Methods for making bicyclic ketone compounds

Abstract

A bicyclic ketone compound of formula (I), or a stereoisomer thereof: TIFF2024513394000133.tif26170 or a pharma- ceutically acceptable salt thereof, 1 , R 2 , R 3 and n is as defined herein], and compounds prepared by these methods are provided herein.

Description

[Technical field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 170,422, filed April 2, 2021, the entire contents of which are incorporated by reference herein.

[0002] Technical Field Provided herein are methods for preparing bicyclic ketone compounds useful for the treatment and / or prevention in mammals, and compounds prepared by the methods. In particular, the bicyclic ketone compounds are chiral RIP1 kinase inhibitors useful for the treatment of diseases and disorders associated with inflammation, cell death, and the like. [Background technology]

[0003] Receptor interacting protein 1 ("RIP1") kinase is a serine / threonine protein kinase. RIP1 is a regulator of cell signaling involved, among other things, in mediating programmed cell death pathways (e.g., necroptosis). The best-studied form of necroptotic cell death is initiated by TNFα (tumor necrosis factor), but necroptosis can also be induced by other members of the TNFα death ligand family (Fas and TRAIL / Apo2L), interferon, Toll-like receptor (TLR) signaling, and viral infection via the DNA sensor DAI (activator of DNA-dependent interferon regulatory factor) [1-3]. Binding of TNFα to TNFR1 (TNF receptor 1) promotes TNFR1 trimerization and the formation of the intracellular complex Complex-I. TRADD (TNF receptor-associated death domain protein) binds to the intracellular death domain of TNFR1 and recruits the protein kinase RIP1 (receptor interacting protein 1) through the death domains present in both proteins [4]. Following initial recruitment to the TNFR1-associated signaling complex, RIP1 translocates to a secondary cytoplasmic complex, complex II [5-7]. Complex II is formed by the proteins FADD (Fas-associated protein), RIP1, caspase-8, and death domain containing cFLIP. If caspase-8 is not fully activated or its activity is blocked, the protein kinase RIP3 is recruited to the complex, forming the necrosome and leading to the initiation of necroptotic cell death [8-10]. Once the necrosome is formed, RIP1 and RIP3 participate in a series of auto- and cross-phosphorylation events that are essential for necroptotic cell death. Necroptosis can be completely blocked by kinase-inactivating mutations in either of the two kinases, or chemically by RIP1 kinase inhibitors (necrostatins) or RIP3 kinase inhibitors [11-13]. Phosphorylation of RIP3 enables the binding and phosphorylation of the pseudokinase MLKL (mixed lineage kinase domain-like), a key component of necroptotic cell death [14, 15].

[0004] Necroptosis has crucial pathophysiological relevance in myocardial infarction, stroke, atherosclerosis, ischemia-reperfusion injury, inflammatory bowel disease, retinal degeneration, and many other common clinical diseases.

[16] Therefore, selective inhibitors of RIP1 kinase activity are desirable as potential treatments for diseases mediated by this pathway and associated with inflammation and / or necroptotic cell death.

[0005] Inhibitors of RIP1 kinase have been described previously. The first published inhibitor of RIP1 kinase activity was necrostatin 1 (Nec-1)

[17] . Modified versions of Nec-1 with varying abilities to block RIP1 kinase activity followed this initial discovery [11, 18]. Recently, additional RIP1 kinase inhibitors that are structurally distinct from the necrostatin class of compounds have been described [20, 21, 22].

[0006] However, the synthesis of inhibitors containing, for example, specific stereoisomers is difficult, and new synthetic methods for making inhibitors are needed.

[0007] The references cited above are as follows, which are incorporated herein by reference in their entireties: 1) Vanden Berghe, T., Linkermann, A., Jouan-Lanhouet, S., Walczak, H. and Vandenabeele, P. (2014) Regulated necrosis: the expanding network of non-apoptotic cell death pathways. Nature reviews. Molecular cell biology. 15, 135-147. 2) Newton, K. (2015) RIPK1 and RIPK3: critical regulators of inflammation and cell death. Trends in cel biology. 25, 347-353. 3) de Almagro, M. C. and Vucic, D. (2015) Necroptosis: Pathway diversity and characteristics. Semin Cell Dev Biol. 39, 56-62. 4) Chen, Z. J. (2012) Ubiquitination in signaling to and activation of IKK. Immunological reviews. 246, 95-106. 5) O’Donnell, M. A., Legarda-Addison, D., Skountzos, P., Yeh, W. C. and Ting, A. T. (2007) Ubiquitination of RIP1 regulates an NF-kappaB-independent cell-death switch in TNF signaling. Curr Biol. 17, 418-424. 6) Feoktistova, M., Geserick, P., Kellert, B., Dimitrova, D. P., Langlais, C., Hupe, M., Cain, K., MacFarlane, M., Hacker, G. and Leverkus, M. (2011) cIAPs block Ripoptosome formation, a RIP1 / caspase-8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Molecular cell. 43, 449-463. 7) Bertrand, M. J., Milutinovic, S., Dickson, K. M., Ho, W. C., Boudreault, A., Durkin, J., Gillard, J. W., Jaquith, J. B., Morris, S. J. and Barker, P. A. (2008) cIAP1 and cIAP2 facilitate cancer cell survival by functioning as E3 ligases that promote RIP1 ubiquitination. Mol Cell. 30, 689-700. 8) Wang, L., Du, F. and Wang, X. (2008) TNF-alpha induces two distinct caspase-8 activation pathways. Cell. 133, 693-703. 9) He, S., Wang, L., Miao, L., Wang, T., Du, F., Zhao, L. and Wang, X. (2009) Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-alpha. Cell.137, 1100-1111. 10) Cho, Y. S., Challa, S., Moquin, D., Genga, R., Ray, T. D., Guildford, M. and Chan, F. K. (2009) Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. Cell.137, 1112-1123. 11) Degterev, A., Hitomi, J., Germscheid, M., Ch’en, I. L., Korkina, O., Teng, X., Abbott, D., Cuny, G. D., Yuan, C., Wagner, G., Hedrick, S. M., Gerber, S. A., Lugovskoy, A. and Yuan, J. (2008) Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol. 4, 313-321. 12) Newton, K., Dugger, D. L., Wickliffe, K. E., Kapoor, N., de Almagro, M. C., Vucic, D., Komuves, L., Ferrando, R. E., French, D. M., Webster, J., Roose-Girma, M., Warming, S. and Dixit, V. M. (2014) Activity of protein kinase RIPK3 determines whether cells die by necroptosis or apoptosis. Science. 343, 1357-1360. 13) Kaiser, W. J., Sridharan, H., Huang, C., Mandal, P., Upton, J. W., Gough, P. J., Sehon, C. A., Marquis, R. W., Bertin, J. and Mocarski, E. S. (2013) Toll-like receptor 3-mediated necrosis via TRIF, RIP3, and MLKL. The Journal of biological chemistry. 288, 31268-31279. 14) Zhao, J., Jitkaew, S., Cai, Z., Choksi, S., Li, Q., Luo, J. and Liu, Z. G. (2012) Mixed lineage kinase domain-like is a key receptor interacting protein 3 downstream component of TNF-induced necrosis. Proceedings of the National Academy of Sciences of the United States of America. 109, 5322-5327. 15) Sun, L., Wang, H., Wang, Z., He, S., Chen, S., Liao, D., Wang, L., Yan, J., Liu, W., Lei, X. and Wang, X. (2012) Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell.148, 213-227. 16) Linkermann, A. and Green, D. R. (2014) Necroptosis. The New England journal of medicine. 370, 455-465. 17) Degterev, A., Huang, Z., Boyce, M., Li, Y., Jagtap, P., Mizushima, N., Cuny, G. D., Mitchison, T. J., Moskowitz, M. A. and Yuan, J. (2005) Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nat Chem Biol.1, 112-119. 18) Takahashi, N., Duprez, L., Grootjans, S., Cauwels, A., Nerinckx, W., DuHadaway, J. B., Goossens, V., Roelandt, R., Van Hauwermeiren, F., Libert, C., Declercq, W., Callewaert, N., Prendergast, G. C., Degterev, A., Yuan, J. and Vandenabeele, P. (2012) Necrostatin-1 analogues: critical issues on the specificity, activity and in vivo use in experimental disease models. Cell Death Dis. 3, e437. 19) Harris, P. A., Bandyopadhyay, D., Berger, S. B., Campobasso, N., Capriotti, C. A., Cox, J. A., Dare, L., Finger, J. N., Hoffman, S. J., Kahler, K. M., Lehr, R., Lich, J. D., Nagilla, R., Nolte, R. T., Ouellette, M. T., Pao, C. S., Schaeffer, M. C., Smallwood, A., Sun, H. H., Swift, B. A., Totoritis, R. D., Ward, P., Marquis, R. W., Bertin, J. and Gough, P. J. (2013) Discovery of Small Molecule RIP1 Kinase Inhibitors for the Treatment of Pathologies Associated with Necroptosis. ACS medicinal chemistry letters. 4, 1238-1243. 20) Najjar, M., Suebsuwong, C., Ray, SS, Thapa, RJ, Maki, JL, Nogusa, S., Shah, S., Saleh, D., Gough, PJ, Bertin, J., Yuan, J., Balachandran, S., Cuny, GD and Degterev, A. (2015) Structure Guided Design of Potent and Selective Ponatinib-Based Hybrid Inhibitors for RIPK1. Cell Rep. 21) International Publication No. 2014 / 125444. 22) International Publication No. 2017 / 004500. Summary of the Invention

[0008] Solutions to these problems and more are provided herein.

[0009] In one embodiment, a compound represented by formula (I) or (II): TIFF2024513394000002.tif33170[In the formula, R 1 is C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 alkyl-N(R N 2) selected from the group consisting of phenyl, benzyl, 4- to 8-membered heterocyclyl, and 5- to 6-membered heteroaryl, where R 1 is attached to the adjacent carbonyl by a carbon atom, and R 1 is F, Cl, Br, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 alkyl-N(R N)2, hydroxyl, hydroxymethyl, cyano, cyanomethyl, cyanoethyl, C(O)C1-C6 alkyl, phenyl, benzyl, CH2-(C3-C6 cycloalkyl), 5- to 6-membered heteroaryl, and CH2-(5- to 6-membered heteroaryl); Each R N are independently selected from the group consisting of H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, and C1-C6 haloalkyl; or two R N may form a 4- to 6-membered ring together with the adjacent N; R 2 is selected from the group consisting of H, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C6 thioalkyl, phenyl, benzyl, CH2-(C3-C6 cycloalkyl), CH2CH2-(C3-C6 cycloalkyl), CH2-(4-6 membered heterocyclyl), CH2CH2-(4-6 membered heterocyclyl), 5-6 membered heteroaryl, and CH2-(5-6 membered heteroaryl); where the phenyl ring, if present, is optionally substituted with 1 to 3 substituents selected from the group consisting of halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, and cyano; R 3 is selected from the group consisting of D, halogen, OH, CN, C1-C4 alkyl, C1-C4 haloalkyl, cyclopropyl, C1-C4 alkoxy and C1-C4 haloalkoxy; n is 1, 2, or 3.

[0010] Provided herein are processes for the preparation of chiral bicyclic ketone compounds of the formula:

[0010] In another aspect, provided herein is a method for the preparation of a chiral bicyclic ketone compound of formula (I), or a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof, wherein R 1 , R 2 , R 3and n is as defined herein, the method comprising: (a) Formula of chiral N-aminolactam p or a stereoisomer thereof: TIFF2024513394000003.tif27170 or a salt thereof in the presence of an acid additive and an alcohol solvent, c Imidate compounds: or a salt thereof, x or a stereoisomer thereof: TIFF2024513394000005.tif26170 or a salt thereof, Pg 1 is an optional hydroxyl protecting group, which may be the same or different in each occurrence; formula x or a stereoisomer thereof is an intermediate compound in the preparation of a chiral bicyclic ketone compound of formula (I) or a stereoisomer thereof.

[0011] In another embodiment, the formula p or a stereoisomer thereof: Provided herein is a method for the preparation of TIFF2024513394000006.tif27170, or a salt thereof, wherein R 2 and n is as defined herein, the method comprising: Formula (a) d or a stereoisomer thereof: TIFF2024513394000007.tif22170 or a salt thereof, e Acid chloride of: or a salt thereof to obtain a compound of the formula f or a stereoisomer thereof: TIFF2024513394000009.tif27170 or a salt thereof; Formula (b) l or a stereoisomer thereof: or a salt thereof in the presence of an acid additive, formula m or a stereoisomer thereof: TIFF2024513394000011.tif26170 or a salt thereof, Pg 2 is optionally substituted with C1-C6 alkyl, C3-C6 cycloalkyl, or aryl; Pg 3 is an optional hydroxyl protecting group, which may be the same or different in each occurrence; Pg 4 is an optional amine protecting group, which may be the same or different in each occurrence; formula f or a stereoisomer thereof, is represented by the formula l or a stereoisomer thereof, formula m or a stereoisomer thereof, has the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0012] In another embodiment, the formula p or a stereoisomer thereof, or a salt thereof, comprising: Formula (a) d or a stereoisomer thereof: TIFF2024513394000012.tif22170 or a salt thereof, e Acid chloride of: or a salt thereof to obtain a compound of the formula f or a stereoisomer thereof: TIFF2024513394000014.tif32170 or a salt thereof, Pg 2 , Pg 3 , and Pg 4 is as defined herein; formula f or a stereoisomer thereof, is represented by the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0013] In another embodiment, the formula p or a stereoisomer thereof, or a salt thereof, comprising: Formula (b) l or a stereoisomer thereof: or a salt thereof in the presence of an acid additive to obtain a compound of the formula m or a stereoisomer thereof: TIFF2024513394000016.tif26170 or a salt thereof, Pg 2 , Pg 3 , and Pg 4 is as defined herein; formula m or a stereoisomer thereof, has the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0014] In another embodiment, the formula b Imidate salt compounds of: Provided herein is a method for the preparation of TIFF2024513394000017.tif19170, the method comprising: Formula (a) a Cyanoformate compounds: TIFF2024513394000018.tif18170 in the presence of an anhydrous acid source in an alcohol solvent to obtain b wherein the anhydrous acid source is TMSCl, the acid is HCl, and Pg 1 is an optional hydroxyl protecting group, which may be the same or different in each occurrence.

[0015] In another embodiment, the compound of formula (III) or (IV): Provided herein is a method for the preparation of a chiral bicyclic ketone compound of formula TIFF2024513394000019.tif28170, or a pharma- ceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , and n are as defined herein.

[0016] In another embodiment, the formula j or a stereoisomer thereof: Provided herein is a method for the preparation of TIFF2024513394000020.tif22170, the method comprising: Formula (a) hh Diketo ester compounds of: TIFF2024513394000021.tif20170 was reacted in the presence of ketoreductase to obtain the compound of formula j or a stereoisomer thereof, 2 , Pg 3 , and n is as defined herein].

[0017] In another aspect, there is provided a compound prepared by the methods described herein.

[0018] The subject process provides unexpectedly good overall yields in the preparation of compounds of formula (I), formula (II), formula (III) or formula (IV), or pharma- ceutically acceptable salts thereof, as well as improved product purity, improved diastereomeric ratios, improved stereoisomeric excess, and improved yields through avoidance of potential racemization events. Additional embodiments and details are provided below. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] definition All chemical formulas and general chemical structures provided herein shall be interpreted to provide appropriate valences and chemically stable bonds between atoms as understood by one of ordinary skill in the art. Where appropriate, substituents may be bonded to adjacent atoms (e.g., alkyl includes methylene where there are two bonds).

[0020] In the chemical formulas provided herein, "halogen" or "halo" refers to fluorine, chlorine, and bromine (i.e., F, Cl, Br).

[0021] "Alkyl" refers to an optionally substituted, straight or branched C-C alkyl group, unless otherwise specifically defined. 12In some embodiments, "alkyl" refers to a C1-C6 alkyl group. Exemplary alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. The substituted alkyl groups provided herein are substituted with one or more substituents selected from the group consisting of halogen, cyano, trifluoromethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, C3-C6 cycloalkyl, phenyl, OH, CO2H, CO2(C1-C4 alkyl), NH2, NH(C1-C4 alkyl), N(C1-C4 alkyl), NH(C=O)C1-C4 alkyl, (C=O)NH(C1-C4 alkyl), (C=O)N(C1-C4 alkyl), S(C1-C4 alkyl), SO(C1-C4 alkyl), SO2(C1-C4 alkyl), SON2NH(C1-C4 alkyl), SON2N(C1-C4 alkyl), and NHSO2(C1-C4 alkyl). In some embodiments, the substituted alkyl group has one or two substituents. In some embodiments, the substituted alkyl group is unsubstituted.

