7 mmol, 1.5 eq.) 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 CH 2 Cl 2 (50 mL), followed by the addition of Et 3 N (3.6 g, 35.4 mmol, 3.0 eq) and pentafluorophenol (2.6 g, 14.1 mmol, 1.2 eq). The resulting mixture was stirred at room temperature for another 2 h and then poured into H 2 O (30 mL). The biphasic solution was extracted with EtOAc (30 mL x 3). The combined organic layers were purified by MgSO 4 After drying, filtering and concentrating under reduced pressure, the residue was purified by silica gel column chromatography to give perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (4.7 g, 9.5 mmol, yield 81%) as a white solid. LCMS (ESI) m / z=419 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )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).

[0202] Step 6: Preparation of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0203] Perfluorophenyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (4.7 g, 9.5 mmol, 1.0 equiv) was added to CH 2 Cl 2 To the solution in 4% parafluorophenyloxy (60 mL), bromotrimethylsilane (12 mL) was added. 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]thiophen-5-yl)methyl)phosphonic acid (3.7 g, 8.4 mmol, 89% yield) as a white solid. LCMS (ESI) m / z=439 [M+H] + .

[0204] 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

[0205]

[0206] Step 1: rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0207] To a cooled (-78 °C) solution of benzyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (2.4 g, 5.73 mmol, 1 eq) in THF (75 mL) and 2-(phenylsulfonyl)-3-phenyloxaziridine (2.97 g, 11.4 mmol, 2 eq) was added a 1 M solution of NaHMDS (11.4 mL, 11.4 mmol, 2 eq) in THF. A dark purple solution was observed upon addition of the base, which turned orange upon complete addition of the base. The mixture was stirred for an additional 10 min followed by addition of saturated NH 4 Cl aqueous solution (50mL). The mixture is warmed to ambient temperature, and EtOAc (75mL) and water (25mL) are added. After stirring for another 30 minutes, each phase is separated. The aqueous layer is extracted with EtOAc (125mL x2). The combined organic extracts are dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Another batch of preparations of equal scale are carried out and combined for purification. The combined material (6.42mmol, 12.15mmol in total) is purified by flash chromatography (20%-100%=EtOAc: heptane) to obtain rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (3.69g, 8.49mmol, 70%) as a white sticky solid. LCMS (ESI) m / z=869.4[2M+H] + ; 1 H NMR (400 MHz, CDCl 3 )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),7.46–7.3 5(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).

[0208] Step 2: rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester

[0209] To rac-5-((diethoxyphosphoryl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (cc) (1.56 g, 3.59 mmol, 1 equiv) was added to CH 2 Cl 2 A cooled (-78 °C) solution (30 mL) in N 2 (g) (diethylamino)sulfur trifluoride (568 μL, 4.30 mmol, 1.2 equiv) was added to the reaction mixture. The reaction was stirred for 15 minutes, followed by the addition of saturated NaHCO 3 After heating to room temperature, the product was washed with CH 2 Cl 2 The crude residue was purified by reverse phase chromatography on a C18 cartridge (eluting with 5-80% acetonitrile / water) to give rac-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (650 mg, 1.48 mmol, 41.6%) as a clear thick oil. LCMS (ESI) m / z=873.2 [2M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ8.12(s,1H),8.02–7.99(m,1H),7.92(d,J=8.7Hz,1H),7.61(d,J=8.7Hz,1H),7.51–7.51(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).

[0210] 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

[0211] Rac-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (650 mg, 1.48 mmol) was subjected to chiral SFC separation (column: Lux i-Amylose 3, 21.2 x 250 mm 5 um column, 75 ml / min, 40% MeOH) to give (R)- or (S)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (304 mg, 0.70 mmol, 46.8% recovery, 99.9% ee) (peak 1) as a clear thick oil and (R)- or (S)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (317 mg, 0.73 mmol, 49% recovery, 99.9% ee) (peak 2) as a clear thick oil. Note: The fastest eluting enantiomer by SFC was arbitrarily designated as (R)-5-(fluoro(phosphono)methyl)benzo[b]thiophene-2-carboxylic acid, and the slowest eluting enantiomer by SFC was arbitrarily designated as (S)-5-(fluoro(phosphono)methyl)benzo[b]thiophene-2-carboxylic acid.

[0212] HPLC method for analysis of enantiomeric excess: Lux Cellulose-3 150 mm 45% H 2 O + 0.05% TFA / 55% MeCN 1 ml / min for 8 min.

[0213] Step 4: Preparation of (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0214] To a mixture of 10% Pd / C (60 mg, 50% wet) and (R)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (peak 1) (60 mg, 0.1374 mmol, 1 eq) in THF (5 mL) was added N 2 (g) Degas for 5 minutes. Bubble H into the mixture. 2 (g) for 5 minutes, and then the reaction was continued under H 2 (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 N 2 (g) 15 minutes, and 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 clear thick oil. LCMS (ESI) m / z = 347.2 [M+H] + .

[0215] The following intermediates in Table 3 were prepared using the procedure outlined above (Step 4) from (S)- or (R)-benzyl 5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylate (Peak 2) and using appropriate reagents.

[0216] Table 3

[0217]

[0218] Representative methods for synthesizing activated linkers

[0219] Method 1: Stepwise Acid Chloride Method for the Synthesis of Hybrid Joints

[0220] Representative Procedure for the Synthesis of 4-Nitrophenyl 5-(((2-(Butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0221]

[0222] Step 1: Preparation of 4-nitrophenyl 5-(difluoro(hydroxy(pyridin-3-yl)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0223] To (difluoro(2-((4-nitrophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (100 mg, 0.2329 mmol, 1 eq.) was added to CH 2 Cl 2 To a cooled (0 °C) heterogeneous solution in 4 mL of 1% paraformaldehyde (4 mL) was added catalytic DMF (2 drops) followed by dropwise addition of oxalyl chloride (198 μL, 2.32 mmol, 10 equiv). The homogeneous reaction mixture was warmed to room temperature and stirred for 2 hours. The reaction was concentrated in vacuo and further dried under high vacuum for 30 minutes to give a yellow solid. The yellow solid was diluted in CH 2 Cl 2 (4 mL) and cooled to -78°C. Pyridin-3-ol (22.0 mg, 232 μmol, 1 eq.) and triethylamine (64.7 μL, 465 μmol, 2 eq.) were slowly added to CH 2 Cl 2 The mixture was stirred at -78 °C for 2 minutes and then allowed to warm to ambient temperature and stirred overnight. After 24 hours, the reaction mixture became heterogeneous and the reaction was concentrated under reduced pressure. The crude product, 4-nitrophenyl 5-(difluoro(hydroxy(pyridin-3-yloxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate, was used directly in the next step without further purification or manipulation. LCMS m / z=507.2 [M+H]+ .

[0224] Step 2: Preparation of 4-nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0225] To nitrophenyl 5-(difluoro(hydroxy(pyridin-3-yl)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (117 mg, 0.2329 mmol, 1 eq.) was added to CH 2 Cl 2 To the cooled (0 ° C) solution in 4% paraformaldehyde (5 mL), 2 drops of DMF were added, followed by dropwise addition of oxalyl chloride (198 μL, 2.32 mmol, 10 equivalents). The reaction was warmed to room temperature and stirred for 1.5 hours. LCMS analysis showed partial conversion to the desired activated intermediate. Additional oxalyl chloride (198 μL, 2.32 mmol, 10 equivalents) was introduced into the reaction mixture, and the mixture was stirred for another 1 hour. The reaction was concentrated in vacuo and further dried under high vacuum for 30 minutes to obtain a yellow solid. The yellow solid was diluted in CH 2 Cl 2 (5 mL) and cooled to -78°C. 1-[(2-hydroxyethyl)sulfonyl]butan-1-one (103 mg, 698 μmol, 3 eq.) diluted in CH 2 Cl 2 (1 mL) [previously treated with anhydrous Na 2 SO 4 Dry], followed by the addition of triethylamine (134 μL, 967 μmol, 2 eq.). After stirring for 2 minutes, the resulting mixture was allowed to warm to ambient temperature and stirred overnight. The mixture was carefully concentrated in vacuo. The crude residue was purified by flash chromatography (gradient elution, 0-60% EtOAc / heptane) to give 4-nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-oxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (22.0 mg, 0.03455 mmol, 14.9% yield) as a clear oil. LCMS m / z=637.2 [M+H] + ; 1 H NMR: (400MHz, DMSO-d 6)δ8.50–8.44(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,2 H),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).

[0226] The following intermediates in Table 4 were prepared using a similar protocol described above for the synthesis of 4-nitrophenyl 5-(((2-(butyrylthio)ethoxy)(pyridin-3-yloxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediates as starting materials.

[0227] Table 4

[0228]

[0229]

[0230]

[0231] Method 2: One-pot acid chloride method for the synthesis of activated linkers

[0232] Representative Procedure for the Synthesis of Perfluorophenyl 5-((Bis(4-((3-methylbutyryl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0233]

[0234] 3 (200 mg, 0.42 mmol, 1.0 equiv) was dissolved in anhydrous CH 2 Cl 2 To a cooled (0 °C) solution of 1% oxalyl chloride (266 mg, 2.10 mmol, 5.0 equiv) (15 mL) and catalytic DMF (3.2 μL, 42.1 μmol, 0.1 equiv) was added dropwise oxalyl chloride (266 mg, 2.10 mmol, 5.0 equiv). The reaction mixture was allowed to warm to 40 °C. After stirring for 2 h, the reaction mixture was concentrated in vacuo and dried (to remove excess oxalyl chloride). The resulting solid was redissolved in anhydrous CH 2 Cl 2(5 mL) and cooled to 0° C. To the cooled solution were added S-(4-hydroxybutyl) 3-methylthiobutanoate (239 mg, 1.26 mmol, 3.0 equiv), DMAP (5.14 mg, 42.1 μmol, 0.1 equiv) and N,N-diisopropylethylamine (217 mg, 1.68 mmol, 4.0 equiv) in anhydrous CH 2 Cl 2 (10 mL) solution. The reaction mixture was allowed to warm to room temperature and stirred for another 18 h. The reaction was concentrated by adding H 2 O (10 mL) and quenched with CH 2 Cl 2 The organic layers were combined and washed with brine (20 mL) and purified by anhydrous Na 2 SO 4 Dry and concentrate in vacuo. The residue was purified by flash column chromatography to give perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (15.0 mg, 18.3 μmol, 4.4% yield). LCMS (ESI) m / z=819 [M+H] + .

[0235] The following intermediates in Table 5 were prepared using a similar protocol described above for the synthesis of perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate higher intermediates as starting materials.

[0236] Table 5

[0237]

[0238] Method 3: One-pot silver salt method for the synthesis of activated linkers

[0239] Representative Procedure for the Synthesis of Perfluorophenyl 5-((Bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0240]

[0241] Step 1: Preparation of silver (I) ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate

[0242] To ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (300 mg, 684 μmol, 1.0 equiv) was added deionized H 2Amberlite was added to the mixture of O (4 mL) and THF (2 mL). Resin (Na + The resulting mixture was stirred at room temperature for 1 hour, and then the suspension was filtered. AgNO was added to the filtrate. 3 (463 mg, 2.73 mmol, 4.0 equiv) in deionized water H 2 O (2mL), and the resulting mixture was stirred at room temperature for another 1 hour. A white precipitate was observed to form, and the solid was collected by filtration. The filter cake was then washed with cold H 2 O (3 x 2 mL), and the solid was dried under reduced pressure to give silver (I) ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate as a dry powder. The silver salt was used without further purification.

[0243] Step 2: Preparation of perfluorophenyl 5-((bis(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0244] To a suspension of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate silver (I) suspended in anhydrous toluene (10 mL) was added iodomethyl 2-methylpropanoate (500 mg, 2.05 mmol, 3.0 equiv) dropwise. After addition, the resulting mixture was stirred at room temperature for another 12 hours. The progress of the reaction was monitored by LCMS, and upon completion, the unreacted silver salt was recovered by filtration. The filtrate was concentrated in vacuo, and the resulting residue was purified by reverse 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-carboxylate (165 mg, 246 μmol, 36% yield) as a white solid. LCMS (ESI) m / z = 671 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ8.28(s,1H),7.96-7.80(m,2H),7.48(d,J=8.5Hz,1H),5.70-5.53(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).

[0245] The following intermediates in Table 6 were prepared using a similar protocol described above for the synthesis of perfluorophenyl 5-((bis(4-((3-methylbutanoyl)thio)butoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediates as starting materials.

[0246] Table 6

[0247]

[0248]

[0249]

[0250] (R)- or (S)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0251]

[0252] Step 1: Preparation of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid

[0253] To (R)- or (S)-5-((diethoxyphosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid benzyl ester (peak 1) (205 mg, 0.4697 mmol) in CH 2 Cl 2 To a cooled (0 °C) solution of 1% iodine (8.0 mL) was added BSTFA (746 μL, 2.81 mmol), followed by trimethylsilyl iodide (1.87 mL, 1.87 mmol) in CH 2 Cl 2 The product was stirred for 1 h at 4 °C for 2 h to obtain a 1.0 M solution in 4% ethyl acetate. After stirring for 1 h, a mixture of acetonitrile (0.66 mL), water (0.33 mL) and 0.1% TFA was added. The solvent was removed under reduced pressure at 0 °C. The crude residue was purified by reverse phase chromatography (C18 cartridge, eluting with 5-40% acetonitrile / water) to give (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid (163 mg, 0.4285 mmol) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )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).

[0254] Step 2: Preparation of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid

[0255] [The following reaction was performed in a foil-covered container and in the absence of ambient light]: To a suspension of (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid (163 mg, 0.4285 mmol, 1 eq) in water (5 mL) was added a solution of aqueous sodium hydroxide (34.2 mg, 857 μmol, 2 eq) in water (2 mL). To the yellow solution was added silver (I) nitrate (181 mg, 1.07 mmol, 2.5 eq) 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 iodomethyl 2,2-dimethylpropanoate (191 μL, 1.28 mmol, 3 eq) was added. After stirring for 20 hours, the reaction mixture was stirred at room temperature for 20 hours. The reaction mixture was filtered and rinsed with toluene. The filtrate was concentrated under reduced pressure. The crude residue was purified (C18 cartridge, eluted with 5-100% acetonitrile / water) to obtain (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 (400 MHz, CDCl 3 )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).

[0256] Step 3: Preparation of (R)- or (S)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid

[0257] In N 2To a solution of (R)-((((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphoryl)bis(oxy))bis(methylene)bis(2,2-dimethylpropionate) (122 mg, 0.200 mmol, 1 eq) in THF (10 mL) was added 10% Pd / C (50% wet, 120 mg, 0.1127 mmol, 0.56 eq). H2O was bubbled into the suspension. 2 (g) for 5 minutes. The reaction mixture was stirred at H 2 (g) (1 atm) and stirred at room temperature. After stirring for 22 hours, the reaction mixture was washed with N 2 (g) Purge and Filter. The filter pad was washed with THF and concentrated under reduced pressure to give (R)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid (0.200 mmol, 99.9%) as a clear thick oil. LC-MS (ESI) m / z [M+H] + =519.1.

[0258] The following intermediates in Table 7 were prepared using a similar protocol described above for the synthesis of (R)- or (S)-5-((bis((pivaloyloxy)methoxy)phosphoryl)fluoromethyl)benzo[b]thiophene-2-carboxylic acid, and utilizing appropriate advanced intermediates as starting materials. The absolute configuration of the starting material (R)- or (S)-((2-((benzyloxy)carbonyl)benzo[b]thiophen-5-yl)fluoromethyl)phosphonic acid was not determined, but the elution peak ("Peak 1" or "Peak 2") of the starting material utilized was as indicated in the table.

[0259] Table 7

[0260]

[0261]

[0262] Method 4: Stepwise silver salt method for the synthesis of mixed linkers

[0263] Representative Procedure for Perfluorophenyl 5-(((2-(Butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0264]

[0265] Step 1: Preparation of silver (I) perfluorophenyl 5-(difluoro(hydroxy(2-(pivaloylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (Difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate was synthesized from (difluoro(2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid using the method described for the synthesis of silver ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonate (I) in Step 1, Method 3.

[0266] To a suspension of ((2-((perfluorophenoxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonatesilver(I) (647 mg, 940 μmol, 1.0 equiv) in anhydrous toluene (10 mL) was added 2,2-dimethylpropanesulfate S-(2-iodoethyl)ester (310 mg, 1.14 mmol, 1.2 equiv) dropwise. After complete addition of the alcohol, the resulting mixture was stirred at room temperature for an additional 12 hours. The heterogeneous mixture was filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by reverse phase chromatography to give perfluorophenyl 5-(difluoro(hydroxy(2-(pivaloylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (240 mg, 388 μmol, 41% yield). LCMS (ESI) m / z=617 [MH] - .

[0267] Step 2: Preparation of perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate

[0268] The synthesis of perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate used a similar method as outlined above. Perfluorophenyl 5-(difluoro(hydroxy(2-(pivaloylthio)ethoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (190 mg, 307 μmol, 1.0 equiv), AgNO 3 (207 mg, 1.22 mmol, 4.0 equiv) and S-(2-iodoethyl)thiobutyrate (94.9 mg, 368 μmol, 1.2 equiv) produced perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate (55.0 mg, 73.4 μmol, 24%) as a white solid. LCMS (ESI) m / z=749 [M+H] + .

[0269] The following intermediates in Table 8 were prepared using a similar protocol described above for the synthesis of perfluorophenyl 5-(((2-(butyrylthio)ethoxy)(2-(pivaloylthio)ethoxy)phosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylate and utilizing the appropriate advanced intermediates as starting materials.

[0270] Table 8

[0271]

[0272] Preparation of 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0273]

[0274] Step 1: Preparation of allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0275] 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (1.0 g, 3.0 mmol, 1.0 equiv) and K 2 CO 3 To a suspension of 3-bromoprop-1-ene (839 mg, 6.1 mmol, 2.0 eq.) in DMF (20 mL) was added 3-bromoprop-1-ene (440 mg, 3.6 mmol, 1.2 eq.). The mixture was stirred at room temperature for 14 h and poured into water (30 mL). The mixture was extracted with EtOAc (25 mL x 3). The combined organic layers were purified by Na 2 SO 4 After drying, filtering and concentrating under reduced pressure, the residue was purified by column chromatography to give allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (0.980 g, 2.7 mmol, 88% yield) as a pale yellow solid. LCMS (ESI) m / z=369 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )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,1H),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).

[0276] Step 2: Preparation of ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid

[0277] To allyl 5-((diethoxyphosphoryl)methyl)benzo[b]thiophene-2-carboxylate (980 mg, 2.7 mmol, 1.0 equiv) was added to CH 2 Cl 2 (15 mL) was added bromotrimethylsilane (3 mL). The mixture was stirred at room temperature for 14 hours and then concentrated under reduced pressure. The resulting residue was washed with H 2 O (5 mL), and the resulting precipitate was filtered. The filter cake was washed with H 2 O (5 mL x 2), and dried under reduced pressure to give ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (0.710 g, 2.3 mmol, yield 86%) as a white solid. LCMS (ESI) m / z=313 [M+H] + ; 1 H NMR (400 MHz, DMSO-d 6 )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).

[0278] Step 3: Preparation of allyl 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0279] To ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (2.80 g, 8.96 mmol, 1 eq) and catalytic DMF (1 drop) was stirred in anhydrous CHCl. 2 Cl 2 (50 mL) in a cooled (0 °C) solution (under constant N 2 (g) oxalyl chloride (3.40 g, 26.8 mmol, 3 eq.) was added to the mixture under a stream of 2% argon (g). After the gas effervescence ceased, the mixture was warmed to 40°C. After 2 hours, the mixture was cooled to room temperature and concentrated in vacuo to give a yellow solid. The solid was then diluted in CH 2 Cl 2 (50 mL) and cooled to 0° C. To the cooled solution were added phenol (0.843 g, 8.96 mmol, 1 eq) and Et 3N (4.53 g, 44.8 mmol, 5 eq.). After complete addition, the mixture was warmed to room temperature and stirred for 1 hour, followed by the introduction of (2S)-2-aminopropionic acid propan-2-yl ester (1.75 g, 13.4 mmol, 1.5 eq.) into the mixture. After stirring for another 2 hours, the mixture was concentrated to dryness.

[0280] The residue was purified by C18 column (elution, 50%-80% acetonitrile in water) to give allyl 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (2.23 g, 4.44 mmol, 49.6% yield) as a white solid. LCMS (ESI) m / z = 502.0 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )δ8.05(d,J=6.9Hz,1H),7.91–7.80(m,2H),7.53–7.43(m,1H),7.29(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).

[0281] Step 4: Preparation of 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0282] Allyl 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (90 mg, 0.1794 mmol, 1 eq), pyrrolidine (12.7 mg, 179 μmol, 1 eq), Pd(PPh 3 ) 4 (10.3 mg, 8.97 μmol, 0.05 eq.) in CH 2 Cl 2 The solution in N 2(g) and stirred. After 2 hours, the reaction was concentrated in vacuo. The residue was purified by C18 column (elution, 30%-70% acetonitrile in water) to give 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (64.0 mg, 0.1386 mmol, 77.4% yield) as a white solid. LCMS (ESI) m / z=462.1[M+H] + .

[0283] The following intermediates in Table 9 were prepared using the synthesis of 5-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid described above and utilizing appropriate starting materials and modifications.

[0284] Table 9

[0285]

[0286] Synthesis scheme: Preparation of 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid

[0287]

[0288] Step 1: Benzyl 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylate

[0289] To a solution of 5-[(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (200 mg, 0.4779 mmol, 1 equivalent) in tetrahydrofuran (10 mL) was added dropwise bis(trimethylsilylamide) sodium solution (1M in THF) (1.43 mL, 1.43 mmol, 3.0 equivalents) at -78 ° C. The reaction was stirred at -78 ° C for 5 minutes, and then 1H-benzotriazole-1-methanol (142 mg, 0.955 mmol, 2 equivalents) was added in one go. The reaction was stirred at -78 ° C for 2 hours. The reaction mixture was quenched with saturated aqueous ammonium chloride solution (10 mL) at -78 ° C, and the ice bath was removed. The product was extracted with ethyl acetate (3x30 mL). The combined extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by reverse phase chromatography on 50 g C 18 Purification was performed on a cartridge with a gradient elution 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 thick oil. LCMS: m / z = 449.2 (M+H)+ ; 1 HNMR (400 MHz, DMSO-d 6 )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,1H),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).

[0290] Step 2: 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid

[0291] A mixture of 10% palladium on carbon (50% wet) (80 mg, 0.03758 mmol, 0.163 equiv) and 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid benzyl ester 5-[1-(diethoxyphosphoryl)-2-hydroxyethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (38 mg, 0.08473 mmol, 1 equiv) 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 was filtered through diatomaceous earth and eluted with MeOH. The filtrate was concentrated under reduced pressure. The crude residue was purified by reverse phase chromatography on 50 g C 18 The product was purified by eluting with 5-80% MeCN / water (containing 0.1% formic acid) on a cartridge, 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 (400 MHz, DMSO-d 6 )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).

[0292] Synthesis Scheme: Synthesis of 5-(1-(diethoxyphosphoryl)ethyl)benzo[b]thiophene-2-carboxylic acid and 5-(2-(diethoxyphosphoryl)propan-2-yl)benzo[b]thiophene-2-carboxylic acid

[0293]

[0294] Step 1: rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester and 5-[2-(diethoxyphosphoryl)propan-2-yl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0295] To a solution of 5-[(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (200 mg, 0.4779 mmol, 1 eq.) and methyl iodide (88.5 μL, 1.43 mmol, 3 eq.) in tetrahydrofuran (5 mL) was added dropwise bis(trimethylsilylamide) sodium solution (1 M in THF) (1.43 mL, 1.43 mmol, 3 eq.) at -78 °C. The mixture was stirred at -78 °C for 2 hours. The ice bath was removed and the reaction was stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated aqueous ammonium chloride solution (10 mL). The product was extracted with ethyl acetate (3 x 30 mL). The combined extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by reverse phase chromatography on 50 g C 18 Purification was performed on a cartridge eluting 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)propan-2-yl]-1-benzothiophene-2-carboxylic acid benzyl ester (22.0 mg, 0.5087 mmol, 10.6%) as a clear oil. rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester LCMS: m / z=433.0 (M+H) + . 1 H NMR (400 MHz, DMSO-d 6 )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).

[0296] Benzyl 5-[2-(diethoxyphosphoryl)propan-2-yl]-1-benzothiophene-2-carboxylate: LCMS: m / z=447.0 (M+H)+ . 1 H NMR (400 MHz, DMSO-d 6 )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).

[0297] Step 2: rac-5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid

[0298] A mixture of 10% palladium on carbon (50% wet) (80 mg, 0.03758 mmol, 0.163 equiv) and 5-[1-(diethoxyphosphoryl)ethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (100 mg, 0.2312 mmol, 1 equiv) 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 was filtered through diatomaceous earth and eluted with THF. The filtrate was concentrated under reduced pressure. The crude residue was purified by reverse phase chromatography on 50 g C 18 Purification was performed on a cartridge with a gradient elution 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) + .

[0299] Step 3: 5-[2-(diethoxyphosphoryl)propan-2-yl]-1-benzothiophene-2-carboxylic acid

[0300] A mixture of 10% palladium on carbon (50% wet) (5mg, 0.002349mmol, equivalent) and 5-[2-(diethoxyphosphoryl)propane-2-yl]-1-benzothiophene-2-carboxylate (33mg, 0.07390mmol, 1 equivalent) in tetrahydrofuran (2mL) was degassed with nitrogen for 5 minutes. Hydrogen was bubbled for 5 minutes, and the reaction was then stirred at room temperature under hydrogen (1atm). The reaction was filtered on a syringe filter and eluted with MeOH. The filtrate was concentrated under reduced pressure. The crude residue was diluted in tetrahydrofuran (2mL). 10% carbon palladium (50% wet) (20mg, 0.009396mmol, 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 (1atm). The reaction was filtered on 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 on carbon (50% wet) (20 mg, 0.009396 mmol, 0.13 equiv) was then added with nitrogen bubbling, followed by hydrogen bubbling for 5 minutes. The reaction was stirred at room temperature under hydrogen (1 atm) for 40 hours. The reaction was filtered on a syringe filter and eluted with MeOH. The filtrate was concentrated under reduced pressure to give crude 5-[2-(diethoxyphosphoryl)propan-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) + .

[0301] Synthesis scheme: Preparation of benzyl 5-{[bis({[(2-methoxyethoxy)carbonyl]oxy}methoxy)phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate

[0302]

[0303] Step 1: 2-Methoxyethyl Chloromethyl Carbonate

[0304] A solution of 2-methoxyethane-1-ol (1 g, 13.1 mmol, 1 eq.) and chloroacetyl chloride (1.68 g, 13.1 mmol, 1 eq.) in diethyl ether (30 mL) was cooled to 0 ° C. under nitrogen. Pyridine (1.04 mL, 13.1 mmol, 1.0 eq.) was added dropwise, and the reaction was then stirred at 0 ° C. for 15 minutes, followed by stirring at room temperature for 16 hours. The white suspension was filtered and rinsed with diethyl ether (30 mL). The filtrate was washed with 1N HCl (20 mL) and water (2 x 20 mL), then 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%) as a clear liquid:1 H NMR (400 MHz, CDCl 3 )δ5.76(s,2H),4.42-4.37(m,2H),3.68-3.64(m,2H),3.42(s,3H).

[0305] Step 2: Benzyl 5-{[bis({[(2-methoxyethoxy)carbonyl]oxy}methoxy)phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate

[0306] Water (2 mL) containing sodium hydroxide (59.9 mg, 1.50 mmol, 2 eq) was added dropwise to a stirred solution of ({2-[(benzyloxy)carbonyl]-1-benzothiophene-5-yl}difluoromethyl)phosphonic acid (300 mg, 0.7531 mmol, 1 eq) in water (15 mL). When the mixture reached a pH of about 8, silver nitrate (382 mg, 2.25 mmol, 3 eq) was added in one go. After standing at room temperature for 2 hours, the gray suspension was cooled to 0 ° C. The precipitate was collected by filtration, washed with water, dissolved in MeCN, dried under reduced pressure, and then dried under high vacuum for 2 hours. The solid was suspended in anhydrous toluene (10 mL), and 2-methoxyethyl chloromethyl carbonate (379 mg, 2.25 mmol, 3 eq) was added. The mixture was stirred at room temperature for 18 hours and then heated to 50 ° C for 5 days. The mixture was adsorbed on silica gel and concentrated under reduced pressure. The crude residue was purified by flash chromatography on a 24 g silica gel cartridge eluting with 0-90% EtOAc / heptane to give benzyl 5-{[bis({[(2-methoxyethoxy)carbonyl]oxy}methoxy)phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate (290 mg, 0.4377 mmol, 58.2%) as a clear oil. LCMS: m / z = 663.2 (M+H) + . 1 H NMR (400 MHz, CDCl 3 )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),5.7 5(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).

[0307] Step 3: Benzyl 5-{[bis({[(2-methoxyethoxy)carbonyl]oxy}methoxy)phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate

[0308] 10% Palladium on carbon (50% wet) (145 mg, 0.1353 mmol, equiv) was added to a mixture of benzyl 5-{[bis({[(2-methoxyethoxy)carbonyl]oxy}methoxy)phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate (145 mg, 0.2188 mmol, 1 equiv) 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 celite, rinsed with 2-MeTHF, and then concentrated under reduced pressure to give crude benzyl 5-{[bis({[(2-methoxyethoxy)carbonyl]oxy}methoxy)phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate (144 mg, 0.2188 mmol, 99%) as a grey solid. LCMS: m / z=573.2 (M+H) + .

[0309] Synthesis scheme: Preparation of 5-{cyano[ethoxy(hydroxy)phosphoryl]methyl}-1-benzothiophene-2-carboxylic acid

[0310] Step 1: 5-[Cyano(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylate

[0311] To a solution of diethyl (cyanomethyl)phosphonate (127 mg, 722 μmol, 1.2 equivalents) in anhydrous 1,2-dimethylethane (5 mL) was added sodium hydride (60% in mineral oil) (50.3 mg, 1.26 mmol, 2.1 equivalents) under nitrogen. The reaction was stirred at room temperature for 10 minutes, 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 hours. The reaction mixture was filtered through diatomaceous earth (eluted with DCM), and the filtrate was concentrated under reduced pressure. The crude residue was purified by reverse phase chromatography on 50 g C 18 Purification was performed on the cartridge using a 5-80% MeCN / water gradient elution. The combined fractions were concentrated to give ethyl 5-[cyano(dioxyphosphoryl)methyl]-1-benzothiophene-2-carboxylate (50.0 mg, 131 μmol, 21.8%) as an orange solid. LCMS: m / z=382.2 (M+H) + . 1 H NMR (400 MHz, DMSO-d 6)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).

[0312] Step 2: 5-{Cyano[ethoxy(hydroxy)phosphoryl]methyl}-1-benzothiophene-2-carboxylic acid

[0313] To a solution of ethyl 5-[cyano(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylate (80 mg, 209 μmol, 1 equiv) in acetonitrile (4 mL) was added hydrochloric acid (6 mL, 3 N). 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 directly purified by reverse phase chromatography using 50 g C 18 The cartridge was purified with a gradient elution of MeCN / water (5% for 3CV then 5 to 100% for 18CV) to give 5-{cyano[ethoxy(hydroxy)phosphoryl]methyl}-1-benzothiophene-2-carboxylic acid (52.0 mg, 159 μmol, 76.5%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6 )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).

[0314] Synthesis scheme: Preparation of rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid

[0315]

[0316] Step 1: rac-5-[(diethoxyphosphoryl)(hydroxy)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0317] A solution of benzyl 5-[(diethoxyphosphoryl)methyl]-1-benzothiophene-2-carboxylate (2.4 g, 5.73 mmol, 1 eq) in tetrahydrofuran (75 mL) was cooled to -78 °C and then 2-(phenylsulfonyl)-3-phenyloxaziridine (2.97 g, 11.4 mmol, 2 eq) was added, followed by dropwise addition of sodium bis(trimethylsilylamide) solution (1.0 M in THF) (11.4 mL, 11.4 mmol, 2 eq) (internal T o = -75°C to -69°C). After the addition of the base, a dark purple solution was observed, which quickly turned orange. The mixture was stirred at -78°C for 10 minutes. The reaction mixture was washed with saturated NH 4 Cl aqueous solution (50mL) quenched, and then the dry ice bath was removed. EtOAc (75mL) and water (25mL) were added, and the mixture was stirred for 30 minutes (until the internal temperature reached 15 ° C). The two phases were separated, and the aqueous layer was then back-extracted with EtOAc (125mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, adsorbed on silica gel, and concentrated under reduced pressure. The crude material was combined with another batch (6.42mmol, a total of 12.15mmol) and purified by flash chromatography on a 330g silica gel cartridge eluted with 20-100% EtOAc / heptane to obtain rac-5-[(diethoxyphosphoryl)(hydroxy)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (3.69g, 8.49mmol, 70%) as a white viscous solid. LCMS: m / z=869.4 (2M+H) + . 1 H NMR (400 MHz, CDCl 3 )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),7.46-7.3 5(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).

[0318] Step 2: rac-5-[(diethoxyphosphoryl)(fluoro)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0319] To a solution of rac-5-[(diethoxyphosphoryl)(hydroxy)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (1.56 g, 3.59 mmol, 1 equivalent) in dichloromethane (30 mL) was added (diethylamino) sulfur trifluoride (568 μL, 4.30 mmol, 1.2 equivalents) at -78 ° C under nitrogen. The reaction was stirred at -78 ° C for 15 minutes. The reaction was quenched by adding saturated aqueous sodium bicarbonate (50 mL), and the product was then 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 purified by reverse phase chromatography on 275 g C 18 Purification was performed on a cartridge eluting with 5-80% MeCN / water to give benzyl rac-5-[(diethoxyphosphoryl)(fluoro)methyl]-1-benzothiophene-2-carboxylate (650 mg, 1.48 mmol, 41.6%) as a clear thick oil. LCMS: m / z = 873.2 (2M+H) + . 1 H NMR (400 MHz, CDCl 3 )δ8.12(s,1H),8.02-7.99(m,1H),7.92(d,J=8.7Hz,1H),7.61(d,J=8.7Hz,1H),7.51-7.51(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).

[0320] Step 3: rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid benzyl ester

[0321] To a solution of 5-[(diethoxyphosphoryl)(fluoro)methyl]-1-benzothiophene-2-carboxylic acid benzyl ester (100 mg, 0.2291 mmol, 1 eq.) and methyl iodide (42.7 μL, 687 μmol, 3 eq.) in tetrahydrofuran (4 mL) was added dropwise bis(trimethylsilylamide) sodium solution (1M in THF) (458 μL, 458 μmol, 2 eq.) at -78 °C. The mixture was stirred at -78 °C for 5 minutes. The reaction was quenched with 1M 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 purified by reverse phase chromatography on 50 g C 18Purification was performed on the cartridge eluting with 5-80% MeCN / water (containing 0.1% formic acid). The fractions were combined, concentrated under reduced pressure, and then freeze-dried to give rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (67.0 mg, 0.1487 mmol, 65.0%) as a pale yellow oil. LCMS: m / z=451.2 (M+H) + . 1 H NMR (400 MHz, CDCl 3 )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).

[0322] Step 4: rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid

[0323] A mixture of 10% palladium on carbon (50% wet) (99 mg, 0.04651 mmol, 0.21 equiv) and 5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid benzyl ester 5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid benzyl ester (99 mg, 0.2197 mmol, 1 equiv) in tetrahydrofuran (6 mL) was degassed with nitrogen for 5 minutes. Hydrogen was bubbled for 5 minutes, and the reaction was then stirred at room temperature under hydrogen (1 atm) for 20 hours. The reaction was filtered through a syringe filter and rinsed with THF. The filtrate was concentrated under reduced pressure to give the crude product rac-5-[1-(diethoxyphosphoryl)-1-fluoroethyl]-1-benzothiophene-2-carboxylic acid (78.9 mg, 0.2197 mmol, 100%) as a clear oil. LCMS: m / z=361.2 (M+H) + .

[0324] Synthesis scheme: Preparation of rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid

[0325]

[0326] Step 1: 7-Bronaphthalene-2-carboxylic acid

[0327] A solution of 2,7-dibromonaphthalene (2 g, 6.99 mmol, 1 eq) in tetrahydrofuran (24 mL) was cooled to -78 °C under nitrogen. n-Butyl lithium solution (1.6 M in hexanes) (4.58 mL, 7.33 mmol, 1.05 eq) was added dropwise. The reaction was stirred at -78 °C for 15 min, then CO was added. 2 The ice bath was removed and the reaction was heated under CO 2 In the case of bubbling, stir for 1 hour.By adding 1N HCl (50mL, pH=2), the reaction is quenched, and then the product is extracted with EtOAc (3x75mL).The combined organic layer is washed with water (50mL), brine (50mL), then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure.The thick residue is ground in heptane, and the gained solid is filtered and dried, to obtain 7- bromonaphthalene -2- carboxylic acid (1.27g, 5.05mmol, 72.5%) as a white solid. 1 H NMR (400 MHz, CDCl 3 )δ13.21(s,1H),8.61(s,1H),8.44(s,1H),8.08-7.96(m,3H),7.81-7.76(m,1H).

[0328] Step 2: tert-Butyl 7-bromonaphthalene-2-carboxylate

[0329] N,N-dimethylformamide dibutyl acetal (3.80mL, 15.9mmol, 4 equivalents) was added to a stirred suspension of 7-bromonaphthalene-2-carboxylic acid (1g, 3.98mmol, 1 equivalent) in toluene (9mL) at 90°C within 15 minutes. The reaction was stirred at 90°C for 2 hours, and then an additional amount of N,N-dimethylformamide dibutyl acetal (0.9mL, 3.38mmol, 1 equivalent) was added dropwise, 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 on silica gel and concentrated under reduced pressure. The crude residue was purified by flash chromatography on a 40g silica gel cartridge with a gradient elution of 0-10% EtOAc / heptane to obtain tert-butyl 7-bromonaphthalene-2-carboxylate (1.11g, 3.61mmol, 90.9%) as a white solid. 1 HNMR (400MHz, CDCl 3 )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).

[0330] Step 3: tert-Butyl 7-methylnaphthalene-2-carboxylate

[0331] By tert-butyl 7-bromonaphthalene-2-formate (1.11g, 3.61mmol, 1 equivalent), trimethylcyclotriboroxane (906mg, 7.22mmol, 2 equivalents), tetrakis (triphenylphosphine) palladium (834mg, 722μmol, 0.2 equivalents) and potassium carbonate (1.99g, 14.4mmol, 4 equivalents) in 1,4-dioxane (20mL) mixture degassed 5 minutes, and then sealed and heated to 110 ° C, for 20 hours. The reaction is diluted with EtOAc (20mL) and adsorbed on silica gel. The crude residue is purified by flash chromatography eluting with 0-10% EtOAc / heptane to obtain tert-butyl 7-methylnaphthalene-2-formate (808mg, 3.33mmol, 92.4%) as a white crystalline solid. 1 H NMR (400 MHz, CDCl 3 )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).

[0332] Step 4: tert-Butyl 7-(bromomethyl)naphthalene-2-carboxylate

[0333] 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 to reflux and stirred at this temperature for 20 hours. The precipitate was filtered out, washed with carbon tetrachloride (10 mL), and the filtrate was then adsorbed on silica gel and concentrated under reduced pressure. The crude material was purified by flash chromatography on 80 g silica gel cartridges eluted with 0-10% EtOAc / heptane to obtain tert-butyl 7-(bromomethyl)naphthalene-2-carboxylate (720 mg, 2.24 mmol, 67.9%) as a white solid containing residual starting material (approximately 25%). 1 H NMR (400 MHz, CDCl 3 )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).

[0334] Step 5: tert-Butyl 7-[(diethoxyphosphoryl)methyl]naphthalene-2-carboxylate Tert-butyl 7-(bromomethyl)naphthalene-2-carboxylate (720 mg, 2.24 mmol) was suspended in triethyl phosphite (2 mL, 11.6 mmol) and the reaction mixture was heated to reflux for 1.5 hours (becoming a solution at 110°C). The reaction was cooled to room temperature and purified by reverse phase chromatography on 50 g C 18 Purification was performed directly on the column using a gradient of 5-80% MeCN / water to give tert-butyl 7-[(diethoxyphosphoryl)methyl]naphthalene-2-carboxylate (580 mg, 1.53 mmol, 68.4%) as a light yellow thick oil. LCMS: m / z = 379.3 (M+H) + . 1 H NMR (400 MHz, CDCl 3 )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).

[0335] Step 6: tert-butyl rac-7-[(diethoxyphosphoryl)(hydroxy)methyl]naphthalene-2-carboxylate

[0336] To a solution of tert-butyl 7-[(diethoxyphosphoryl)methyl]naphthalene-2-carboxylate (200mg, 0.5285mmol, 1 equivalent) in tetrahydrofuran (5mL) was added dropwise bis(trimethylsilylamide)sodium solution (1M in THF) (792μL, 792μmol, 1.5 equivalents) at -78°C. The mixture was stirred for 2 minutes, and then 2-(benzenesulfonyl)-3-phenyloxaziridine (274mg, 1.05mmol, 2.0 equivalents) was added in batches. The dark red solution was stirred at -78°C for 20 minutes. The reaction mixture was quenched with saturated aqueous ammonium chloride solution (50mL) at -78°C, and then the ice bath was removed. EtOAc (25mL) and water (25mL) were added, and the mixture was stirred for 30 minutes. Each phase was separated, and then the water layer was stripped with EtOAc (2x 25mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography on a 24 g silica gel cartridge eluting 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%) as a white sticky solid. LCMS: m / z=394.6 (M+H) + . 1 H NMR (400 MHz, CDCl 3 )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).

[0337] Step 7: tert-Butyl rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylate

[0338] To a solution of tert-butyl 7-[(diethoxyphosphoryl)(hydroxy)methyl]naphthalene-2-carboxylate (140 mg, 0.3549 mmol, 1 equivalent) in dichloromethane (5 mL) was added dropwise a solution of (diethylamino) sulfur trifluoride (56.1 μL, 425 μmol, 1.2 equivalents) at -78 ° C under nitrogen. The reaction was stirred at -78 ° C for 20 minutes. The reaction was quenched by adding saturated aqueous sodium bicarbonate (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 purified by reverse phase chromatography on 50 g C 18 Purification was performed on a cartridge eluting with a gradient of 5-80% MeCN / water to give tert-butyl rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylate (80.0 mg, 0.2018 mmol, 57.1%) as a clear oil. LCMS: m / z = 397.4 (M+H) + . 1 H NMR (400 MHz, CDCl 3 )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),7.7 2(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).

[0339] Step 8: rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylic acid

[0340] To a solution of tert-butyl rac-7-[(diethoxyphosphoryl)(fluoro)methyl]naphthalene-2-carboxylate (80 mg, 0.2018 mmol) in dichloromethane (4 mL) was added trifluoroacetic acid (1 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, 1 eq) as a clear oil. LCMS: m / z=341.2 (M+H) + .

[0341] Synthesis scheme: Preparation of 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid

[0342]

[0343] Step 1: (2S)-2-Methyl 1-chloromethyl pyrrolidine-1,2-dicarboxylate

[0344] Chloroacetyl chloride (777 mg, 6.03 mmol) was added to a solution of (2S)-pyrrolidine-2-carboxylic acid methyl ester hydrochloride (1.00 g, 6.03 mmol) and N, N-diisopropylethylamine (818 mg, 6.33 mmol, 1.05 equivalents) in dichloromethane (15 mL) at 0 ° C under a nitrogen atmosphere. After stirring for 0.5 hours, the reaction mixture was washed with 1N aqueous hydrochloric acid solution (50 mL), water (50 mL) and brine in sequence. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The product was purified by flash chromatography using a 25 g silica gel cartridge eluted with a gradient of ethyl acetate / heptane (0 to 50%). The fractions were combined and concentrated under reduced pressure to obtain (2S)-pyrrolidine-1,2-dicarboxylic acid 1-chloromethyl 2-methyl ester (701 mg, 3.16 mmol, 52.7%). 1 HNMR:(400MHz,CDCl 3 )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).

[0345] Step 2: (2S)-pyrrolidine-1,2-dicarboxylic acid 1-{[({2-[(benzyloxy)carbonyl]-1-benzothiophen-5-yl}difluoromethyl)({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]oxy}methyl ester 2-methyl

[0346] Sodium hydroxide (59.9 mg, 1.50 mmol) in water (2 mL) was added dropwise to a stirred solution of ({2-[(benzyloxy)carbonyl]-1-benzothiophen-5-yl}difluoromethyl)phosphonic acid (300 mg, 753 μmol, 1 eq) in water (15 mL), followed by the addition of silver nitrate (382 mg, 2.25 mmol). After standing at room temperature for 2 h, the suspension was cooled to 0 °C. The precipitate was collected by filtration, washed with water, evaporated with toluene (2x), and dried under high vacuum. The powder was suspended in anhydrous toluene (10 mL), and (2S)-2-methyl 1-chloromethyl pyrrolidine-1,2-dicarboxylate (582 mg, 2.63 mmol, 3.5 eq) was added. The mixture was stirred at room temperature for 18 h. The crude product was directly purified by normal phase chromatography using a 12 g silica gel cartridge eluting with an ethyl acetate / heptane gradient (0-100%, 18 CV) to afford 2-methyl (2S)-pyrrolidine-1,2-dicarboxylate 1-{[({2-[(benzyloxy)carbonyl]-1-benzothiophen-5-yl}difluoromethyl)({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]oxy}methyl ester (90.0 mg, 117 μmol, 15.5%) as a white solid. 1 H NMR: (400MHz, CDCl 3 )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).

[0347] Step 3: 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid

[0348] Palladium on carbon (10% loading, 50% wet support) (62.1 mg, 58.5 μmol, 0.5 eq) was added to a mixture of 1-{[({2-[(benzyloxy)carbonyl]-1-benzothiophen-5-yl}difluoromethyl)({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]oxy}methyl 2-methyl ester (90 mg, 117 μmol, 1 eq) in anhydrous tetrahydrofuran (10 mL). Hydrogen was bubbled into the suspension for 2 minutes, and the reaction mixture was then stirred under 1 atm (balloon) hydrogen for 16 hours. The reaction mixture was filtered over celite and rinsed with Me-THF. The filtrate was concentrated in vacuo to give crude 5-{[bis({[(2S)-2-(methoxycarbonyl)pyrrolidine-1-carbonyloxy]methoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid (95.0 mg, 140 μmol, 119%) as a colorless oil. 1 H NMR: (400MHz, CDCl 3 )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).

[0349] Preparation of 5-(((Benzylamino)(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid:

[0350]

[0351] Step A: Allyl 5-(((Benzylamino)(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate

[0352] Oxalyl chloride (814.4 mg, 6.4 mmol, 10 equiv) was added dropwise to a solution of ((2-((allyloxy)carbonyl)benzo[b]thiophen-5-yl)methyl)phosphonic acid (200 mg, 0.64 mmol, 1.0 equiv) in anhydrous DCM (6 mL) and DMF (1 drop) at 0°C. The reaction was heated to 40°C and then stirred for an additional 1 to 2 hours. The reaction was monitored by pipetting a small amount of the crude sample and quenching with MeOH to ensure that 5-((dichlorophosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid bisCl phosphoryl chloroallyl ester (bismethoxyphosphonate observed by LCMS) had been completely formed. Upon completion, excess oxalyl chloride and solvent were removed under reduced pressure and the residue was redissolved in anhydrous DCM (5 mL). A solution containing BnNH was then added to the solution at -40°C. 2 (64.8 mg, 0.64 mmol, 1.0 equiv) in anhydrous DCM (2 mL) and Et 3 N (194.32 mg, 1.92 mmol, 3.0 equiv). The reaction was monitored by pipetting a small amount of the crude sample and quenching with MeOH to ensure that the majority of the product was allyl 5-(((benzylamino)chlorophosphonyl)methyl)benzo[b]thiophene-2-carboxylate (monomethoxyphosphonate observed by LCMS). Anhydrous DCM (2 mL) containing L-alanine isopropyl ester (107.2 mg, 0.64 mmol, 1.0 equiv) was then added to the solution at -40 °C. The reaction was allowed to warm to room temperature and stirred for an additional 2 hours. Upon completion, the reaction was removed by adding H 2 O (10 mL) and extracted with DCM (10 mL x 3). The organic layers were combined and washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The mixture was dried and concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography to give allyl 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (100 mg, 194.33 μmol, 30%). LCMS (ESI): m / z=515.2 [M+H] + .

[0353] Step B: 5-(((Benzylamino)(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid

[0354] Allyl 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylate (100 mg, 194 μmol, 1 eq), Pd(PPh 3 )4 A solution of (22.47 mg, 19.4 μmol, 0.1 eq) and pyrrolidine (13.8 mg, 194 μmol, 1 eq) 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 flash column chromatography to give 5-(((benzylamino)(((S)-1-isopropoxy-1-oxopropyl-2-yl)amino)phosphoryl)methyl)benzo[b]thiophene-2-carboxylic acid (70 mg, 147.5 μmol, 76%). LCMS(ESI):m / z=475.1[M+H] + .

[0355] Synthesis scheme: Preparation of 4-nitrophenyl 5-[(bis{[(propoxycarbonyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylate

[0356]

[0357] Step 1: Propyl chloromethyl carbonate

[0358] A solution of propan-1-ol (1.24 mL, 16.5 mmol, 1 eq.) and chloroacetyl chloride (1.46 mL, 16.5 mmol, 1 eq.) in diethyl ether (50 mL) was cooled to 0 ° C. under nitrogen. Pyridine (1.32 mL, 16.5 mmol, 1.0 eq.) was added dropwise, and the reaction mixture was then stirred at 0 ° C. for 15 minutes, followed by stirring at room temperature for 3 hours. The white suspension was filtered and rinsed with diethyl ether (20 mL). The filtrate was washed with 1N HCl (20 mL) and water (2 x 15 mL), and then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give chloromethyl carbonate propyl ester (1.26 g, 8.25 mmol, 50.1%) as a clear oil: 1 H NMR (400 MHz, CDCl 3 )δ5.74(s,2H),4.25-4.17(m,2H),1.74(sxt,J=7.1Hz,2H),1.02-0.94(m,3H).

[0359] Step 2: 4-Nitrophenyl 5-[(bis{[(propoxycarbonyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylate

[0360] A solution of H containing sodium hydroxide (37.1 mg, 930 μmol, 2 eq.) was added. 2To a stirred suspension of [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid (200 mg, 465 μmol, 1 eq.) in water (4 mL) was added dropwise 1% 4-nitropropene (2-[(4-nitrophenoxy)carbonyl]-1-benzothiophene-5-yl})methyl]phosphonic acid (200 mg, 465 μmol, 1 eq.) O (1 mL) was added dropwise to a stirred suspension of [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophene-5-yl})methyl]phosphonic acid (200 mg, 465 μmol, 1 eq.) in water (4 mL). When the mixture reached a pH of about 8, the product began to precipitate, and silver nitrate (236 mg, 1.39 mmol, 3 eq.) was added in one portion. After standing at 0° C. for 2 hours, the yellow precipitate was collected by filtration, washed with water, and dried in vacuo. The powder was suspended in anhydrous toluene (10 mL), and propyl chloromethyl carbonate (282 mg, 1.85 mmol, 4 eq.) 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 an EtOAc / heptane gradient (0 to 60% in 18 CV) to give 4-nitrophenyl 5-[(bis{[(propoxycarbonyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylate (10 mg, 15.1 μmol, 3.25%) as a white semisolid: LCMS: m / z=662.2 (M+H) + . 1 H NMR (400 MHz, DMSO-d 6 )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).

[0361] Synthesis scheme: 4-nitrophenyl 5-({bis[({[(1-methoxy-2-methylpropan-2-yl)oxy]carbonyl}oxy)methoxy]phosphoryl}difluoromethyl)-1-benzothiophene-2-carboxylate

[0362]

[0363] Step 1: 1-methoxy-2-methylpropan-2-yl chloromethyl carbonate

[0364] A solution of 1-methoxy-2-methylpropan-2-ol (2g, 19.2mmol, 1 equivalent) and chloroacetyl chloride (1.7mL, 19.2mmol, 1 equivalent) in diethyl ether (60mL) was cooled to 0°C under nitrogen. Pyridine (1.53mL, 19.2mmol, 1.0 equivalent) was added dropwise, and the reaction mixture was then stirred at 0°C for 15 minutes, then at room temperature for 3 hours. The white suspension was filtered and rinsed with diethyl ether (20mL). The filtrate was washed with 1N HCl (30mL) and water (2x 30mL), and then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain chloromethyl carbonate 1-methoxy-2-methylpropan-2-yl ester (860mg, 4.37mmol, 22.8%) as a clear oil. 1 HNMR (400MHz, CDCl 3 )δ7.20-7.17(m,2H),5.04-5.00(m,2H),4.90(s,3H),3.03-2.99(m,6H).

[0365] Step 2: 4-Nitrophenyl 5-[(bis{[(propoxycarbonyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylate

[0366] A solution of H containing sodium hydroxide (37.1 mg, 930 μmol, 2 eq.) was added. 2 To a stirred suspension of [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid (200 mg, 465 μmol, 1 eq.) in water (5 mL) was added dropwise 1 mL of 4-nitropropene (1 mL). When the mixture reached a pH of about 8, the product began to precipitate, and silver nitrate (236 mg, 1.39 mmol, 3 eq.) was added in one portion. After standing at 0 °C for 2 hours, the white precipitate was collected by filtration, washed with water, and dried in vacuo. The powder was suspended in anhydrous toluene (10 mL), and chloromethyl carbonate 1-methoxy-2-methylprop-2-yl ester (318 mg, 1.62 mmol, 3.5 eq.) was added. The mixture was stirred at 50 °C for 18 hours. The crude mixture was directly purified by normal phase chromatography using a 24 g silica gel cartridge eluting with an EtOAc / heptane gradient (0 to 60% in 18 CV) to afford 4-nitrophenyl 5-({bis[({[(1-methoxy-2-methylpropan-2-yl)oxy]carbonyl}oxy)methoxy]phosphoryl}difluoromethyl)-1-benzothiophene-2-carboxylate (51.0 mg, 68.0 μmol, 14.6%) as a white semisolid. 1 H NMR (400 MHz, CDCl 3)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).

[0367] Synthesis scheme: Preparation of 4-nitrophenyl 5-[(bis{[(dipropylcarbamoyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylate

[0368]

[0369] Step 1: N,N-dipropylcarbamate

[0370] Dipropylamine (5.85 g, 57.9 mmol, 2.5 eq) was added to a solution of chloroacetyl chloride (2.06 mL, 23.2 mmol, 1 eq) in hexane (30 mL) under nitrogen at 0 ° C and stirred at the same temperature for 30 minutes. The reaction mixture was then diluted with ethyl acetate (50 mL). The phases were separated and the organic layer was washed with 1N hydrochloric acid (50 mL), water and brine in sequence. The organic layer was dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure. The product was purified by flash chromatography using an 80 g silica gel cartridge with a gradient elution of 0-10% ethyl acetate / heptane. The combined pure fractions were dried under reduced pressure to give N,N-dipropylcarbamic acid chloromethyl ester (2.89 g, 14.9 mmol, 64.3%) as a colorless oil. LCMS: m / z=194.2 (M+H) + . 1 H NMR (400 MHz, CDCl 3 )δ5.75(s,2H),3.21-3.11(m,4H),1.59-1.47(m,4H),0.87-0.82(m,6H).

[0371] Step 2: 4-Nitrophenyl 5-[(bis{[(dipropylcarbamoyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylate

[0372] A solution of H containing sodium hydroxide (37.1 mg, 930 μmol, 2 eq.) was added. 2 O (1 mL) was added dropwise to [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid (200 mg, 465 μmol, 1 eq) in H 2O (5mL) in a stirred suspension. When the mixture reaches pH of about 8, the product is precipitated, and silver nitrate (236mg, 1.39mmol, 3 equivalents) is added at one time. After standing at 0 ° C for 2 hours, the yellow precipitate is collected by filtration, washed with water, and dried in a vacuum. The powder is suspended in anhydrous toluene (10mL), and N, N-dipropylcarbamic acid chloromethyl ester (313mg, 1.62mmol, 3.5 equivalents) is added. The mixture is stirred at room temperature for 18 hours. The crude product is directly purified by normal phase chromatography using 12g silica gel cartridges with EtOAc / heptane gradient (0 to 60%, in 18CV) elution to obtain 5-[(bis{[(dipropylcarbamoyl)oxy]methoxy}phosphoryl)difluoromethyl]-1-benzothiophene-2-carboxylic acid 4-nitrophenyl ester (110mg, 147μmol, 31.8%) as a clear oil. 1 H NMR (400 MHz, DMSO-d 6 )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).