[0022] "Cycloalkyl" means, unless otherwise specifically defined, an optionally substituted C-C 12It refers to a cycloalkyl group, including fused, spirocyclic, and bridged bicyclic groups, where the substituents are selected from the group consisting of halogen, cyano, trifluoromethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, C3-C6 cycloalkyl, phenyl, OH, CO2H, CO2(C1-C4 alkyl), NH2, NH(C1-C4 alkyl), N(C1-C4 alkyl)2, NH(C=O)C1-C4 alkyl, (C=O)NH(C1-C4 alkyl), (C=O)N(C1-C4 alkyl)2, S(C1-C4 alkyl), SO(C1-C4 alkyl), SO2(C1-C4 alkyl), SON2NH(C1-C4 alkyl), SON2N(C1-C4 alkyl)2, and NHSO2(C1-C4 alkyl). In some embodiments, cycloalkyl refers to a C3-C6 cycloalkyl group. In some embodiments, the C3-C6 cycloalkyl group is optionally substituted with 1 to 3 halogen atoms. In some embodiments, the C3-C6 cycloalkyl group is optionally substituted with 1 to 3 fluorine atoms. Exemplary C3-C6 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Exemplary C3-C6 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. 12 Cycloalkyl groups further include bicyclo[3.1.0]hexyl, bicyclo[2.1.1]hexyl, cycloheptyl, bicyclo[4.1.0]heptyl, spiro[4.2]heptyl, cyclooctyl, spiro[4.3]octyl, spiro[5.2]octyl, bicyclo[2.2.1]heptanyl, bicyclo[2.2.2]octanyl, adamantanyl, decalinyl, and spiro[5.4]decanyl. If desired, the cycloalkyl group may be fused to other groups such that there are multiple chemical bonds between the cycloalkyl group and another ring system (e.g., the C ring of formula I). ​​In some embodiments, the cycloalkyl group is unsubstituted.

[0023] "Haloalkyl" means, unless otherwise specifically defined, a straight or branched C-C alkyl group in which one or more hydrogen atoms are replaced by halogen. 12In some embodiments, "haloalkyl" refers to a C1-C6 haloalkyl group. In some embodiments, 1 to 3 hydrogen atoms of the haloalkyl group are replaced by halogen. In some embodiments, all hydrogen atoms of the haloalkyl group are replaced by halogen (e.g., trifluoromethyl). In some embodiments, haloalkyl is as defined herein, where in each instance, halogen is fluorine. Exemplary haloalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, and pentafluoroethyl.

[0024] "Alkoxy" refers to any straight or branched C-C alkyl group, in each instance where one or more oxygen atoms are present between two carbon atoms, unless otherwise specifically defined. 12 It refers to an alkyl group. In some embodiments, "alkoxy" refers to a C1-C6 alkoxy group. In some embodiments, the C1-C6 alkoxy groups provided herein have one oxygen atom. Exemplary alkoxy groups include methoxy, ethoxy, CHOCH, CHCHOCH, CHOCHCH, CHCHOCHCH, CHOCHCHCH, CHOCHCHCH, CHOCH(CH), CHOC(CH), CH(CH)OCH, CHCH(CH)OCH, CH(CH)OCH, CH(CH)OCHCH, CHOCHOCH, CHCHOCHCHOCH, and CHOCH2OCH2OCH.

[0025] "Cycloalkoxy" refers to a C-C alkyl group, as defined above, which is cyclic and contains one oxygen atom, unless otherwise specifically defined. 10 or a C4-C6 alkoxy group. Exemplary cycloalkoxy groups include oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl.

[0026] "Haloalkoxy" refers to a C1-C6 haloalkyl group as defined above, in which, unless otherwise specifically defined, one or two oxygen atoms are present between two carbon atoms in each instance. In some embodiments, the C1-C6 haloalkoxy group provided herein has one oxygen atom. Exemplary haloalkoxy groups include OCF3, OCHF2, and CH2OCF3.

[0027] "Thioalkyl" means, unless otherwise specifically defined, a C1-C6 alkyl group as defined above in which an oxygen atom is replaced by a sulfur atom. 12 Or C1-C6 alkoxy groups. In some embodiments, thioalkyl groups may contain sulfur atoms replaced by one or more oxygen atoms (i.e., alkylsulfones and alkylsulfoxides). Exemplary thioalkyl groups are those exemplified in the definition of alkoxy above, in which each oxygen atom is replaced by a sulfur atom in each instance.

[0028] "Thiocycloalkyl" refers to a C-C alkyl group, as defined above, which is cyclic and contains one sulfur atom, unless otherwise specifically defined. 10 Or C4-C6 thioalkyl groups. In some embodiments, the sulfur atom of the thiocycloalkyl group is replaced by one or two oxygen atoms (i.e., cyclic sulfone or sulfoxide). Exemplary thiocycloalkyl groups include thietanyl, thiolanyl, thianyl, 1,1-dioxothiolanyl, and 1,1-dioxothianyl.

[0029] "Heterocyclyl" refers to a single saturated or partially unsaturated 4-8 membered ring, unless otherwise specifically defined, having at least one atom other than carbon in the ring, where the atom is selected from the group consisting of oxygen, nitrogen and sulfur. The term also includes multiple condensed ring systems having at least one such saturated or partially unsaturated ring, the multiple condensed ring systems having 7-12 atoms, as detailed below. Thus, the term includes a single saturated or partially unsaturated ring (e.g., 3-, 4-, 5-, 6-, 7- or 8-membered ring) from about 1-7 carbon atoms in the ring and about 1-4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur. The ring may be C-branched (i.e., substituted with C1-C4 alkyl). The ring may be substituted with one or more (e.g., 1, 2 or 3) oxo groups, and the sulfur and nitrogen atoms may be present in their oxidized forms. Exemplary heterocycles include, but are not limited to, azetidinyl, tetrahydrofuranyl, and piperidinyl. The rings of a multiple condensed ring system may be connected to each other through fused bonds, spiro bonds, and bridge bonds, if valence requirements permit. The individual rings of a multiple condensed ring system may be connected to each other in any order. Additionally, the point of attachment of a multiple condensed ring system (as defined above for heterocycles) may be at any position of the multiple condensed ring system. Additionally, the point of attachment of a heterocycle or heterocycle multiple condensed ring system may be at any suitable atom of the heterocyclyl group, including carbon and nitrogen atoms.Exemplary heterocycles include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, homopiperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, tetrahydrofuran, dihydrooxazolyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,2,3,4-tetrahydroquinolyl, benzoxazinyl, dihydrooxazolyl, chromanyl, 1,2-dihydropyridinyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxolyl, 1,4-benzodioxanyl, spiro[cyclopropane-1,1'-isoindolinyl]-3'-one, isoindolinyl-1-one, 2-oxa-6-azaspiro[3.3]heptanyl, imidazolidin-2-one. These include N-methylpiperidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, 1,4-dioxane, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, pyran, 3-pyrroline, thiopyran, pyrone, tetrahydrothiophene, quinuclidine, tropane, 2-azaspiro[3.3]heptane, (1R,5S)-3-azabicyclo[3.2.1]octane, (1s,4s)-2-azabicyclo[2.2.2]octane, (1R,4R)-2-oxa-5-azabicyclo[2.2.2]octane, and pyrrolidin-2-one.

[0030] In some embodiments, heterocyclyl is a C-C heteroatom having 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. 10 Heterocyclyl. In some embodiments, the heterocyclyl group is not bicyclic or spirocyclic. In some embodiments, the heterocyclyl is a C5-C6 heterocyclyl having 1-3 heteroatoms, where if there are 3 heteroatoms, at least 2 are nitrogen.

[0031] "Aryl", unless otherwise specifically defined, refers to a single all-carbocyclic aromatic ring or a multiple condensed all-carbocyclic ring system, where at least one of the rings is aromatic, and the aryl group has 6-20 carbon atoms, 6-14 carbon atoms, 6-12 carbon atoms, or 6-10 carbon atoms. Aryl includes phenyl groups. Aryl also includes multiple condensed ring systems (e.g., ring systems containing 2, 3, or 4 rings) having about 9-20 carbon atoms, where at least one ring is aromatic and the other rings may or may not be aromatic (i.e., carbocyclic). Such multiple condensed ring systems may be substituted with one or more (e.g., 1, 2, or 3) oxo groups on any carbocyclic portion of the multiple condensed ring system. The rings of the multiple condensed ring system may be connected to each other through fused bonds, spiro bonds, and bridge bonds, where valency requirements permit. The point of attachment of the multiple condensed ring system may be at any position of the ring system, including the aromatic or carbocyclic portions of the rings, as defined above. Exemplary aryl groups include phenyl, indenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, and the like.

[0032] "Heteroaryl" refers to a 5-6 membered aromatic ring having at least one atom other than carbon in the ring, where the atom is selected from the group consisting of oxygen, nitrogen and sulfur, unless otherwise specifically defined; "heteroaryl" also includes multiple condensed ring systems having 8-16 atoms with at least one such aromatic ring, which multiple condensed ring systems are described in detail below. Thus, "heteroaryl" includes a single aromatic ring consisting of about 1-6 carbon atoms and about 1-4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur. The sulfur and nitrogen atoms may also be present in oxidized form when the ring is aromatic. Exemplary heteroaryl ring systems include, but are not limited to, pyridyl, pyrimidinyl, oxazolyl or furyl. "Heteroaryl" also includes multiple condensed ring systems (e.g., ring systems containing 2 or 3 rings), where the heteroaryl group is fused to one or more rings selected from heteroaryl (e.g., to form naphthyridinyl, such as 1,8-naphthyridinyl), heterocycle (e.g., to form 1,2,3,4-tetrahydronaphthyridinyl, such as 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycle (e.g., to form 5,6,7,8-tetrahydroquinolyl), and aryl (e.g., to form indazolyl) as defined above to form a multiple condensed ring system. Thus, a heteroaryl (single aromatic ring or multiple condensed ring system) has 1 to 15 carbon atoms and about 1 to 6 heteroatoms in the heteroaryl ring. Such multiple condensed ring systems may be substituted with one or more (e.g., 1, 2, 3, or 4) oxo groups on the carbocyclic or heterocyclic portions of the fused rings. The rings of a multiple condensed ring system may be connected to each other through fused bonds, spiro bonds, and bridge bonds, if valence requirements permit. The individual rings of a multiple condensed ring system may be connected to each other in any order. The point of attachment of a multiple condensed ring system (as defined above for heteroaryl) may be at any position of the multiple condensed ring system, including the heteroaryl, heterocyclic, or carbocyclic portions of the multiple condensed ring system. The point of attachment of a heteroaryl or heteroaryl multiple condensed ring system may be at any suitable atom of the heteroaryl or heteroaryl multiple condensed ring system, including carbon atoms and heteroatoms (e.g., nitrogen).Exemplary heteroaryls include, but are not limited to, pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furyl, oxadiazolyl, thiadiazolyl, quinolyl, isoquinolyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalyl, quinazolyl, 5,6,7,8-tetrahydroisoquinolinylbenzofuranyl, benzimidazolyl, thianaphthenyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl-4(3H)-one, triazolyl, 4,5,6,7-tetrahydro-1H-indazole, and 3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclo-penta[1,2-c]pyrazole.

[0033] As used herein, the term "chiral" refers to a molecule that has the property of not being superimposable on its mirror image partner, while the term "achiral" refers to a molecule that is superimposable on its mirror image partner.

[0034] As used herein, the term "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

[0035] As used herein, crossing a bond in a chemical structure TIFF2024513394000022.tif6170 shows the point of attachment of the bond in the chemical structure that is crossed by a wavy bond to the rest of the molecule.

[0036] As used herein, the term "C-linked" means that the group it describes is attached to the remainder of the molecule through a ring carbon atom.

[0037] As used herein, the term "N-linked" means that the group it describes is attached to the remainder of the molecule through a ring nitrogen atom.

[0038] "Diastereomer" refers to a stereoisomer with two or more chiral centers and whose molecules are not mirror images of one another. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers can separate under high resolution analytical procedures such as electrophoresis and chromatography.

[0039] "Enantiomers" refers to two stereoisomers of a compound which are non-superimposable mirror images of one another.

[0040] Stereochemical definitions and conventions used herein generally follow SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds," John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric or chiral centers and therefore exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, and atropisomers, and mixtures thereof, such as racemic mixtures, are intended to form part of the present invention. Many organic compounds exist in optically active forms, i.e., they can rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule around its chiral center. The prefixes d and l or (+) and (-) are used to indicate the sign of rotation of plane polarized light by a compound, with (-) or 1 meaning that the compound is levorotatory. Compounds with the prefix (+) or d are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer is also referred to as an enantiomer, and a mixture of such isomers may be called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or racemate, which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar compound of two enantiomeric species, devoid of optical activity.

[0041] When a bond in a compound formula herein is drawn non-stereochemically (e.g., planar), the atom to which the bond is attached includes all stereochemical possibilities. When a bond in a compound formula herein is drawn in a fixed stereochemical manner (e.g., bold, bold-wedge, dashed, or dashed-wedge), it is understood that the atom to which the stereochemical bond is attached is enriched in the absolute stereoisomer depicted, unless otherwise indicated. In one embodiment, the compound may be at least 51% of the absolute stereoisomer depicted. In another embodiment, the compound may be at least 80% of the absolute stereoisomer depicted. In another embodiment, the compound may be at least 90% of the absolute stereoisomer depicted. In another embodiment, the compound may be at least 95% of the absolute stereoisomer depicted. In another embodiment, the compound may be at least 97% of the absolute stereoisomer depicted. In another embodiment, the compound may be at least 98% of the absolute stereoisomer depicted. In another embodiment, the compound may be at least 99% of the absolute stereoisomer depicted.

[0042] The term "tautomer" or "tautomeric form" as used herein refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions via rearrangement of some of the bonding electrons.

[0043] As used herein, the term "solvate" refers to the association or complex of one or more solvent molecules and the compound of the present invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water. In some embodiments, the hydrate of the compound provided herein is a ketone hydrate.

[0044] As used herein, the term "protective group" or "protecting group" refers to a substituent that is commonly used to block or protect a particular functional group on a compound. For example, an "amino-protecting group" is a substituent added to an amino group that blocks or protects the amino functionality in the compound. Suitable amino-protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a "hydroxy-protecting group" refers to a substituent of the hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A "carboxy-protecting group" refers to a substituent of the carboxy group that blocks or protects the carboxy functionality. Common carboxy protecting groups include phenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl, etc. For a general overview of protecting groups and their uses, see PGM Wuts and TW Greene, Greene's Protective Groups in Organic Synthesis 4 th See, for example, U.S. Pat. No. 6,313,316, 2006.

[0045] As used herein, the term "mammal" includes, but is not limited to, humans, mice, rats, guinea pigs, monkeys, dogs, cats, horses, cows, pigs, and sheep.

[0046] As used herein, the term "pharmaceutical acceptable salts" is meant to include salts of active compounds prepared with relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of salts derived from pharmaceutical acceptable inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganous, manganous, potassium, sodium, zinc, etc. Salts derived from pharma- ceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted, cyclic, naturally occurring amines, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When a compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharma- ceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and the like, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like.Also included are salts of amino acids, such as arginates, and salts of organic acids, such as glucuronic or galacturic acids (see, e.g., Berge, SM, et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19).Certain compounds of the invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0047] The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for purposes of this invention.

[0048] In addition to salt forms, the present invention provides compounds that are in prodrug form. As used herein, the term "prodrug" refers to a compound that readily undergoes chemical changes under physiological conditions to provide a compound of the present invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0049] The prodrugs of the present invention include compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., 2, 3, or 4) amino acid residues, is covalently linked to a free amino, hydroxy, or carboxylic acid group of a compound of the present invention through an amide or ester bond. The amino acid residues include, but are not limited to, the 20 naturally occurring amino acids, usually represented by their three-letter symbols, and also include phosphoserine, phosphothreonine, phosphotyrosine, 4-hydroxyproline, hydroxylysine, decomosine, isodecomosine, gamma-carboxyglutamate, hippuric acid, octahydroindole-2-carboxylic acid, statin, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, penicillamine, ornithine, 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine, methyl-alanine, para-benzoylphenylalanine, phenylglycine, propargylglycine, sarcosine, methionine sulfone, and tert-butylglycine.