[0373] Synthesis scheme: 4-nitrophenyl 5-{[bis({1-[(propoxycarbonyl)oxy]ethoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate

[0374]

[0375] Step 1: 1-Chloroethyl Propyl Carbonate

[0376] A solution of propan-1-ol (2g, 33.2mmol, 1 equivalent) and 1-chloroethyl chloroformate (4.74g, 33.2mmol, 1 equivalent) in diethyl ether (100mL) was cooled to 0°C under nitrogen. Pyridine (2.66mL, 33.2mmol, 1.0 equivalent) was added dropwise, and the reaction mixture was then stirred at 0°C for 15 minutes, then at room temperature for 3 hours. The white suspension was filtered and rinsed with diethyl ether (20mL). The filtrate was washed with 1N HCl (20mL) and water (2x 15mL), and then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 1-chloroethyl propyl carbonate (4.62g, 27.7mmol, 83.5%) as a clear oil. 1 H NMR (400 MHz, CDCl 3)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).

[0377] Step 2: 4-Nitrophenyl 5-{[bis({1-[(propyloxycarbonyl)oxy]ethoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylate

[0378] A solution of H containing sodium hydroxide (37.1 mg, 930 μmol, 2 eq.) was added. 2 O (1 mL) was added dropwise to [difluoro({2-[(4-nitrophenoxy)carbonyl]-1-benzothiophen-5-yl})methyl]phosphonic acid (200 mg, 465 μmol, 1 eq) in H 2 O (4mL) in stirring suspension. When the mixture becomes clear (pH is about 8), silver nitrate (236mg, 1.39mmol, 2 equivalents) is added once. After standing for 2 hours at 0 ° C, the yellow precipitate is collected by filtration and dried in a vacuum. The powder is suspended in anhydrous toluene (10mL), and 1-chloroethyl propyl carbonate (269mg, 1.62mmol, 3.5 equivalents) is added. The mixture is stirred at room temperature for 18 hours. The crude product is directly purified by normal phase chromatography using 12g silica gel cartridges with EtOAc / heptane gradient (0 to 60%, in 18CV) elution to obtain 5-{[bis({1-[(propoxycarbonyl)oxy]ethoxy})phosphoryl]difluoromethyl}-1-benzothiophene-2-carboxylic acid 4-nitrophenyl ester (113mg, 163μmol, 35.3%) as a clear oil. LCMS: m / z=712.2 (M+Na) + .

[0379] Preparation of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylic acid

[0380]

[0381] Step A: tert-Butyl 2-(diethoxyphosphoryl)propionate

[0382] A mixture of tert-butyl 2-bromopropionate (3 g, 14.3 mmol, 1 eq.) and diethyl phosphite-2-yl cation-1-yl ester (2.82 g, 17.1 mmol, 1.2 eq.) was stirred at 110 °C for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography to give tert-butyl 2-(diethoxyphosphoryl)propionate (3.0 g, 11.3 mmol, 79%) as a white solid. LCMS (ESI): m / z=267 [M+H] + .

[0383] Step B: (E)-tert-butyl 3-(3-iodophenyl)-2-methacrylate

[0384] To a solution of tert-butyl 2-(diethoxyphosphoryl)propanoate (1.14 g, 4.30 mmol, 1 eq) in THF (10 mL) was added n-BuLi (1.72 mL, 4.30 mmol, 1.0 eq) at -78 °C under nitrogen, 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 eq) in THF (5 mL) was added, and after addition, the reaction mixture was allowed to warm to room temperature and stirred for 14 h. After completion, the reaction mixture was heated to room temperature by adding H 2 The reaction mixture was quenched with 1% 4% dHO (20 mL) and then extracted with EtOAc (15 mL x 3). The organic layers were combined and washed with brine (15 mL) and dried over anhydrous Na 2 SO 4 Dry and concentrate under reduced pressure. The residue was purified by silica gel flash column chromatography to give (E)-tert-butyl 3-(3-iodophenyl)-2-methacrylate (739.6 mg, 2.15 mmol, 50%) as a white solid. LC-MS (ESI) m / z=345 [M+H] + .

[0385] Step C: (E)-tert-butyl 3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylate

[0386] A solution of diethyl (bromodifluoromethyl)phosphonate (1.08 g, 4.06 mmol, 2 eq) and zinc (265 mg, 4.06 mmol, 2 eq) in DMAc (7 mL) was stirred at 60° C. for 1 hour under a nitrogen atmosphere. CuBr (582 mg, 4.06 mmol, 2 eq) was then added, and the mixture was stirred at 60° C. for another 1 hour. (E)-tert-butyl 3-(3-iodophenyl)-2-methacrylate (700 mg, 2.03 mmol, 1 eq) was added, and the mixture was stirred at 60° C. for another 12 hours. Upon completion, the suspension was filtered through The product was filtered through a pad of 4% paraformaldehyde and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography to give (E)-tert-butyl 3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylate (180 mg, 445 μmol, 22%) as a yellow oil. LCMS (ESI): m / z=405 [M+H] + .

[0387] Step D: (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylic acid

[0388] A solution of (E)-tert-butyl 3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)-2-methacrylate (180 mg, 445 μmol, 1 eq.) in a mixture of DCM (8 mL) and TFA (4 mL) was stirred at room temperature for 1 hour. Upon 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 was used directly in the next step without further purification. LCMS (ESI): m / z=349 [M+H] + .

[0389] Preparation of (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid

[0390]

[0391] Step A: (E)-tert-butyl 3-(3-iodophenyl)but-2-enoate

[0392] n-BuLi (1.72 mL, 4.06 mmol, 1.0 eq) was added to THF (20 mL) containing tert-butyl 2-(diethoxyphosphoryl)acetate (1.02 g, 4.06 mmol, 1 eq) at -78 ° C, and the mixture was stirred at -78 ° C for 0.5 hours. Then a solution of 1-(3-iodophenyl)ethan-1-one (1 g, 4.06 mmol, 1 eq) in THF (5 mL) was added dropwise to the reaction. After addition, the reaction mixture was allowed to warm to room temperature and stirred for 18 hours. After completion, the reaction mixture was heated to room temperature by adding H 2 O (20 mL) and extracted with EtOAc (20 mL x 3). The organic layers were combined and washed with brine (15 mL) and dried over anhydrous Na 2 SO 4Dry and concentrate under reduced pressure.The residue was purified by silica gel flash column chromatography to give (E)-tert-butyl 3-(3-iodophenyl)but-2-enoate (800 mg, 2.32 mmol, 58%) as an oil. 1 H NMR (400 MHz, CDCl 3 )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).

[0393] Step B: (E)-tert-butyl 3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoate

[0394] To a stirred suspension of Zn (301mg, 4.64mmol, 2 equivalents) in anhydrous DMAc (2mL) was slowly added a solution of diethyl (bromodifluoromethyl)phosphonate (1.23g, 4.64mmol, 2 equivalents) in DMAc (2mL) under a nitrogen atmosphere. The reaction mixture was stirred at 45 ° C for 2 hours, and then CuBr (665mg, 4.64mmol, 2 equivalents) was added, and the resulting mixture was stirred at room temperature for 45 minutes. A suspension of (E)-3-(3-iodophenyl) but-2-enoic acid tert-butyl ester (800mg, 2.32mmol, 1 equivalent) in DMAc (3mL) was added to the reaction mixture. The mixture was stirred at 45 ° C for 24 hours under a nitrogen atmosphere. The reaction mixture was then distributed between water and ether. The mixture was passed through diatomaceous earth and extracted with ether. The organic extract was washed with brine and anhydrous Na 2 SO 4 The solvent was removed in vacuo and the residue was purified by flash column chromatography on silica to give (E)-tert-butyl 3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoate (350 mg, 0.87 mmol, 37%) as an oil. 1 H NMR (400 MHz, CDCl 3 )δ7.67(d,J=13.2Hz,1H),7.63-7.54(m,2H),7.45(t,J=7.7Hz,1H),6.07(d,J=1.2H z, 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).

[0395] Step C: (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid

[0396] To a solution of tert-butyl (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoate (350 mg, 0.87 mmol, 1 eq) in DCM (4 mL) was added TFA (6.13 g, 53.8 mmol, 62.2 eq) at room temperature, and the resulting mixture was stirred at room temperature for 18 hours. Upon completion, the reaction mixture was concentrated under reduced pressure to give (E)-3-(3-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid (250 mg, 0.72 mmol, 83%) as an oil, which was used in the next step without further purification. LC-MS (ESI) m / z=347.2 [MH] -

[0397] Preparation of perfluorophenyl 5-((ethoxy(hydroxy)phosphoryl)difluoromethyl)-6-fluorobenzo[b]thiophene-2-carboxylate

[0398]

[0399] Step A: Methyl 4-amino-2-chloro-5-iodobenzoate

[0400] Methyl 4-amino-2-chlorobenzoate (5.0 g, 26.9 mmol, 1 eq.) and CaCO were added at room temperature. 3 To a suspension of 5-nitropropene (5.4 g, 53.8 mmol, 2.0 equiv) in MeOH (54 mL) was added a solution of ICl (4.8 g, 29.5 mmol, 1.1 equiv) in DCM (25 mL). The reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was The filtrate was concentrated under reduced pressure to give a residue. The residue was triturated with petroleum ether:ethyl acetate=3:1 (30 mL) to give methyl 4-amino-2-chloro-5-iodobenzoate (6.1 g, 19.5 mmol, 73%) as a yellow solid. LCMS (ESI) m / z=312 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ8.10(s,1H),6.80(s,1H),6.24(s,2H),3.75(s,3H).

[0401] Step B: (4-amino-2-chloro-5-iodophenyl)methanol

[0402] DIBAL-H (48.7 mL, 48.7 mmol, 2.5 equiv) was slowly added to a solution of 4-amino-2-chloro-5-iodobenzoic acid methyl ester (6.1 g, 19.5 mmol, 1 equiv) in DCM (120 mL) and THF (30 mL) at -78 ° C under nitrogen for 30 minutes. After addition, the reaction mixture was stirred at room temperature for 30 minutes. After completion, the reaction mixture was quenched with saturated sodium potassium tartrate solution (100 mL) at 0 ° C and stirred at room temperature for 2 hours. The mixture was extracted with DCM (200 mL). The organic phase was washed with brine (50 mL) and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give (4-amino-2-chloro-5-iodophenyl)methanol (5.0 g, 17.6 mmol, 91%) as a yellow solid. LCMS (ESI) m / z=284 [M+H] + .

[0403] Step C: 4-Amino-2-chloro-5-iodobenzaldehyde

[0404] To (4-amino-2-chloro-5-iodophenyl)methanol (4.0 g, 14.1 mmol, 1 eq.) was added CHCl at room temperature. 3 MnO was added to the solution (50 mL) 2 (7.4 g, 84.6 mmol, 6.0 equiv.). After addition, the mixture was stirred at 60 °C for 5 hours. After completion, the reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure to give 4-amino-2-chloro-5-iodobenzaldehyde (3.80 g, 13.5 mmol, 96%) as a yellow solid. LCMS: m / z=282 [M+H] + .

[0405] Step D: Ethyl 6-amino-5-iodobenzo[b]thiophene-2-carboxylate

[0406] 4-Amino-2-chloro-5-iodobenzaldehyde (900 mg, 3.19 mmol, 1 eq.) and K were added at room temperature. 2 CO 3 To a solution of ethyl 2-sulfylacetate (881 mg, 6.38 mmol, 2.0 equiv) in DMA (10 mL) was added ethyl 2-sulfylacetate (420 mg, 3.50 mmol, 1.1 equiv). After addition, the reaction mixture was stirred at 60 °C for 16 hours. The reaction mixture was cooled to room temperature and poured into water (50 mL). The mixture was extracted with EtOAc (50 mL x 3). The combined organic phases were washed with brine (50 mL) and washed with anhydrous Na 2 SO 4Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by silica gel flash column chromatography to give ethyl 6-amino-5-iodo-1-benzothiophene-2-carboxylate (5, 410 mg, 1.18 mmol, 37%) as a yellow solid. LCMS (ESI): m / z=348 [M+H] + .

[0407] Step E: Ethyl 6-fluoro-5-iodobenzo[b]thiophene-2-carboxylate

[0408] To a solution of ethyl 6-amino-5-iodo-1-benzothiophene-2-carboxylate (410 mg, 1.18 mmol, 1 eq.) in THF (4 mL) was added 48% HBF at 0 °C. 4 (2.4 g, 12.9 mmol, 11.0 equiv). Then NaNO was added dropwise 2 (73mg, 1.06mmol, 0.9 equivalent) in water (4mL), and the resulting yellow suspension was stirred at 0°C for 1 hour. The solid was collected by vacuum filtration and dried to obtain 400mg of diazonium cation salt. Diazonium benzene tetrafluoroborate was heated to 160°C under nitrogen for 3 hours. The remaining material was purified by silica gel flash column chromatography to obtain 6-fluoro-5-iodobenzo[b]thiophene-2-carboxylic acid ethyl ester (210mg, 599μmol, 51%) as a yellow oil. 1 H NMR (400 MHz, CDCl 3 )δ8.26(d,J=5.8Hz,1H),7.93(s,1H),7.55(d,J=7.6Hz,1H),4.41(q,J=7.1Hz,2H),1.41(t,J=7.1Hz,3H).

[0409] Step F: Ethyl 5-((diethoxyphosphoryl)difluoromethyl)-6-fluorobenzo[b]thiophene-2-carboxylate

[0410] A mixture of diethyl (bromodifluoromethyl)phosphonate (317 mg, 1.19 mmol, 2.0 equiv) and activated Zn powder (67 mg, 1.19 mmol, 2.0 equiv) in DMA (4 mL) was stirred at 60 ° C for 1 hour. Then, CuBr (170 mg, 1.19 mmol, 2.0 equiv) was added. The mixture was stirred at 50 ° C for 1 hour. A solution of ethyl 6-fluoro-5-iodobenzo[b]thiophene-2-carboxylate (210 mg, 599 μmol, 1 equiv) in DMA (2 mL) was added. After addition, the reaction mixture was stirred at 60 ° C under nitrogen overnight. The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with water (10 mL) and extracted with EtOAc (20 mL x3). The combined organic phase was washed with brine (20 mL) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by silica gel flash column chromatography to give ethyl 5-((diethoxyphosphoryl)difluoromethyl)-6-fluoro-1-benzothiophene-2-carboxylate (94.0 mg, 229 μmol, 38%) as a yellow oil. LCMS (ESI): m / z=411 [M+H] + .

[0411] Step G: 5-((Ethoxy(hydroxy)phosphoryl)difluoromethyl)-6-fluorobenzo[b]thiophene-2-carboxylic acid

[0412] 5-[(diethoxyphosphoryl)difluoromethyl]-6-fluoro-1-benzothiophene-2-carboxylate (7, 94 mg, 229 μmol, 1 eq.) was dissolved in THF (2 mL) and H 2 LiOH.H O (0.5 mL) was added to the mixture. 2 O (48 mg, 1.14 mmol, 5.0 equiv). After addition, the reaction mixture was stirred at room temperature overnight. After completion, the reaction mixture was carefully neutralized with 5N HCl aqueous solution until the pH was adjusted to pH = 3. The mixture was extracted with EtOAc (10 mL x 2). The organic phase was washed with brine (10 mL) and purified by Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give 5-{[ethoxy(hydroxy)phosphoryl]difluoromethyl}-6-fluoro-1-benzothiophene-2-carboxylic acid (50.0 mg, 141 μmol, 62%) as a yellow solid. LCMS (ESI): m / z=355 [M+H] + .

[0413] Step H: Perfluorophenyl 5-((ethoxy(hydroxy)phosphoryl)difluoromethyl)-6-fluorobenzo[b]thiophene-2-carboxylate

[0414] To a solution of 5-{[ethoxy(hydroxy)phosphoryl]difluoromethyl}-6-fluoro-1-benzothiophene-2-carboxylic acid (50 mg, 141 μmol, 1 eq.), pentafluorophenol (26 mg, 141 μmol, 1 eq.) and DMAP (2 mg, 14.1 μmol, 0.1 eq.) in THF (10 mL) was added DCC (41 mg, 197 μmol, 1.4 eq.) at 0 °C. After addition, the reaction mixture was stirred at room temperature overnight. After completion, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL x 3). The organic phase was washed with brine (10 mL) and dried over anhydrous Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by silica gel flash column chromatography to give perfluorophenyl 5-((ethoxy(hydroxy)phosphoryl)difluoromethyl)-6-fluorobenzo[b]thiophene-2-carboxylate (60.0 mg, 115 μmol, 82%) as a white solid. LCMS (ESI): m / z=520 [M+H] + .

[0415] Bis- and mono-SATE esters of di-F-naphthyl PFP:

[0416] Preparation of 7-((bis(2-(butyrylthio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoic acid perfluorophenyl ester and 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid perfluorophenyl ester:

[0417]

[0418] Step A: Perfluorophenyl 7-((bis(2-(butyrylthio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoate and perfluorophenyl 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoate

[0419]

[0420] Oxalyl chloride (266 mg, 2.13 mmol, 1.0 equiv) was added dropwise to a solution of (difluoro(7-((perfluorophenoxy)carbonyl)naphthalene-2-yl)methyl)phosphonic acid (1 g, 2.10 mmol, 1 equiv) in anhydrous DCM (18 mL) at 25 ° C, followed by DMF (155 mg, 2.13 mmol, 1.0 equiv). The reaction mixture was warmed to 40 ° C and then stirred at reflux for another 1.2 hours. The reaction was monitored by pipetting a small amount of crude sample and quenching with MeOH to ensure complete formation of bis-Cl phosphoryl chloride (bis-methoxyphosphonate was observed by LCMS). After completion, excess oxalyl chloride and solvent were removed under reduced pressure. The residue was redissolved in anhydrous DCM (14 mL), and then a mixture of S-(2-hydroxyethyl)thiobutyrate (641 mg, 4.33 mmol, 2.2 eq) and triethylamine (198 mg, 1.97 mmol, 1.0 eq) in anhydrous DCM (15 mL) was added at 0°C. The reaction was warmed to room temperature and stirred for another 12 hours. The progress of the reaction was monitored by LCMS, and upon completion, the reaction was concentrated by adding H 2 O (10mL) quench the reaction and concentrate under reduced pressure. The residue is purified by C18 column chromatography to obtain 7-((bis(2-(butyrylthio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoic acid perfluorophenyl ester (2A, 680 mg, 1.13 mmol, 58%) and 7-(((2-(butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid perfluorophenyl ester (2B, 65.0 mg, 89.2 μmol) as a white solid. 2A: LCMS (ESI): m / z=729 [M+H] + . 2B: LCMS(ESI):m / z=599[M+H] + .

[0421] Step B: Perfluoro-7-(((2-(Butyrylthio)ethoxy)(((isopropoxycarbonyl)oxy)methoxy)phosphoryl)difluoromethyl)-2-naphthoate

[0422]

[0423] Perfluorophenyl 7-(((2-(Butyrylthio)ethoxy)(hydroxy)phosphoryl)difluoromethyl)-2-naphthoate (300 mg, 501 μmol, 1 eq) was added to deionized H 2 Amberlite was added to the mixture of O (8 mL) and THF (2 mL). Resin (Na +The resulting mixture was stirred at room temperature for 1 hour, and then the excess resin was removed by filtration. 3 (127 mg, 751 μmol, 1.5 eq.) deionized H 2 O (2 mL) was added to the resulting solution. After addition, the resulting mixture was stirred at room temperature for another 1 hour. During this period, the silver salt formed a white precipitate and was collected by filtration. The filter cake was washed with cold H 2 O (2 mL x 3), and the silver salt was further dried under reduced pressure to obtain a dry powder which was pure enough for the next step without further purification.

[0424] The isolated mono-Ag salt was suspended in ACN (8 mL), and isopropyl iodomethyl carbonate (244 mg, 1.00 mmol, 2 equiv.) was added dropwise. After addition, the resulting mixture was stirred at 40 ° C for another 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)phosphoryl)difluoromethyl)-2-naphthoic acid perfluorophenyl ester (30 mg, 41.9 μmol, 8%) as a colorless oil. LCMS (ESI) m / z=737[M+Na] + .

[0425] Preparation of 5-((diethoxyphosphoryl)difluoromethyl)thieno[2,3-b]pyridine-2-carboxylic acid:

[0426]

[0427] Step 1: Ethyl 5-bromothienyl[2,3-b]pyridine-2-carboxylate.

[0428] To a solution of 5-bromo-2-fluoropyridine-3-carboxaldehyde (10 g, 49 mmol, 1 eq.) in dimethylformamide (70 mL) was added potassium carbonate (27 g, 0.2 mol, 4 eq.) and ethyl 2-sulfonylacetate (6.5 g, 54 mmol, 1.1 eq.). The mixture was stirred at 60 °C for 16 hours. The reaction was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 2), the combined organic layers were washed with saturated brine (200 mL x 2), and the mixture was purified by Na 2 O 4 '-trifluoroacetic acid (100 mL x 2). 2 SO 4Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give ethyl 5-bromothieno[2,3-b]pyridine-2-carboxylate (13 g, yield 96%) as a yellow solid. LCMS: m / z (M+H) + =287.9

[0429] Step 2: Ethyl 5-iodothienyl[2,3-b]pyridine-2-carboxylate.

[0430] To a solution of ethyl 5-bromothieno[2,3-b]pyridine-2-carboxylate (13 g, 45 mmol, 1 eq) and sodium iodide (14 g, 91 mmol, 2 eq) in dioxane (90 mL) was added N 1 ,N 2 -dimethylethane-1,2-diamine (0.4 g, 4.5 mmol, 0.1 eq.) and CuI (0.4 g, 2.3 mmol, 0.05 eq.) were added, and the mixture was then heated under microwave conditions at N 2 The mixture was stirred at 100 °C for 5 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give ethyl 5-iodothieno[2,3-b]pyridine-2-carboxylate (13 g, yield 89%) as a yellow solid. LCMS: m / z (M+H) + =333.8.

[0431] Step 3: 5-iodothienyl[2,3-b]pyridine-2-carboxylic acid.

[0432] To ethyl 5-iodothieno[2,3-b]pyridine-2-carboxylate (12 g, 36 mmol, 1 eq.) was added H 2 Lithium hydroxide (2.6 g, 0.1 mol, 3 equivalents) was added to a solution of O:THF:MeOH=1:1::1 (90 mL). The mixture was stirred at 25 ° C for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was then diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The aqueous phase was adjusted to pH = 5 to 6 with FA (1 M) and extracted with ethyl acetate (50 mL x 2). 2 SO 4 Dry, filter, and concentrate under reduced pressure to give crude 5-iodothieno[2,3-b]pyridine-2-carboxylic acid (8 g, crude) as a yellow oil. LCMS: m / z (M+H) + =305.9.

[0433] Step 4: Benzyl 5-iodothienyl[2,3-b]pyridine-2-carboxylate.

[0434] To a solution of 5-iodothieno[2,3-b]pyridine-2-carboxylic acid (8 g, 26 mmol, 1 eq) in DMF (50 mL) was added potassium carbonate (7.2 g, 52 mmol, 2 eq) and (bromomethyl)benzene (4.9 g, 29 mmol, 1.1 eq). The mixture was stirred at 25 °C for 16 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 2), the combined organic layers were washed with saturated brine (100 mL x 2), and the mixture was purified by HPLC. 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give benzyl 5-iodothieno[2,3-b]pyridine-2-carboxylate (4.4 g, 43% yield) as a yellow solid. LCMS: m / z (M+H) + =396.1. 1 H NMR (400 MHz, DMSO-d 6 )δ9.03-8.63(m,1H),8.12(s,1H),7.62-7.08(m,6H),5.55-5.02(m,2H).

[0435] Step 5: Benzyl 5-((diethoxyphosphoryl)difluoromethyl)thieno[2,3-b]pyridine-2-carboxylate.

[0436] To a suspension of zinc (0.4 g, 5.7 mmol, 2 eq.) in degassed anhydrous THF (10 mL) was added trimethylsilyl chloride (0.1 g, 1.2 mmol, 0.4 eq.), and the suspension was heated to 50° C. for 1.5 h with vigorous stirring. A solution of diethyl (bromodifluoromethyl)phosphonate (1.6 g, 6.1 mmol, 2 eq.) in THF (15 mL) was added dropwise (temperature maintained between 50-55° C.). The reaction mixture was stirred at 50° C. for 2 h, and λ1-copper (1+) bromide (0.4 g, 3.0 mmol, 1.0 eq.) and dimethylacetamide (6.7 mL, 73 mmol, 24 eq.) were added, and the mixture was stirred at room temperature for 30 min. A solution of benzyl 5-iodothieno[2,3-b]pyridine-2-carboxylate (1.2 g, 3.0 mmol, 1 eq.) in N,N-dimethylacetamide (10 mL) was slowly added to the reaction mixture and then heated to 50°C for 12 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), and the combined organic layers were washed with saturated brine (50 mL x 2) and purified by Na 2 SO 4Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by reverse phase (column: 120g flash column Welch Ultimate XB_C18 20-40μm; 120A, mobile phase: H2O+ACN; gradient B%: 10-65% 15 minutes; 65% 3 minutes) to lyophilize to give 5-((diethoxyphosphoryl)difluoromethyl)thieno[2,3-b]pyridine-2-carboxylic acid benzyl ester (0.6 g, yield 42%) as a yellow oil. LCMS: m / z (M+H) + =456.3. 1 H NMR (400 MHz, METHANOL-d 4 )δ8.84(s,1H),8.59(s,1H),8.24(s,1H),7.57-7.31(m,5H),5.43(s,2H),4.38-4.16(m,4H),1.33(t,J=7.2Hz,6H)

[0437] Step 6: 5-((Diethoxyphosphoryl)difluoromethyl)thieno[2,3-b]pyridine-2-carboxylic acid.

[0438] In N 2 To a solution of benzyl 5-((diethoxyphosphoryl)difluoromethyl)thieno[2,3-b]pyridine-2-carboxylate (0.3 g, 0.7 mmol, 1 eq) in methanol (1.5 mL) and tetrahydrofuran (1.5 mL) was added dry Pd / C (30 mg). The mixture was heated at 25 °C under H 2 (15 psi) for 4 hours. The reaction mixture was filtered, and the filter was concentrated to give crude 5-((diethoxyphosphoryl)difluoromethyl)thieno[2,3-b]pyridine-2-carboxylic acid (0.2 g, crude) as a yellow oil. LCMS: m / z (M+H)+=365.9.

[0439] Preparation of 6-(difluoro(phosphono)methyl)-2-naphthoic acid:

[0440]

[0441] Step 1: Methyl 6-iodo-2-naphthoate.

[0442] To a solution of methyl 6-bromonaphthalene-2-carboxylate (5.0 g, 19 mmol, 1 eq.) and sodium iodide (5.6 g, 38 mmol, 2 eq.) in dioxane (50 mL) was added N 1 ,N 2-dimethylethane-1,2-diamine (0.17 g, 1.9 mmol, 0.1 eq.) and CuI (0.18 g, 0.94 mmol, 0.05 eq.) were added, and the mixture was then heated under microwave conditions at N 2 The mixture was stirred at 100° C. for 5 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic layers were washed with saturated brine (50 mL×2) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by RP-Flash (column: 330 g flash column, Welch Ultimate XB_C18 20-40 μm; 120A; mobile phase: 5-80% 35 minutes; 80% 10 minutes) to freeze-dry to give methyl 6-iodo-2-naphthoate (4.2 g, yield 70%) as a white solid. LCMS: m / z (M+H) + =313.0

[0443] Step 2: Methyl 6-((diethoxyphosphoryl)difluoromethyl)-2-naphthoate.