[0050] Additional types of prodrugs are also included. For example, free carboxyl groups of the compounds of the invention can be derivatized as amides or alkyl esters. As another example, compounds of the invention containing free hydroxyl groups can be derivatized as prodrugs by converting the hydroxyl group to groups such as, but not limited to, phosphate esters, hemisuccinates, dimethylaminoacetates, or phosphoryloxymethyloxycarbonyl groups, as reviewed in Fleisher, D. et al., (1996) Improved oral drug delivery: solubility limitations overcome by the use of prodrugs Advanced Drug Delivery Reviews, 19:115. Carbamate prodrugs of hydroxy and amino groups are also included, as are carbonate prodrugs, sulfonate esters, and sulfate esters of hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers, where the acyl group can be an alkyl ester that may be substituted with groups including, but not limited to, ether, amine, and carboxylic acid functionalities, or where the acyl group is an amino acid ester as described above, are also included. This type of prodrug is described in J. Med. Chem., (1996), 39:10. More specific examples include prodrugs in which the hydrogen atom of the alcohol group is replaced with (C 1-6 ) alkanoyloxymethyl, 1-((C 1-6 )alkanoyloxy)ethyl, 1-methyl-1-((C 1-6 )alkanoyloxy)ethyl, (C 1-6 ) alkoxycarbonyloxymethyl, N-(C 1-6 ) alkoxycarbonylaminomethyl, succinoyl, (C 1-6 ) Alkanoyl, alpha-amino (C 1-4 ) alkanoyl, arylacyl, and alpha-aminoacyl or alpha-aminoacyl-alpha-aminoacyl (each alpha-aminoacyl group is independently selected from the naturally occurring L-amino acids, P(O)(OH), -P(O)(O(C 1-6) alkyl) 2 or glycosyl (the radical obtained from removal of the hydroxyl group of the hemiacetate form of a carbohydrate).

[0051] For additional examples of prodrug derivatives, see, for example, a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol. 42, p. 309-396 edited by K. Widder, et al. (Academic Press, 1985); b) A Textbook of Drug Design and Developmen, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 "Design and Application of Prodrugs", H. Bundgaard p. 113-191(1991); c) H. Bundgaard, Advanced Drug Delivery Reviews, 8:1-38(1992); d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77:285(1988); and e) N. Kakeya, et al., Chem. Pharm. Bull., 32:692 (1984), each of which is expressly incorporated herein by reference.

[0052] The present invention further provides metabolites of the compounds of the present invention. As used herein, "metabolite" refers to a product produced through metabolism in the body of a particular compound or its salt. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc., of the administered compound.

[0053] The metabolites are typically radiolabeled (e.g. 14 C or 3H) isotopes, parenterally administering it to animals such as rats, mice, guinea pigs, monkeys, etc., or to humans in detectable doses (e.g., greater than about 0.5 mg / kg), allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours), and isolating the conversion products from urine, blood, or other biological samples. Such products are easily isolated because they are labeled (others are isolated by using antibodies that can bind to epitopes remaining in the metabolites). The structures of the metabolites are determined in conventional manner, for example, by MS, LC / MS, or NMR analysis. In general, the analysis of the metabolites is performed in the same manner as conventional drug metabolism tests well known to those skilled in the art. The metabolites are useful in diagnostic assays for therapeutic administration of the compounds of the invention, unless they are otherwise found in vivo.

[0054] Certain compounds of the present invention may exist in solvated forms, including hydrated forms, as well as unsolvated forms. In general, solvated forms are equivalent to unsolvated forms and are intended to be included within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0055] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds. The racemates, diastereomers, geometric isomers, positional isomers, and individual isomers (e.g., separate optical isomers) are all intended to be encompassed within the scope of the present invention.

[0056] The term "composition," as used herein, is intended to encompass not only a product containing the specified ingredients in the specified amounts, but also any product that results directly or indirectly as a result of combining the specified ingredients in the specified amounts. "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0057] The terms "treat" and "treatment" refer to both therapeutic and / or prophylactic treatment or preventative measures, the purpose of which is to prevent or slow down (reduce) undesirable physiological changes or disorders, such as the onset or spread of cancer. In the context of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, whether detectable or not, reduction in the extent of the disease or disorder, stabilization (i.e., not worsening) of the disease or disorder, delay or slowing of disease progression, improvement or palliation of the condition or disorder, and remission (whether partial or total). "Treatment" can also mean prolonging survival as compared to expected survival in the absence of treatment. Those in need of treatment include those already with the disease or disorder, as well as those susceptible to the disease or disorder or those with the disease or disorder to be prevented.

[0058] The term "therapeutically effective amount" or "effective amount" refers to an amount of a compound of the present invention that (i) treats or prevents a particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of a particular disease, condition or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition or disorder described herein. With respect to cancer treatment, efficacy can be measured, for example, by assessing the time to disease progression (TTP) and / or determining the response rate (RR).

[0059] The term "bioavailability" refers to the systemic availability (i.e., blood / plasma levels) of a given amount of drug administered to a patient. Bioavailability is an absolute term that indicates that it measures both the time (rate) and total amount (extent) of drug that reaches the systemic circulation from an administered dosage form.

[0060] "Ketoreductase" and "KRED" are used interchangeably herein to refer to a polypeptide having the enzymatic ability to reduce a carbonyl group to the corresponding alcohol. For example, certain enzymes belonging to the ketoreductase (KRED) or carbonyl reductase class (EC 1.1.1.184) have been found to be useful for stereoselectively converting prostereoisomeric aldehyde or ketone substrates to the corresponding chiral alcohol products. KREDs usually convert ketone or aldehyde substrates to the corresponding alcohol products, but may also catalyze the reverse reaction, i.e., oxidation of an alcohol substrate to the corresponding ketone / aldehyde product, in which case they may be referred to as, for example, alcohol dehydrogenases (ADHs). Alcohol dehydrogenases (EC 1.1.1.1) belong to a group of enzymes that facilitate the conversion of alcohols to aldehydes or ketones. The reduction of ketones and aldehydes and the oxidation of alcohols by enzymes such as KREDs require a cofactor, most commonly reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH) for the oxidation reaction, and nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP) for the oxidation reaction. NADH and NADPH function as electron donors, while NAD and NADP function as electron acceptors. KREDs are increasingly being used for the stereoselective conversion of ketones and aldehydes to chiral alcohol compounds, which are used in the production of key pharmaceutical compounds.

[0061] Examples of the use of KREDs to produce useful compounds include the asymmetric reduction of 4-chloroacetoacetate (e.g., Zhou et al., J. Am. Chem. Soc. (1983), 105(18):5925-5926; Santaniello et al., J. Chem. Res., Synop. (1984), 4:132-133; U.S. Patent Nos. 5,559,030; 5,700,670 and 5,891,685), reduction of dioxocarboxylic acids (e.g., U.S. Patent No. 6,399,339), reduction of tert-butyl (S)-chloro-5-hydroxy-3-oxohexanoate (e.g., U.S. Patent No. 6,645,746 and WO 01 / 40450), reduction of pyrrolotriazine compounds (e.g., U.S. Patent Application Publication No. 2006 / 0286646), reduction of substituted acetophenones (e.g., U.S. Patent No. 6,800,477), and reduction of ketothiolanes (e.g., WO 2005 / 054491 and ).

[0062] In some embodiments, the keto reduction can be carried out in the presence of an alcohol, such as isopropanol, to provide a substrate for the reverse oxidation reaction (alcohol dehydrogenation). In some embodiments, the NADH / NADPH consumed in the keto reduction reaction is regenerated by the reverse oxidation reaction. In some embodiments, the KRED can stereoselectively reduce 2,4-dioxo-4-phenyl-butanoate to the corresponding alcohol, (-)-ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate. In some embodiments, the KRED utilizes the cofactor reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleophosphate (NADPH) as the reducing agent. In other embodiments, the NADH / NADPH consumed in the keto reduction reaction is regenerated by a coenzyme, such as glucose dehydrogenase or formate dehydrogenase. For example, the substrate for glucose dehydrogenase is glucose, and the substrate for formate dehydrogenase is formate. As used herein, ketoreductases and coenzymes include naturally occurring (wild-type) ketoreductases and coenzymes as well as engineered ketoreductases and coenzymes. Examples of engineered ketoreductases are described, for example, in U.S. Patent No. 7,820,421.

[0063] In some embodiments, when the method is carried out using whole cells of the host organism, the whole cells provide the KRED, coenzyme, and / or cofactor, either naturally or recombinantly. In some embodiments, the engineered ketoreductase and / or coenzyme are added to the reaction mixture in the form of purified enzymes, whole cells transformed with genes encoding the enzymes, and / or cell extracts and / or lysates of such cells. The genes encoding the engineered ketoreductase and / or coenzyme can be transformed into the host cells separately or together into the same host cell. For example, one set of host cells can be transformed with genes encoding the engineered ketoreductase and another set with genes encoding the coenzyme. Both sets of transformed cells can be utilized together in the reaction mixture in the form of whole cells or in the form of lysates or extracts obtained therefrom. In other embodiments, the host cells can be transformed with genes encoding both the engineered ketoreductase and the coenzyme.

[0064] In some embodiments, whole cells transformed with gene(s) encoding an engineered ketoreductase or coenzyme, or cell extracts and / or lysates thereof, are employed in a variety of different forms, including solid (e.g., lyophilized, spray dried, etc.) or semi-solid (e.g., crude paste). For example, cell extracts or cell lysates may be partially purified by precipitation (ammonium sulfate, polyethyleneimine, heat treatment, etc.), followed by a desalting procedure before lyophilization (e.g., ultrafiltration, dialysis, etc.). Any cell preparation may be stabilized by cross-linking, using well-known cross-linking agents such as glutaraldehyde, or immobilization to a solid phase (e.g., Eupergit C, etc.).

[0065] In some embodiments, the enzymes of the present invention are used in various forms, including purified enzymes, crude enzymes, microbial cultures, bacterial cells, and their processed products. Examples of processed products used herein include freeze-dried bacterial cells, acetone-dried bacterial cells, crushed bacterial cells, autolyzed material of bacterial cells, sonicated bacterial cells, bacterial cell extracts, or alkaline-treated bacterial cells. In other embodiments, enzymes of various purities or forms as described above may be used after immobilization by well-known methods, including, for example, adsorption to inorganic carriers such as silica gel, ceramics, cellulose, and ion exchange resins, polyacrylamide method, sulfur-containing polysaccharide gel method (e.g., carrageenan gel method), alginic acid gel method, agar gel method, and the like. To immobilize the enzyme on the carrier, any enzyme immobilization means generally known in the art may be used. For example, the enzyme may be directly bound to a resin membrane or granules having one or more functional groups, or may be bound to the resin via a crosslinking compound having one or more functional groups, such as glutaraldehyde. Such enzyme immobilization reactions are described, for example, in Microbial Enzymes and Biotechnology (Elsevier Applied Science 1990; Ed. WM Fogarty and CT Kelly), 2nd Edition, pp. 369-394.

[0066] "Naturally-occurring" or "wild-type" refers to a form found in nature. For example, a naturally-occurring or wild-type polypeptide or polynucleotide sequence is a sequence present in an organism that can be isolated from a source in nature and that has not been intentionally modified by human manipulation.

[0067] As used herein, "engineered ketoreductase" refers to a ketoreductase having a variant sequence produced by artificial manipulation (e.g., a sequence produced by directed evolution of a naturally occurring parent enzyme, or a sequence produced by directed evolution of a variant previously derived from a naturally occurring enzyme).

[0068] As used herein, "highly stereoselective" refers to a ketoreductase that can convert or reduce a substrate to the corresponding product (e.g., ethyl 2,4-dioxo-4-phenyl-butanoate to (-)-ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate) in at least about 99% stereoisomeric excess. In some embodiments, a KRED is highly stereoselective. In some embodiments, "stereoisomeric excess" as used herein refers to enantiomeric excess. "Enantiomeric excess" or "ee" are used interchangeably herein to refer to the degree to which a sample contains one enantiomer compared to the corresponding non-superimposable mirror compound. A racemic mixture has an ee of 0%, whereas a sample containing only one enantiomer has an ee of 100%.

[0069] Methods for producing RIP1 kinase inhibitors Provided herein are methods for the preparation of compounds useful in the treatment of diseases and disorders associated with inflammation, cell death, neurological disorders, and other diseases. In some embodiments, the prepared compounds include inhibitors of RIP1 kinase useful in the treatment of such diseases and disorders. In some embodiments, the prepared compounds include, for example, compounds exemplified in U.S. Patent Application Publication No. 2019 / 0100530, the contents of which are incorporated herein by reference in their entirety. The methods described herein improve, for example, product purity, diastereomeric ratio (dr), stereoisomeric excess, and / or yield of the final product and key intermediates in its synthesis. The methods described herein will be more fully understood with reference to several reaction schemes below. In some embodiments, the methods unexpectedly provide improved product purity, improved diastereomeric ratio, improved stereoisomeric excess, and / or improved yield. Improved product purity includes, for example, improved chiral purity of the reaction product.

[0070] In some embodiments, the compound of Formula (I) or Formula (II): or a pharma- ceutically acceptable salt thereof, wherein: R 1 is C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 alkyl-N(R N 2) selected from the group consisting of phenyl, benzyl, 4- to 8-membered heterocyclyl, and 5- to 6-membered heteroaryl, where R 1 is attached to the adjacent carbonyl by a carbon atom, and R 1 is F, Cl, Br, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 alkyl-N(R N )2, hydroxyl, hydroxymethyl, cyano, cyanomethyl, cyanoethyl, C(O)C1-C6 alkyl, phenyl, benzyl, CH2-(C3-C6 cycloalkyl), 5- to 6-membered heteroaryl, and CH2-(5- to 6-membered heteroaryl); Each R N are independently selected from the group consisting of H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, and C1-C6 haloalkyl; or two R N may form a 4- to 6-membered ring together with the adjacent N; R 2 is selected from the group consisting of H, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C6 thioalkyl, phenyl, benzyl, CH2-(C3-C6 cycloalkyl), CH2CH2-(C3-C6 cycloalkyl), CH2-(4-6 membered heterocyclyl), CH2CH2-(4-6 membered heterocyclyl), 5-6 membered heteroaryl, and CH2-(5-6 membered heteroaryl); where the phenyl ring, if present, is optionally substituted with 1 to 3 substituents selected from the group consisting of halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, and cyano; R 3 is selected from the group consisting of D, halogen, OH, CN, C1-C4 alkyl, C1-C4 haloalkyl, cyclopropyl, C1-C4 alkoxy and C1-C4 haloalkoxy; n is 1, 2, or 3.

[0071] In another embodiment, provided herein is a process for the preparation of a chiral bicyclic ketone compound of formula (I), or a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 and n is as defined herein, the method comprising: (a) Chiral N-aminolactam formula p or a stereoisomer thereof: TIFF2024513394000024.tif27170 or a salt thereof in the presence of an acid additive and an alcohol solvent, c Imidate compounds: or a salt thereof, x or a stereoisomer thereof: TIFF2024513394000026.tif26170 or a salt thereof, Pg 1 is an optional hydroxyl protecting group, which may be the same or different in each occurrence; formula x or a stereoisomer thereof is an intermediate compound in the preparation of a chiral bicyclic ketone compound of formula (I) or a stereoisomer thereof.