[0444] To a solution of diethyl (bromodifluoromethyl)phosphonate (5.4 g, 20 mmol, 3.5 eq.) in DMF (50 mL) was added cadmium (2.3 g, 21 mmol, 3.6 eq.) and stirred at 25 °C under N 2 Stir for 8 hours at N to obtain Solution A. To a solution of methyl 6-iodo-2-naphthoate (1.8 g, 5.8 mmol, 1 eq.) in dimethylformamide (20 mL) was added CuCl (1.1 g, 11 mmol, 2.0 eq.) and stirred at N 2 The mixture was stirred at 25°C for 2 hours to obtain solution B. Solution A was then filtered, and the filtrate was added to solution B, and the mixture was stirred at N 2 The mixture was stirred at 25°C for 16 hours. The reaction mixture was filtered, and the filtrate was quenched by 100 mL of water and extracted with 240 mL (80 mL x 3) of ethyl acetate. The combined organic layers were washed with 240 mL (80 mL x 3) of brine and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by RP-Flash (column: 330 g flash column, Welch Ultimate XB_C18 20-40 μm; 120A; mobile phase: 5-60% 30 minutes; 60% 5 minutes) to give methyl 6-((diethoxyphosphoryl)difluoromethyl)-2-naphthoate (1.0 g, yield 47%) as a white solid. LCMS: m / z (M+1) + =373.0

[0445] Step 3: 6-((Ethoxy(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid.

[0446] To a solution of methyl 6-((diethoxyphosphoryl)difluoromethyl)-2-naphthoate (0.35 g, 0.94 mmol, 1 eq) in tetrahydrofuran (3 mL) and water (1 mL) was added lithium hydroxide (1+) hydrate (79 mg, 1.9 mmol, 2 eq), and the mixture was stirred at 25 °C for 2 h.

[0447] The reaction mixture was concentrated under reduced pressure to give 6-((ethoxy(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid (0.29 g, crude) as a white solid. LCMS: m / z (M+H) + =331.0. 1 H NMR (400 MHz, DMSO-d 6 )δ8.38(s,1H),8.05-8.00(m,2H),7.91(d,J=8.8Hz,1H),7.84(d,J=8.4Hz,1H),7.64(d,J=8.8Hz,1H),3.80-3.76(m,2H),1.07(t,J=7.2Hz,3H)

[0448] Step 4: 6-(Difluoro(phosphono)methyl)-2-naphthoic acid.

[0449] To a solution of 6-((ethoxy(hydroxy)phosphoryl)difluoromethyl)-2-naphthoic acid (0.10 g, 0.30 mmol, 1 eq) in dichloromethane (1 mL) at 0° C. was added BSTFA (0.46 g, 1.8 mmol, 6 eq) and the mixture was stirred at 0° C. for 10 min, then TMSI (0.24 g, 1.2 mmol, 4 eq) in dichloromethane (1 mL) was added at 0° C. and stirred at 0° C. for 20 min. The reaction was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 x 25 x 10 um; mobile phase: [water (0.225% FA)-ACN]; B%: 10%-40%, 11 minutes) to give 6-(difluoro(phosphono)methyl)-2-naphthoic acid (20 mg, yield 25%) as a yellow oil. LCMS: m / z (M+H) + =302.9

[0450] Preparation of 6-(dimethoxyphosphoryl)-2-naphthoic acid

[0451]

[0452] Step 1: Methyl 6-(dimethoxyphosphoryl)-2-naphthoate.

[0453] In N 2 To a solution of methyl 6-bromonaphthalene-2-carboxylate (5.0 g, 19 mmol, 1 eq.) in dioxane (50 mL) was added K 3 PO 4 (8.0 g, 38 mmol, 2 eq), NiCl 2 (dppp) (1.0 g, 1.9 mmol, 0.1 eq.) was added dropwise to a solution of dimethyl phosphonate (4.1 g, 38 mmol, 2 eq.) in dioxane (50 mL). The reaction mixture was stirred at 100 °C for 12 hours. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 2), the combined organic layers were washed with saturated brine (200 mL x 2), and the mixture was purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography to give methyl 6-(dimethoxyphosphoryl)-2-naphthoate (1.5 g, yield 27%) as a white solid. LCMS: m / z (M+H) + =295.3. 1 H NMR (400MHz, CD 3 OD)δ8.66(s,1H),8.45(d,J=15.6Hz,1H),8.19-8.08(m,3H),7.81(m,1H),3.98(s,3H),3.85-3.76(m,6H)

[0454] Step 2: 6-(Dimethoxyphosphoryl)-2-naphthoic acid

[0455] To a solution of methyl 6-(dimethoxyphosphoryl)-2-naphthoate (0.50 g, 1.7 mmol, 1 eq) in methanol (13 mL), tetrahydrofuran (13 mL) and water (13 mL) was added lithium hydroxide (1+) hydrate (0.14 g, 3.4 mmol, 2 eq). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), and HCl (1 M) was added to adjust the aqueous solution to pH = 4 to 6. The solution was then extracted with ethyl acetate (50 mL x 2) and washed with Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give crude 6-(dimethoxyphosphoryl)-2-naphthoic acid (0.30 g, crude) as a white solid.

[0456] Preparation of 7-(dimethoxyphosphoryl)-2-naphthoic acid

[0457]

[0458] Step 1: 7-Bromo-2-naphthoic acid.

[0459] A solution of 2,7-dibromonaphthalene (20 g, 69 mmol, 1 eq.) in tetrahydrofuran (200 mL) was cooled to -76 °C and n-BuLi (4.5 g, 70 mmol, 1 eq.) was added dropwise. 2 (gas) was bubbled through the reaction mixture. After 15 minutes, the cooling bath was removed and the mixture was heated to 400 °C by continuing to bubble CO 2 The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1.5 hours. 4 The mixture was quenched with HCl (200 mL) and extracted with ethyl acetate (200 mL x 2). The combined organic layers were washed with saturated brine (200 mL x 2) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give 7-bromo-2-naphthoic acid (14 g, crude) as a yellow solid. LCMS: m / z (MH) - =250.0. 1 H NMR (400 MHz, DMSO-d 6 )δ8.60(d,J=0.8Hz,1H),8.43(d,J=2.0Hz,1H),8.07-8.01(m,2H),8.01-7.95(m,1H),7.77(dd,J=2.2,8.7Hz,1H)

[0460] Step 2: Methyl 7-bromo-2-naphthoate.

[0461] 7-Bromo-2-naphthoic acid (4.3 g, 17 mmol, 1 eq.) was dissolved in methanol (40 mL) and H 2 SO 4 The solution in 4% paraformaldehyde (8 mL) was stirred at 70 ° C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), and the combined organic layer was washed with saturated brine (50 mL x 2) and washed with Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography to give methyl 7-bromo-2-naphthoate (3.6 g, yield 84%) as a yellow solid. 1 H NMR (400 MHz, METHANOL-d 4)δ8.58-8.53(m,1H),8.22(d,J=2.0Hz,1H),8.08-8.03(m,1H),7.95(d,J=8.4Hz,1H),7.87(d,J=8.4Hz,1H),7.73-7.67(m,1H),3.97(s,3H)

[0462] Step 3: Methyl 7-(dimethoxyphosphoryl)-2-naphthoate.

[0463] To a solution of methyl 7-bromo-2-naphthoate (3.3 g, 12 mmol, 1 eq.) in dioxane (30 mL) were added potassium phosphate (5.3 g, 24 mmol, 2 eq.) and dimethyl phosphonate (2.7 g, 25 mmol, 2 eq.), followed by N 2 Add NiCl 2 (dppp) (0.67 g, 1.2 mmol, 0.1 eq.) was added and the mixture was stirred at 100°C for 12 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 2). The combined organic layers were washed with saturated brine (100 mL x 2) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by column chromatography to give methyl 7-(dimethoxyphosphoryl)-2-naphthoate (0.60 g, 16% yield) as a white solid. LCMS: m / z (M+H) + =295.1

[0464] Step 4: 7-(Dimethoxyphosphoryl)-2-naphthoic acid.

[0465] To a solution of methyl 7-(dimethoxyphosphoryl)-2-naphthoate (0.6 g, 2.0 mmol, 1 eq) in water (2 mL), tetrahydrofuran (2 mL) and methanol (2 mL) was added lithium hydroxide hydrate (85 mg, 2.0 mmol, 1 eq) and the mixture was stirred at 25 °C for 3 hours. The reaction mixture was concentrated under reduced pressure to remove THF and methanol to give a suspension. The suspension was purified by preparative HPLC (column: Phenomenex Luna C18 150 x 25 mm x 10 um, mobile phase: water (0.225% FA)-ACN; B%: 13%-43%, 11.5 min) and lyophilized to give 7-(dimethoxyphosphoryl)-2-naphthoic acid (0.45 g, 79% yield) as a white solid. 1 H NMR (400 MHz, METHANOL-d 4)δ8.77(s,1H),8.55(d,J=15.6Hz,1H),8.27-8.18(m,1H),8.16-8.10(m,1H),8.09-8.04(m,1H),7.93-7.82(m,1H),3.85(d,J=11.2Hz,6H)

[0466] Preparation of ((2-(((5S,8S,10aR)-3-acetyl-8-(methyl(phenyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[d]thiazol-5-yl)difluoromethyl)phosphonic acid

[0467]

[0468] Step 1: 5-iodobenzo[d]thiazole.

[0469] To a solution of benzo[d]thiazole-5-amine (10 g, 67 mmol, 1 eq) in acetonitrile (50 mL) was added HCl (7.3 g, 200 mmol, 3 eq) at 0 °C, followed by water (50 mL) containing sodium nitrite (5.5 g, 80 mmol, 1.2 eq). The solution was stirred at 0 °C for 30 minutes. Then water (50 mL) containing potassium iodide (17 g, 0.1 mol, 1.5 eq) was added at 0 °C. The reaction mixture was stirred at 0 °C for 1 hour to obtain a black suspension. The reaction mixture was poured into water (100 mL). The aqueous phase was extracted with ethyl acetate (200 mL x 2). The combined organic phase was washed with brine (200 mL x 2) and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate in vacuo to obtain a residue. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 100 / 1, 5 / 1) to obtain 5-iodobenzo[d]thiazole (9.0 g, 34 mmol, yield 52%) as a yellow solid. LCMS: m / z (M+H) + =243.9

[0470] Step 2: Ethyl 5-iodobenzo[d]thiazole-2-carboxylate.

[0471] Then in N 2To a solution of 5-iodobenzo[d]thiazole (1 g, 3.8 mmol, 1 eq.) and ethyl cyanoformate (0.76 g, 7.7 mmol, 2 eq.) in THF (15 mL) at -78 °C was added lithium bis(trimethylsilyl)azide (1.6 g, 9.6 mmol, 2.5 eq.), and the reaction mixture was stirred at -78 °C for 1 hour to give a yellow solution. The reaction solution was poured into water (45 mL). The aqueous phase was extracted with ethyl acetate (45 mL x 2). The combined organic phase was washed with brine (45 mL x 2) and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate in vacuo to give a residue. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel) to give ethyl 5-iodobenzo[d]thiazole-2-carboxylate (1 g, 3.0 mmol, yield 79%) as a yellow oil. LCMS: m / z (M-28+H) + =305.8. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.97 (s, 1H), 8.52 (d, J = 0.8 Hz, 1H), 7.73 (d, J = 2.0 Hz, 1H), 4.57 (d, J = 7.2 Hz, 2H), 1.50 (t, J = 7.2 Hz, 3H).

[0472] Step 3: Ethyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[d]thiazole-2-carboxylate.

[0473] In N 2 To a suspension of zinc (0.22 g, 3.4 mmol, 2.5 eq) in THF (5 mL) was added trimethylsilyl chloride (59 mg, 0.54 mmol, 0.4 eq) at 30 °C, and the reaction mixture was stirred at 55 °C for 1.5 hours. The contents were cooled to 30 °C, and a solution of diethyl (bromodifluoromethyl)phosphonate (0.72 g, 2.7 mmol, 2 eq) in THF (5 mL) was added dropwise (temperature maintained between 50-55 °C). The reaction mixture was stirred at 30 °C for 2 hours (the turbid suspension became a solution), and a solution containing copper bromide (1 +) (0.19 g, 1.4 mmol, 1.0 eq) in DMA (2 mL), and the mixture was stirred at 20 ° C for 10 minutes. A solution of ethyl 5-iodobenzo[d]thiazole-2-carboxylate (0.45 g, 1.4 mmol, 1 eq) in DMA (2 mL) was added, and the reaction mixture was stirred at 55 ° C for 16 hours to give a green suspension. The reaction solution was poured into ice water (w / w=1 / 1) (35 mL). The aqueous phase was extracted with ethyl acetate (35 mL x 2). The combined organic phase was washed with brine (35 mL x 2) and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate in vacuo to give a residue. The residue was purified by silica gel chromatography to give ethyl 5-((diethoxyphosphoryl)difluoromethyl)benzo[d]thiazole-2-carboxylate (0.1 g, 0.25 mmol, 19% yield) as a yellow solid. LCMS: m / z (M+H) + =394.4. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.49 (s, 1H), 8.08 (d, J = 8.8 Hz, 1H), 7.83 (d, J = 8.8 Hz, 1H), 4.58 (q, J = 7.2 Hz, 2H), 4.30-4.19 (m, 4H), 1.51 (t, J = 7.2 Hz, 3H), 1.34 (t, J = 7.2 Hz, 6H).

[0474] Preparation of ((2-(((5S,8S,10aR)-3-acetyl-8-(cinnolin-6-yl(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)quinolin-7-yl)difluoromethyl)phosphonic acid

[0475]

[0476] Step 1: Benzyl 7-bromoquinoline-2-carboxylate.

[0477] To a solution of 7-bromoquinoline-2-carboxylic acid (2.0 g, 7.9 mmol, 1 eq.) in dimethylformamide (10 mL) was added (bromomethyl)benzene (1.3 g, 7.9 mmol, 1 eq.) and potassium carbonate (2.2 g, 16 mmol, 2 eq.). The mixture was stirred at 25 °C for 16 hours to give a white suspension. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 2), the combined organic layers were washed with saturated brine (30 mL x 2), and the mixture was purified by Na 2 O 4 '-trifluoroacetic acid (10 mL x 2). 2 SO 4 The residue was purified by flash silica gel chromatography (40 g The product was purified by silica gel flash column, elution with a gradient of 0 to 25% petroleum ether in ethyl acetate at 50 mL / min) to give benzyl 7-bromoquinoline-2-carboxylate (2.1 g, 77% yield) as a yellow solid. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.44 (s, 1H), 8.25-8.04 (m, 2H), 7.68-7.62 (m, 2H), 7.54-7.41 (m, 2H), 7.37-7.24 (m, 3H), 5.45 (s, 2H).

[0478] Step 2: Benzyl 7-iodoquinoline-2-carboxylate.

[0479] To a solution of benzyl 7-bromoquinoline-2-carboxylate (2.0 g, 5.8 mmol, 1 eq) and sodium iodide (1.7 g, 12 mmol, 2 eq) in dioxane (10 mL) was added methyl[2-(methylamino)ethyl]amine (51.4 mg, 584 μmol, 0.1 eq) and λ1-copper(1+) iodide (56 mg, 0.29 mmol, 0.05 eq). The mixture was heated in a microwave reactor at 150 °C under N 2 The mixture was stirred for 5 hours to obtain a black suspension. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography (40 g Silica gel flash column, elution with a gradient of 0 to 25% petroleum ether / ethyl acetate at 12 mL / min), collected and concentrated in vacuo to give benzyl 7-iodoquinoline-2-carboxylate (1.3 g, 57% yield) as a white solid. LCMS: m / z (M+H) + =390.0. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.20-8.15 (m, 1H), 8.13-8.08 (m, 1H), 7.85-7.82 (M, 1H), 7.54-7.41 (m, 3H), 7.37-7.27 (m, 4H), 5.45 (s, 2H).

[0480] Step 3: Benzyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate.

[0481] To a suspension of zinc (0.44 g, 6.7 mmol, 2 eq) in degassed anhydrous tetrahydrofuran (30 mL) was added trimethylsilyl chloride (0.14 mg, 1.3 mmol, 0.4 eq) and the suspension was heated to 60° C. for 1.5 hours with vigorous stirring. The contents were cooled to 30° C. and a solution of diethyl (bromodifluoromethyl)phosphonate (1.1 g, 4.0 mmol, 1.2 eq) in tetrahydrofuran (10 mL) was added dropwise (temperature maintained between 50-55° C.). The reaction mixture was stirred at 30° C. for 2 hours (the turbid suspension became a solution), then copper (1+) bromide (479 mg, 3.34 mmol, 1.0 eq) and dimethylacetamide (7.0 g, 80 mmol, 24 eq) were added and the mixture was stirred at room temperature for 10 minutes. A solution of benzyl 7-iodoquinoline-2-carboxylate (1.3 g, 3.4 mmol, 1 eq.) in tetrahydrofuran (10 mL) was slowly added to the reaction mixture and heated to 50 °C for 12 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), and the combined organic layers were washed with saturated brine (50 mL x 2) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by RP-flash (TFA system) and freeze-dried to give benzyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate (0.3 g, yield 19%) as a yellow oil. LCMS: m / z (M+H) + =450.1. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.59 (s, 1H), 8.43-8.33 (m, 1H), 8.32-8.22 (m, 1H), 8.04-7.97 (m, 1H), 7.96-7.89 (m, 1H), 7.61-7.53 (m, 2H), 7.47-7.33 (m, 3H), 5.57 (s, 2H), 4.39-4.15 (m, 4H), 1.35 (t, J = 7.2, 6H).

[0482] Step 4: Benzyl 7-iodoquinoline-2-carboxylate.

[0483] To a solution of benzyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate (0.32 mg, 0.71 mmol, 1 eq) in methanol (3 mL) was added wet Pd / C (0.1 g, 2.9 mmol) and the mixture was degassed and heated with H 2 The mixture was purged three times under H 2The mixture was stirred at 25° C. for 3 hours under pressure (15 psi) to give a black suspension. The reaction mixture was filtered and the filtrate was concentrated to give benzyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate (0.25 g, crude). LCMS: m / z (M+H) + =360.1.

[0484] Step 5: 4-Nitrophenyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate.

[0485] To a solution of benzyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate (50 mg, 0.14 mmol, 1 eq) in dichloromethane (1 mL) was added 4-dimethylaminopyridine (25 mg, 0.21 mmol, 1.5 eq), 4-nitrophenol (25 mg, 0.18 mmol, 1.3 eq) and dicyclohexylcarbodiimide (43 mg, 0.21 μmol, 1.5 eq). The mixture was stirred at 25 °C for 1 hour to give a yellow suspension. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (4 g Silica gel flash column, elution with a gradient of 0 to 60% petroleum ether / ethyl acetate at 30 mL / min) to afford 4-nitrophenyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate (30 mg, 45% yield). LCMS: m / z (M+H) + =481.1. 1 H NMR (400MHz, chloroform-d) δ8.65(s,1H),8.49-8.45(m,1H),8.42-8.36(m,2H),8.16-8.11(m,1H),8.08-8.03( m,1H),8.00-7.94(m,1H),7.59-7.47(m,1H),6.97-6.84(m,1H),4.40-4.05(m,4H),1.42-1.31(m,6H).

[0486] Step 6: (Difluoro(2-((4-nitrophenoxy)carbonyl)quinolin-7-yl)methyl)phosphonic acid.

[0487] To a solution of 4-nitrophenyl 7-((diethoxyphosphoryl)difluoromethyl)quinoline-2-carboxylate (30 mg, 62 μmol, 1 eq) in dichloromethane (1 mL) was added (1E)-2,2,2-trifluoro-N-trimethylsilylethylimidate (96 mg, 0.37 mmol, 6 eq) and trimethylsilyl iodide (50 mg, 0.25 mmol, 4 eq). The mixture was stirred at 0 °C for 2 hours to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give the crude product (difluoro(2-((4-nitrophenoxy)carbonyl)quinolin-7-yl)methyl)phosphonic acid (30 mg, crude). LCMS: m / z (M+H) + =425.1

[0488] Preparation of ((6-(((5S,8S,10aR)-3-acetyl-8-(cinnolin-6-yl(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)isoquinolin-3-yl)difluoromethyl)phosphonic acid

[0489]

[0490] Step 1: 3-Chloroisoquinoline-6-carboxylic acid.

[0491] 6-Bromo-3-chloroisoquinoline (5.0 g, 21 mmol, 1 eq.) was dissolved in THF (45 mL) and H 2 Add bis((cyclopenta-2,4-diyn-1-yl)biphenylphosphate)palladium iron dichloride (2 + ) (1.5 g, 2.1 mmol, 0.1 eq.) and triethylamine (4.2 g, 41 mmol, 2 eq.). The mixture was stirred at 70 °C for 16 hours under CO (50 psi) to obtain a black solution. The reaction mixture was quenched with water (50 mL) and extracted with 150 mL (50 mL x 3) of ethyl acetate, and the combined aqueous layers were collected. The combined aqueous layers were adjusted to pH 3 to 4 with 20 mL of 1 N formic acid, and then extracted with 150 mL (50 mL x 3) of ethyl acetate. The combined organic layers were washed with 450 mL (150 mL x 3) of saturated brine and purified by Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give 3-chloroisoquinoline-6-carboxylic acid (3.5 g, 76% yield) as a white solid. LCMS: m / z (M+H) + =208.1. 1 HNMR (400 MHz, DMSO-d 6)δ13.94-13.02(m,1H),9.33(s,1H),8.64(s,1H),8.33-8.22(m,2H),8.13(dd,J=1.6,8.4Hz,1H).

[0492] Step 2: Benzyl 3-chloroisoquinoline-6-carboxylate.

[0493] To a solution of 3-chloroisoquinoline-6-carboxylic acid (3.5 g, 17 mmol, 1 eq.) and (bromomethyl)benzene (4.3 g, 25 mmol, 1.5 eq.) in DMF (40 mL) was added dipotassium carbonate (4.6 g, 34 mmol, 2 eq.). The mixture was stirred at 25 °C for 2 hours to give a grey solution. The reaction mixture was quenched with 50 mL of water and extracted with 150 mL (50 mL x 3) of ethyl acetate. The combined organic layers were washed with brine (50 mL x 3) and purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash silica gel flash column, 0 to 100% ethyl acetate / petroleum ether gradient eluent at 40 ml / min) to give 3-chloroisoquinoline-6-carboxylic acid benzyl ester (3.6 g, yield 71%) as a white solid. LCMS: m / z (M+H) + =297.9. 1 H NMR (400 MHz, CHLOROFORM-d) δ 9.17 (s, 1H), 8.55 (s, 1H), 8.23 ​​(d, J = 8.4 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.85 (s, 1H), 7.59-7.35 (m, 5H), 5.47 (s, 2H).

[0494] Step 3: Benzyl 3-iodoisoquinoline-6-carboxylate.

[0495] To a solution of benzyl 3-chloroisoquinoline-6-carboxylate (1.0 g, 3.4 mmol, 1 eq.) and sodium iodide (1.0 g, 6.7 mmol, 2 eq.) in dioxane (10 mL) was added copper iodide (1 + ) (32 mg, 0.17 mmol, 0.05 eq.) and methyl[2-(methylamino)ethyl]amine (30 mg, 0.34 mmol, 0.1 eq.), and the mixture was then heated in a microwave reactor under N 2 The mixture was stirred at 150°C for 8 hours to obtain a gray suspension. The reaction mixture was filtered, and the filtrate was quenched by 20 mL of water and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layers were washed with 60 mL (20 mL x 3) of brine and purified by Na2 SO 4 Dry, filter, and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash silica gel flash column, 0 to 100% ethyl acetate / petroleum ether gradient eluent at 40 ml / min) to give 3-iodoisoquinoline-6-carboxylic acid benzyl ester (0.80 g, 22% yield) as a white solid. LCMS: m / z (M+H) + =389.9

[0496] Step 4: Benzyl 3-((diethoxyphosphoryl)difluoromethyl)isoquinoline-6-carboxylate

[0497] To a solution of zinc (0.27 g, 4.1 mmol, 2 eq.) in THF (8 mL) was added trimethylsilyl chloride (0.11 g, 1.0 mmol, 0.5 eq.), and the mixture was then stirred under N 2 The mixture was stirred at 55° C. for 1.5 hours. Then a solution of diethyl (bromodifluoromethyl)phosphonate (1.1 g, 4.1 mmol, 2 eq.) in THF (8 mL) was added and stirred at N 2 The mixture was stirred at 55°C for 2 hours. Then, copper bromide (1 + ) (0.59 g, 4.1 mmol, 2 eq.) and DMA (4 mL) were added and stirred at 55 °C for 10 min. Then, a solution of benzyl 3-iodoisoquinoline-6-carboxylate (0.8 g, 2.1 mmol, 1 eq.) in DMA (4 mL) was added and stirred at N 2 The mixture was stirred at 55°C for 16 hours to obtain a light yellow solution. The reaction mixture was filtered and the filtrate was purified by H 2 O (20 mL) and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layers were washed with 60 mL (20 mL x 3) of brine and purified by Na 2 SO 4 The residue was purified by preparative HPLC (column: Phenomenex LunaC18 150 x 25 x 10 um; mobile phase: [water (10 mM NH 4 HCO 3 )-ACN]; B%: 50%-75%, 18 minutes). The solution was lyophilized to give benzyl 3-((diethoxyphosphoryl)difluoromethyl)isoquinoline-6-carboxylate (0.30 g, yield 32%) as a yellow solid. LCMS: m / z (M+H) + =450.0. 1H NMR (400 MHz, CHLOROFORM-d) δ 9.33 (s, 1H), 8.63 (s, 1H), 8.26 (dd, J = 1.2, 8.8 Hz, 1H), 8.14 (s, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.46-7.40 (m, 2H), 7.39-7.25 (m, 3H), 5.38 (s, 2H), 4.38-4.10 (m, 4H), 1.31 (t, J = 7.2 Hz, 6H).

[0498] Step 5: 3-((Diethoxyphosphoryl)difluoromethyl)isoquinoline-6-carboxylic acid.