[0072] In another embodiment, the formula p or a stereoisomer thereof: Provided herein is a method for the preparation of TIFF2024513394000027.tif27170, or a salt thereof, wherein R 2and n is as defined herein, the method comprising: Formula (a) d or a stereoisomer thereof: TIFF2024513394000028.tif22170 or a salt thereof, e Organic acid chlorides: or a salt thereof to obtain a compound of the formula f or a stereoisomer thereof: forming TIFF2024513394000030.tif27170 or a salt thereof; Formula (b) l or a stereoisomer thereof: or a salt thereof in the presence of an acid additive to obtain a compound of the formula m or a stereoisomer thereof: TIFF2024513394000032.tif26170 or a salt thereof, Pg 2 is optionally substituted with C1-C6 alkyl, C3-C6 cycloalkyl, or aryl; Pg 3 is an optional hydroxyl protecting group, which may be the same or different in each occurrence; Pg 4 is an optional amine protecting group, which may be the same or different in each occurrence; formula f or a stereoisomer thereof, is represented by the formula l or a stereoisomer thereof, formula m or a stereoisomer thereof, has the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0073] In another embodiment, the formula p or a stereoisomer thereof, or a salt thereof, comprising: Formula (a) d or a stereoisomer thereof: TIFF2024513394000033.tif22170 or a salt thereof, e Organic acid chlorides: or a salt thereof to obtain a compound of the formula f or a stereoisomer thereof: TIFF2024513394000035.tif27170 or a salt thereof, Pg 2 , Pg 3 , and Pg 4 is as defined herein; formula f or a stereoisomer thereof, is represented by the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0074] In another embodiment, the formula p or a stereoisomer thereof, or a salt thereof, comprising: Formula (b) l or a stereoisomer thereof: or a salt thereof in the presence of an acid additive to obtain a compound of the formula m or a stereoisomer thereof: TIFF2024513394000037.tif26170 or a salt thereof, Pg 2 , Pg 3 , and Pg 4 is as defined herein; formula m or a stereoisomer thereof, has the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0075] In some embodiments, the formula b Imidate salt compounds of: Provided herein is a method for the preparation of TIFF2024513394000038.tif19170, the method comprising: Formula (a) a Cyanoformate compounds: TIFF2024513394000039.tif18170 in the presence of an anhydrous acid source in an alcohol solvent to obtain b wherein the anhydrous acid source is TMSCl, the acid is HCl, and Pg 1 is an optional hydroxyl protecting group, which may be the same or different in each occurrence.

[0076] In some embodiments, Provided herein is a method for the preparation of a compound selected from the group consisting of: TIFF2024513394000040.tif76170, or a pharma- ceutically acceptable salt thereof, wherein R 1 , R 3 and n is as defined herein; Each R 4 is selected from the group consisting of H, F, Cl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, and C1-C6 haloalkoxy; m is 0, 1, 2, or 3.

[0077] In some embodiments,

[0023] Provided herein is a method for the preparation of a compound having the formula: 1 , R 4, m, and n are as defined herein.

[0078] In some of the embodiments described herein, R 1 is selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 haloalkyl, phenyl, benzyl, oxtetanyl, oxabicyclo[3.1.0]hexan-6-yl, thienyl, and pyrazolyl; 1 is optionally substituted by (i) one substituent selected from the group consisting of F, Cl, methyl, hydroxyl, hydroxymethyl, cyano, and trifluoromethyl, or (ii) two F substituents. In some embodiments, R 1 is optionally substituted with one or two substituents selected from the group consisting of F, Cl, methyl, ethyl, hydroxyl, hydroxymethyl, methoxymethyl, cyano, trifluoromethyl, difluoromethoxy, and trifluoromethoxy. In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 1 is C1-C4 alkyl. In some embodiments, R 1 is C-C cycloalkyl. In some embodiments, R 1 is C-C cycloalkyl. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is CF3CH2. In some embodiments, R 1 is 2-propyl. In some embodiments, R 1 is tert-butyl. In some embodiments, R 1 is (2-hydroxy)-2-propyl. In some embodiments, R 1 is (2-cyano)-2-propyl. In some embodiments, R 1 is C1-C6 haloalkyl. In some embodiments, R 1 is C1-C4 haloalkyl. In some embodiments, R1 is preferably cyclopropyl. In some embodiments, R 1 is monofluorocyclopropyl or difluorocyclopropyl. In some embodiments, R 1 is 1-fluorocyclopropyl. In some embodiments, R 1 is 2-fluorocyclopropyl. In some embodiments, R 1 In some embodiments, R 1 is 1-(trifluoromethyl)cyclopropyl. In some embodiments, R 1 is 1-methylcyclopropyl. In some embodiments, R 1 is 1-(hydroxymethyl)cyclopropyl. In some embodiments, R 1 is cyclobutyl. In some embodiments, R 1 is cyclopentyl. In some embodiments, R 1 is phenyl. In some embodiments, R 1 is benzyl. In some embodiments, R 1 is oxetan-3-yl. In some embodiments, R 1 is 3-methyloxetan-3-yl. In some embodiments, R 1 is oxabicyclo[3.1.0]hexan-6-yl. In some embodiments, R 1 is 2-pyridyl. In some embodiments, R 1 In some embodiments, R 1 is 2-thienyl.

[0079] In some of the embodiments described herein, each R N is independently selected from the group consisting of H and C1-C6 alkyl. In some embodiments, each R N is C1-C4 alkyl. In some embodiments, each R N is methyl.

[0080] In some of the embodiments described herein, R 2 is preferably phenyl. In some embodiments, R 2 is monofluorophenyl or difluorophenyl. In some embodiments, R 2 is monochlorophenyl or dichlorophenyl. In some embodiments, R 2 is pyridinyl. In some embodiments, R 2 is a chloro-substituted pyridinyl. In some embodiments, R 2 is fluoro-substituted pyridinyl. In some embodiments, R 2 In some embodiments, R 2 is 1-methyl-1H-pyrazol-4-yl. In some embodiments, R 2 is 4-chloro-1-methyl-1H-pyrazol-3-yl.

[0081] In some of the embodiments described herein, R 3 is H. In some embodiments, R 3 is preferably F. In some embodiments, R 3 is Cl. In some embodiments, R 3a and R 3b are each methyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is OH. In some embodiments, R 3 is CN. In some embodiments, R 3 is D.

[0082] In some of the embodiments described herein, R 4 is selected from the group consisting of H, F, Cl, CH3, CH2CH3, OCH3, CF3, OCF3, CF2H, and OCF2H. 4 is preferably F.

[0083] In some of the embodiments described herein, m is preferably 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3.

[0084] In some of the embodiments described herein, n is preferably 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0085] Protecting groups are depicted generically in some of the reaction schemes described herein, and one of skill in the art will recognize that in many cases a variety of different protection and deprotection schemes can be used alternatively, as described in "Greene's Protective Groups in Organic Synthesis", 5th Edition, 2014, John Wiley & Sons, Inc. In some embodiments, an amine or hydroxyl substituent is represented by the variable R 1 From R 4 and R N It should be understood that may be present and appropriate protecting groups may be utilized in association with such substituents.

[0086] In some embodiments, a process (P1) for the preparation of a chiral bicyclic ketone compound of formula (I), or a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof, is provided, 1 , R 2 , R 3 and n is as defined herein] includes: (a) Chemical formula of chiral N-aminolactam p or a stereoisomer thereof: or a salt thereof in the presence of an acid additive and an alcohol solvent, c Imidate compounds: or a salt thereof, x or a stereoisomer thereof: TIFF2024513394000044.tif26170 or a salt thereof, Pg 1 is an optional hydroxyl protecting group, which may be the same or different in each occurrence; formula x or a stereoisomer thereof is an intermediate compound in the preparation of a chiral bicyclic ketone compound of formula (I) or a stereoisomer thereof.

[0087] In a preferred embodiment, the chiral N-aminolactam formula p is (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one. In a preferred embodiment, c is ethyl-2-ethoxy-2-iminoacetate. In a preferred embodiment, the imidate compound of the formula x The chiral bicyclic triazole compound is ethyl (5S,7R)-7-hydroxy-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylate.

[0088] In some embodiments, the acid additive is a carboxylic acid, a sulfonic acid, or an inorganic acid. In some embodiments, the acid additive is acetic acid, oxalic acid, succinic acid, benzoic acid, isobutyric acid, pivalic acid, salicylic acid, oxamic acid, 2-picolinic acid, trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid, formic acid, hydrochloric acid, or trimethylsilyl chloride. In a preferred embodiment, the acid additive is acetic acid. In some embodiments, the acid additive is isobutyric acid. In some embodiments, the acid additive is salicylic acid.

[0089] In some embodiments, the formula of step (a) of method (P1) cThe imidate compound of is replaced with another reagent. In some embodiments, the replacement reagent is ethyl thiooxamate, ethyl cyanoformate, methyl cyanoformate, or triethyl 1,3,5-triazine-2,4,6-tricarboxylate. In some embodiments, the replacement reagent is ethyl thiooxamate. In some embodiments, the replacement reagent is triethyl 1,3,5-triazine-2,4,6-tricarboxylate. In some embodiments, the replacement reagent is thiooxamate and the acid additive is isobutyric acid. In some embodiments, the replacement reagent is triethyl 1,3,5-triazine-2,4,6-tricarboxylate and the acid additive is salicylic acid.

[0090] In a preferred embodiment, the formula in step (a) of method (P1) x The yield of the chiral bicyclic triazole compound is at least 80%. In particularly preferred embodiments, the yield is at least 85%. In some embodiments, the yield is at least 90%. In some embodiments, the yield is at least 95%. In some embodiments, the yield is at least 98%.

[0091] In a preferred embodiment, the alcohol solvent in step (a) of method (P1) is EtOH. In a particularly preferred embodiment, the acid additive and alcohol solvent are a mixture of EtOH and acetic acid. In some embodiments, step (a) of method (P1) further comprises maintaining the temperature at about 60° C. before cooling the temperature to about 25±10° C. In some embodiments, step (a) of method (P1) further comprises adding water. In some embodiments, step (a) of method (P1) comprises reacting a compound of the formula x The method further comprises adding a seed of a chiral bicyclic triazole compound of formula (I).

[0092] In some embodiments, the method (P1) comprises: Formula (b) x or a stereoisomer thereof, in the presence of a halogenating agent, to obtain a chiral bicyclic triazole compound of the formula yor a stereoisomer thereof: TIFF2024513394000045.tif26170, or a salt thereof, wherein X is a halogen.

[0093] In a preferred embodiment, the formula y The chiral halogenated bicyclic compound of is cyclopropyl-[(5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl]methanone. In some embodiments, the halogenating agent is a fluorinating agent. Examples of fluorinating agents are described by MK Nielsen et al. in J. Am. Chem. Soc. 140(15):5004-5008 (2018). In some embodiments, the halogenating agent is a sulfonyl chloride. In a preferred embodiment, the halogenating agent is PBSF. In some embodiments, the halogenating agent is PyFluor (2-pyridinesulfonyl fluoride). In some embodiments, the halogenating agent is diethylaminosulfur trifluoride (DAST). In some embodiments, the halogenating agent is bis(2-methoxyethyl)aminosulfur trifluoride (Deoxo-Fluor or BAST).

[0094] In some embodiments, step (b) of method (P1) is carried out in the presence of an organic base and an organic solvent. In a preferred embodiment, the organic base is N,N-diisopropylethylamine and the organic solvent is acetonitrile. In some embodiments, an additive is present. In a preferred embodiment, the additive is triethylamine trihydrofluoride. In some embodiments, the additive is N,N-diisopropylethylamine trihydrofluoride. In some embodiments, the additive acts as a fluoride source.

[0095] In some embodiments, step (b) of method (P1) further comprises slowly adding the reagent over at least 1 hour at RT to reduce evaporation.y is at least 80%. In particularly preferred embodiments, the yield is at least 85%. In some embodiments, the yield is at least 90%. In some embodiments, the yield is at least 95%. In some embodiments, the yield is at least 98%.

[0096] In some embodiments, the method (P1) comprises: (c) formula y or a stereoisomer thereof, in the presence of an ethereal solvent / water mixture with an acid to produce a chiral halogenated bicyclic compound of the formula z or a stereoisomer thereof: TIFF2024513394000046.tif26170, or a salt thereof.

[0097] In a preferred embodiment, the formula z is (5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylic acid. In a preferred embodiment, the ethereal solvent / water mixture is a THF / water mixture and the acid is HCl. In some embodiments, step (c) of method (P1) further comprises maintaining the temperature at about 50° C. before cooling the temperature to about 35±10° C. In some embodiments, step (c) of method (P1) further comprises cooling the temperature to about 20° C.±10° C. and adding water followed by a solution of KOH. In some embodiments, step (c) of method (P1) further comprises maintaining the temperature at about 30° C. after adding the water and the solution of KOH.

[0098] In some embodiments, the method (P1) comprises: Formula (d) z or a stereoisomer thereof, aa Compounds of: or a salt thereof in the presence of a coupling agent to obtain a compound of the formula bb or a stereoisomer thereof: or a salt thereof, wherein each Pg 5 is an amine protecting group and may be the same or different in each occurrence.

[0099] In a preferred embodiment, the formula aa is N,O-dimethylhydroxylamine hydrochloride. In a preferred embodiment, the coupling agent is EDCI. In a preferred embodiment, the compound of formula bb is (5S,7S)-7-fluoro-N-methoxy-N-methyl-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide. In some embodiments, step (d) of method (P1) further comprises maintaining the temperature at about 65° C. In some embodiments, step (d) of method (P1) is carried out in the presence of an additive. In some embodiments, the additive is NMI.

[0100] In some embodiments, step (d) of method (P1) comprises reacting a compound of formula bb In a preferred embodiment, the method further comprises adding a seed of a chiral bicyclic amide compound of the formula bb The addition of the chiral bicyclic amide compound seed of is carried out in the presence of CPME. In some embodiments, step (d) of method (P1) further comprises adding an anti-solvent before cooling the temperature to about 0° C. In a preferred embodiment, the anti-solvent is heptane.

[0101] In some embodiments, the method (P1) comprises: Formula (e) bb or a stereoisomer thereof, cc Compounds of: or a salt thereof, dd or a stereoisomer thereof: TIFF2024513394000050.tif28170, or a salt thereof.

[0102] In a preferred embodiment, the formula cc is cyclopropylmagnesium bromide. In a preferred embodiment, the compound of formula dd is cyclopropyl-[(5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl]methanone. In some embodiments, the alkylation may be carried out in an organic solvent. In some embodiments, the organic solvent is THF during the alkylation step. In some embodiments, step (e) of method (P1) further comprises maintaining a temperature at about -5°C ± 10°C. In some embodiments, step (e) of method (P1) further comprises reacting a compound of formula dd In some embodiments, the method further comprises adding a seed of a chiral bicyclic ketone compound of the formula dd The addition of the chiral bicyclic ketone compound seed is carried out in the presence of an organic solvent. In a preferred embodiment, the organic solvent is EtOH. In a particularly preferred embodiment, the organic solvent is an aqueous solution of EtOH.

[0103] In some embodiments, the chiral bicyclic ketone compound of formula (I) is TIFF2024513394000051.tif76170, or a pharma- ceutically acceptable salt thereof; During the ceremony, Each R 4 is selected from the group consisting of H, F, Cl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, and C1-C6 haloalkoxy; m is 0, 1, 2, or 3.

[0104] In some embodiments, the chiral bicyclic ketone compound of formula (I) is TIFF2024513394000052.tif31170 or a pharma- ceutically acceptable salt thereof.

[0105] In a particularly preferred embodiment, R 1 is cyclopropyl. In particularly preferred embodiments, m is 0. In particularly preferred embodiments, n is 1.

[0106] In some embodiments, the chiral bicyclic ketone compound of formula (I) is a compound of formula (II): TIFF2024513394000053.tif28170 or a pharma- ceutically acceptable salt thereof.

[0107] In some embodiments, the compound of formula (II) is TIFF2024513394000054.tif75170, or a pharma- ceutically acceptable salt thereof; During the ceremony, Each R 4 is selected from the group consisting of H, F, Cl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, and C1-C6 haloalkoxy; m is 0, 1, 2, or 3.

[0108] In some embodiments, the compound of formula (II) is TIFF2024513394000055.tif31170 or a pharma- ceutically acceptable salt thereof.

[0109] In a preferred embodiment, R 1 is cyclopropyl. In a preferred embodiment, m is 0. In a preferred embodiment, n is 1.

[0110] In some embodiments, the compound of Formula (II), Formula (III), or Formula (IV): Provided herein is a method for the preparation of a chiral bicyclic ketone compound of formula TIFF2024513394000056.tif28170, or a pharma- ceutically acceptable salt thereof, wherein R 1 , R2 , R 3 and n are as defined herein. In a preferred embodiment, R 1 is cyclopropyl. In a preferred embodiment, R 2 is phenyl. In a preferred embodiment, R 3 is F. In a preferred embodiment, n is 1. The compounds of formula (III) and formula (IV) are, for example, diastereomers of the compound of formula (I) and diastereomers of the compound of formula (II), respectively. Those skilled in the art will readily recognize various changes and modifications to these methods based on the embodiments described herein.