[0499] To a solution of benzyl 3-((diethoxyphosphoryl)difluoromethyl)isoquinoline-6-carboxylate (0.27 g, 0.60 mmol, 1 eq) in MeOH (5 mL) was added wet Pd / C (0.15 g). The mixture was stirred under H 2 The mixture was stirred at 25° C. for 3 hours under pressure (15 psi). The reaction mixture was filtered and concentrated under reduced pressure to give crude benzyl 3-((diethoxyphosphoryl)difluoromethyl)isoquinoline-6-carboxylate (0.2 g, crude). LCMS: m / z (M+H) + =359.9

[0500] Preparation of ((7-(((5S,8S,10aR)-3-acetyl-8-(cinnolin-6-yl(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)quinolin-2-yl)difluoromethyl)phosphonic acid

[0501]

[0502] Step 1: Benzyl 2-chloroquinoline-7-carboxylate

[0503] To a solution of ethyl 2-chloroquinoline-7-carboxylate (0.50 g, 2.1 mmol, 1 eq) in benzyl alcohol (5 mL) were added tripotassium phosphate (45 mg, 0.21 mmol, 0.1 eq) and potassium carbonate (0.30 g, 2.1 mmol, 1 eq), and the mixture was stirred at 25 °C for 12 hours. The residue was diluted with water (20 mL) and extracted with ethyl acetate (40 mL x 2), and the combined organic layers were washed with saturated brine (20 mL x 2), and the mixture was stirred for 12 hours at 25 °C. 2 SO 4 The residue was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2, petroleum ether / ethyl acetate=100:0 to 90:10) to give benzyl 2-chloroquinoline-7-carboxylate (0.39 g, yield 62%) as a white solid. 1 H NMR (400 MHz, chloroform-d) δ 8.80 (s, 1H), 8.24-8.12 (m, 2H), 7.89 (d, J = 8.4 Hz, 1H), 7.52-7.47 (m, 3H), 7.46-7.35 (m, 3H)

[0504] Step 2: Benzyl 2-iodoquinoline-7-carboxylate

[0505] To a solution of benzyl 2-chloroquinoline-7-carboxylate (0.3 g, 0.87 mmol, 1 eq.) and sodium iodide (0.26 g, 1.7 mmol, 2 eq.) in dioxane (38 mL) was added copper iodide (1 + ) (8.3 mg, 44 μmol, 0.05 eq.), and then the mixture was stirred at 150 °C under microwave for 5 hours as a gray suspension. The reaction mixture was filtered and the filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 10: 1 to 3: 1) to give benzyl 2-iodoquinoline-7-carboxylate (0.1 g, yield 29%) as a white solid. LCMS: m / z (M+H) + =389.9

[0506] Step 3: Benzyl 2-((diethoxyphosphoryl)difluoromethyl)quinoline-7-carboxylate

[0507] In N 2 To a suspension of zinc (47 mg, 0.72 mmol, 2 eq) in tetrahydrofuran (2 mL) was added trimethylsilyl chloride (16 mg, 0.14 mmol, 0.4 eq) and the reaction mixture was stirred at 55 °C for 1.5 hours before a solution of diethyl (bromodifluoromethyl)phosphonate (0.19 g, 0.72 mmol, 2 eq) in tetrahydrofuran (2 mL) was added dropwise (temperature maintained between 50-55 °C). The reaction mixture was stirred at 55 °C for 2 hours (the turbid suspension became a solution) and a solution containing copper bromide (1 +) (51 mg, 0.36 mmol, 1 eq) in dimethylacetamide (2 mL), and the mixture was stirred at 55 ° C for 10 minutes. A solution of 2-iodoquinoline-7-carboxylic acid benzyl ester (0.1 g, 0.36 mmol, 1 eq) in dimethylacetamide (2 mL) was added, and the reaction mixture was stirred at 55 ° C for 16 hours to give a green suspension. The reaction mixture was concentrated under reduced pressure to remove THF. The mixture was purified by preparative HPLC (column: Phenomenex Luna C18 150 x 25 mm x 10 um, mobile phase: water (0.225% FA)-ACN; B%: 54%-84%, 10 minutes) and lyophilized to give 2-((diethoxyphosphoryl)difluoromethyl)quinoline-7-carboxylic acid benzyl ester (40 mg, yield 30%) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 8.95 (s, 1H), 8.36 (d, J = 8.4 Hz, 1H), 8.31-8.22 (m, 1H), 7.95 (d, J = 8.8 Hz, 1H), 7.53-7.49 (m, 2H), 7.46-7.35 (m, 4H), 5.46 (s, 2H), 4.40-4.31 (m, 4H), 1.39 (t, J = 7.2 Hz, 6H)

[0508] Step 4: 2-((diethoxyphosphoryl)difluoromethyl)quinoline-7-carboxylic acid

[0509] In N 2 To a solution of benzyl 2-((diethoxyphosphoryl)difluoromethyl)quinoline-7-carboxylate (40 mg, 89 μmol, 1 eq) in methanol (1 mL) and tetrahydrofuran (1 mL) was added wet Pd / C (40 mg), and the mixture was stirred under H 2 (15 psi) at 25° C. for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated to give 2-((diethoxyphosphoryl)difluoromethyl)quinoline-7-carboxylic acid (15 mg, crude) as a yellow oil. LCMS: m / z (M+H) + =360.1

[0510] Synthesis of building blocks

[0511] Synthesis of 6-phenyl-4-azaspiro[2.4]heptane

[0512]

[0513] Step 1: Preparation of methyl 3-cyano-2-phenylpropionate

[0514] To a cooled (-78°C) solution of methyl 2-phenylacetate (5.0 g, 33.3 mmol, 1.0 eq) in anhydrous THF (50 mL) was slowly added a solution of 2M LDA (20 mL, 40.0 mmol, 1.2 eq) in THF. After 1 hour, 2-bromoacetonitrile (4.2 g, 35.0 mmol, 1.1 eq) was slowly added dropwise and the reaction was further aged at -78°C for another 1 hour. Saturated NH 4 Cl aqueous solution (5 mL) was added and the mixture was warmed to room temperature. The mixture was washed with EtOAc (20 mL x 3). The organic layers were combined, washed with brine (150 mL), and washed with anhydrous Na 2 SO 4 Dry and concentrate under reduced pressure. The residue was purified by silica gel flash column chromatography to give methyl 3-cyano-2-phenylpropanoate (5.0 g, 26.5 mmol, 79%) as a white solid. LCMS (ESI) m / z=190.1 [M+H] + .

[0515] Step 2: Preparation of 6-phenyl-4-azaspiro[2.4]heptan-5-one

[0516] To methyl 3-cyano-2-phenylpropanoate (5.0 g, 26.5 mmol, 1.0 equiv) and Ti(OiPr) 4 To a cooled (0 °C) solution of 4-(9.0 g, 31.7 mmol, 1.2 eq.) in anhydrous THF (100 mL) was slowly added 3M EtMgBr solution (20 mL, 59.6 mmol, 2.25 eq.) while maintaining the internal temperature between -5 °C and 0 °C. After complete addition of EtMgBr, the mixture was stirred at 0 °C for another 1 hour. The reaction mixture was quenched by addition of 2N HCl aqueous solution (60 mL). The resulting acidic solution was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (20 mL x 2) and washed with anhydrous Na 2 SO 4 Dry and concentrate under reduced pressure. The residue was purified by flash column chromatography to give 6-phenyl-4-azaspiro[2.4]heptan-5-one (2.1 g, 11.2 mmol, 42% yield) as a white solid. LCMS (ESI) m / z=188 [M+H] + ; 1 HNMR (400MHz, CDCl 3)δ7.39-7.24(m,6H),3.86(dd,J=9.4,7.6Hz,1H),2.51(dd,J=12.9,9.4Hz,1H),2.2 9(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).

[0517] Step 3: Preparation of 6-phenyl-4-azaspiro[2.4]heptane

[0518] To a cooled (10 °C) solution of 6-phenyl-4-azaspiro[2.4]heptan-5-one (2.1 g, 11.2 mmol, 1.0 equiv) in anhydrous THF (50 mL) was added NaBH in portions. 4 (2.1 g, 56.0 mmol, 5.0 equiv.) BF was added dropwise to the mixture. 3 ·Et 2 O (6.7 mL, 56.0 mmol, 5.0 equiv). The reaction mixture was heated at 60 ° C for 16 hours. The mixture was cooled to ambient temperature and 2N HCl aqueous solution (30 mL) was slowly introduced. The acidic mixture was extracted with EtOAc (30 mL x 3). The organic layers were combined, washed with brine (20 mL), and washed with anhydrous Na 2 SO 4 The residue was dried and concentrated under reduced pressure. Purification by C18 column gave 6-phenyl-4-azaspiro[2.4]heptane (1.5 g, 8.7 mmol, 78%) as a colorless oil. LCMS (ESI) m / z=174 [M+H] + ; 1 H NMR (400 MHz, CDCl 3 )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).

[0519] Pyrrolidine building block synthesis:

[0520] Synthesis of rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile

[0521]

[0522] Step 1: Preparation of (E)-3-cyclohexylacrylonitrile

[0523] To a solution of cyclohexylcarboxaldehyde (500 mg, 4.45 mmol, 1.0 eq) in THF (15 mL) was added t-BuOK (998 mg, 8.90 mmol, 2.0 eq) and diethyl (cyanomethyl)phosphonate (788 mg, 4.45 mmol, 1.0 eq) at room temperature. The solution was stirred at room temperature for 1 h. After completion, the mixture was heated to 40 ℃ and then heated to 30 ℃. 2 The reaction mixture was quenched with 1% 4% dHO (10 mL), and then extracted with EtOAc (10 mL x 3). The organic layers were combined and washed with brine (10 mL), and then dried over anhydrous Na 2 SO 4 Dry, filter and concentrate under reduced pressure. The residue was purified by silica gel flash column chromatography to give (E)-3-cyclohexylacrylonitrile (300 mg, 2.21 mmol, yield 50%) as a colorless oil. LC-MS (ESI) m / z=136 [M+H] + .

[0524] Step 2: Preparation of rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carbonitrile

[0525] (E)-3-Cyclohexylacrylonitrile (300 mg, 2.21 mmol, 1.0 equiv) was dissolved in CH 2 Cl 2 N-benzyl-1-methoxy-N-((trimethylsilyl)methyl)methanamine (524 mg, 2.21 mmol, 1.0 eq) and TFA (25.1 mg, 221 μmol, 0.1 eq) were added to a solution in 4% ethyl acetate (5 mL). The reaction mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was washed with CH 2 Cl 2 (10 mL) and washed with saturated NaHCO 3 The mixture was washed with aqueous solution (5 mL), the organic layer was separated and concentrated under reduced pressure to give crude rel-(trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carbonitrile (700 mg) as a colorless oil. LC-MS (ESI) m / z=269 [M+H] + .

[0526] Step 3: Preparation of rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile

[0527] To a solution of (trans)-1-benzyl-4-cyclohexylpyrrolidine-3-carbonitrile (700 mg) in anhydrous 1,2-dichloroethane (15 mL) was added 1-chloroethyl chloroformate (3.71 g, 26.0 mmol, 10.0 equiv). The resulting mixture was stirred at 70 ° C for 12 hours and then concentrated under reduced pressure, and the crude product was dissolved in MeOH (5 mL) and stirred at 70 ° C for 1 hour. After completion, the reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography to give rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile (211 mg, 1.18 mmol, 46% yield) as a colorless oil. LC-MS (ESI) m / z=179 [M+H] + .

[0528] The following intermediates in Table 10 were prepared according to the representative procedure described for rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile (steps 1 to 3) using appropriate starting materials and modifications. The compounds were prepared as racemates with trans stereochemical configurations relative to the C3 and C4 stereocenters within the pyrrolidine ring.

[0529] Table 10

[0530]

[0531] Rel-(trans)-tert-butyl 3-cyano-4-phenylpyrrolidine-1-carboxylate and tert-butyl (trans)-3-cyano-4-(2-oxo-1,2-dihydropyrrolidine-4-yl)pyrrolidine-1-carboxylate were prepared according to the method described above for the synthesis of rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile. The racemic mixture of the trans isomers was purified under SFC conditions and the absolute stereochemistry was arbitrarily assigned as shown.

[0532]

[0533] Preparative separation methods:

[0534] Instrument: Waters Thar 80 Preparative SFC; Column: ChiralPak C-IG, 100 x 4.6 mm ID, 5 μm; Mobile phase: A for CO 2 and B represents methanol (0.05% diethylamine); gradient: 10% to 40% B in 8 minutes; flow rate: 2.5 ml / min; back pressure: 100 bar; column temperature: 40° C.; wavelength: 210 nm; cycle time: 2 minutes

[0535] The following intermediates in Table 11 were prepared according to the procedure described for rel-(trans)-4-cyclohexylpyrrolidine-3-carbonitrile using appropriate starting materials and modifications.

[0536] Table 11

[0537]

[0538] Synthesis of 2-methyl-2-(4-(methylamino)phenyl)propionitrile

[0539]

[0540] 2-(4-aminophenyl)-2-methylpropionitrile (360 mg, 2.24 mmol, 1 equivalent), PPh 3 A solution of 4-(4-(methylamino)phenyl)propionitrile (50.0 mg, 516 μmol, 23%), 4,5-dichloro-3,6-dioxocyclohexa-1,4-diene-1,2-dicarbonitrile (587 mg, 2.24 mmol, 1 eq.), and MeOH (71.6 mg, 2.24 mmol, 1.0 eq.) in DCM (10 mL) was stirred at room temperature under nitrogen for 1 hour. After completion, the reaction was filtered and concentrated to give a residue, which was purified by silica gel flash column chromatography to give 2-methyl-2-(4-(methylamino)phenyl)propionitrile (90.0 mg, 516 μmol, 23%) as a white solid. LCMS (ESI): m / z=175.3 [M+H] + .

[0541] The following intermediates in Table 12 were prepared according to the procedure described for 2-methyl-2-(4-(methylamino)phenyl)propionitrile using appropriate starting materials and modifications.

[0542] Table 12

[0543]

[0544] Synthesis of 2-methyl-2-(4-(methylamino)phenyl)propan-1-ol

[0545]

[0546] A solution of 2-(4-aminophenyl)-2-methylpropan-1-ol (230 mg, 1.39 mmol, 1 eq.), formaldehyde (112 mg, 1.39 mmol, 1 eq.), sodium cyanoborohydride (87.3 mg, 1.39 mmol, 1 eq.) in MeOH (20 mL) was stirred overnight at room temperature under nitrogen. After completion, the reaction was filtered and concentrated. The residue was purified by C18 column to give 2-methyl-2-[4-(methylamino)phenyl]propan-1-ol (60.0 mg, 334 μmol, 24%) as a white solid. LCMS (ESI): m / z=180.2[M+H] + .

[0547] The following intermediates in Table 13 were prepared according to the procedures described for 2-methyl-2-(4-(methylamino)phenyl)propan-1-ol using appropriate starting materials and modifications.

[0548] Table 13

[0549]

[0550] Synthesis of intermediate: (rel-trans)-5-phenylpiperidine-3-carbonitrile

[0551]

[0552] Step A: 5-phenylnicotinate methyl ester

[0553] Under nitrogen, Na 2 CO 3 (1.96 g, 18.5 mmol, 2.0 equiv) in H 2 An aqueous solution of 5-bromonicotinate (2.0 g, 9.25 mmol, 1 eq.) and a solution of phenylboronic acid (1.34 g, 11.0 mmol, 1.2 eq.) in MeOH (5 mL) were added to methyl 5-bromonicotinate (2.0 g, 9.25 mmol, 1 eq.) and Pd(PPh 3 ) 4 (320 mg, 277 μmol, 0.03 equiv) in a stirred solution in toluene (20 mL). The stirred mixture was stirred at 80 ° C for 4 hours. The reaction mixture was cooled to room temperature and extracted with EtOAc (60 mL). The solvent was removed under reduced pressure. The residue was purified by flash chromatography on silica gel to give 5-phenylnicotinate methyl ester (1.30 g, 6.09 mmol, 61%) as a white solid. 1 H NMR (400 MHz, CDCl 3)δ9.19(d,J=1.9Hz,1H),9.01(d,J=2.3Hz,1H),8.49(t,J=2.1Hz,1H),7.68-7.57(m,2H),7.56-7.35(m,3H),3.99(s,3H).

[0554] Step B: Methyl 5-phenylpiperidine-3-carboxylate

[0555] To 5-phenylnicotinate methyl ester (1.0 g, 4.68 mmol, 1 eq.) and H 2 SO 4 To a solution of PtO (0.05 mL) in MeOH (10 mL) was added 2 (100 mg, 10%). The suspension was degassed under vacuum and washed with H 2 The mixture was purged three times under H 2 (balloon) at room temperature for 16 hours. The reaction mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give methyl 5-phenylpiperidine-3-carboxylate (1 g, 4.56 mmol, 98%) as a white solid. LCMS (ESI) m / z=220 [M+H] + .

[0556] Step C: (rel-trans)-5-phenylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester

[0557] To a solution of 5-phenylpiperidine-3-carboxylic acid methyl ester (1.0 g, 4.56 mmol, 1 equiv) in THF (10 mL) was added TEA (1.37 g, 13.6 mmol, 3.0 equiv) and di-tert-butyl dicarbonate (1.49 g, 6.84 mmol, 1.5 equiv) at room temperature. After addition, the reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel flash chromatography (cis and trans isomers separable) to give (trans)-5-phenylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) ester 3-methyl ester (800 mg, 2.50 mmol, 55%) as a colorless oil. LCMS (ESI) m / z=320 [M+H] + . 1 H NMR (400 MHz, CDCl 3 )δ7.34-7.23(m,5H),4.46-3.96(m,2H),3.72(s,3H),3.29-2.89(m,3H) ,2.81-2.64(m,1H),2.44-2.24(m,1H),1.98-1.83(m,1H),1.45(s,9H).

[0558] Step D: (rel-trans)-1-(tert-Butyloxycarbonyl)-5-phenylpiperidine-3-carboxylic acid

[0559] To (rel-trans)-5-phenylpiperidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (120 mg, 375 μmol, 1 eq.) in THF (4 mL), H 2 To the solution in a mixture of 2H2O (1 mL) and MeOH (1 mL) was added LiOH-H 2 O (78.4 mg, 1.87 mmol, 5.0 equiv.). After addition, the reaction mixture was stirred at room temperature overnight. The reaction mixture was cooled to 0°C and acidified to pH = 3 with 2.0 M HCl aqueous solution. The mixture was extracted with EtOAc (30 mL). The organic phase was purified by Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give (rel-trans)-1-(tert-butoxycarbonyl)-5-phenylpiperidine-3-carboxylic acid (110 mg, 360 μmol, 96.4%) as a colorless oil. LCMS (ESI): m / z=304 [MH] - .

[0560] Step E: (rel-trans)-tert-butyl 3-carbamoyl-5-phenylpiperidine-1-carboxylate

[0561] A solution of (rel-trans)-1-(tert-butoxycarbonyl)-5-phenylpiperidine-3-carboxylic acid (110 mg, 360 μmol, 1 eq) and CDI (64 mg, 396 μmol, 1.1 eq) in EtOAc (4 mL) was stirred at room temperature for 2 h. NH 3 .H 2 The mixture was stirred at 45 ° C for 2 hours. The reaction mixture was cooled to room temperature and extracted with EtOAc (15 mL). The mixture was washed with water (5 mL). The organic phase was concentrated under reduced pressure to obtain a residue. The residue was purified by C18 column to obtain (rel-trans)-3-carbamoyl-5-phenylpiperidine-1-carboxylic acid tert-butyl ester (80 mg, 262 μmol, 73%) as a white solid. LCMS (ESI): m / z=305.2[M+H] + .

[0562] Step F: (rel-trans)-3-cyano-5-phenylpiperidine-1-carboxylic acid tert-butyl ester

[0563] To a solution of (rel-trans)-3-carbamoyl-5-phenylpiperidine-1-carboxylic acid tert-butyl ester (80 mg, 262 μmol, 1 equiv) in DCM (2 mL) at room temperature was added TEA (66 mg, 655 μmol, 2.5 equiv). The mixture was cooled to 0 ° C, and TFAA (71 mg, 340 μmol, 1.3 equiv) was added dropwise. After addition, the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with DCM (15 mL). The mixture was washed with water (5 mL). The organic phase was concentrated under reduced pressure to give a residue. The residue was purified by C18 column to give (rel-trans)-3-cyano-5-phenylpiperidine-1-carboxylic acid tert-butyl ester (57 mg, 199 μmol, 76%) as a white solid. LCMS (ESI): m / z=309[M+Na] + .

[0564] Step G: (rel-trans)-5-phenylpiperidine-3-carbonitrile

[0565] A solution of (rel-trans)-3-cyano-5-phenylpiperidine-1-carboxylic acid tert-butyl ester (57 mg, 199 μmol, 1 equiv) in 4.0 M HCl / dioxane (2 mL) was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to give (rel-trans)-5-phenylpiperidine-3-carbonitrile (35 mg, 187 μmol, 95%) as a white solid, which was used in the next step without purification. LCMS (ESI): m / z=187 [M+H] + .

[0566] Preparation of (3S,5R)-5-Benzylpyrrolidine-3-carbonitrile

[0567]

[0568] Step A: (S)-2-(Benzylamino)-3-phenylpropan-1-ol

[0569] Benzoyl-L-phenylalanine (1 g, 3.71 mmol, 1 eq.) was added dropwise to a solution containing LiAlH 4 (421 mg, 11.1 mmol, 3 eq) was added to THF (30 mL) and the resulting mixture was stirred at 70 °C for 18 h. After completion, the mixture was purified by Na 2 SO 4 -10H 2The reaction was quenched, filtered, and the filter cake was washed with DCM (50 mL). The filtrate was concentrated in vacuo. The residue was purified by silica gel flash column chromatography to give (S)-2-(benzylamino)-3-phenylpropan-1-ol (700 mg, 2.90 mmol, 78%) as a white solid. LCMS (ESI): m / z=242.2 [M+H] + .

[0570] Step B: (S)-3-(Benzyl(1-hydroxy-3-phenylpropan-2-yl)amino)propionitrile

[0571] To 1,4-dioxane (15 mL) containing (S)-2-(benzylamino)-3-phenylpropan-1-ol (650 mg, 2.69 mmol, 1 eq) and prop-2-enenitrile (9.97 g, 188 mmol, 70 eq) was added AcOH (322 mg, 5.38 mmol, 2 eq) at room temperature, and the solution was stirred at 80 ° C for 48 hours. After completion, the solvent was removed under reduced pressure, and the residue was purified by silica gel flash column chromatography to give (S)-3-(benzyl(1-hydroxy-3-phenylpropan-2-yl)amino)propionitrile (560 mg, 1.90 mmol, 71%) as a white solid. LCMS (ESI): m / z=295.2 [M+H] + .

[0572] Step C: (S)-3-(Benzyl(1-chloro-3-phenylpropan-2-yl)amino)propionitrile

[0573] At room temperature, SOCl 2 (553 mg, 4.65 mmol, 2.5 eq) was added to (S)-3-(benzyl(1-hydroxy-3-phenylpropan-2-yl)amino)propionitrile (550 mg, 1.86 mmol, 1 eq) in DCM (10 mL), and the solution was stirred at 70 °C for 0.5 h. After completion, the reaction was cooled to room temperature and washed with NaHCO 3 The mixture was quenched with (aqueous solution) (2 mL), extracted with EtOAc (10 mL x 2), the combined organic layers were washed with brine, and purified by Na 2 SO 4 Dry, filter, and concentrate under reduced pressure to give crude (S)-3-(benzyl(1-chloro-3-phenylpropan-2-yl)amino)propionitrile (550 mg, 1.75 mmol, 95%) as an oil. LCMS (ESI): m / z=313.2 [M+H] + .

[0574] Step D: (3R,5S)-1,5-dibenzylpyrrolidine-3-carbonitrile

[0575] To (S)-3-(benzyl(1-chloro-3-phenylpropan-2-yl)amino)propionitrile (550 mg, 1.75 mmol, 1 eq) in THF (20 mL) was added NaHMDS (3 mL, 1.2 eq) under nitrogen at room temperature, and the solution was stirred at room temperature for 0.5 h. After completion, the reaction was washed with NH 4 The solution was quenched with Cl(aqueous solution) (0.5 mL) and extracted with EtOAc (5 mL x 2). The combined organic layers were washed with brine and purified by Na 2 SO 4 The mixture was dried, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography to give (3S,5R)-1,5-dibenzylpyrrolidine-3-carbonitrile (150 mg, 0.54 mmol, 31%) and (3R,5R)-1,5-dibenzylpyrrolidine-3-carbonitrile (250 mg, 0.90 mmol, 52%) as oils. LCMS (ESI): m / z=277.2 [M+H] + .

[0576] Step E: (3R,5S)-5-Benzylpyrrolidine-3-carbonitrile

[0577] To a solution of (3S, 5R)-1,5-dibenzylpyrrolidine-3-carbonitrile (150 mg, 0.5427 mmol, 1 eq) in DCE (10 mL) was added 1-chloroethyl chloroformate (387 mg, 2.71 mmol, 5 eq) at room temperature, and then stirred at 80 ° C for 18 hours. The solution was concentrated, and the residue was then dissolved in MeOH (10 mL) and stirred at 70 ° C for 2 hours. After completion, the solution was concentrated, and the residue was purified by silica gel flash column chromatography to give (3S, 5R)-5-benzylpyrrolidine-3-carbonitrile (50.0 mg, 0.27 mmol, 50%). LCMS (ESI): m / z=187.2[M+H] + . 1 H NMR (400 MHz, CDCl 3 )δ7.34-7.16(m,5H),3.35-3.24(m,2H),3.19-3.10(m,1H),2.96-2.72(m,3H),2.32-2.20(m,1H),1.79-1.63(m,1H).

[0578] Step E (cis): (3R,5R)-5-benzylpyrrolidine-3-carbonitrile

[0579] 1-Chloroethyl chloroformate (516 mg, 3.61 mmol, 5 eq) was added to a solution of (3R, 5R)-1,5-dibenzylpyrrolidine-3-carbonitrile (200 mg, 0.72 mmol, 1 eq) in DCM (10 mL) at room temperature and then stirred at 80 ° C for 18 hours. After completion, the solution was concentrated, and the residue was then dissolved in MeOH (10 mL), and the solution was stirred at 70 ° C for 2 hours. The solution was concentrated under reduced pressure, and the residue was purified by silica gel flash column chromatography to give (3R, 5R)-5-benzylpyrrolidine-3-carbonitrile (30.0 mg, 0.16 mmol, 22%). LCMS (ESI): m / z=187.2[M+H] + . 1 H NMR (400 MHz, CDCl 3 )δ7.34-7.16(m,5H),3.56-3.43(m,1H),3.42-3.30(m,1H),3.11-3.01(m,1H), 2.99-2.88(m,1H),2.76(d,J=6.7Hz,2H),2.22-2.10(m,1H),1.85-1.73(m,1H).

[0580] Preparation of (rel-trans)-4-phenylpiperidine-3-carbonitrile

[0581]

[0582] Step A: (rel-trans)-tert-butyl 3-carbamoyl-4-phenylpiperidine-1-carboxylate

[0583] A solution of (rel-trans)-1-(tert-butoxycarbonyl)-4-phenylpiperidine-3-carboxylic acid (400 mg, 1.30 mmol, 1 eq) and CDI (231 mg, 1.43 mmol, 1.1 eq) in EtOAc (5 mL) was stirred at room temperature for 2 h. NH 3 .H 2 O (2 mL). The reaction mixture was stirred at 45 °C for 2 h. After completion, the reaction mixture was cooled to room temperature and washed with H 2 O (5 mL), extracted with EtOAc (10 mL x 3). The organic layers were combined and washed with brine (10 mL), and dried over anhydrous Na 2 SO 4 Dry and concentrate under reduced pressure. The residue was purified by C18 column to give (rel-trans)-3-carbamoyl-4-phenylpiperidine-1-carboxylic acid tert-butyl ester (390 mg, 1.28 mmol, 99%) as a white solid. LCMS (ESI) m / z=305 [M+H]+ .

[0584] Step B: (rel-trans)-tert-butyl 3-cyano-4-phenylpiperidine-1-carboxylate

[0585] To a solution of (rel-trans)-tert-butyl 3-carbamoyl-4-phenylpiperidine-1-carboxylate (390 mg, 1.28 mmol, 1 eq) in DCM (5 mL) was added TEA (323 mg, 3.20 mmol, 2.5 eq) at room temperature. The mixture was cooled to 0 ° C, and TFAA (348 mg, 1.66 mmol, 1.3 eq) was added dropwise. After addition, the reaction mixture was stirred at room temperature for 16 hours. After completion, the reaction mixture was washed with H 2 O (5 mL) and extracted with DCM (5 mL x 3). The organic layers were combined and washed with brine (10 mL) and dried over anhydrous Na 2 SO 4 Dry and concentrate under reduced pressure. The residue was purified by C18 column to give (rel-trans)-tert-butyl 3-cyano-4-phenylpiperidine-1-carboxylate (160 mg, 558 μmol, 44%) as a white solid. LCMS (ESI) m / z=309 [M+Na].

[0586] Step C: (rel-trans)-4-phenylpiperidine-3-carbonitrile hydrochloride

[0587] A solution of (rel-trans)-3-cyano-4-phenylpiperidine-1-carboxylate (80 mg, 279 μmol, 1 eq.) in 4.0 M HCl / dioxane (2 mL) was stirred at room temperature for 1 hour. After completion, the reaction mixture was concentrated under reduced pressure to give (rel-trans)-4-phenylpiperidine-3-carbonitrile hydrochloride (52 mg, 279 μmol, 100%) as a white solid, which was used directly in the next step without purification. LCMS: m / z=187 [M+H] + .

[0588] Preparation of (rel-trans)-3-(difluoromethyl)-4-phenylpyrrolidine

[0589]

[0590] Step A: (rel-trans)-tert-butyl 3-formyl-4-phenylpyrrolidine-1-carboxylate

[0591] at -78 °C in N 2DIBAL-H (3.66 mL, 3.66 mmol, 2 eq) solution was added to a solution of (rel-trans)-3-cyano-4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (500 mg, 1.83 mmol, 1 eq) in THF (10 mL) via syringe over a 10-minute period. The mixture was then stirred at -78 °C for 1 hour. Upon completion, once the reaction mixture reached -60 °C to -20 °C, NH 4 Cl (1 mL, aqueous solution) solution was added dropwise to the reaction mixture. The residue was partitioned between EtOAc (20 mL) and 1N HCl aqueous solution (10 mL). The separated organic layer was washed with brine (10 mL) and purified by Na 2 SO 4 Dry and evaporate to dryness. The crude product was purified by silica gel flash column chromatography to give (rel-trans)-3-formyl-4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (300 mg, 1.08 mmol, 60%) as a colorless oil. LCMS (ESI): m / z=221.2 [M+H-56] + .

[0592] Step B: (rel-trans)-tert-butyl 3-(difluoromethyl)-4-phenylpyrrolidine-1-carboxylate

[0593] To a solution of (rel-trans)-3-formyl-4-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (100 mg, 363 μmol, 1 eq.) in anhydrous DCM (2 mL) was added DAST (117 mg, 726 μmol, 2 eq.) in several batches at 0 ° C., and the mixture was stirred at room temperature for 14 hours. The reaction was then quenched with MeOH (2.0 mL) and extracted with dichloromethane (10 mL x 3). The organic layers were combined and washed with brine (10 mL x 2) and washed with anhydrous Na 2 SO 4 Dried and concentrated under reduced pressure.The residue was purified by silica gel column chromatography to give tert-butyl (rel-trans)-3-(difluoromethyl)-4-phenylpyrrolidine-1-carboxylate (100 mg, 336 μmol, 93%) as a colorless oil.

[0594] Step C: (rel-trans)-3-(difluoromethyl)-4-phenylpyrrolidine

[0595] To a solution of tert-butyl (rel-trans)-3-(difluoromethyl)-4-phenylpyrrolidine-1-carboxylate (110 mg, 369 μmol, 1 eq) in DCM (2 mL) was added TFA (419 mg, 3.68 mmol, 10 eq). The reaction mixture was stirred at room temperature for 1 hour. After the reaction, the reaction was cooled in an ice bath and then treated with NaHCO 3 (aqueous solution) was carefully neutralized until the pH was adjusted to pH = 8-9. The resulting mixture was extracted with DCM (10 mL x 3), and the combined organic layers were washed with brine (10 mL x 2), washed with anhydrous Na 2 SO 4 Drying and then concentration under reduced pressure gave a crude product (rel-trans)-3-(difluoromethyl)-4-phenylpyrrolidine (73 mg, 369 μmol, quantitative) as a white solid, which was used directly in the next step without further purification. LCMS (ESI): m / z=198.2 [M+H] + .

[0596] Synthesis of 2-(4-amino-[1,1'-biphenyl]-3-yl)acetonitrile

[0597]

[0598] Step 1: 2-(2-amino-4-bromophenyl)acetonitrile

[0599] To a solution of 2-(2-aminophenyl)acetonitrile (2.0 g, 15 mmol, 1 eq.) in dimethylformamide (20 mL) was added N-bromosuccinimide (3.0 g, 17 mmol, 1.1 eq.). The mixture was stirred at 25 °C for 12 hours. The mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 2), the combined organic layers were washed with saturated brine (200 mL x 2), and the mixture was purified by Na 2 O 4 '-trifluoroacetic acid (THF) reaction. 2 SO 4 Dry, filter and concentrate under reduced pressure to give 2-(2-amino-4-bromophenyl)acetonitrile (3.2 g, crude) as a white solid. LCMS: m / z (M+H)+=210.9.

[0600] Step 2: 2-(4-amino-[1,1'-biphenyl]-3-yl)acetonitrile

[0601] To a solution of 2-(2-amino-4-bromophenyl)acetonitrile (1 g, 4.7 mmol, 1 eq) in water (2 mL) and dioxane (8 mL) were added tetrakis(triphenylphosphine)palladium (0.27 g, 0.24 mmol, 0.05 eq), potassium carbonate (1.9 g, 14 mmol, 3 eq) and phenylboronic acid (0.65 g, 5.2 mmol, 1.1 eq). The mixture was degassed and heated with N 2 The mixture was purged 3 times and then stirred at 85°C for 12 hours. The reaction mixture was filtered and the filtrate was concentrated to give a residue. The residue was extracted with water (100 mL) and ethyl acetate (100 mL x 2). The combined organic layers were washed with saturated brine (100 mL x 2) and purified by Na 2 SO 4 The residue was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate=1:0 to 2:1) to give 2-(4-amino-[1,1′-biphenyl]-3-yl)acetonitrile (0.50 g, crude) as a white solid. LCMS: m / z (M+1)=209.0.

[0602] Synthesis of 2-(4-amino-[1,1'-biphenyl]-3-yl)acetamide

[0603]

[0604] Step 1: 2-(5-Bromo-2-nitrophenyl)acetyl chloride

[0605] A solution of 2-(5-bromo-2-nitrophenyl)acetic acid (0.30 g, 1.2 mmol, 1 eq.) 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) as a white solid.

[0606] Step 2: 2-(5-bromo-2-nitrophenyl)acetamide

[0607] 2-(5-bromo-2-nitrophenyl)acetyl chloride (0.30 g, 1.1 mmol, 1 equivalent) was dissolved in tetrahydrofuran (3 mL) and NH 3 .H 2 The solution in 0 (4 mL) was stirred at 25° C. for 30 minutes. The reaction mixture was concentrated under reduced pressure to give 2-(5-bromo-2-nitrophenyl)acetamide (0.30 g, crude) as a white solid. LCMS: m / z (M+H)=258.9.

[0608] Step 3: 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide

[0609] To a solution of 2-(5-bromo-2-nitrophenyl)acetamide (0.10 g, 0.39 mmol, 1 eq) in dioxane (1 mL) and water (0.2 mL) were added sodium carbonate (0.12 g, 1.2 mmol, 3 eq), phenylboronic acid (52 mg, 0.42 mmol, 1.1 eq), bis(cyclopenta-2,4-diyn-1-ylbiphenyl-λ4-phosphonate)dichloropalladium iron (2+) (14 mg, 0.019 mmol, 0.05 eq), and the mixture was stirred at 100° C. for 12 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2), the combined organic layers were washed with saturated brine (20 mL x 2), and the mixture was purified by purifying with Na 2 SO 4 Dry, filter and concentrate 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 (60 mg, 60% yield) as a white solid. LCMS: m / z (M+23)+=279.0. 1 H NMR (400MHz, CD 3 OD)δ8.17(d,J=8.4Hz,1H),7.79-7.69(m,4H),7.52-7.40(m,3H),4.06(s,2H)

[0610] Step 4: 2-(4-amino-[1,1'-biphenyl]-3-yl)acetamide

[0611] To a solution of 2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (60 mg, 0.23 mmol, 1 eq) in ethanol (0.8 mL) and water (0.2 mL) was added ammonium chloride (0.038 g, 0.70 mmol, 3 eq) and iron (0.13 g, 2.4 mmol, 10 eq). 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) as a white solid. LCMS: m / z (M+H) = 227.0. 1 H NMR (400MHz, DMSO) δ7.53 (d, J = 7.6 Hz, 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).

[0612] Synthesis of 3-(thiazol-2-ylmethyl)-[1,1'-biphenyl]-4-amine

[0613]

[0614] Step 1: 4-Nitro-[1,1'-biphenyl]-3-carbaldehyde

[0615] In N 2 To a solution of 5-bromo-2-nitrobenzaldehyde (5 g, 22 mmol, 1 eq) in dioxane (30 mL) and water (10 mL) was added phenylboronic acid (5.3 g, 43 mmol, 2 eq) and potassium carbonate (12 g, 87 mmol, 4 eq), tetrakis(triphenylphosphine)palladium (1.2 g, 1.1 mmol, 0.05 eq). The reaction was stirred at 90 °C for 16 hours to give a black solution. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), the combined organic layers were washed with saturated brine (50 mL x 2), and the mixture was purified by Na 2 O 4 '-triphenylphosphine (10 mL x 2). 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give 4-nitro-[1,1'-biphenyl]-3-carbaldehyde (3.5 g, 71% yield) as a yellow solid. 1 H NMR (400 MHz, chloroform-d) δ 10.52 (s, 1H), 8.22 (d, J = 8.4 Hz, 1H), 8.14 (d, J = 2.0 Hz, 1H), 7.94 (d, J = 6.4 Hz, 1H), 7.69-7.63 (m, 2H), 7.56-7.46 (m, 3H)

[0616] Step 2: (4-nitro-[1,1'-biphenyl]-3-yl)(thiazol-2-yl)methanol

[0617] Monobutyl lithium hydride (6.3 mL, 16 mmol, 1.2 eq) was added dropwise to a solution of 2-bromo-1,3-thiazole (2.2 g, 13 mmol, 1 eq) in tetrahydrofuran (30 mL) at -60 ° C and stirred at -60 ° C for 30 minutes. A solution of 4-nitro-[1,1'-biphenyl]-3-carboxaldehyde (3 g, 13 mmol, 1 eq) in THF (10 mL) was then added dropwise at -60 ° C. The reaction mixture was warmed to 25 ° C and stirred for 2 hours to obtain a black solution. The reaction mixture was quenched with saturated ammonium chloride (80 mL). The mixture was diluted with water (150 mL) and extracted with ethyl acetate (150 mL x 2), and the combined organic layers were washed with saturated brine (150 mL x 2) and washed with Na 2 SO 4Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give (4-nitro-[1,1'-biphenyl]-3-yl)(thiazol-2-yl)methanol (1.9 g, 46% yield) as a yellow solid. 1 H NMR (400 MHz, chloroform-d) δ8.17 (d, J = 8.4 Hz, 1H), 8.00 (d, J = 1.6 Hz, 1H), 7.79-7.67 (m, 2H), 7.67-7.57 (m, 2H), 7.53-7.42 (m, 3H), 7.35 (d, J = 3.2 Hz, 1H), 6.76 (s, 1H)

[0618] Step 3: 3-(Thiazol-2-ylmethyl)-[1,1'-biphenyl]-4-amine

[0619] In H 2 To a solution of (4-nitro-[1,1'-biphenyl]-3-yl)(thiazol-2-yl)methanol (0.1 g, 0.3 mmol, 1 equiv) in methanol (1 mL) and tetrahydrofuran (1 mL) was added HCl (0.5 mL, 1 M) and Pd / C (0.1 g). The mixture was stirred at 60 ° C for 6 hours under 50 psi. The reaction mixture was filtered and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography to give 3-(thiazol-2-ylmethyl)-[1,1'-biphenyl]-4-amine (70 mg, 82% yield) as a red oil. LCMS: (ES+) m / z (M+H)+=266.9 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.60 (d, J = 3.2 Hz, 1H), 7.47 (dd, J = 1.2, 8.4 Hz, 2H), 7.36-7.26 (m, 4H), 7.22-7.17 (m, 1H), 7.12 (d, J = 3.2 Hz, 1H), 6.69 (d, J = 8.0 Hz, 1H), 4.24 (s, 2H).

[0620] Synthesis of 6-phenyl-1,2,3,4-tetrahydroquinoline-4-carboxamide

[0621]

[0622] Step 1: 6-phenylquinoline-4-carboxylic acid

[0623] Potassium carbonate (0.55 g, 4.0 mmol) was added to a solution of 6-bromoquinoline-4-carboxylic acid (0.50 g, 2.0 mmol) and phenylboronic acid (0.29 g, 2.4 mmol), bis(triphenylphosphine)palladium dichloro (2+) (0.14 g, 0.20 mmol) in dioxane (6 mL) and water (2 mL). The reaction mixture was then stirred at 100 ° C for 3 hours to obtain a black suspension. The reaction mixture was filtered, and the filtrate was collected, poured into water (20 mL), and adjusted to pH = 12 with NaOH (2 M), and washed with EtOAc (30 mL x 2), and then the aqueous phase was adjusted to pH = 5 with HCl (1 M) to obtain a white suspension. The suspension was extracted with ethyl acetate (30 mL x 2). The combined organic phase was washed with saturated brine (30 mL) and washed with anhydrous Na 2 SO 4 Dry and then filter, concentrate in vacuo to give 6-phenylquinoline-4-carboxylic acid (0.6 g, crude) as a white solid. LCMS: m / z (M+H)+=251.0

[0624] Step 2: Ethyl 5-iodobenzo[d]thiazole-2-carboxylate

[0625] In N 2 6-Phenylquinoline-4-carboxylic acid (1.2 g, 4.8 mmol) was dissolved in AcOH (5 mL) and MeOH (10 mL) and then heated under N 2 Pd / C (0.15 g, dry) was added at 15°C, and the reaction mixture was then stirred under H 2 (15 psi) at 15 ° C. for 16 hours to give a black suspension. The reaction mixture was filtered and the filtrate was concentrated to give a residue. The residue was purified by silica gel chromatography to give 6-phenyl-1,2,3,4-tetrahydroquinoline-4-carboxylic acid (0.45 g, 37% yield) as a white solid. LCMS: m / z (M+H) + = 254.0

[0626] Step 3: 6-phenyl-1,2,3,4-tetrahydroquinoline-4-carboxamide

[0627] 6-Phenyl-1,2,3,4-tetrahydroquinoline-4-carboxylic acid (0.3 g, 1.2 mmol) was dissolved in DCM (5 mL), carbonyldiimidazole (0.21 g, 1.3 mmol) was added at 0 °C for 30 min, and then NH 3(15 psi, balloon), and then the reaction mixture was stirred at 15 ° C for 4 hours to give a yellow solution. The mixture was poured into water (35 mL), and the aqueous phase was extracted with ethyl acetate (35 mL x 3). The combined organic phase was washed with brine (35 mL x 2) and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate in vacuo to give a residue. The residue was purified by silica gel chromatography to give 6-phenyl-1,2,3,4-tetrahydroquinoline-4-carboxamide (0.15 g, 50% yield) as a yellow oil. 1 HNMR (400 MHz, CHLOROFORM-d) δ7.55-7.50 (m, 2H), 7.44-7.34 (m, 3H), 7.34-7.28 (m, 2H), 6.64 (d, J = 8.4 Hz, 1H), 5.82-5.38 (m, 2H), 3.81-3.69 (m, 1H), 3.44-3.28 (m, 2H), 2.52 (qd, J = 3.2, 13.2 Hz, 1H), 2.11-2.00 (m, 1H).

[0628] Synthesis of 2-(4-amino-[1,1'-biphenyl]-3-yl)-N-methylacetamide

[0629]

[0630] Step 1: 2-(5-Bromo-2-nitrophenyl)-N-methylacetamide

[0631] To a solution of 2-(5-bromo-2-nitrophenyl)acetic acid (0.10 g, 0.38 mmol, 1 eq) in dichloromethane (1 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (0.062 g, 0.38 mmol, 1 eq) and N,N-diisopropylethylamine (0.13 g, 1.5 mmol, 3 eq) and the mixture was stirred at 25 °C for 15 minutes. Methylamine (0.014 g, 0.46 mmol, 1.2 eq) was then added to the mixture. The mixture was stirred at 25 °C for 12 hours. The mixture was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 1:0 to 0:1) and concentrated in vacuo to give 2-(5-bromo-2-nitrophenyl)-N-methylacetamide (0.10 g, yield 96.1%) as a white solid. LCMS: m / z (M+H) + =272.8. 1 H NMR (400 MHz, METHANOL-d 4)δ7.98(d,J=9.2Hz,1H),7.73-7.63(m,2H),3.91(s,2H),2.73(s,3H).

[0632] Step 2: N-methyl-2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide

[0633] To a solution of 2-(5-bromo-2-nitrophenyl)-N-methylacetamide (0.10 g, 0.37 mmol, 1 eq) in dioxane (1 mL) and water (0.2 mL) was added sodium carbonate (0.12 g, 1.1 mmol, 3 eq), phenylboronic acid (0.054 g, 0.44 mmol, 1.2 eq) and bis(cyclopenta-2,4-diyn-1-yldiphenyl-λ4-phosphonate)dichloropalladium iron (2+) (0.013 g, 0.018 mmol, 0.05 eq), and the mixture was stirred at 100 ° C for 12 hours. The reaction mixture was filtered to obtain a filtrate. The filtrate was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2), the combined organic layer was washed with saturated brine (20 mL x 2), and the mixture was stirred at 100 ° C for 12 hours. 2 SO 4 The residue was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 10: 1 to 0: 1) to give N-methyl-2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (0.10 g, yield 91.0%) as a white solid. LCMS: m / z (M+H) + =271.0

[0634] Step 3: 2-(4-amino-[1,1'-biphenyl]-3-yl)-N-methylacetamide

[0635] To a solution of N-methyl-2-(4-nitro-[1,1'-biphenyl]-3-yl)acetamide (0.10 g, 0.37 mmol, 1 eq) in ethanol (0.8 mL) and water (0.2 mL) was added ammonium chloride (0.059 g, 1.1 mmol, 3 eq) and iron (0.21 g, 3.7 mmol, 10 eq). 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)-N-methylacetamide (0.10 g, crude) as a white solid. LCMS: m / z (M+H) + =241.2.

[0636] Synthesis of (rel-trans)-2-phenylazetidine-3-carbonitrile

[0637]

[0638] Step 1: (E)-N-Benzylidene-1-(4-methoxyphenyl)methanamine

[0639] To a solution of benzaldehyde (5.0 g, 36 mmol, 1.5 eq.) and 1-(4-methoxyphenyl)methylamine (9.7 g, 71 mmol, 1.5 eq.) in DCM (15 mL) was added magnesium sulfate (4.4 g, 36 mmol, 1.5 eq.) at 15 °C, and the reaction mixture was stirred at 15 °C for 16 hours to give a white suspension. The mixture was poured into water (50 mL). The aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined organic phases were washed with brine (50 mL x 2) and washed with anhydrous Na 2 SO 4 Dry, filter and concentrate in vacuo to give a residue. The residue was purified by silica gel chromatography to give (E)-N-benzylidene-1-(4-methoxyphenyl)methanamine (5.6 g, 82% yield) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.27 (s, 1H), 7.71-7.63 (m, 2H), 7.46-7.28 (m, 3H), 7.16 (d, J=8.8 Hz, 2H), 6.83-6.75 (m, 2H), 4.67 (s, 2H), 3.69 (s, 3H).

[0640] Step 2: (rel-trans)-3-chloro-1-(4-methoxybenzyl)-4-phenylazetidin-2-one

[0641] To a solution of (E)-N-benzylidene-1-(4-methoxyphenyl)methanamine (5.6 g, 25 mmol, 1 eq) and lutidine (6.7 g, 62 mmol, 2.5 eq) in toluene (30 mL) was added 2-chloroacetyl chloride (4.2 g, 37 mmol, 1.5 eq) at 15°C, and the reaction mixture was stirred at 80°C for 16 hours to give a yellow solution. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 2), and the combined organic layers were washed with saturated brine (100 mL x 2), and the mixture was purified by Na 2 O 4 '-trifluoroacetyl chloride (400 mL x 2). 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by silica gel chromatography to give (rel-trans)-3-chloro-1-(4-methoxybenzyl)-4-phenylazetidine-2-one (4.5 g, 60% yield) as a yellow oil. LCMS: m / z (M+H)+=302.1. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.50-7.39 (m, 3H), 7.27 (s, 2H), 7.11 (s, 2H), 6.89-6.82 (m, 2H), 4.89-4.81 (m, 1H), 4.57 (d, J = 0.8 Hz, 1H), 4.38 (d, J = 1.2 Hz, 1H), 3.83 (s, 3H), 3.80 (d, J = 15.2 Hz, 1H).

[0642] Step 3: (rel-trans)-3-chloro-1-(4-methoxybenzyl)-2-phenylazetidine

[0643] To a solution of aluminum chloride (5.9 g, 4.9 mmol, 3 eq.) in THF (45 mL) was added lithium (1+) (1.7 g, 4.9 mmol, 3 eq.) of aluminum tetrahydride at 0°C, and the reaction mixture was stirred at 0°C for 10 minutes and at 70°C for 30 minutes. (rel-trans)-3-chloro-1-(4-methoxybenzyl)-4-phenylazetidine-2-one (4.5 g, 1.7 mmol, 1 eq.) was slowly added in THF (45 mL), and the reaction was stirred at 70°C for 4 hours to give a white suspension. 5 mL of water was added to the reaction at 0°C and stirred for 30 minutes, then the reaction mixture was filtered, and the filtrate was collected and concentrated in vacuo to give a yellow solid. The residue was purified by silica gel chromatography to give (rel-trans)-3-chloro-1-(4-methoxybenzyl)-2-phenylazetidine (2.5 g, 59% yield) as a beige oil. LCMS: m / z (M+H)+=288.1.

[0644] Step 4: (rel-trans)-1-(4-methoxybenzyl)-2-phenylazetidine-3-carbonitrile

[0645] To a solution of (rel-trans)-3-chloro-1-(4-methoxybenzyl)-2-phenylazetidine (1.5 g, 5.2 mmol, 1 eq) in dimethyl sulfoxide (20 mL) was added potassium cyanide (0.67 g, 10 mmol, 2 eq) and the mixture was stirred at 100 °C for 4 hours to give a brown solution. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 2), the combined organic layers were washed with saturated brine (100 mL x 2), and the mixture was purified by HPLC. 2 SO 4 Dry, filter and concentrate under reduced pressure to obtain a residue. The aqueous phase was adjusted to pH = 14 with 1N NaOH, then NaOCl aqueous solution (200 mL) was added and allowed to stand for 48 hours, and then discarded after passing the professional test. The residue was purified by column chromatography (SiO2 , petroleum ether / ethyl acetate=100:1 to 90:10) to give (rel-trans)-1-(4-methoxybenzyl)-2-phenylazetidine-3-carbonitrile (0.65 g, yield 45%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 )δ7.49-7.44(m,2H),7.41-7.30(m,3H),7.20-7.15(m,2H),6.86-6.81(m,2H),4.31(d,J=8.4Hz,1 H), 3.71 (s, 3H), 3.66 (d, J = 12.8Hz, 1H), 3.49-3.42 (m, 2H), 3.40-3.34 (m, 1H), 3.10-3.04 (m, 1H).

[0646] Step 5: (rel-trans)-2-phenylazetidine-3-carbonitrile

[0647] To a solution of (rel-trans)-1-(4-methoxybenzyl)-2-phenylazetidine-3-carbonitrile (0.1 g, 0.36 mmol, 1 eq) in acetonitrile (3 mL) and water (3 mL) was added bis(nitrate)diamine cerium tetranitrate (4+) (1.2 g, 2.2 mmol, 6 eq) to give a red solution, and the mixture was stirred at 60° C. for 12 hours to give a yellow solution. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), and the combined organic layers were washed with saturated brine (50 mL x 2), and the mixture was purified by Na 2 O 4 '-trifluoroacetic acid (400 mg / ml) and purified by HPLC. 2 SO 4 The residue was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2 , DCM:MeOH=100:1 to 10:1, UV=220 nm) to give (rel-trans)-2-phenylazetidine-3-carbonitrile (80 mg, crude) as a yellow oil. LCMS: m / z (M+H) + =159.6.

[0648] Synthesis of (3R,5R)-5-phenylpyrrolidine-3-carbonitrile and (3R,5S)-5-phenylpyrrolidine-3-carbonitrile

[0649]

[0650] Step 1: (4R)-tert-butyl 4-cyano-2-phenylpyrrolidine-1-carboxylate

[0651] In a 40 mL oven-dried vial equipped with a magnetic stir bar, iodobenzene (0.20 g, 0.98 mmol, 1 eq), (3R,4R)-1-[(tert-butyloxy)carbonyl]-4-cyanopyrrolidine-3-carboxylic acid (0.31 g, 1.3 mmol, 1.3 eq), Ir[dF(Me)ppy] 2 (dtbpy)(PF 6 )(8.0 mg, 0.01 equivalent), NiCl 2 .dtbbpy (12mg, 0.05 equivalent) and cesium carbonate (0.48g, 1.5mmol, 1.5 equivalent) DMA (10mL). The vial is placed in nitrogen and sealed. The reaction is stirred and irradiated with a 10W blue LED lamp (outside 3cm), and the reaction temperature is maintained at 25°C with cold water for 12 hours. The mixture is purified by preparative HPLC (column: Phenomenex Luna C18 150x 25mm x 10um, mobile phase: water (0.1% TFA)-ACN; B%: 38%-68%, 11.5 minutes), to freeze-dry, to obtain (4R)-4-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (0.15g, yield 55%) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 7.30-7.23 (m, 2H), 7.22-7.13 (m, 2H), 7.06 (d, J = 7.2 Hz, 1H), 5.14-4.60 (m, 1H), 3.90-3.80 (m, 1H), 3.15-3.01 (m, 1H), 2.80-2.69 (m, 1H), 2.65-2.48 (m, 1H), 2.21-2.13 (m, 1H), 1.57-0.93 (m, 9H)

[0652] Step 2: SFC separation of (2R,4R)-4-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester and (2S,4R)-4-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester

[0653] SFC showed two peaks of (4R)-4-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (rt = 0.8481.136 minutes). The residue was purified by SFC (column: REGIS (S, S) WHELK-O1 (250 mm x 25 mm, 10 um), mobile phase: 0.1% NH 3 H 2O MeOH; B%: 15%-15%) and concentrated under reduced pressure to give tert-butyl (2R,4R)-4-cyano-2-phenylpyrrolidine-1-carboxylate (40 mg, yield 27%, rt = 0.848 min) as a white solid and tert-butyl (2S,4R)-4-cyano-2-phenylpyrrolidine-1-carboxylate (80 mg, yield 54%, rt = 1.136 min) as a white solid. tert-butyl (2R,4R)-4-cyano-2-phenylpyrrolidine-1-carboxylate: 1 H NMR (400 MHz, chloroform-d) δ7.38-7.31 (m, 2H), 7.30-7.19 (m, 3H), 4.85-4.52 (m, 1H), 4.20 (s, 1H), 3.78-3.69 (m, 1H), 3.17

[0654] -3.03 (m, 1H), 2.87-2.76 (m, 1H), 2.28-2.11 (m, 1H), 1.51-1.01 (m, 9H) (2S, 4R)-4-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester: 1 H NMR (400 MHz, chloroform-d) δ 7.30-7.22 (m, 2H), 7.22-7.15 (m, 1H), 7.06 (d, J = 7.2 Hz, 2H), 5.18-4.84 (m, 1H), 3.91-3.64 (m, 2H), 3.08 (m, 1H), 2.56 (m, 1H), 2.24-2.07 (m, 1H), 1.43-1.03 (m, 9H)

[0655] Step 3: Synthesis of (3R,5R)-5-phenylpyrrolidine-3-carbonitrile and (3R,5S)-5-phenylpyrrolidine-3-carbonitrile

[0656] For a solution of (4R)-4-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (1 equivalent) in dichloromethane (0.1 mL) and trifluoroacetic acid (0.3 mL), the mixture was stirred at 25° C. for 1 hour to give a yellow solution. The reaction mixture was concentrated under reduced pressure to give (3R,5R)-5-phenylpyrrolidine-3-carbonitrile (40 mg, crude) and (3R,5S)-5-phenylpyrrolidine-3-carbonitrile (80 mg, crude) as yellow oils. (3R,5R)-5-phenylpyrrolidine-3-carbonitrile: LCMS: m / z (M+H)+=173.0 (3R,5S)-5-phenylpyrrolidine-3-carbonitrile: LCMS: m / z (M+H)+=173.0

[0657] Synthesis of (rel-trans)-2-phenylpyrrolidine-3-carbonitrile

[0658]

[0659] Step 1: Ethyl 2-(cyclohexa-1,5-dien-1-yl)-1H-pyrrole-3-carboxylate

[0660] To a solution of ethyl 2-cyano-4,4-diethoxybutyrate (8.0 g, 35 mmol, 1.0 eq.) in acetic acid (12 mL) and water (12 mL) were added phenylboronic acid (5.1 g, 42 mmol, 1.2 eq.) and bipyridine (0.27 g, 1.7 mmol, 0.05 eq.) and the mixture was stirred with N 2 Purge 3 times. Then at N 2 The mixture was added with palladium (2+) acetate (0.29 g, 1.7 mmol, 0.05 eq) under a 3-H atmosphere and stirred at 80° C. for 16 hours to obtain a black sludge solution. The reaction was diluted with water (500 mL) and extracted with ethyl acetate (500 mL x 2). The combined organic layers were washed with saturated brine (500 mL x 2) and purified by Na 2 SO 4 . 2 SO 4 The residue was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (80 g Silica gel flash column, elution with a gradient of 0 to 10% ethyl acetate / petroleum ether at 80 mL / min) to give ethyl 2-(cyclohexa-1,5-dien-1-yl)-1H-pyrrole-3-carboxylate (3.00 g, 40% yield) as a white solid. LCMS: m / z (M+H) + =215.9. 1H NMR (400 MHz, CHLOROFORM-d) δ8.54 (s, 1H), 7.66-7.55 (m, 2H), 7.45-7.32 (m, 3H), 6.75 (d, J=2.4 Hz, 2H), 4.21 (q, J=7.2 Hz, 2H), 1.26 (t, J=7.2 Hz, 3H).