[0111] In another embodiment, the formula p or a stereoisomer thereof: TIFF2024513394000057.tif27170 or a salt thereof (P2) 2 and n is as defined herein] includes: Formula (a) d or a stereoisomer thereof: TIFF2024513394000058.tif22170 or a salt thereof, e Organic acid chlorides: or a salt thereof to obtain a compound of the formula f or a stereoisomer thereof: forming TIFF2024513394000060.tif27170 or a salt thereof; Formula (b) l or a stereoisomer thereof: or a salt thereof in the presence of an acid additive to obtain a compound of the formula m or a stereoisomer thereof: TIFF2024513394000062.tif26170 or a salt thereof, Pg 2 is optionally substituted with C1-C6 alkyl, C3-C6 cycloalkyl, or aryl; Pg 3 is an optional hydroxyl protecting group, which may be the same or different in each occurrence; Pg 4 is an optional amine protecting group, which may be the same or different in each occurrence; formula f or a stereoisomer thereof, is represented by the formula l or a stereoisomer thereof, formula m or a stereoisomer thereof, has the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0112] In some embodiments, the formula p or a stereoisomer thereof, or a salt thereof, includes the following: Formula (a) d or a stereoisomer thereof: TIFF2024513394000063.tif22170, or a salt thereof, with an organic acid chloride of formula e: or a salt thereof to obtain a compound of the formula f or a stereoisomer thereof: TIFF2024513394000065.tif27170 or a salt thereof, Pg 2 , Pg 3 , and Pg 4 is as defined herein; formula f or a stereoisomer thereof, is represented by the formula por an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0113] In some embodiments, the formula p or a stereoisomer thereof, or a salt thereof, includes the following: Formula (b) l or a stereoisomer thereof: or a salt thereof in the presence of an acid additive to obtain a compound of the formula m or a stereoisomer thereof: TIFF2024513394000067.tif26170 or a salt thereof, Pg 2 , Pg 3 , and Pg 4 is as defined herein; formula m or a stereoisomer thereof, has the formula p or an intermediate in the preparation of a chiral N-aminolactam compound of the formula:

[0114] In a preferred embodiment, the formula d The chiral hydroxydicarboxylic acid compound of formula (I) is D-malic acid. e In a preferred embodiment, the acid chloride solvent of formula f The chiral cyclic carboxylic acid anhydride compound is (S)-(-)-2-acetoxy-succinic anhydride. In some embodiments, step (a) of method (P2) further comprises adding i-PrOAc. In some embodiments, step (a) of method (P2) further comprises adding n-heptane.

[0115] In a preferred embodiment, the formula lThe protected hydrazone compound of formula (I) is tert-butyl (R,E)-2-(4-ethoxy-3-hydroxy-4-oxo-1-phenylbutylidene)hydrazine-1-carboxylate. In a preferred embodiment, the protected hydrazone compound of formula (I) is tert-butyl (R,E)-2-(4-ethoxy-3-hydroxy-4-oxo-1-phenylbutylidene)hydrazine-1-carboxylate. m is tert-butyl 2-((1S,3R)-4-ethoxy-3-hydroxy-4-oxo-1-phenylbutyl)hydrazine-1-carboxylate. In some embodiments, the acid additive is a carboxylic acid. In some embodiments, the acid additive is oxalic acid, succinic acid, benzoic acid, isobutyric acid, pivalic acid, salicylic acid, oxamic acid, 2-picolinic acid, trifluoroacetic acid, formic acid, hydrochloric acid, or acetic acid. In a preferred embodiment, the acid additive is acetic acid. In some embodiments, the acid additive is p-toluenesulfonic acid. In some embodiments, the acid additive is methanesulfonic acid. In some embodiments, the acid additive is hydrochloric acid in ethanol. In some embodiments, the acid additive is trimethylsilyl chloride in ethanol. In some embodiments, step (b) of method (P2) is carried out in the presence of tetramethylammonium triacetoxyborohydride or sodium triacetoxyborohydride.

[0116] In a preferred embodiment, the formula in step (a) of method (P2) f The yield of the chiral cyclic carboxylic acid anhydride compound is at least 80%. In some embodiments, the yield is at least 85%. In some embodiments, the yield is at least 90%. In some embodiments, the yield is at least 92%. In particularly preferred embodiments, the yield is at least 93%.

[0117] Step (b) of method (P2) m The yield of the chiral hydroxyester hydrazine compound is at least 70%. In some embodiments, the yield is at least 80%. In particularly preferred embodiments, the yield is at least 85%. In some embodiments, the yield is at least 90%.

[0118] In a preferred embodiment, the formula in step (b) of method (P2) m The diastereomeric ratio (dr) of the chiral hydroxyester hydrazine compound to its diastereomer is at least 10:1. In some embodiments, dr is at least 11:1. In some embodiments, dr is at least 12:1. In some embodiments, dr is at least 13:1. In particularly preferred embodiments, dr is at least 14:1.

[0119] In some embodiments, the method (P2) comprises: (c) formula f or a stereoisomer thereof, with a reactive arene compound of the formula h or a stereoisomer thereof: TIFF2024513394000068.tif26170, or a salt thereof.

[0120] In some embodiments, the reactive arene compound is benzene. In some embodiments, the reactive arene compound is represented by the formula h is (R)-2-acetoxy-4-oxo-4-phenylbutanoic acid. In some embodiments, the contacting step (c) is carried out in the presence of a Lewis acid in an organic solvent. In a preferred embodiment, the Lewis acid is AlCl3 and the organic solvent is CH2Cl2. In some embodiments, the organic solvent comprises n-heptane.

[0121] In some embodiments, the method (P2) comprises: Formula (d) h or a stereoisomer thereof, i : The reaction was carried out in an alcohol solvent of the formula TIFF2024513394000069.tif8170. j or a stereoisomer thereof: TIFF2024513394000070.tif21170 or a salt thereof.

[0122] In a preferred embodiment, the formulai The alcohol solvent of formula (d) is EtOH. In some embodiments, reacting step (d) is carried out in the presence of an acid. In some embodiments, the acid is H2SO4. In a preferred embodiment, j The compound is (-)-ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate.

[0123] In some embodiments, the method (P2) comprises: Formula (e) j or a stereoisomer thereof, k Hydrazine compounds: or a salt thereof, l or a stereoisomer or a salt thereof.

[0124] In some embodiments, the contacting step (e) is carried out in the presence of an acid additive. In some embodiments, the acid additive is formic acid. In preferred embodiments, the compound of the formula k The hydrazine compound is NH2NHBoc.

[0125] In some embodiments, the method (P2) comprises: (f) formula l or a stereoisomer thereof to give a compound of the formula n or a stereoisomer thereof. or a salt thereof; Formula (g) n or a stereoisomer thereof to give a chiral protected N-aminolactam compound of the formula o Salt compounds of: forming TIFF2024513394000073.tif26170 or a stereoisomer thereof; (h) formula o or a stereoisomer thereof in the presence of a base to obtain a compound of the formula por a stereoisomer thereof. Further includes:

[0126] In a preferred embodiment, the formula n The chiral protected N-aminolactam compound of formula (I) is tert-butyl ((3R,5S)-3-hydroxy-2-oxo-5-phenylpyrrolidin-1-yl)carbamate. In a preferred embodiment, the compound of formula (I) is o is (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one hydrochloride. In a preferred embodiment, the salt compound of formula p is (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one. In some embodiments, step (g) of method (P2) further comprises adding an acid in an organic solvent. In some embodiments, the acid is HCl and the organic solvent is n-propanol. In some embodiments, step (g) of method (P2) further comprises maintaining the temperature at about 20-25° C. In some embodiments, step (h) of method (P2) further comprises adding an aqueous solution of a base. In some embodiments, the base is sodium hydroxide.

[0127] In a preferred embodiment, the base is NaOH in aqueous base solution. In some embodiments, the optional amine protecting group Pg 4 is Boc. In some embodiments, n is 1.

[0128] In some embodiments, the formula b Imidate salt compounds of: The method for preparation of TIFF2024513394000074.tif19170 (P3) includes: Formula (a) a Cyanoformate compounds: TIFF2024513394000075.tif18170 in the presence of an anhydrous acid source in an alcohol solvent to obtain bwherein the anhydrous acid source is TMSCl, the acid is HCl, and Pg 1 is an optional hydroxyl protecting group, which may be the same or different in each occurrence.

[0129] In some embodiments, the formula of step (a) of method (P3) a is replaced with another reagent. In some embodiments, the replacement reagent is ethyl thiooxamate, ethyl cyanoformate, methyl cyanoformate, or triethyl 1,3,5-triazine-2,4,6-tricarboxylate. In some embodiments, the replacement reagent is ethyl thiooxamate. In some embodiments, the replacement reagent is triethyl 1,3,5-triazine-2,4,6-tricarboxylate. In some embodiments, the replacement reagent is ethyl thiooxamate. In some embodiments, the replacement reagent is triethyl 1,3,5-triazine-2,4,6-tricarboxylate.

[0130] In a preferred embodiment, the alcohol solvent is EtOH in MTBE.

[0131] In a preferred embodiment, the formula in step (a) of method (P3) b The yield of the imidate salt compound is at least 65%. In some embodiments, the yield is at least 70%. In some embodiments, the yield is at least 75%. In particularly preferred embodiments, the yield is at least 78%.

[0132] In some embodiments, the formula j or a stereoisomer thereof: The method for preparation of TIFF2024513394000076.tif22170 (P4) includes: Formula (a) hh Diketo ester compounds of: TIFF2024513394000077.tif20170 was reacted in the presence of ketoreductase (KRED) to obtain the compound of formula j or a stereoisomer thereof, 2 , Pg 3 , and n is as defined herein].

[0133] In some embodiments, the method (P4) comprises: Formula (b) j or a stereoisomer thereof to obtain a hydroxyketoester compound of the formula l or a stereoisomer thereof: TIFF2024513394000078.tif28170, or a salt thereof.

[0134] In a preferred embodiment, the KRED is an engineered ketoreductase. In a more preferred embodiment, the engineered ketoreductase is ADH-114 (c-LEcta GmbH, Germany) or 1-200-0-16 (Porton Pharma Solutions Ltd, China).

[0135] In a preferred embodiment, the formula j The hydroxyketoester compound of formula (I) is (-)-ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate. In a preferred embodiment, the hydroxyketoester compound of formula (I) is (-)-ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate. hh The diketoester compound of formula (I) is ethyl 2,4-dioxo-4-phenyl-butanoate. In a preferred embodiment, the diketoester compound of formula (I) is ethyl 2,4-dioxo-4-phenyl-butanoate. l The protected hydrazone compound is tert-butyl (R,E)-2-(4-ethoxy-3-hydroxy-4-oxo-1-phenylbutylidene)hydrazine-1-carboxylate.

[0136] In preferred embodiments, the KRED is highly stereoselective. In some embodiments, the stereoisomeric excess of the hydroxyketoester compound of formula j in step (a) of method (P4) is at least 80%. In some embodiments, the stereoisomeric excess is at least 85%. In preferred embodiments, the stereoisomeric excess is at least 90%. In more preferred embodiments, the stereoisomeric excess is at least 95%. In particularly preferred embodiments, the stereoisomeric excess is at least 99%.

[0137] In a preferred embodiment, in step (a) of the method (P4), j The yield of hydroxyketoester compound is at least 80%. In some embodiments, the yield is at least 85%. In some embodiments, the yield is at least 90%. In some embodiments, the yield is at least 92%. In preferred embodiments, the yield is at least 93%. In particularly preferred embodiments, the yield is at least 95%.

[0138] In a preferred embodiment, the formula in step (b) of method (P4) l The yield of the protected hydrazone compound is at least 80%. In some embodiments, the yield is at least 85%. In some embodiments, the yield is at least 90%. In some embodiments, the yield is at least 92%. In preferred embodiments, the yield is at least 93%. In particularly preferred embodiments, the yield is at least 95%.

[0139] In some embodiments, the method (P4) further comprises the presence of an alcohol. For example, the alcohol is used to regenerate the cofactor. In a preferred embodiment, the method (P4) comprises the presence of an alcohol without the presence of a cofactor. In some embodiments, the alcohol is a secondary alcohol. For example, secondary alcohols include lower secondary alkanols and aryl-alkyl carbinols. Examples of lower secondary alcohols include isopropanol, 2-butanol, 3-methyl-2-butanol, 2-pentanol, 3-pentanol, and the like. In a preferred embodiment, the secondary alcohol is isopropanol. Examples of aryl-alkyl carbinols include unsubstituted and substituted 1-arylethanol. In some embodiments, the secondary alcohol is the R-enantiomer of a chiral secondary alcohol. In other embodiments, the secondary alcohol is the S-enantiomer of a chiral secondary alcohol. In some embodiments, the method (P4) further comprises the presence of a cofactor. In a preferred embodiment, the cofactor is glucose dehydrogenase. In a more preferred embodiment, the glucose dehydrogenase is GDH-105 (Codexis, Inc., California, USA) or 1-030-0-05 (Porton Pharma Solutions Ltd, China). In some embodiments, the KRED is provided in immobilized or whole cell form.

[0140] In some embodiments, any step of the method (P1-P4) is scalable. In preferred embodiments, at least one step of the method (P1-P4) is scalable to at least kilogram scale. TIFF2024513394000079.tif20170

[0141] Scheme 1 Referring now to Scheme 1, the intermediate imidate compound c The synthesis of Pg is shown. 1 is as defined herein. In step 1 of scheme 1, a cyanoformate compound aundergoes imidate salt formation, and the imidate salt compound b The salt formation in step 1 can be carried out using an anhydrous acid source, e.g., TMSCl for HCl, in the presence of an alcohol, e.g., EtOH, in an ethereal solvent, e.g., MTBE. In some embodiments, the dry source used in this step is TMSCl and the acid is HCl.

[0142] In step 2, the imidate salt is converted to the free base compound using an organic base. c In some embodiments, the organic base in this step is triethylamine. In some embodiments, the conversion can be carried out in the presence of a drying agent in an organic solvent. In some embodiments, the drying agent in this step is Na2SO4. In some embodiments, the organic solvent in this step is MTBE. Compound e can then be used as shown in Schemes 3A or 3B below. TIFF2024513394000080.tif133170

[0143] Scheme 2A Scheme 2A shows a chiral N-aminolactam compound p Here, Pg 2 , Pg 3 , Pg 4 , R 2 and n is as defined herein.

[0144] In step 1 of Scheme 2A, a chiral hydroxydicarboxylic acid compound is d undergoes dehydration, cyclodehydration and esterification to give organic acid chlorides e Among them, chiral carboxylic acid dihydrofurandione compounds f is obtained.

[0145] In step 2 of Scheme 2A, the compound f is a reactive arene compound g The compound undergoes a Friedel-Crafts reaction in the presence of hIn some embodiments, the Friedel-Crafts reaction is carried out in the presence of a Lewis acid in an organic solvent. g is benzene. In some embodiments, the Lewis acid in this step is AlCl. In some embodiments, the organic solvent in this step is CH2Cl2.

[0146] In step 3 of Scheme 2A, the compound h undergoes transesterification to form an alcohol solvent. i Among the compounds j In some embodiments, the transesterification can be carried out in the presence of an acid. In some embodiments, the transesterification can be carried out in the presence of an alcohol solvent. i is EtOH in this step. In some embodiments, the acid is H2SO4 in this step.

[0147] In step 4 of Scheme 2A, the compound j undergoes hydrazone formation to give the protected hydrazone compound l In some embodiments, the hydrazone formation can be carried out in the presence of an acid additive in this step. In some embodiments, the acid additive in this step is formic acid. In some embodiments, the protecting group Pg 4 is Boc in this step.

[0148] In step 5 of Scheme 2A, a hydrazone compound is l can be subjected to diastereoselective reduction using a reducing agent to give the chiral hydrazine compound m In some embodiments, the reduction can be carried out in the presence of an acid additive in an organic solvent. In some embodiments, the reducing agent in this step is Me4NBH(OAc)3 or NaBH(OAc)3. In some embodiments, the acid additive in this step is AcOH. In some embodiments, the organic solvent in this step is CH2Cl2.

[0149] In step 6 of Scheme 2A, a chiral hydrazine compound is mis cyclized to give the chiral protected N-aminolactam compound n In some embodiments, the cyclization may be carried out in an alcohol solvent upon heating. In some embodiments, the alcohol solvent is EtOH for this step.

[0150] In step 7 of Scheme 2A, the chiral protected N-aminolactam compound n is deprotected to give the target compound p Salt o In some embodiments, the salt o is the HCl salt in this process.