[0661] Step 2: 1-tert-Butyl 3-ethyl 2-phenyl-1H-pyrrole-1,3-dicarboxylate

[0662] To a solution of ethyl 2-(cyclohexa-1,5-dien-1-yl)-1H-pyrrole-3-carboxylate (3.0 g, 14 mmol, 1.0 eq) in DCM (30 mL) was added di-tert-butyl dicarbonate (9.1 g, 42 mmol, 3.0 eq), 4-dimethylaminopyridine (0.17 g, 1.4 mmol, 0.1 eq) and triethylamine (2.1 g, 21 mmol, 1.5 eq) and the mixture was stirred at 25 °C for 1 hour to give a black clear solution. The reaction was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 2), the combined organic layers were washed with saturated brine (200 mL x 2), washed with Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give 1-tert-butyl 3-ethyl 2-phenyl-1H-pyrrole-1,3-dicarboxylate (3.5 g, yield 80%) as a light yellow oil. LCMS: m / z (M+H) + =316.0. 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.32-7.27 (m, 3H), 7.25 (dd, J = 1.2, 3.6 Hz, 1H), 7.23-7.17 (m, 2H), 6.60 (dd, J = 1.2, 3.6 Hz, 1H), 4.00 (q, J = 7.2 Hz, 2H), 1.16 (s, 9H), 1.00 (t, J = 7.2 Hz, 3H).

[0663] Step 3: 1-tert-Butyl 3-ethyl 2-phenylpyrrolidine-1,3-dicarboxylate

[0664] To a solution of tert-butyl 3-ethyl 2-phenyl-1H-pyrrole-1,3-dicarboxylate (3.5 g, 11 mmol, 1.0 equiv) in EtOH (30 mL) was added dioxopalatin (7.5 g, 33 mmol, 3.0 equiv). The mixture was stirred under H 2 The mixture was stirred at 25° C. for 3 hours to give a colorless solution. The reaction mixture was filtered, and the filtrate was concentrated to give 1-tert-butyl 3-ethyl 2-phenylpyrrolidine-1,3-dicarboxylate (1.8 g, crude) as a white solid. LCMS: m / z (M-Boc+H) + =219.9. 1H NMR (400 MHz, CHLOROFORM-d) δ = 7.26-7.19 (m, 3H), 7.16-7.09 (m, 2H), 5.27-4.99 (m, 1H), 3.95-3.73 (m, 3H), 3.57-3.37 (m, 2H), 2.56-2.35 (m, 1H), 2.13-2.01 (m, 1H), 1.57 (s, 9H), 1.17 (s, 3H).

[0665] Step 4: 1-(tert-Butyloxycarbonyl)-2-phenylpyrrolidine-3-carboxylic acid

[0666] To a solution of 1-tert-butyl 3-ethyl 2-phenylpyrrolidine-1,3-dicarboxylate (1.8 g, 5.6 mmol, 1.0 equiv) in THF (6.0 mL), water (6.0 mL) and MeOH (6.0 mL) was added lithium hydroxide (1+) hydrate (0.47 g, 11 mmol, 2.0 equiv) and the mixture was stirred at 25 °C for 1 hour to give a yellow clear solution. The reaction mixture was filtered and the filtrate was concentrated to give a residue. The residue was purified by preparative HPLC (basic conditions; column: Welch Xtimate C18 150x25mm x 5um, mobile phase: water (NH 3 H 2 O)-ACN; B%: B%: 5%-35%, 8 minutes) was purified and freeze-dried to give 1-(tert-butoxycarbonyl)-2-phenylpyrrolidine-3-carboxylic acid (0.8 g, 2.7 mmol, yield 49%) as a white solid. LCMS: m / z (MH) - =290.1

[0667] Step 5: tert-Butyl 3-carbamoyl-2-phenylpyrrolidine-1-carboxylate

[0668] To a solution of 1-(tert-butoxycarbonyl)-2-phenylpyrrolidine-3-carboxylic acid (0.8 g, 2.7 mmol, 1.0 eq.) in THF (10 mL) was added carbonyldiimidazole (1.3 g, 8.2 mmol, 3 eq.). The mixture was stirred at 25 °C for 30 min to give a colorless clear solution. Then NH 3 ·H 2 O (3.0 mL), and the reaction mixture was stirred for 10 minutes to obtain a colorless clear solution. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography (12 g Silica gel flash column, 0 to 10% DCM:MeOH eluent at 22 mL / min) to give tert-butyl 3-carbamoyl-2-phenylpyrrolidine-1-carboxylate (0.8 g, 2.1 mmol, 75% yield) as a white solid. LCMS: m / z (M-Boc+H) + =191.1. 1 H NMR (400 MHz, CHLOROFORM-d) δ 7.32-7.11 (m, 5H), 5.70 (s, 1H), 5.43 (s, 1H), 4.87 (s, 1H), 3.88 (s, 1H), 3.63-3.52 (m, 1H), 2.78 (d, J = 6.8 Hz, 1H), 2.26-2.07 (m, 2H), 1.45-1.05 (m, 9H).

[0669] Step 6: (Rel-trans)-3-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester

[0670] To a solution of tert-butyl 3-carbamoyl-2-phenylpyrrolidine-1-carboxylate (0.6 g, 2.1 mmol, 1.0 eq) in DCM (6.0 mL) was added trimethylamine (0.5 g, 5.2 mmol, 2.5 eq), the mixture was cooled to 0 °C, and trifluoroacetic anhydride (0.5 g, 2.6 mmol, 1.3 eq) was added. The reaction was allowed to warm to 25 °C and stirred for 1 hour to give a colorless solution. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (FA conditions; column: Phenomenex Luna C18 100 x 30 mm x 5 um; mobile phase: water (FA)-ACN; B%: B: 42%-72%, 8 minutes) to lyophilize to give tert-butyl (2S,3S)-3-cyano-2-phenylpyrrolidine-1-carboxylate (0.2 g, 0.73 mmol, 36% yield) as a white solid. The rel-trans stereochemistry was assigned by 2D NMR. LCMS: m / z (Mt-bu+H) + =217.1. 1H NMR (400MHz, chloroform-d) δ7.40-7.28 (m, 3H), 7.20 (d, J=7.2Hz, 2H), 5.02 (s, 1H), 3.87 (s, 1H), 3.76-3.68 (m, 1H), 3.05-2.93 (m, 1H), 2.36-2.16 (m, 2H), 1.49-1.13 (m, 9H)

[0671] Step 7: (rel-trans)-2-phenylpyrrolidine-3-carbonitrile

[0672] (rel-trans)-3-cyano-2-phenylpyrrolidine-1-carboxylic acid tert-butyl ester (50 mg, 0.18 mmol, 1.0 equiv) was dissolved in a solution of DCM (1.0 mL) and TFA (0.5 mL). The reaction was stirred at 25 ° C for 0.5 hours to give a colorless solution. The reaction mixture was concentrated under reduced pressure to give (2S, 3S)-2-phenylpyrrolidine-3-carbonitrile (50 mg, crude) as a light yellow oil and used in the next step without further purification.

[0673] Synthesis of (5S,8S,10aR)-3-acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid

[0674]

[0675] Step 1: (5S,8S,10aR)-3-Acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctate-8-carboxylic acid benzyl ester

[0676] To a solution of (5S,8S,10aR)-3-acetyl-5-{[(tert-butoxy)carbonyl]amino}-6-oxo-decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid (1.0 g, 2.7 mmol, 1.0 equiv) in dimethylformamide (10 mL) was added K 2 CO 3 (0.56 g, 4.1 mmol, 1.5 eq.), then (bromomethyl)benzene (0.5 g, 3.0 mmol, 1.1 eq.) was added at 0 °C. The mixture was stirred at 25 °C for 2 hours. The reaction was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 2), the combined organic layers were washed with saturated brine (100 mL x 2), and the mixture was purified by Na 2 SO 4 The mixture was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give (5S,8S,10aR)-3-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid benzyl ester (0.5 g, 40% yield) as a white solid. LCMS: m / z (M+H) + =460.1.

[0677] Step 2: (5S,8S,10aR)-3-Acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctate-8-carboxylic acid benzyl ester

[0678] A mixture of (5S,8S,10aR)-3-acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctate-8-carboxylic acid benzyl ester (0.5 g, 1.1 mmol, 1.0 equiv) in dichloromethane (2.5 mL) and HCl / dioxane (2.5 mL) was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure to give (5S,8S,10aR)-3-acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctate-8-carboxylic acid benzyl ester (0.5 g, crude) as a white solid. LCMS: m / z (M+H) + =360.3.

[0679] Step 3: (5S,8S,10aR)-3-Acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctate-8-carboxylic acid benzyl ester

[0680] To a solution of 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (0.34 g, 0.93 mmol, 1.0 equiv) in dimethylformamide (1.0 mL) was added N,N-diisopropylethylamine (0.48 g, 3.7 mmol, 4.0 equiv) and HATU (0.53 g, 1.39 mmol, 1.5 equiv). The mixture was stirred at 25 °C for 10 minutes. Then a solution of (5S,8S,10aR)-3-acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid benzyl ester (0.4 g, 1.1 mmol, 1.2 equiv) in dimethylformamide (1.0 mL) and N,N-diisopropylethylamine (0.48 g, 3.7 mmol, 4.0 equiv) was added. The reaction mixture was stirred at 25°C for 1 hour. The reaction was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2), and the combined organic layers were washed with saturated brine (40 mL x 2) and purified by Na 2 SO 4The mixture was dried, filtered and concentrated under reduced pressure to give a residue. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography to give (5S, 8S, 10aR) -3-acetyl -5- (5- ((diethoxyphosphoryl) difluoromethyl) benzo [b] thiophene -2-carboxamido) -6-oxodecahydropyrrolo [1, 2-a] [1, 5] diazacyclooctate -8- carboxylic acid benzyl ester (0.3 g, 46% yield) as a yellow solid. LCMS: m / z (M + H) + =706.4.

[0681] Step 4: (5S,8S,10aR)-3-Acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid

[0682] A solution of benzyl (5S,8S,10aR)-3-acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctate-8-carboxylate (0.4 g, 0.56 mmol, 1.0 equiv) in methanol (20 mL) was degassed and heated to 40 °C with N 2 Purge 3 times. Pd / C (0.4 g) was carefully added to the mixture, and the system was placed under H 2 Atmosphere, and stirred at 25 ° C for 2 hours.The reaction mixture is filtered, and the filter is concentrated to obtain a filtrate.The filtrate is concentrated under reduced pressure to obtain (5S, 8S, 10aR) -3- acetyl -5- (5- ((diethoxyphosphoryl) difluoromethyl) benzo [b] thiophene -2- formamido) -6- oxodecahydropyrrolo [1,2-a] [1,5] diazacyclooctatetraene -8- carboxylic acid (0.3g, crude product) in off-white oil, which can be used in the next step without further purification.

[0683] Representative solutions:

[0684] Representative procedure of general scheme 1: Synthesis of 1

[0685]

[0686] Step A: tert-Butyl ((5S,8S,10aR)-3-acetyl-6-oxo-8-(6-phenyl-4-azaspiro[2.4]heptane-4-carboxyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate

[0687] A mixture of 6-phenyl-4-azaspiro[2.4]heptane (168 mg, 0.97 mmol, 1.2 eq), (5S,8S,10aR)-3-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid (300 mg, 0.812 mmol, 1.0 eq), HATU (463 mg, 1.218 mmol, 1.5 eq) and N,N-diisopropylethylamine (246.8 mg, 2.43 mmol, 3.0 eq) in DCM (10 mL) was stirred in a sealed tube at room temperature for 3 h. After completion, the mixture was concentrated by adding H 2 The reaction mixture was quenched with 1% O (5 mL), extracted with EtOAc (5 mL x 3), and the combined organic layers were washed with brine (5 mL x 2), dried over anhydrous Na 2 SO 4 The residue was purified by C18 column to give tert-butyl ((5S,8S,10aR)-3-acetyl-6-oxo-8-(6-phenyl-4-azaspiro[2.4]heptane-4-carbonyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate (80 mg, 152.5 μmol, 19%) as a white solid. LCMS (ESI): m / z=525.4 [M+H] + .

[0688] Step B: (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

[0689] A solution of tert-butyl ((5S,8S,10aR)-3-acetyl-6-oxo-8-(6-phenyl-4-azaspiro[2.4]heptane-4-carbonyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate (80 mg, 152.5 μmol, 1 eq) in DCM (3 mL) and TFA (2 mL) was stirred at room temperature for 1 h. After the reaction, the reaction was cooled with an ice bath and then treated with NaHCO 3 (aqueous solution) was carefully neutralized until the pH was adjusted to pH = 8-9. The resulting mixture was extracted with DCM (10 mL x 3), and the combined organic layers were washed with brine (10 mL x 2), washed with anhydrous Na 2 SO 4The reaction mixture was dried and then concentrated under reduced pressure to give a crude product of tert-butyl ((5S,8S,10aR)-3-acetyl-6-oxo-8-(6-phenyl-4-azaspiro[2.4]heptane-4-carbonyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate (60 mg, 141.3 μmol) as a white solid, which was used directly in the next step without further purification. LCMS (ESI): m / z=425.3 [M+H] + .

[0690] Step C: Bis(3-methylthiobutanoic acid) S,S'-(((((7-(((5S,8S,10aR)-3-acetyl-6-oxo-8-(6-phenyl-4-azaspiro[2.4]heptane-4-carbonyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)naphthalen-2-yl)difluoromethyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl))

[0691] To 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 (30 mg, 70.7 μmol, 1 eq) and DIEA (27.4 mg, 212 μmol, 3 eq) in DMF (3 mL) was added perfluorophenyl 7-((bis(2-((3-methylbutanoyl)thio)ethoxy)phosphoryl)difluoromethyl)-2-naphthoate (53.5 mg, 70.7 μmol, 1 eq). The resulting mixture was stirred at room temperature for 2 h and then heated to room temperature by adding H 2 The reaction was quenched with 1% CO (10 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL x 2) and purified by anhydrous Na 2 SO 4 The mixture was dried and then concentrated under reduced pressure. The residue was purified by C18 column to give S,S'-(((((7-(((5S,8S,10aR)-3-acetyl-6-oxo-8-(6-phenyl-4-azaspiro[2.4]heptane-4-carbonyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)naphthalen-2-yl)difluoromethyl)phosphoryl)bis(oxy))bis(ethane-2,1-diyl))bis(3-methylthiobutanoate (15.4 mg, 15.4 μmol, 22%) as a white solid. 1 H NMR (400 MHz, CDCl 3)δ8.51-8.45(m,1H),8.29-8.19(m,1H),8.04-7.89(m,4H),7.83-7.76(m,1H),7.43-7.34(m,3H),7.34-7.30 (m,2H),4.95-4.80(m,1H),4.66-4.45(m,1H),4.33-4.16(m,7H),4.07-3.86(m,1H),3.72-56(m,1H),3.41-3. 24(m,2H),3.22-3.13(m,4H),2.55-2.49(m,3H),2.47-2.41(m,5H),2.40-2.22(m,3H),2.21-2.18(m,1H),2.1 8-2.12(m,2H),2.09-2.01(m,2H),1.96-1.79(m,3H),1.67(s,1H),0.96(d,J=6.6Hz,12H),0.83-0.39(m,2H). LCMS(ESI):m / z=997.4[M+H] + .

[0692] Representative procedure of general scheme 2: Synthesis of 2

[0693]

[0694]

[0695] Step A: (5S,8S,10aR)-3-Acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester

[0696] (5S,8S,10aR)-3-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid (3 g, 8.12 mmol, 1 equivalent) and K 2 CO 3 To a solution of 3-bromoprop-1-ene (2.25 g, 16.2 mmol, 2 eq.) in DMF (2 mL) was added 3-bromoprop-1-ene (1.17 g, 9.74 mmol, 1.2 eq.). The reaction mixture was stirred at 20 °C for 16 hours. Upon completion, the reaction mixture was poured into water (10 mL) and then extracted with EtOAc (20 mL x 2). The organic layers were combined and washed with brine (150 mL) and dried over anhydrous Na 2 SO 4The residue was purified by C18 column to give (5S,8S,10aR)-3-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester (2.10 g, 5.12 mmol, 63%) as a white solid. LCMS (ESI): m / z=410.2 [M+H] + .

[0697] Step B: (5S,8S,10aR)-3-Acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester

[0698] A solution of (5S,8S,10aR)-3-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester (2.1 g, 5.12 mmol, 1 eq) in HCl / EtOAc (15 mL) was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated under reduced pressure to give (5S,8S,10aR)-3-acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester (HCl salt) (1.30 g, 4.20 mmol, 82%) as a white solid, which was used in the next step without further purification. LCMS (ESI): m / z=310.1 [M+H] + .

[0699] Step C: (5S,8S,10aR)-3-Acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester

[0700] To a solution of (5S,8S,10aR)-3-acetyl-5-amino-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester (1.3 g, 4.20 mmol, 1 eq), 5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxylic acid (1.68 g, 4.62 mmol, 1.1 eq) and TEA (1.27 g, 12.6 mmol, 3 eq) in DCM (20 mL) were added EDCI (885 mg, 4.62 mmol, 1.1 eq) and HOBt (574 mg, 4.62 mmol, 1.1 eq) and the resulting mixture was stirred under nitrogen at room temperature overnight. After completion, the reaction was filtered and concentrated to give a residue, and the residue was purified by C18 column to give (5S,8S,10aR)-3-acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid allyl ester (1.80 g, 2.74 mmol, 65%) as a white solid. LCMS (ESI): m / z=656.2[M+H] + .

[0701] Step D: (5S,8S,10aR)-3-Acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid

[0702] Allyl (5S,8S,10aR)-3-acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylate (1.8 g, 2.74 mmol, 1 equivalent), Pd(PPh 3 ) 4 A solution of (158 mg, 137 μmol, 0.05 eq) and pyrrolidine (194 mg, 2.74 mmol, 1 eq) in DCM (20 mL) was stirred overnight at room temperature under nitrogen. The reaction was filtered and concentrated to give a residue, which was purified by a C18 column to give (5S, 8S, 10aR) -3-acetyl-5- (5- ((diethoxyphosphoryl) difluoromethyl) benzo [b] thiophene-2-carboxamido) -6-oxodecahydropyrrolo [1, 2-a] [1, 5] diazacyclooctatetraene-8-carboxylic acid (1.28 g, 2.07 mmol, 76%) as a white solid. LCMS (ESI): m / z = 616.1 [M + H] + .

[0703] Step E: Diethyl ((2-(((5S,8S,10aR)-3-acetyl-8-((4-(2-cyanopropan-2-yl)phenyl)(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonate

[0704] A solution of (5S,8S,10aR)-3-acetyl-5-(5-((diethoxyphosphoryl)difluoromethyl)benzo[b]thiophene-2-carboxamido)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid (60 mg, 97.4 μmol, 1 eq), 2-chloro-1-methylpyridinium-1-cation iodide (29.6 mg, 116 μmol, 1.2 eq), 2-methyl-2-(4-(methylamino)phenyl)propionitrile (18.6 mg, 107 μmol, 1.1 eq) and TEA (29.5 mg, 292 μmol, 3 eq) in DCM (2 mL) was stirred at 35 °C for 30 h. After completion, the reaction was filtered and concentrated to give a residue, and the residue was purified by silica gel flash column chromatography to give diethyl ((2-(((5S,8S,10aR)-3-acetyl-8-((4-(2-cyanopropan-2-yl)phenyl)(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonate (25.0 mg, 32.3 μmol, 33%) as a white solid. LCMS (ESI): m / z=772[M+H] + .

[0705] Step F: ((2-(((5S,8S,10aR)-3-acetyl-8-((4-(2-cyanopropan-2-yl)phenyl)(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid

[0706] To a solution of diethyl ((2-(((5S,8S,10aR)-3-acetyl-8-((4-(2-cyanopropan-2-yl)phenyl)(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonate (23 mg, 29.7 μmol, 1 eq) in DCM (2 mL) was added TMSBr (226 mg, 1.48 mmol, 50 eq) and the resulting mixture was stirred at 35 °C for 30 h. Upon completion, the reaction was filtered and concentrated to give a residue, and the residue was purified by C18 column to afford ((2-(((5S,8S,10aR)-3-acetyl-8-((4-(2-cyanopropan-2-yl)phenyl)(methyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)benzo[b]thiophen-5-yl)difluoromethyl)phosphonic acid (20.0 mg, 27.9 μmol, 94%) as a white solid. 1 H NMR (400MHz, CD 3 OD)δ8.06-7.85(m,3H),7.70-7.62(m,1H),7.53-7.33(m,4H),5.20-5.10(m,1H),4.42-3 .66(m,6H),3.25-3.15(m,3H),2.34-2.21(m,3H),2.12-1.76(m,6H),1.65-1.49(m,6H). LCMS(ESI):m / z=716.1[M+H] + .

[0707] Representative procedure of general scheme 3: Synthesis of 3

[0708]

[0709]

[0710] Step A: 3-Benzyl 8-methyl (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-3,8(4H)-dicarboxylate

[0711] Methyl (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylate (1 g, 2.92 mmol, 1 eq) and NaHCO were added at room temperature. 3To a solution of 1.21 g, 14.5 mmol, 5 equivalents) in a mixture of THF (10 mL) and water (2 mL) was added benzyl chloroformate (996 mg, 5.84 mmol, 2 equivalents). After completion, the reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was poured into water and then extracted with EtOAc (100 mL x 3). The organic layers were combined and washed with brine (50 mL) and washed with anhydrous Na 2 SO 4 The reaction mixture was dried and concentrated under reduced pressure to give crude (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-3,8(4H)-dicarboxylic acid 3-benzyl ester 8-methyl ester (1.10 g, 2.31 mmol, 80%) as a yellow oil. LCMS (ESI): m / z=476.2 [M+H] + .

[0712] Step B: (5S,8S,10aR)-3-((benzyloxy)carbonyl)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid

[0713] To a solution of 3-benzyl 8-methyl (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-3,8(4H)-dicarboxylate (1.3 g, 2.73 mmol, 1 eq) in a mixture of MeOH (5 mL) and water (1.5 mL) was added lithium hydroxide (544 mg, 13.6 mmol, 5 eq) and the resulting mixture was stirred at room temperature for 12 hours. Upon completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by Et 2 O (30 mL) to give (5S,8S,10aR)-3-((benzyloxy)carbonyl)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid (1.15 g, 2.49 mmol, 92%) as a white solid. LCMS (ESI): m / z=462.2 [M+H] + .

[0714] Step C: (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-8-(methyl(phenyl)carbamoyl)-6-oxooctahydropyrrolo[1,2-a][1,5]diazacyclooctate-3(4H)-carboxylic acid benzyl ester

[0715] To a solution of (5S,8S,10aR)-3-((benzyloxy)carbonyl)-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxylic acid (400 mg, 866 μmol, 1 eq), TEA (262 mg, 2.59 mmol, 3 eq) and N-methylaniline (185 mg, 1.73 mmol, 2 eq) in DCE (15 mL) was added T. 3 P (550 mg, 1.73 mmol, 2 eq.) was added and the resulting mixture was heated at 90 °C for 20 hours. After completion, the reaction mixture was poured into water (50 mL) and then extracted with EtOAc (100 mL x 3). The organic layers were combined and washed with brine (50 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by C18 column to give (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-8-(methyl(phenyl)carbamoyl)-6-oxooctahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-3(4H)-carboxylic acid benzyl ester (400 mg, 726 μmol, 84%) as a white solid. LC-MS (ESI) m / z=551.2 [M+H] + .

[0716] Step D: tert-butyl ((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate

[0717] To a solution of benzyl (5S,8S,10aR)-5-((tert-butoxycarbonyl)amino)-8-(methyl(phenyl)carbamoyl)-6-oxooctahedralpyrrolo[1,2-a][1,5]diazacyclooctatetraene-3(4H)-carboxylate (540 mg, 980 μmol, 1 eq) in THF (20 mL) was added Pd / C (100 mg) under nitrogen. The suspension was degassed under vacuum and heated to 40 ℃ for 2 h. 2 The resulting mixture was stirred at room temperature overnight. The mixture was filtered through a pad of 4% paraformaldehyde (20 mL) and the filter cake was washed with THF (20 mL). The combined filtrates were concentrated to dryness to give tert-butyl ((5S, 8S, 10aR)-8-(methyl (phenyl) carbamoyl)-6-oxodecahydropyrrolo [1, 2-a] [1, 5] diazacyclooctatetraen-5-yl) carbamate (400 mg, 960 μmol, 98%) as a white solid. LCMS (ESI): m / z = 417 [M + H] + .

[0718] Step E: tert-butyl ((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate

[0719] To a solution of tert-butyl ((5S, 8S, 10aR) -8- (methyl (phenyl) carbamoyl) -6-oxodecahydropyrrolo [1, 2-a] [1, 5] diazacyclooctatetraen-5-yl) carbamate (80 mg, 192 μmol, 1 eq) and TEA (58.2 mg, 576 μmol, 3 eq) in DCM (3 mL) was added 2-phenylacetyl chloride (44.5 mg, 288 μmol, 1.5 eq) at 20 ° C, and the resulting mixture was stirred at 20 ° C for 1 hour. After completion, the reaction mixture was poured into water (10 mL) and then extracted with DCM (20 mL x 3). The organic layers were combined and washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The reaction mixture was dried and concentrated under reduced pressure to give crude tert-butyl ((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamate (90.0 mg, 168 μmol, 88%) as a yellow solid, which was used directly in the next step without further purification. LC-MS (ESI) m / z=573 [M+K] + .

[0720] Step F: (5S,8S,10aR)-5-amino-N-methyl-6-oxo-N-phenyl-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxamide

[0721] A solution of tert-butyl ((5S, 8S, 10aR) -8- (methyl (phenyl) carbamoyl) -6- oxo -3- (2- phenylacetyl) decahydropyrrolo [1, 2-a] [1, 5] diazacyclooctatetraen-5-yl) carbamate (90 mg, 168 μmol, 1 eq) in HCl / dioxane (2 mL, 4M) was stirred at 20 ° C for 1 hour. After completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by a C18 column to give (5S, 8S, 10aR) -5-amino-N-methyl-6-oxo-N-phenyl-3- (2-phenylacetyl) decahydropyrrolo [1, 2-a] [1, 5] diazacyclooctatetraen-8-carboxamide (70.0 mg, 161 μmol, 96%) as a white solid. LC-MS (ESI) m / z = 435.2 [M + H]+ .

[0722] Step G: Diethyl (difluoro(2-(((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonate

[0723] To a solution of (5S,8S,10aR)-5-amino-N-methyl-6-oxo-N-phenyl-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraene-8-carboxamide (69.8 mg, 161 μmol, 1 eq), TEA (81.3 mg, 805 μmol, 5 eq) and HATU (91.8 mg, 241 μmol, 1.5 eq) in DMF (1 mL) was added 8 (61.4 mg, 177.1 μmol, 1.1 eq) and the resulting mixture was stirred at 20 °C for 16 hours. Upon completion, the reaction mixture was diluted with water (10 mL) and then extracted with EtOAc (20 mL x 3). The organic layers were combined and washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The mixture was dried and concentrated under reduced pressure. The residue was purified by C18 column to give diethyl (difluoro(2-(((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonate (120 mg, 157 μmol, 69%) as a white solid. LC-MS (ESI) m / z=764.1 [M+H] + .

[0724] Step H: (Difluoro(2-(((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid

[0725] To a solution of diethyl (difluoro(2-(((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonate (90 mg, 0.12 mmol, 1.0 equiv) in DCM (1 mL) was added TMSBr (179 mg, 1.17 mmol, 10 equiv) at room temperature. After addition, the reaction mixture was stirred at room temperature overnight. Upon completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by RP-preparative HPLC to give (difluoro(2-(((5S,8S,10aR)-8-(methyl(phenyl)carbamoyl)-6-oxo-3-(2-phenylacetyl)decahydropyrrolo[1,2-a][1,5]diazacyclooctatetraen-5-yl)carbamoyl)-1H-indol-5-yl)methyl)phosphonic acid (3) (50.0 mg, 70.6 μmol, 60%) as a yellow solid. LCMS (ESI): m / z=708.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ12.37-11.72(m,1H),8.89-8.29(m,1H),7.98-7.82(m,1H),7.57-7.13(m,13H),5.09-4. 91(m,1H),4.32-4.20(m,2H),4.08-4.00(m,1H),3.96-3.89(m,2H),3.81-3.75(m,1H),3.42 -3.31(m,2H),3.20(s,3H),2.20-1.53(m,6H).

[0726] Representative procedure of general scheme 4: Synthesis of 4

[0727]

[0728] Step A: Methyl (5S,8S,10aR)-2-acetyl-5-((tert-butoxycarbonyl)amino)-6-oxodecahydropyrrolo[1,2-a][1,4]diazacyclooctate-8-carboxylate

[0729] To a solution of (5S, 8S, 10aR) -5- ((tert-butoxycarbonyl) amino) -6- oxodecahydropyrrolo [1, 2-a] [1, 4] diazacyclooctatetraene-8-carboxylate (400 mg, 1.17 mmol, 1 eq) and DIEA (453 mg, 3.51 mmol, 3 eq) in DMF (6 mL) was added acetyl acetate (178 mg, 1.75 mmol, 1.5 eq) and the resulting mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel flash column chromatography...

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; X is selected from S, SO2, -S(=O)=NH and NR 8 ; R 1 is selected from 8- to 10-membered fused bicyclic heteroaryl, the 8- to 10-membered fused bicyclic heteroaryl being substituted with: -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; 8- to 10-membered fused bicyclic heterocyclic group, the 8- to 10-membered fused bicyclic heterocyclic group is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; aryl, said aryl being 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ], wherein the aryl group may be further optionally substituted by 1 or 2 groups independently selected from cyano, (C1-C4)alkoxy and halo; -(C1-C4)alkyl(aryl), wherein the aryl portion of -(C1-C4)alkyl(aryl) is 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 ]、-P(O)OR 1b OR 2b 、 -[P(O)[NHR Ty ][NH(AA)C(O)OR T ], -[P(O)[NH(AA)C(O)ORT][NH(AA)C(O)ORT] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; and -(C2-C4)alkenyl(aryl), wherein the aryl portion of -(C2-C4)alkenyl(aryl) is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; 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)alkyl]-SC(O)-[(C1-C4)alkyl] 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]-OC(O)N[(C1-C4)alkyl] ]2 and aryl, wherein the 5- to 6-membered heteroaryl and aryl are each optionally and independently substituted with 1 to 2 groups selected from halo, cyano and (C1-C4) alkyl when valence permits, and wherein the 5- to 7-membered heterocyclyl in [(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 Each is independently selected from hydrogen, halo, (C1-C4) alkyl, halo (C1-C4) alkyl, hydroxy (C1-C4) alkyl, - (C1-C4) alkylphenyl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, - (C1-C4) alkyl (C1-C4) alkoxy, hydroxy, cyano, -NR a R b , phenyl, (C3-C6) cycloalkyl, 5- to 6-membered heteroaryl and 4- to 6-membered heterocyclyl, wherein the phenyl, (C3-C6) cycloalkyl, 5- to 6-membered heteroaryl and 4- to 6-membered heterocyclyl are each optionally substituted with 1 to 3 selected from R where valence permits S The group substitution; 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 the (C1-C4)alkyl is optionally substituted with 1 to 3 selected from R when valence permits. Y The phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic group and 5- to 10-membered monocyclic or bicyclic heteroaryl group are each optionally substituted with 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, where valence permits, by 1 to 3 selected from R Q The group substitution; R 8 is selected from hydrogen, (C1-C4) alkyl, halo (C1-C4) alkyl, (C3-C6) cycloalkyl, 5- to 7-membered heterocyclyl, -(C1-C4) [5- to 7-membered heterocyclyl], 5- to 10-membered heteroaryl, -(C1-C4) [5- to 10-membered heteroaryl], phenyl, -(C1-C4) alkylphenyl, -C(O)R Ha 、-C(O)OR Ha 、-C(O)NR Ha R Hb 、-C(O)OR Ha 、-SOR Ha R Hb and-SO2R Ha wherein the (C3-C6)cycloalkyl, 5- to 7-membered heterocyclyl, 5- to 10-membered heteroaryl, phenyl, the phenyl on the (C1-C4)alkylphenyl, the 5- to 7-membered heterocyclyl on -(C1-C4)[5- to 7-membered heterocyclyl], and the 5- to 6-membered heteroaryl on -(C1-C4)[5- to 6-membered heteroaryl] are each optionally substituted, where valence permits, with 1 to 3 selected from R U The group substitution; R Ha Selected from (C1-C 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 )alkynyl, phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, and 4- to 10-membered monocyclic or bicyclic heterocyclic group, wherein said (C1-C 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 ) alkynyl groups are each optionally substituted, where valence permits, with 1 to 2 selected from R O wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 10-membered monocyclic or bicyclic heterocyclyl are each optionally substituted, where valence permits, by 1 to 3 groups selected from the group consisting of halo, (C1-C4) alkyl, halo(C1-C4) alkyl, (C1-C4) alkoxy, halo(C1-C4) alkoxy, oxo, cyano, and 4- to 6-membered heterocyclyl optionally substituted by C1-C4 alkyl; R O Selected from halo, (C1-C4) alkoxy, OH, phenyl, NH2, -NH(C1-C 10 )alkyl, -N[(C1-C 10 )alkyl], (C 3-6 cycloalkyl), 4- to 10-membered monocyclic or fused bicyclic heterocyclic radicals, and 5- to 10-membered monocyclic or bicyclic heteroaryl radicals, wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl radicals and the 4- to 10-membered monocyclic or bicyclic heterocyclic radicals are each optionally substituted, as valence permits, with 1 to 3 groups selected from the group consisting of halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, oxo, and cyano; R Hb 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, 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 and R U Each is independently selected from halo, (C2-C4) alkenyl, (C1-C4) alkyl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, cyano, phenyl, hydroxy, 4- to 9-membered monocyclic or bicyclic heterocyclic group, 5- to 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 (C1-C4)alkyl are each optionally and independently substituted, where valence permits, by 1 to 3 selected from R M wherein the phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C3-C6) cycloalkyl and 4- to 9-membered monocyclic or bicyclic heterocyclyl are each optionally and independently substituted with 1 to 3 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 , hydroxy, phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl are each optionally substituted with 1 to 3 selected from R where valence permits. 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)=NH(C1-C4)alkyl、-S(O)2NR e R f , hydroxy, phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the phenyl, 4- to 6-membered heterocyclyl and 5- to 10-membered monocyclic or bicyclic heteroaryl are each optionally substituted with 1 to 3 selected from R where valence permits. X The group substitution; R F , R S , R X and R Z Each is independently selected from halo, cyano, (C1-C4) alkyl, (C3-C6 cycloalkyl), halo (C1-C4) alkyl, -(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, -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 、-C(O)OR e 、-NR c C(O)R e 、-(C1-C4 alkyl)C(O)R g 、-C(O)R g 、-(C1-C4 alkyl)C(O)NR c R d 、-C(O)NR c R d , -NO2 and -NR a R b wherein the (C1-C4)alkyl is optionally substituted by cyano, wherein the phenyl, the 4-membered to 6-membered heterocyclyl and the phenyl of -(C1-C4)alkylphenyl are each optionally and independently substituted by 1 to 3 groups selected from the following groups when valence permits: halo, cyano, oxo, (C1-C 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 the (C1-C 10 )alkyl, (C2-C 10 )alkenyl and (C2-C 10 ) alkynyl is each optionally substituted, where valence permits, by 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 with a 5- to 7-membered heterocyclyl optionally substituted with 1 to 2 oxo; and R a , R b , R c , R d , R e , R f 、 R g and R h Where valence permits, each 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, where valence permits, by 1 to 3 selected from R J The phenyl, (C3-C6)cycloalkyl, 4- to 6-membered heterocyclyl and 5- to 6-membered heteroaryl groups are each independently optionally substituted by 1 to 3 groups selected from the following groups: halo, cyano, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, hydroxy, phenyl and benzyl, where valence permits.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from 8- to 10-membered fused bicyclic heteroaryl, the 8- to 10-membered fused bicyclic heteroaryl being substituted with: -CR 1a R 2a P(O)OR 1b OR 2b , -CR 1a R 2a P(O)[OR 1b ][NH(AA)C(O)OR T ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; 8- to 10-membered fused bicyclic heterocyclic group, the 8- to 10-membered fused bicyclic heterocyclic group is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; aryl, said aryl being 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; -(C1-C4)alkyl(aryl), wherein the aryl portion of -(C1-C4)alkyl(aryl) is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; and -(C2-C4)alkenyl(aryl), wherein the aryl portion of -(C2-C4)alkenyl(aryl) is 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 ]、-P(O)OR 1b OR 2b 、-[P(O)[NHR Ty ][NH(AA)C(O)OR T ] or -P(O)[OR 1b ][NH(AA)C(O)OR T ]; R 8 is selected from hydrogen, (C1-C4) alkyl, halo (C1-C4) alkyl, (C3-C6) cycloalkyl, 5- to 7-membered heterocyclyl, -(C1-C4) [5- to 7-membered heterocyclyl], 5- to 6-membered heteroaryl, -(C1-C4) [5- to 6-membered heteroaryl], phenyl, -(C1-C4) alkylphenyl, -C(O)R Ha 、-C(O)OR Ha 、-C(O)NR Ha R Hb 、-C(O)OR Ha 、-SOR Ha R Hb and-SO2R Ha wherein the (C3-C6)cycloalkyl, 5- to 7-membered heterocyclyl, 5- to 6-membered heteroaryl, phenyl, the phenyl on the (C1-C4)alkylphenyl, the 5- to 7-membered heterocyclyl on -(C1-C4)[5- to 7-membered heterocyclyl], and the 5- to 6-membered heteroaryl on -(C1-C4)[5- to 6-membered heteroaryl] are each optionally substituted, where valence permits, with 1 to 3 selected from R U The group substitution; R Ha Selected from (C1-C 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 )alkynyl, phenyl, 5- to 10-membered monocyclic or bicyclic heteroaryl, and 4- to 10-membered monocyclic or bicyclic heterocyclic group, wherein said (C1-C 10 )alkyl, (C2-C 10 )alkenyl, (C2-C 10 ) alkynyl groups are each optionally substituted, where valence permits, with 1 to 2 selected from R O wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 10-membered monocyclic or bicyclic heterocyclic group are each optionally substituted, where valence permits, by 1 to 3 groups selected from the group consisting of halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, oxo, and cyano; and R O Selected from halo, (C1-C4) alkoxy, OH, phenyl, NH2, -NH(C1-C 10 )alkyl, -N[(C1-C 10 )alkyl], a 4- to 10-membered monocyclic or fused bicyclic heterocyclic group, and a 5- to 10-membered monocyclic or bicyclic heteroaryl, wherein the 5- to 10-membered monocyclic or bicyclic heteroaryl and the 4- to 10-membered monocyclic or bicyclic heterocyclic group are each optionally substituted, as valence permits, with 1 to 3 groups selected from the group consisting of halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, oxo, and cyano.

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 p is 1.

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein R 2 It's hydrogen.

6. The compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, wherein R 3 It's hydrogen.

7. A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R 4 It's hydrogen. 8 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein q is 1, and t is 1. 9 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein q is 0 and t is 2.

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

11. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, wherein R 5 It's hydrogen.

12. A compound according to any one of claims 1 to 11 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 replace.

13. A compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from benzothienyl, indolyl, naphthyl, thienopyridinyl, benzothiazolyl, quinolyl, isoquinolyl, 4,5,6,7-tetrahydropyrrolopyridinyl, 5,6,7,8-tetrahydroimidazopyrazinyl, each of which is replaced by -CR 1a R 2a P(O)OR 1b OR 2b replace.

14. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, wherein R 1 15. A compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, wherein R 1 yes 16. A compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, wherein R 1a is hydrogen and R 2a is fluorine, or R 1a is fluorine and R 2a It's fluorine.

17. A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, wherein R 1a and R 2a It's fluorine.

18. A compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b each independently selected from hydrogen, (C1-C4)alkyl, -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)O-[(C1-C4)alkyl], -[(C1-C4)alkyl]-SC(O)-[(C1-C4)alkyl], -[(C1-C4)alkyl]-SC(O)-[halo(C1-C4)alkyl], -[(C1-C4)alkyl]-OC(O)NR 2c R 2d ], [(C1-C4) alkyl]-OC(O)O-[5- to 7-membered heterocyclyl] and phenyl, wherein the 5- to 7-membered heterocyclyl of [(C1-C4) alkyl]-OC(O)O-[5- to 7-membered heterocyclyl] is optionally substituted by C(O)OR 2c Substituted, wherein any of the (C1-C)4 alkyl groups is optionally substituted with 1 or 2 (C1-C4)alkyl groups.

19. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b Each is -[(C1-C4)alkyl]-OC(O)-[(C1-C4)alkyl].

20. A compound according to any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, wherein R 1b and R 2b It's hydrogen.

21. A compound according to any one of claims 1 to 15 and 18, or a pharmaceutically acceptable salt thereof, wherein -CR 1a R 2a P(O)OR 1b OR 2b Selected from or a pharmaceutically acceptable salt thereof.

22. A compound according to any one of claims 1 to 15, 18 and 21, or a pharmaceutically acceptable salt thereof, wherein -CR 1a R 2a P(O)OR 1b OR 2b yes 23. A compound according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R 7 is selected from (C1-C4)alkyl, phenyl, 4- to 6-membered monocyclic heterocyclyl, 9- or 10-membered fused bicyclic heterocyclyl, 5- or 6-membered monocyclic heteroaryl and 9- or 10-membered fused bicyclic heteroaryl, wherein the (C1-C4)alkyl is optionally substituted, where valence permits, by 1 to 3 selected from R Y The phenyl, 5-membered or 6-membered monocyclic heterocyclic group, 9-membered or 10-membered fused bicyclic heterocyclic group, 5-membered or 6-membered monocyclic heteroaryl group and 9-membered or 10-membered fused bicyclic heteroaryl group are each optionally substituted with 1 to 3 groups selected from R Z The group is substituted.

24. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R 7 is selected from (C1-C4)alkyl, phenyl, pyridyl, pyrimidinyl, naphthyl, cinnolinyl, quinoxalinyl, quinazolinyl, quinolyl, isoquinolyl, chromanyl, pyrazolyl, indazolyl, benzisoxazolyl, imidazo[1,2-a]pyridinyl, pyrrolidinyl, cyclopentyl, cyclohexyl, azetidinyl, piperidinyl and dihydropyridinyl, and wherein the (C1-C4)alkyl is optionally substituted, where valence permits, by 1 to 3 selected from R Y substituted with a group, and the phenyl, pyridyl, pyrimidinyl, naphthyl, cinnolinyl, quinoxalinyl, quinazolinyl, quinolyl, isoquinolyl, chromanyl, pyrazolyl, indazolyl, benzisoxazolyl, imidazo[1,2-a]pyridinyl, pyrrolidinyl, cyclopentyl, cyclohexyl, azetidinyl, piperidinyl and dihydropyridinyl are each optionally substituted with 1 to 3 selected from R where valence permits Z The group is substituted.

25. A compound according to any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, wherein R Y Selected from cyano, (C1-C4) alkyl, -NR a R b and hydroxyl groups.

26. A compound according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, wherein R Z is selected from halo, cyano, (C1-C4) alkyl, halo (C1-C4) alkyl, hydroxy (C1-C4) alkyl, -(C1-C4) alkylphenyl, -(C1-C4) alkylheteroaryl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, -OR e , oxo, phenyl, 4- to 6-membered heterocyclic group, -S(O)2R f 、-(C1-C4 alkyl)C(O)R g 、-C(O)R g 、-(C1-C4 alkyl)C(O)NR c R d 、-C(O)NR c R d , -NO2 and -NR a R b , wherein the (C1-C4)alkyl is optionally substituted by cyano, and wherein the phenyl and the 4- to 6-membered heterocyclyl are each optionally and independently substituted by 1 to 3 groups selected from cyano and oxo, where valence permits.

27. A compound according to any one of claims 1 to 26, or a pharmaceutically acceptable salt thereof, wherein R Z Select halo, cyano, (C1-C4) alkyl, halo (C1-C4) alkyl, hydroxy (C1-C4) alkyl, - (C1-C4) alkylphenyl, - (C1-C4) alkylheteroaryl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, -OR e , oxo, -S(O)2R f 、-(C1-C4 alkyl)C(O)R g 、-C(O)R g 、-(C1-C4 alkyl)C(O)NR c R d 、-C(O)NR c R d 、-NO2、-NR a R b , phenyl and dihydropyridinyl, wherein the (C1-C4)alkyl is optionally substituted by cyano, and wherein the phenyl and the dihydropyridinyl are each optionally and independently substituted by 1 to 3 groups selected from cyano and oxo where valence permits.

28. A compound according to any one of claims 1 to 22, 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 6-membered monocyclic heterocyclic group, a 7- to 13-membered spiro bicyclic heterocyclic group, or a 9- to 10-membered fused bicyclic heterocyclic group, each of which is optionally substituted, where valence permits, by 1 to 3 selected from R Q The group is substituted.

29. A compound according to any one of claims 1 to 22 and 28, or a pharmaceutically acceptable salt thereof, wherein R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms an azetidinyl, spiro[indoline-3,3'-pyrrolidinyl], pyrrolidinyl, 3,4-dihydrobenzoxazinyl, 1,2,3,4-tetrahydroquinolinyl, octahydrothiopyranopyrrolyl, octahydro-1H-thieno[3',4':3,4]cyclobutane[1,2-c]pyrrolyl, 2,6-diazabicyclo[3.2.0]heptanyl, piperazinyl, morpholinyl, 1,4,5,6-tetrahydropyrrolopyrrolyl, oxazolyl, 4-azaspiro[2.4]heptyl, 2-oxa-5-azabicyclo[2.2.1]heptyl, 7-oxa-4-azaspiro[2.5]octanyl, 8-oxa-3-azabicyclo[3.2.1]octanyl, 4,6-diazaspiro[2.4]heptyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl or piperidinyl, each of which is optionally substituted, where valence permits, by 1 to 3 selected from R Q The group is substituted.

30. A compound according to any one of claims 1 to 22 and 28, or a pharmaceutically acceptable salt thereof, wherein R 6 and R 7 together with the nitrogen atom to which it is attached, form an azetidinyl, spiro[indoline-3,3'-pyrrolidinyl], pyrrolidinyl, 3,4-dihydrobenzoxazinyl, 1,2,3,4-tetrahydroquinolinyl, octahydrothiopyranopyrrolyl, octahydro-1H-thieno[3',4':3,4]cyclobuta[1,2-c]pyrrolyl, 2,6-diazabicyclo[3.2.0]heptanyl, piperazinyl, 1,4,5,6-tetrahydropyrrolopyrazolyl, 5-oxa-2,6-diazaspiro[3.4]oct-6-enyl or piperidinyl, each of which is optionally substituted, as valence permits, by 1 to 3 selected from R Q The group is substituted.

31. A compound according to any one of claims 1 to 22, 28, 29 and 30, 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 pyrrolidinyl and an azetidinyl group, each of which is optionally substituted, where valence permits, with 1 to 3 selected from R Q The group is substituted.

32. A compound according to any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof, wherein R Q Selected from halo, (C1-C4) alkyl, halo (C1-C4) alkyl, -S(O)2R f , cyano, phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic group, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C3-C6) cycloalkyl, oxo, -OR e 、-C(O)R g 、-C(O)NR c R d 、-C(O)OR e 、-NR c C(O)R e wherein the (C1-C4)alkyl groups are each optionally and independently substituted, where valence permits, by 1 to 3 groups selected from R M wherein the phenyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl and (C3-C6) cycloalkyl are each optionally and independently substituted with 1 to 3 selected from R F The group is substituted.

33. A compound according to any one of claims 1 to 31 or a pharmaceutically acceptable salt thereof, wherein R Q is selected from halo, (C1-C4) alkyl, halo (C1-C4) alkyl, cyano, phenyl, 4- to 9-membered monocyclic or bicyclic heterocyclic group, 5- to 10-membered monocyclic or bicyclic heteroaryl, (C3-C6) cycloalkyl, oxo, -OR e 、-C(O)R g 、-C(O)NR c R d 、-C(O)OR e 、-NR c C(O)R e wherein the (C1-C4)alkyl groups are each optionally and independently substituted, where valence permits, by 1 to 3 groups selected from R M wherein the phenyl, 4- to 6-membered heterocyclyl, 5- to 10-membered monocyclic or bicyclic heteroaryl and (C3-C6) cycloalkyl are each optionally and independently substituted with 1 to 3 selected from R F The group is substituted.

34. A compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, wherein R Q Selected from cyano, (C1-C4) alkyl, halo (C1-C4) alkyl, -S(O)2R f , oxo, -C(O)R g 、-OR e 、-C(O)OR e 、-C(O)NR c R d 、-NR c C(O)R e , cyclobutyl, cyclopropyl, cyclohexyl, phenyl, benzisothiazolyl, pyridyl, pyrimidinyl, dihydropyridyl, pyrazolyl, piperidinyl, tetrahydroimidazopyridinyl and isothiazolidinyl, wherein the (C1-C4)alkyl is optionally and independently replaced by 1 to 3 selected from R where valence permits M wherein the cyclobutyl, cyclopropyl, cyclohexyl, phenyl, benzisothiazolyl, pyridyl, pyrimidinyl, dihydropyridyl, pyrazolyl, piperidinyl, tetrahydroimidazopyridinyl and isothiazolidinyl are each optionally and independently substituted with 1 to 3 groups selected from R where valence permits F The group is substituted.

35. A compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, wherein R Q is selected from cyano, (C1-C4)alkyl, halo (C1-C4)alkyl, oxo, -C(O)R g 、-OR e 、-C(O)OR e 、-C(O)NR c R d 、-NR c C(O)R e , cyclobutyl, cyclopropyl, cyclohexyl, phenyl, pyridyl, pyrimidinyl, dihydropyridyl, pyrazolyl, piperidinyl, tetrahydroimidazopyridinyl and isothiazolidinyl, wherein the (C1-C4)alkyl is optionally and independently replaced by 1 to 3 selected from R where valence permits M wherein the cyclobutyl, cyclopropyl, cyclohexyl, phenyl, pyridyl, pyrimidinyl, dihydropyridyl, pyrazolyl, piperidinyl, tetrahydroimidazopyridinyl and isothiazolidinyl are each optionally and independently substituted with 1 to 3 selected from R where valence permits F The group is substituted.

36. A compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, wherein R M phenyl and 5- to 6-membered monocyclic heteroaryl, wherein the phenyl and 5- to 6-membered monocyclic heteroaryl and the phenyl are each optionally substituted by 1 to 3 selected from R X The group is substituted.

37. A compound according to any one of claims 1 to 36, or a pharmaceutically acceptable salt thereof, wherein R M is selected from phenyl and pyridyl, wherein the phenyl and pyridyl are each optionally substituted, where valence permits, with 1 to 3 selected from R X The group is substituted.

38. A compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof, wherein R X Selected from halo and (C1-C4)alkoxy.

39. A compound according to any one of claims 1 to 38, or a pharmaceutically acceptable salt thereof, wherein R F Selected from halo, cyano, (C1-C4) alkoxy, oxo and -NR c C(O)R e .

40. A compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt thereof, wherein R c and R d Each independently selected from hydrogen and (C1-C4)alkyl as valence permits, and wherein the (C1-C4)alkyl is optionally substituted with 1 to 3 selected from R J The group is substituted.

41. A compound according to any one of claims 1 to 40, or a pharmaceutically acceptable salt thereof, wherein R e independently selected from hydrogen, phenyl, 5- to 6-membered heteroaryl and (C1-C4)alkyl, and wherein the (C1-C4)alkyl is optionally substituted, where valence permits, by 1 to 3 selected from R J The group is substituted.

42. A compound according to any one of claims 1 to 41 or a pharmaceutically acceptable salt thereof, wherein R J independently selected from phenyl and 5- to 6-membered heteroaryl, and wherein the 5- to 6-membered heteroaryl is optionally substituted with (C3-C6)cycloalkyl.

43. A compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, wherein R J It is phenyl.

44. A compound according to any one of claims 1 to 43, or a pharmaceutically acceptable salt thereof, wherein R g is selected from piperidinyl, morpholinyl and piperazinyl.

45. A compound according to any one of claims 1 to 9 or 11 to 44, or a pharmaceutically acceptable salt thereof, wherein X is NR 8 .

46. ​​A compound according to any one of claims 1 to 45, wherein: R 8 is hydrogen, -C(O)R Ha 、-C(O)OR Ha 、(C1-C4)alkyl、-SO2R Ha 、-C(O)NR Ha R Hb , halo(C1-C4)alkyl, phenyl, -(C1-C4)[4- to 10-membered monocyclic or bicyclic heterocyclic group], -(C1-C4)[5- to 10-membered monocyclic or bicyclic heteroaryl], -(C3-C6)cycloalkyl or 4- to 10-membered monocyclic or bicyclic heterocyclic group, wherein the phenyl group on the -(C1-C4)alkylphenyl group is optionally substituted by 1 or 2 independently selected from -(C 1-3 alkoxy) and a 5- to 6-membered heteroaryl group optionally substituted by -(C1-C4)alkyl, and wherein the (C1-C4)alkyl is optionally substituted by 1 to 3 phenyl groups; R Ha is (C1-C4)alkyl, -(C1-C4)[5- to 6-membered heteroaryl], -(C1-C4)[phenyl], -(C1-C4)[(C3-C6)cycloalkyl], phenyl, 4- to 10-membered monocyclic or bicyclic heterocyclyl, wherein the phenyl and 4- to 10-membered monocyclic or bicyclic heterocyclyl are each optionally substituted with 1 to 3 groups independently selected from halo, (C1-C4)alkyl, (C1-C4)alkoxy, and 4- to 6-membered monocyclic heterocyclyl optionally substituted with C1-C4alkyl; and R Hb It's H.

47. The compound according to claim 46, wherein: R 8 is -H, -CH2CH3, -CH2CH(CH3)2, -CH2CF2, -CH2CF3, cyclobutyl, -CH2-quinuclidine, -CH2-imidazo[1,5-a]pyridinyl, -CH2-pyrimidinyl, -CH2-phenyl, phenyl, pyrimidinyl, pyridinyl, isoquinolinyl, -C(O)R Ha 、-C(O)OR Ha 、-SO2R Ha or -C(O)NR Ha R Hb , or R 8 Represented by the following structure: and R Ha is methyl, ethyl, propyl, isopropyl, -CH2-quinolyl, -CH2-phenyl, -CH2CH2-phenyl, -CH(CH3)-phenyl, -CH(CH3)CH2-phenyl, -CH2CH(CH3)-phenyl, -CH2CH2-pyrimidinyl, -CH2C(CH3)2-cyclohexyl, quinolyl, indazolyl, benzisoxazolyl, imidazo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, 5,6,7,8-tetrahydroimidazo[1,5-a]pyridinyl, 4,5,6,7-tetrahydrobenzo[d]isoxazolyl or 6,7-dihydro-4H-pyrano[3,4-d]isoxazolyl, and wherein the quinolyl, indazolyl, benzisoxazolyl, imidazo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, 5,6,7,8-tetrahydroimidazo[1,5-a]pyridinyl, 4,5,6,7-tetrahydrobenzo[d]isoxazolyl, 6,7-dihydro-4H-pyrano[3,4-d]isoxazolyl are each optionally substituted by 1 to 3 groups independently selected from the following: -Cl, -CH3, -CH2CH3, -OCH3 or N-methylpiperazinyl.

48. The compound according to claim 1, wherein the compound is selected from any one of Compounds 1 to 441; or a pharmaceutically acceptable salt thereof.

49. A pharmaceutically acceptable composition comprising a compound according to any one of claims 1 to 48 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

50. 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 48, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition according to claim 49.

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