[0151] In step 8 of Scheme 2A, a salt o is converted to a free base to produce an N-aminolactam compound in the presence of a base. p In some embodiments, the base in this step is NaOH. In some embodiments, the reaction of step 8 can be carried out in an aqueous base solution. TIFF2024513394000081.tif29170

[0152] Scheme 2A' Scheme 2A' shows a hydroxyketoester compound j Here, Pg 3 , R 2 and n is as defined herein; Pg 3’ is an optional hydroxyl protecting group, which may be the same or different at each occurrence. In some embodiments, Pg 3 Pg 3 It is.

[0153] In step 1 of Scheme 2A', an oxalic acid diester compound ff is condensed to give an aryl methyl ketone compound gg In the presence of a diketo ester compound hh In some embodiments, the aryl methyl ketone compound gg is acetophenone in this process.

[0154] In step 2 of Scheme 2A', a diketoester compound hh undergoes enzymatic ketone reduction to give the hydroxyketoester compound j In some embodiments, the enzymatic reduction is carried out in the presence of a ketoreductase (KRED). In some embodiments, the KRED is highly stereoselective. In some embodiments, the KRED is an engineered ketoreductase. In more preferred embodiments, the engineered ketoreductase is ADH-114 (c-LEcta GmbH, Germany) or 1-200-0-16 (Porton Pharma Solutions Ltd, China).

[0155] In some embodiments of step 2 of Scheme 2A', the enzymatic reduction is further carried out in the presence of a cofactor. In some embodiments, the cofactor is NAD, NADH, NADP, or NADPH. In some embodiments of step 2 of Scheme 2A', the enzymatic reduction is further carried out in the presence of a coenzyme. In some embodiments, the coenzyme is glucose dehydrogenase. In some embodiments, the glucose dehydrogenase is GDH-105 (Codexis, Inc., California, USA) or 1-030-0-05 (Porton Pharma Solutions Ltd, China). In some embodiments of step 2 of Scheme 2A', the enzymatic reduction is further carried out in the presence of an alcohol. In some embodiments, the alcohol is ethanol. In some embodiments of step 2 of Scheme 2A', the enzymatic reduction is carried out in the presence of an alcohol in the absence of a coenzyme.

[0156] In some embodiments, step 2 of Scheme 2A' further comprises the presence of D-(+)-glucose. In some embodiments, step 2 of Scheme 2A' further comprises the presence of an additive and / or an organic co-solvent. In some embodiments, the organic co-solvent in this step is ethanol or acetonitrile. In some embodiments, the additive in this step is NaOAc, MgCl2·6H2O, Na2CO3, or NaOH. In some embodiments, step 2 of Scheme 2A' is carried out in a buffer solution at a controlled pH and temperature. In some embodiments, the buffer solution comprises K2HPO4, KH2PO4, and water. In some embodiments, the controlled pH is in the range of 6-9. In preferred embodiments, the controlled pH is 6.5 or 7. In some embodiments, the temperature is in the range of 20-40°C. In preferred embodiments, the temperature is 25°C or 30°C. TIFF2024513394000082.tif135170

[0157] Scheme 2B Scheme 2B shows chiral N-aminolactam compounds w Here, Pg 2 , Pg 3 , Pg 4 , R 2 and n are as defined herein. Steps 1 to 7 of Scheme 2B are the same as steps 1 to 7 of Scheme 2A, and step 1 is the reaction of a chiral hydroxydicarboxylic acid compound o Starting from the chiral compound p , q , r , s , t , u , v , and chiral N-aminolactam compounds w is obtained. TIFF2024513394000083.tif85170

[0158] Scheme 3A Scheme 3A shows a chiral bicyclic ketone compound. ddHere, Pg 1 , Pg 5 , R 1 , R 2 , X and n are as defined herein. In some embodiments, the chiral bicyclic ketone compound dd is a chiral 6,7-dihydro-5H-pyrrolo[1,2-b][1,2,5]triazole ketone.

[0159] In step 1 of Scheme 3A, the compound p and c are combined to form a triazole, resulting in a chiral bicyclic triazole compound x In some embodiments, the triazole formation may be carried out in the presence of an acid additive and an alcohol solvent. In some embodiments, the acid additive in this step is acetic acid. In some embodiments, the alcohol solvent in this step is ethanol.

[0160] In step 2 of Scheme 3A, a chiral bicyclic triazole compound is x undergoes deoxyhalogenation in the presence of a halogenating agent to give chiral halogenated bicyclic compounds yIn some embodiments, the deoxyhalogenation can be carried out in the presence of an organic base in an organic solvent. In some embodiments, the deoxyhalogenation comprises deoxyfluorination in the presence of a fluorinating agent. In a preferred embodiment, the fluorinating agent in this step is PBSF. In some embodiments, the fluorinating agent in this step is PyFluor (2-pyridinesulfonyl fluoride). In a preferred embodiment, the organic base in this step is N,N-diisopropylethylamine. In some embodiments, the organic solvent is acetonitrile. In a preferred embodiment, an additive is present. In a particularly preferred embodiment, the additive is triethylamine trihydrofluoride. In some embodiments, the additive is N,N-diisopropylethylamine trihydrofluoride. In some embodiments, the additive acts as a fluoride source. In some embodiments, the deoxyhalogenation is carried out by slow addition of the reagent at room temperature over a period of at least 1 hour to reduce evaporation.

[0161] In step 3 of Scheme 3A, a chiral halogenated bicyclic compound is y undergoes ester hydrolysis and acidification to give halogenated bicyclic carboxylic acid compounds z In some embodiments, the ester hydrolysis and acidification can be carried out in the presence of an ethereal solvent / water mixture containing an acid. In some embodiments, the solvent / water mixture in this step is a THF / water mixture. In some embodiments, the acid in this step is HCl.

[0162] In step 4 of Scheme 3A, a halogenated bicyclic carboxylic acid compound is z is an amide aa Following Weinreb amide formation with bb In some embodiments, the Weinreb amide formation can be carried out in the presence of a coupling agent. In some embodiments, the amide formation can be carried out in the presence of an additive in an organic solvent. In some embodiments, the amide aais N,O-dimethylhydroxylamine in this step. In some embodiments, the coupling reagent is EDCI in this step. In some embodiments, the additive is N-methylimidazole and the organic solvent is CH2Cl2 in this step.

[0163] In step 5 of Scheme 3A, a chiral bicyclic amide is bb is an organometallic reagent cc to form a chiral target compound by alkylation in the presence of dd In some embodiments, the alkylation can be carried out in an organic solvent. In some embodiments, the organometallic reagent cc is an alkyl magnesium bromide in this step. In some embodiments, the alkyl magnesium bromide is a cyclopropyl magnesium bromide in this step. In some embodiments, the organic solvent is THF in this step. In some embodiments, this step comprises reacting a compound of the formula dd In some embodiments, the method further comprises adding a seed of a chiral bicyclic ketone compound of the formula dd The addition of the chiral bicyclic ketone compound seed is carried out in the presence of an organic solvent. In a preferred embodiment, the organic solvent is EtOH. In a particularly preferred embodiment, the organic solvent is an aqueous solution of EtOH. TIFF2024513394000084.tif86170

[0164] Scheme 3B Scheme 3B shows a chiral bicyclic ketone compound ii Here, Pg 1 , Pg 5 , R 1 , R 2 , X and n are as defined herein. Steps 1 to 5 of Scheme 3B are the same as steps 1 to 5 of Scheme 3A, and Step 1 is to obtain the compound w and c Starting from the chiral compound eee , ff , gg , gg and chiral bicyclic ketone compounds ii is obtained. TIFF2024513394000085.tif70170

[0165] Scheme 4 Scheme 4 depicts syntheses for preparing additional bicyclic diversity of compounds of Formulas (I)-(IV) using a variety of nucleophiles, including but not limited to halide and cyanide sources.

[0166] Many variations of the reactions in Schemes 1 to 4 are possible and will be obvious to those skilled in the art. For example, in many embodiments, the order of the reactions can be changed. In some instances, the reactions involve stereoisomers of the compounds depicted in the schemes described herein. In some cases, the reaction products do not need to be isolated and can be used in situ in the next reaction. The chemical nature of the amine and hydroxyl protecting groups, as well as the timing of the protection and deprotection events, can be varied, for example, from the specific embodiments described herein. EXAMPLES

[0167] The present invention will be more fully understood by reference to the following examples, which should not be construed as limiting the scope of the invention.

[0168] These examples serve to provide guidance to those of ordinary skill in the art for preparing and using the compounds, compositions and methods of the present invention. Although specific embodiments of the present invention have been described, those of ordinary skill in the art will recognize that various changes and modifications can be made without departing from the spirit and scope of the invention.

[0169] The chemical reactions of the described examples can be easily adapted to prepare some other compounds of the present invention, and alternative methods for preparing the compounds of the present invention are considered to be within the scope of the present invention. The synthesis of non-exemplary compounds according to the present invention can be successfully carried out by modifications obvious to those skilled in the art, such as, for example, by appropriately protecting interfering groups using other suitable reagents known in the art other than those described, and / or by making routine modifications to reaction conditions.

[0170] In the following examples, all temperatures are in degrees Celsius unless otherwise indicated. Commercially available reagents were purchased from suppliers such as Aldrich Chemical Company, Lancaster, TCI, or Maybridge and were used without further purification unless otherwise indicated. Reactions described below were generally carried out under a positive pressure of nitrogen or argon, or using drying tubes in anhydrous solvents (unless otherwise indicated), and reaction flasks were typically fitted with rubber septa for the introduction of substrates and reagents via syringe. Glassware was oven-dried and / or heat-dried. 1 H NMR spectra were obtained in deuterated CDCl3, d6-DMSO, CH3OD or d6-acetone solvent solutions (reported in ppm) using trimethylsilane (TMS) or residual non-deuterated solvent peaks as the reference standard. When peak multiplicities are reported, the following abbreviations are used: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet, br (broad), dd (double doublet), dt (double triplet). Coupling constants, when given, are reported in Hz (Hertz).

[0171] All abbreviations used to describe reagents, reaction conditions, or equipment are intended to be consistent with the definitions set forth in the following list of abbreviations: Chemical names for the individual compounds of the invention were generally obtained using the structure naming feature of the ChemDraw naming program.

[0172] Abbreviation ACN Acetonitrile AcOH Acetic acid Boc tert-butoxycarbonyl CPME Cyclopentyl methyl ether DAST Diethylaminosulfur trifluoride DCE 1,2-dichloroethane DCM Dichloromethane DIPEA or i-Pr2NEt N,N-Diisopropylethylamine DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide DPPH 2,2-diphenyl-1-picrylhydrazyl EDCI 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide ET external temperature Et3N·3HF Triethylamine trihydrofluoride EtOH Ethanol GC Gas Chromatography HPLC High Performance Liquid Chromatography IT internal temperature KF Karl Fischer titration KRED Ketoreductase LCMS Liquid Chromatography Mass Spectrometry 2-MeTHF 2-Methyltetrahydrofuran MTBE Methyl tert-butyl ether NAD Nicotinamide Adenine Dinucleotide NADH reduced nicotinamide adenine dinucleotide NADP Nicotinamide adenine dinucleotide phosphate NADPH reduced nicotinamide adenine dinucleotide phosphate NMI N-Methylimidazole NMR nuclear magnetic resonance PBS Phosphate buffer PBSF Perfluorobutanesulfonyl fluoride PCC Pyridinium Chlorochromate RP Reverse Phase RT or R T retention time SEM 2-(trimethylsilyl)-ethoxymethyl SFC Supercritical Fluid Chromatography TBAH Tetrabutylammonium triacetoxyborohydride TFA Trifluoroacetic acid THF Tetrahydrofuran TMSCl Trimethylsilyl chloride

[0173] Example 1: Preparation of Ethyl-2-ethoxy-iminoacetate Hydrochloride A3 TIFF2024513394000086.tif29170

[0174] Scheme 5 The synthesis of ethyl-2-ethoxy-iminoacetate A3 is shown in Scheme 5.

[0175] Ethyl-2-ethoxy-iminoacetate hydrochloride (A2) : A reactor was charged with MTBE (28.0 kg, 6V, KF: 360 ppm), ethyl cyanoformate A1 (6.3 kg, 63.6 mol, 1.0 equiv), and TMSCl (21.4 kg, 197.1 mol, 3.1 equiv) under N2 atmosphere at RT (ca. 30 °C). The mixture was cooled to 0-5 °C. EtOH (12.0 kg, 4.1 equiv, KF: 200 ppm) was added dropwise at 0-5 °C over 30 min. After the addition was complete, the mixture was warmed to 5-10 °C and then stirred for 23 h. The reaction was monitored by GC (ethyl cyanoformate A1 < 5 A%). The mixture was filtered under N2 atmosphere and the cake was washed with MTBE (17 kg x 3, 3.7 V x 3). The cake was dried under N2 flow at 25-30 °C for 4 h to give the product A2 (9.0 kg, 78% yield) as a white powder.

[0176] Ethyl-2-ethoxy-iminoacetate hydrochloride (A3) : A reactor was charged with Na2SO4 (9.0 kg, 100 wt%), A2 (9.0 kg, 49.5 mol, 1.0 equiv), and MTBE (953.0 kg, 9.0 V) under N2 atmosphere at RT (ca. 30 °C). The mixture was cooled to 0 °C. A solution of Et3N (5.4 kg, 1.08 equiv, KF: 360 ppm) in MTBE (6.7 kg, 1 V, KF: 300 ppm) was added dropwise over 2 h at 0-5 °C. After the addition was complete, the mixture was warmed to 20 °C and then stirred for 4 h. The reaction was monitored by GC (cpd.2 < 0.5 A%). The mixture was filtered under N2 atmosphere and the cake was washed with MTBE (7.5 kg, 1.1 V). The filtrate was concentrated under vacuum and dried under vacuum at 20 °C to give the product A3 as a liquid, which was combined with other batches (ca. 60% yield).

[0177] 1 H NMR (400 MHz, CDCl3) δ 8.77 (s, 1H), 4.30 (dq, J = 9.6, 7.1 Hz, 4H), 1.36 (dt, J = 8.2, 7.1 Hz, 6H). 13 C NMR (100 MHz, CDCl3) δ 159.4, 158.0, 63.4, 63.0, 14.0, 13.9.

[0178] Example 2: Preparation of (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one A12 TIFF2024513394000087.tif117170

[0179] Scheme 6 The synthesis of (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one A12 is shown in Scheme 6.

[0180] (S)-(-)-2-Acetoxysuccinic anhydride (A5) : D-Malic acid A4 (12.0 kg, 89.5 mol, 1.0 equiv.) in acetyl chloride (52.8 kg, 4 V) was heated to 40-53 °C (IT) and stirred for 16 h. The reaction was monitored by HPLC (D-Malic acid A4 < 0.5 A%). The reaction mixture was concentrated under vacuum at 40-60 °C (ET) to remove volatiles. The crude oil was dissolved in i-PrOAc (10.6 kg, 1.0 V). The inert solvent n-heptane (27.7 kg, 3.3 vol.) was added at 15 °C. After completion of the addition of n-heptane, the solid was allowed to precipitate. The suspension was stirred at 0 °C for another hour, after which the solid was filtered and flushed with n-heptane (8.3 kg x 2, 2 V x 2). The cake was dried under vacuum at 40-45° C. to give (S)-(−)-2-acetoxysuccinic anhydride A5, which was consistent with commercial samples and literature reports (e.g., Shiuey, SJ; Partridge, JJ; Uskokovic, MRJ Org. Chem. 1988, 53, 1040-1046).

[0181] 1 H NMR (400 MHz, CDCl3) δ 5.53 (dd, J = 9.6, 6.3 Hz, 1H), 3.38 (dd, J = 18.9, 9.6 Hz, 1H), 3.02 (dd, J = 18.9, 6.3 Hz, 1H), 2.19 (s, 3H).

[0182] (R)-2-Acetoxy-4-oxo-4-phenylbutanoic acid (A6) : A suspension of anhydrous aluminum chloride (25.3 kg, 189.8 mol, 2.5 equiv) in CHCl (320.0 kg, 20 V) was stirred under N at −10°C to −5°C. (S)-(−)-2-acetoxysuccinic anhydride A5 (12.0 kg, 75.9 mol, 1.0 equiv) was added over 15 min. The mixture was stirred under N at −10°C to −5°C for 10 min. Benzene (18.7 kg, 239.1 mol, 3.15 equiv) was added to the mixture under N at −10°C to −5°C over 30–90 min. The mixture was stirred under N at −5°C to 0°C for 18 h. The reaction was monitored by HPLC (anhydride <0.5 A%). The mixture was quenched with aqueous HCl (90.0 kg, 3.0 M, 3.5–3.6 equiv). The organic phase was allowed to settle for 2 hours and then separated from the suspension. The aqueous phase was extracted with i-PrOAc (52.8 kg x 2, 5 V x 2). All the organic phases were combined and washed with brine (68 kg x 2, 5 V x 2). The organic phase was concentrated under vacuum at 35-40 °C. The crude was dissolved in i-PrOAc (105.6 kg, 10 V) at 40 °C for 30 min. The suspension was filtered through celatom (2.4 kg, 20 wt%) and the cake was washed with i-PrOAc (10.6 kg, 1 V). The filtrate was concentrated under vacuum at 35-40 °C. The crude was slurried in a mixture of CHCl / n-heptane (15.6 kg / 42.0 kg, 2V / 10V), filtered, washed with n-heptane (10.6 kg, 1V), and then dried at 30-35° C. to give (R)-2-acetoxy-4-oxo-4-phenylbutanoic acid A6 (96.1 kg, 70% yield) as a white powder, which is consistent with commercial samples and literature reports (see, e.g., Wilkins, TD; Tucker, KD Process for Producing Optically Active 2-Hydroxy-4-Arylbutyric Acid or its Ester, U.S. Patent No. 5,959,139, Sep. 28, 1999).

[0183] 1H NMR (400 MHz, CDCl3) δ 11.26 (br s, 1H), 8.01 - 7.92 (m, 2H), 7.61 (t, J = 7.4 Hz, 1H), 7.49 (t, J = 7.7 Hz, 2H), 5.74 (dd, J = 7.8, 3.5 Hz, 1H), 3.66 (dd, J = 17.8, 7.8 Hz, 1H), 3.54 (dd, J = 17.8, 3.6 Hz, 1H), 2.11 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 194.7, 174.8, 170.1, 136.0, 133.8, 128.8, 128.2, 67.3, 39.6, 20.5. C 12 H 13 O5 [M+H] + The calculated HRMS(APCI) for is 237.0763, the measured value is 237.0790.

[0184] (-)-Ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate (A7) : A solution of concentrated sulfuric acid (14.6 kg, 148.17 mol, 2.5 equiv) in ethanol (33.4 k, 3.0 V) was added over 35 min with stirring at 12-14 °C (IT) to a solution of A6 (14.0 kg, 59.3 mol, 1.0 equiv) in ethanol (22.2 kg, 2.0 V), heated to 20-25 °C (IT) and stirred for 16-20 h. The reaction was monitored by HPLC (cpd. A6 < 0.5 A%). The reaction mixture was poured into ice water (210 kg, 1500 wt%) with stirring at 0-10 °C and the aqueous phase was extracted with i-PrOAc (123.2 kg x 2, 10 V x 2). The combined organic phase was washed with saturated sodium bicarbonate (72.2 kg, 5 V) and brine (72.2 kg, 5 V). The organic phase was concentrated (without drying) to remove volatiles and dried at 40° C. (ET) to yield a yellow oil. The crude oil was dissolved in MTBE (10.0 kg, 1 V) with stirring and filtered at 40° C. n-heptane (49.0 kg, 5 V) was added over 1 h at 0° C. to 5° C. (IT). The suspension was stirred at 0° C. to 5° C. (IT) for 1 h and the solid was filtered. The cake was washed with n-heptane (19.6 kg×2, 2 V×2). The white solid was dried under reduced pressure to give the desired product (−)-ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate A7 (10.5 kg, 83% yield) as a white solid. This is consistent with commercial samples and literature reports (see, e.g., Li, W.; Lu, B.; Xie, X.; Zhang, Z. Org. Lett. 2019, 21, 5509-5513).

[0185] Chiral HPLC>99% ee. 1H NMR (400 MHz, CDCl3) δ 7.98 - 7.90 (m, 2H), 7.62 - 7.53 (m, 1H), 7.45 (dd, J = 10.5, 4.8 Hz, 2H), 4.66 (dd, J = 6.0, 3.9 Hz, 1H), 4.25 (q, J = 7.1 Hz, 2H), 3.53 (dd, J = 17.5, 3.9 Hz, 1H), 3.45 (dd, J = 17.5, 6.0 Hz, 1H), 3.42 (br s, 1H), 1.26 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 197.5, 173.7, 136.4, 133.5, 128.6, 128.1, 67.2, 61.8, 42.1, 14.1. C 12 H 14 NaO4[M+Na] + The calculated HRMS(ESI) for is 245.0790, the measured value is 245.0786.

[0186] tert-Butyl (R,E)-2-(4-ethoxy-3-hydroxy-4-oxo-1-phenylbutylidene)hydrazine-1-carboxylate (A8) : Compound A7 (70.0 kg, 315.0 mol, 1.0 equiv) and NHNHBoc (54.1 kg, 409.4 mol, 1.3 equiv) in ethanol (552 kg, 10 V) and formic acid (3.6 kg, 78.2 mol, 0.25 equiv) were heated to 55-60 °C (IT) and stirred under N for 16 h. The reaction was monitored by HPLC (cpd. A7 < 2.0 A%). The reaction mixture was concentrated under vacuum at 40-50 °C (ET, jacket temperature) to remove volatiles. Azeotropic distillation with n-heptane (93 kg x 2, 2 V x 2) was performed. More n-heptane (186 kg, 4 V) was added to the suspension and the mixture was stirred at 45-50 °C for 12 h. The slurry was then treated with MTBE (280 kg, 5.4 V) and heptane (210 kg, 4.5 V) and stirred at 0 °C for an additional 2 h, after which the solid was filtered and rinsed with n-heptane (93 kg x 2, 2 V x 2). The cake was dried under vacuum at 40-45 °C for 16 h to give tert-butyl (R,E)-2-(4-ethoxy-3-hydroxy-4-oxo-1-phenylbutylidene)hydrazine-1-carboxylate A8 (95.6 kg, 92% yield) as a white solid.

[0187] HPLC: 97:3 E / Z hydrazone ratio. 1 H NMR (600 MHz, DMSO-d6) δ 9.88 (s, 1H), 7.71 - 7.64 (m, 2H), 7.43 - 7.29 (m, 3H), 6.14 (d, J = 5.4 Hz, 1H), 4.22 (dt, J = 8.1, 5.0 Hz, 1H), 4.03 - 3.86 (m, 2H), 3.17 - 3.00 (m, 2H), 2.50 - 2.43 (m, 1H), 1.44 (s, 9H), 1.09 (t, J = 7.1 Hz, 3H). 13 C NMR (150 MHz, DMSO-d6) δ 173.0, 153.5, 148.9, 138.0, 129.3, 128.8, 126.8, 80.1, 68.8, 61.0, 31.9, 28.6, 14.4. C 17 H 25 N2O5[M+H] +The calculated HRMS(ESI) for is 337.1758, the measured value is 337.1767.

[0188] (3R,5S)-1-Amino-3-hydroxy-5-phenylpyrrolidin-2-one hydrochloride (A11) : To a suspension of TBAH (123.2 kg, 468 mol, 1.75 equiv) in CHCl (712 kg, 10 V) was added acetic acid (675 kg, 7.5 V) at 10-20 °C under N. After the addition was complete, the mixture was stirred at 20-25 °C for 1 h. To the resulting solution, compound A8 (90 kg, 268 mol, 1.0 equiv) was added as a solution in CHCl (90 kg, 1.5 V) after cooling to -5 °C. The mixture was stirred at -5 °C (IT) for 16 h under N. The reaction was monitored by HPLC (cpd. A8 < 2.0 A%). The mixture was then quenched with ethanol (137 kg, 2 V) at 15-20 °C. The resulting solution was stirred under N2 at 25-30 °C for 16 h to give tert-butyl 2-((1S,3R)-4-ethoxy-3-hydroxy-4-oxo-1-phenylbutyl)hydrazine-1-carboxylate A9. The reaction was monitored by HPLC (A9 < 0.5A%). The reaction mixture was quenched with water (450 kg) and then extracted with CHCl (600 kg x 2, 10 V x 2). The organic phase was washed with water (450 kg x 2, 5 V x 2), 10% aqueous sodium carbonate (500 kg x 3, 5 V x 3; pH = ca. 10), and brine (520 kg x 2, 5 V x 2; pH = ca. 7). The organic phase was concentrated in vacuo to yield a solution of crude tert-butyl ((3R,5S)-3-hydroxy-2-oxo-5-phenylpyrrolidin-1-yl)carbamate A10 in CH2Cl2.

[0189] A solution of crude compound A10 (1.0 equiv.) in CHCl (320 kg) was cooled to 0-5°C, and then a solution of 6 M HCl in n-propanol (200 L, 6.0 equiv.) was added at 60°C. The mixture was stirred at 20-25°C for 16 h. The reaction was monitored by HPLC (cpd. A10 < 0.5 A%). The resulting mixture was filtered and the cake was washed with CHCl (200 kg x 2, 2 V x 2). The cake was dried in a filter drier under N flow at 40-45°C to give (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one hydrochloride A11 (45.3 kg, 85% yield, 3 steps) as a white powder.

[0190] 1 H NMR (400 MHz, CD3OD) δ 7.52 - 7.32 (m, 5H), 5.07 (dd, J = 8.2, 4.3 Hz, 1H), 4.62 (dd, J = 8.0, 6.1 Hz, 1H), 2.55 (ddd, J = 14.1, 8.2, 6.1 Hz, 1H), 2.49 - 2.34 (m, 1H). 13 C NMR (101 MHz, D2O) δ 175.3, 140.1, 131.9, 131.5, 129.1, 69.1, 63.9, 38.9. 10 H 13 N2O2[M+H] + The calculated HRMS(ESI) for is 193.0977, the measured value is 193.0973.

[0191] (3R,5S)-1-Amino-3-hydroxy-5-phenylpyrrolidin-2-one (A12) : Crude compound A11 (21.6 kg, 1.0 equiv.) was dissolved in water (16.2 kg, 0.75 V) at 30-35 °C and the solution was filtered through a polish filter (PP, 1 um). The filtrate was cooled to 10-20 °C and then a solution of sodium hydroxide (1.93 kg, 52.8 mol, 0.51 equiv.) in water (2.8 kg, 0.13 V) (previously filtered through a polish filter) was added. The mixture was stirred for 60 min at 10-20 °C. Finally, a solution of sodium hydroxide (1.93 kg, 52.8 mol, 0.51 equiv.) in water (2.8 kg, 0.13 V) (previously filtered through a polish filter) was added and the mixture was stirred for another 60 min at 10-20 °C. The resulting mixture was filtered and the cake was washed with chilled water (4.32 kg x 1, 2.16 kg x 1, 0.2 V x 1, 0.1 V x 1; 10-15 °C) and chilled n-PrOH (4.24 kg x 2, 0.25 V x 2; 10-15 °C). The cake was dried in a filter drier under N2 flow at 40-45 °C to give (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one A12 (13.2 kg, 73% yield) as a white powder.

[0192] HPLC: >99.5:0.5 dr. Chiral HPLC: >99.5% ee. 1 H NMR (600 MHz, DMSO-d6) δ 7.35 (t, J = 7.5 Hz, 2H), 7.28 (t, J = 7.5 Hz, 1H), 7.18 (d, J = 7.6 Hz, 2H), 5.69 (d, J = 4.9 Hz, 1H), 4.66 (dd, J = 8.4, 3.6 Hz, 1H), 4.35 - 4.22 (m, 1H), 2.24 - 2.15 (m, 1H), 2.07 (ddd, J = 12.7, 8.1, 3.6 Hz, 1H). 13 C NMR (150 MHz, DMSO-d6) δ 171.8, 141.5, 128.6, 127.4, 126.3, 66.5, 61.4, 36.9. 10 H 13 N2O2[M+H] +Calculated HRMS (ESI) for is 193.0972, observed value is 193.0978.

[0193] Example 3: Cyclopropyl-[(5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl]methanone (A17): TIFF2024513394000088.tif168170

[0194] Scheme 7 The synthesis of cyclopropyl-[(5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl]methanone A1 is shown in Scheme 7.

[0195] Ethyl (5S,7R)-7-hydroxy-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylate (A13) : TIFF2024513394000089.tif38170 Compound A12 (5.02 kg, 26.11 mol, 1.00 equiv) was charged to a 100 L reactor 1 under nitrogen. EtOH (15.95 kg, 20.22 L, 4.0 vol), compound A3 (5.60 kg, 38.58 mol, 1.47 equiv), EtOH (15.85 kg, 20.09 L, 4.0 vol), AcOH (4.60 kg, 4.38 L, 76.60 mol, 2.93 equiv), and EtOH (8.15 kg, 10.33 L, 2.1 vol) were charged to the reactor to give a suspension. The internal temperature was adjusted to 80±10° C. and the reaction was stirred for 21 h. During this time the reaction clarified before becoming a suspension again. The internal temperature was adjusted to 60±15°C over 30 minutes. The Aurora filter was heated to a jacket temperature of 60±15°C and a portion of the contents of Reactor 1 was passed through the Aurora filter and approximately 70 L of filtrate was collected in the clean 100 L reactor 2. Insoluble solids that are not soluble in certain organic solvents are retained. For example, the solids are oligomers or polymers of compound A3 (ethyl-2-ethoxy-2-iminoacetate) as confirmed by solid-state NMR spectroscopy.

[0196] The first portion of the filtrate from reactor 2 was concentrated under reduced pressure over 1 hour to approximately 55 L (11 vol), while maintaining an internal temperature below 60° C. EtOH (16.30 kg, 20.66 L, 3.2 vol) was added to reactor 1 and the remaining reactor 1 contents were passed through an Aurora filter and approximately 20 L of additional filtrate was collected in reactor 2, giving a total volume of approximately 75 L. The contents of reactor 2 were concentrated under reduced pressure over 1 hour to a volume of approximately 30 L (6 vol), while maintaining an internal temperature below 60° C. As the volume of the solution decreases, solids may be observed to form. Prior to the addition of the seeds, a suspension may be observed. In some batches, the early solids formation event does not appear to affect the product quality. The reactor was cooled to an internal temperature of 25±10° C. Water (15.10 kg, 15.10 L, 3.0 vol) was added to reactor 2 over 60±30 minutes. Compound A13 seeds (25 g, 0.5 wt%) and water (150 mL) were combined in a glass bottle and charged to reactor 2. A suspension was observed. Additional water (49.15 kg, 49.15 L, 9.8 vol) was added to reactor 2 over 60±30 minutes.

[0197] Reactor 2 was stirred for 3 hours. The internal temperature was adjusted to 0±5°C over 3 hours and stirred for 10 hours. The slurry was transferred to a filter dryer and the filtrate was collected in a glass carboy. Reactor 2 was rinsed with water (23.30 kg, 23.30 L, 4.6 vol) and stirred for a minimum of 5 minutes, the contents were transferred to a filter dryer and the filtrate was collected in a glass carboy. The solid cake in the filter dryer was rinsed with water (20.55 kg, 20.55 L, 4.1 vol). For example, minimizing AcOH in solid samples is necessary to prevent competition with the fluoride nucleophile in the alcohol in the subsequent deoxyfluorination step. In some instances, high levels of acetic acid have led to the formation of acetic acid products.

[0198] The solid cake was dried under vacuum with a nitrogen sweep for 27 hours with intermittent stirring at a jacket temperature of 50±5° C. The filter dryer was cooled to a jacket temperature of 20±5° C. and the product compound A13 (6.36 kg, 23.27 mol, 86% yield) was discharged into a sealed bag as a crystalline off-white solid.

[0199] HPLC 99.9 A%, >99.9:0.1 dr. 1 H NMR (600 MHz, DMSO-d6) δ 7.44 - 7.32 (m, 3H), 7.25 (d, J = 7.3 Hz, 2H), 6.18 (d, J = 5.9 Hz, 1H), 5.76 (t, J = 6.6 Hz, 1H), 5.33 - 5.25 (m, 1H), 4.37 - 4.22 (m, 1H), 2.99 - 2.86 (m, J = 6.3 Hz, 2H), 1.27 (t, J = 7.1 Hz, 3H). 13 C NMR (150 MHz, DMSO-d6) δ 163.6, 159.5, 158.8, 138.6, 128.9, 128.5, 127.0, 62.6, 61.0, 60.2, 46.5, 14.0. C 14 H 16 N3O3[M+H] + The calculated HRMS(ESI) for is 274.1186, the measured value is 274.1195.

[0200] Other acid additives were evaluated including acetic acid, oxalic acid, succinic acid, benzoic acid, isobutyric acid, pivalic acid, salicylic acid, oxamic acid, 2-picolinic acid, trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid, formic acid, hydrochloric acid in ethanol, and trimethylsilyl chloride in ethanol. In some instances, the yields with other acid additives were in the range of 14-81%. In some instances, the yield is 98% when the acid additive is acetic acid.

[0201] Other alternative reagents to compound A3 were evaluated, such as ethyl thioxamate, ethyl cyanoformate, methyl cyanoformate, and triethyl 1,3,5-triazine-2,4,6-tricarboxylate. In some cases, the HLPC loading and yield of these reagents were in the range of 90-100% and 40-76%, respectively. In other cases, the yield of these reagents was less than 81%.

[0202] In one example, the acid additive is isobutyric acid and the reagent reacted with compound A12 is ethyl thioxamate, with a yield of 72.3%. In another example, the acid additive is salicylic acid and the reagent is triethyl 1,3,5-triazine-2,4,6-tricarboxylate, with a yield of 81%. In yet another example, the acid additive is formic acid and the reagent is ethyl cyanoformate, with a yield of 14%.

[0203] (5S,7S)-7-Fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxylic acid (A15) : TIFF2024513394000090.tif49170Under nitrogen, a 100 L reactor 1 was charged with compound A13 (4.51 kg, 15.97 mol, 1.00 equiv), CH3CN (17.90 kg, 22.77 L, 5.0 vol), and i-Pr2NEt (12.95 kg, 100.20 mmol, 6.27 equiv). Et3N·3HF (5.30 kg, 32.88 mmol, 2.59 equiv) was added slowly over 1 h via peristaltic pump while maintaining the internal temperature below 30 °C with jacket cooling (addition of Et3N·3HF is exothermic). CH3CN (0.36 kg, 0.45 L, 0.1 vol) was added through the same peristaltic pump tubing. PBSF (7.50 kg, 24.83 mmol, 1.55 equiv) was added slowly over 1 h, maintaining the temperature below 30° C. with jacket cooling (the addition of PBSF is exothermic). CH3CN (0.36 kg, 0.45 L, 0.1 vol) was added through the same peristaltic pump tubing. The contents of Reactor 1 were stirred for 1 h.

[0204] 2-MeTHF (31.00 kg, 36.05 L, 8.0 vol) was added to reactor 1 and the mixture was stirred for 20 minutes. The contents of reactor 1 were transferred to 200 L reactor 2 containing a stirred solution of K2HPO4 (7.88 kg) in water (32.40 kg, 32.40 L, 7.2 vol) with jacket cooling to maintain the internal temperature below 30°C. 2-MeTHF (17.40 kg, 20.23 L, 4.5 vol) was added to reactor 1 and the rinse was transferred to reactor 2 with jacket cooling to maintain the internal temperature below 30°C. The stirring was stopped and the layers were allowed to settle for 30 minutes. The three-phase mixture was separated and the two bottom layers were collected in separate glass carboys (the bottom layer is referred to as the dense layer and may contain fluorinated by-products, the middle layer is referred to as the aqueous layer). A solution of NaCl (14.49 kg) in water (63.35 kg, 63.35 L, 14.0 vol) was prepared. A portion of this NaCl solution (25.72 kg) was transferred to Reactor 2 and the contents were stirred for a minimum of 5 minutes. The stirring was stopped and the layers were allowed to settle for at least 30 minutes (actual time: 30 minutes).

[0205] The biphasic mixture was separated and the aqueous phase was collected in a glass carboy. Another portion of the NaCl solution (25.72 kg) was transferred to reactor 2 and the contents were stirred for 10 minutes. The stirring was stopped and the layers were allowed to settle for 15 minutes. The biphasic mixture was separated and the aqueous phase was collected in a glass carboy. Another portion of this NaCl solution (25.72 kg) was transferred to reactor 2 and the contents were stirred for a minimum of 5 minutes. The stirring was stopped and the layers were allowed to settle for 30 minutes. The biphasic mixture was separated and the aqueous phase was collected in a glass carboy. A crude solution of compound A14 in 2-MeTHF and CH3CN was obtained.

[0206] A portion of the solution in reactor 2 (approximately 60 L) was transferred to a clean 100 L reactor 3. The contents of reactor 3 were distilled under reduced pressure to approximately 30 L (6.7 vol) while maintaining an internal temperature below 50°C. THF (120.15 L, 135.15 L, 29.9 vol) was added to reactor 2. The contents of reactor 2 were continuously transferred to reactor 3 to maintain a target volume of 27-45 L (6.0-10.0 vol). Distillation was carried out over 4 hours, stopped for 16 hours, and then resumed over 1 hour, reaching a final volume of approximately 40 L (8.9 vol). Distillation was continued until the volume was approximately 23 L (5.1 vol). The solution was cooled to an internal temperature of 35±10°C. The reduction in 2-MeTHF and CH3CN content may facilitate ester hydrolysis.

[0207] Reactor 3 was cooled to an internal temperature of 20±10° C. and water (22.85 kg, 22.85 L, 5.1 vol) was added followed by a solution of KOH (3.25 kg, 57.92 mmol, 4.38 equiv) in water (11.90 kg, 11.90 L, 2.6 vol) (5 M aq KOH) while maintaining the temperature below 30° C. The reaction mixture was stirred for 1 hour. Water (22.90 kg, 22.90 L, 5.1 vol) was charged to reactor 3 and the contents transferred to reactor 2. A solution of concentrated hydrochloric acid (6.75 kg) in water (5.80 kg, 5.80 L, 1.3 vol) (6 M aq HCl) was added slowly while maintaining the internal temperature below 30° C., passing the vapors through a NaOH scrubber solution with phenolphthalein indicator. Smoky vapors may be HCl, and if produced, are neutralized by a NaOH scrubber. The reaction was stirred at 20±10° C. for 13 hours. The pH range (e.g., target: 0≦pH≦2) ensures, e.g., complete protonation to the carboxylic acid and minimal loss of aqueous wash during filtration.

[0208] The suspension was transferred to a filter dryer and the filtrate was collected in a glass carboy. The filter cake was rinsed with a solution of water (40.30 kg, 40.30 L, 8.9 vol) and THF (3.85 kg, 4.33 L, 1.0 vol) and the filtrate was collected in a glass carboy. A combined organic / aqueous wash is used to purge any residual perfluorobutanesulfonate that may still be present after acidification. For example, in a usage test where this wash was not performed, the isolated material had a pH of 10. 19 F NMR analysis showed an extra fluorine signal. The filter cake was rinsed with water (45.12 kg, 45.12 L, 10 vol) and the filtrate was collected in a glass carboy. The jacket temperature was increased to 50±5° C. and the wet cake was dried for 4 days. The filter dryer was cooled to a jacket temperature of 20±5° C. and the product compound A15 (3.76 kg, 14.66 mol, 92% yield) was discharged into a sealed bag as a light brown solid.

[0209] HPLC 99.8 A%, >99.95:0.05 dr. 1 H NMR (600 MHz, DMSO-d6) δ 7.45 - 7.40 (m, 2H), 7.40 - 7.35 (m, 1H), 7.24 (d, J = 7.5 Hz, 2H), 6.21 (ddd, J = 56.6, 7.2, 1.9 Hz, 1H), 5.69 (ddd, J = 9.1, 6.7, 3.0 Hz, 1H), 3.72 (ddd, J = 25.8, 15.4, 7.8, 7.8 Hz, 1H), 2.70 (ddd, J = 26.8, 15.3, 2.4, 2.4 Hz, 1H). 13 C NMR (150 MHz, DMSO-d6) δ 160.5, 158.8, 138.4, 129.0, 128.6, 126.7, 126.7, 83.2, 60.0, 42.9. 19 F NMR (565 MHz, DMSO-d6) δ -167.0. C 12 H 11 FN3O2[M+H] + The calculated HRMS(ESI) for is 248.0830, the measured value is 248.0837. TIFF2024513394000091.tif34170

[0210] In another example, toluene (70 mL, 10 vol.), DBU (7 g, 0.11 mol, 2.0 equiv.) and compound A12 (7 g, 0.11 mol, 1.0 equiv.) were charged to a 250 mL flask at rt (ca. 24 °C) under N2 atmosphere. The solution was cooled to 0-10 °C. PyFluor (7 g, 0.11 mol, 1.2 equiv.) was added dropwise to the above solution at 0-10 °C. The solution was stirred at 0-10 °C for 2 h. After completion of the reaction, the mixture was quenched with saturated aqueous NH4Cl at 0-20 °C. The mixture was extracted with TBME (70 mL, 10 vol.). The organic phase was washed with brine (70 mL, 10 vol.) and then dried over Na2SO4 (ca. 5 g). The mixture was filtered and the cake was washed with TBME (20 mL). The filtrate was concentrated under vacuum at 35° C. to give approximately 10 g of crude compound A14 in quantitative yield with HPLC purity of 95 A%.

[0211] (5S,7S)-7-Fluoro-N-methoxy-N-methyl-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide (A16) : TIFF2024513394000092.tif43170To a 100 L reactor 1 under nitrogen was added compound A15 (4.22 kg, 17.03 mol, 1.00 equiv), N,O-dimethylhydroxylamine HCl (2.09 kg, 21.00 mol, 1.23 equiv), CHCl (16.80 kg, 12.68 L, 3.0 vol), 1-methylimidazole (1.25 kg, 15.22 mol, 0.89 equiv), CHCl (2.85 kg, 2.15 L, 0.5 vol), EDCI (4.36 kg, 21.83 mol, 1.28 equiv), and CHCl (8.75 kg, 6.60 L, 1.6 vol). Process optimization may lead to cleaner reaction profiles and faster reaction rates when the charge on 1-methylimidazole is reduced, and reducing the charge on EDCI (1.3 equivalents in this modified procedure compared to 1.5 equivalents in the telescoping step 4 / 5 procedure) may not adversely affect the reaction. The contents of Reactor 1 were stirred for 2 hours.

[0212] A 6M aqueous HCl solution was prepared by dissolving concentrated hydrochloric acid (6.34 kg) in water (15.75 kg, 15.75 L). This 6M HCl solution (21.17 kg) was slowly charged to reactor 1 while maintaining an internal temperature below 30°C. The contents of reactor 1 were stirred for 10 minutes. The stirring was stopped and the phases were allowed to settle for 15 minutes. The aqueous phase was drained into a glass carboy and the organic phase was returned to reactor 1. A solution of K2HPO4 (2.11 kg) in water (19.01 kg, 19.01 L) was prepared. A portion of this aqueous K2HPO4 solution (21.13 kg) was transferred to reactor 1 while maintaining an internal temperature below 30°C. The contents of reactor 1 were stirred for 10 minutes. The stirring was stopped and the phases were allowed to settle for 15 minutes. The aqueous phase was drained into a glass carboy and the organic phase was returned to reactor 1. Water (21.13 kg, 21.13 L, 5.0 vol) was charged to Reactor 1. Agitation was stopped and the phases were allowed to settle for 15 minutes.

[0213] The aqueous phase was drained into a glass carboy and the organic phase was returned to Reactor 1. The contents of Reactor 1 were distilled to approximately 12 L (2.8 vol) while maintaining an internal temperature below 50° C. CPME (18.20 kg, 21.16 L, 5.0 vol) was added to Reactor 1. HTE solubility screening showed that CPME, for example, is a unique solvent that exhibits high solubility at high temperatures and low solubility at low temperatures. The contents of Reactor 1 were distilled under reduced pressure to a target volume of approximately 25 L (5.9 vol) while maintaining the temperature below 65° C. CPME (7.27 kg, 8.45 L, 2.0 vol) was added to Reactor 1. The contents of Reactor 1 were distilled under reduced pressure to a target volume of approximately 33 L (7.8 vol) while maintaining the temperature below 65° C.

[0214] The reactor temperature was adjusted to 80±5° C. The reaction remained in solution. The reactor temperature was adjusted to 60±5° C. over 40 min. A slurry of (5S,7S)-7-fluoro-N-methoxy-N-methyl-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide A16 seeds (20.5 g, 0.071 mmol, 0.5 wt%) in CPME (84.5 g, 98 mL) was charged to reactor 1. The reaction began to form a suspension. The reactor contents were stirred for 30 min. Heptane (8.66 kg, 12.66 L, 3.0 vol) was added to reactor 1 over 30 min. Heptane was added as an anti-solvent, for example to further reduce mother liquor losses in crystallization. Reactor 1 was adjusted to 0±5° C. over 3 h. The reactor contents were stirred for 16 hours. Heptane (14.45 kg, 21.13 L, 5.0 vol) was charged to reactor 3, cooled to 0° C., and stirred for 4.5 hours. The reactor 1 contents were transferred to a filter dryer maintained at 20° C., and the filtrate was collected in reactor 1. The reactor 1 contents were transferred to a filter dryer, and the filtrate was again collected in reactor 1.

[0215] The contents of Reactor 3 were transferred to a filter dryer and the filtrate was collected in Reactor 1. The contents of the filter dryer were dried under vacuum with a nitrogen sweep at ambient temperature for 21 hours. The filter dryer jacket temperature was increased to 50±5° C. and the contents were dried for 3 days with intermittent stirring of the wet cake. The filter dryer temperature was adjusted to 20±10° C. and the product (5S,7S)-7-fluoro-N-methoxy-N-methyl-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide A16 (4.62 kg, 15.88 mol, 93% yield) was discharged into a sealed bag as a light brown solid.

[0216] HPLC 99.8 A%, >99.95:0.05 dr. 1H NMR (600 MHz, DMSO-d6) δ 7.47 - 7.32 (m, J = 7.7, 6.5, 1.4 Hz, 3H), 7.26 - 7.19 (m, 2H), 6.21 (ddd, J = 56.7, 7.1, 1.8 Hz, 1H), 5.74 - 5.65 (m, 1H), 3.83 - 3.65 (m, 1H), 3.69 (s, 3H), 3.30 (s, 3H), 2.78 - 2.62 (m, 1H). 13 C NMR (150 MHz, DMSO-d6) δ 161.9, 158.0, 138.5, 129.0, 128.6, 126.5, 99.5, 83.2, 61.6, 59.8, 43.1, 32.3. 19 F NMR (565 MHz, DMSO-d6) δ -166.6.C 14 H 16 FN4O2[M+H] + The calculated HRMS(ESI) for is 291.1252, the measured value is 291.1265.

[0217] Cyclopropyl-[(5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl]methanone (A17) : TIFF2024513394000093.tif48170Under nitrogen, a 100 L reactor 1 was charged with compound A16 (4.52 kg, 15.54 mol, 1.00 equiv) and THF (47.95 kg, 10.6 vol). The internal temperature of reactor 1 was adjusted to -5°C ± 10°C. Cyclopropylmagnesium bromide solution (0.69 M in 2-MeTHF) (27.84 kg, 18.80 mol, 1.21 equiv) was charged slowly to reactor 1 over 2 hours while maintaining the internal temperature at -5°C ± 10°C and titrated with 2-butanol titration according to Sigma Aldrich Quality Control SOP. The titer of the commercial Grignard solution from Sigma Aldrich decreased from 0.89 M to 0.69 M over a 3 month period. The contents of Reactor 1 were stirred at -5°C ± 10°C for 30 minutes.

[0218] A 6M aqueous HCl solution was prepared by dissolving concentrated hydrochloric acid (6.55 kg) in water (16.94 kg, 17.10 L). A portion of the previously prepared 6M aqueous HCl solution (23.55 kg) and 2-MeTHF (38.15 kg, 44.3 L, 9.8 vol) were charged to reactor 2. Reactor 2 was cooled to -15°C. The contents of reactor 1 were slowly transferred to reactor 2 while maintaining the internal temperature of reactor 2 below 10°C. The quench is exothermic and a slow transfer is recommended. 2-MeTHF (4.29 kg, 4.98 L, 1.1 vol) was charged to reactor 1 and rinsed into reactor 2 while maintaining the internal temperature of reactor 2 below 10°C. The temperature of reactor 2 was adjusted to 20±5°C. The contents of reactor 2 were stirred for 15 minutes. The stirring was stopped and the phases were allowed to settle for 30 minutes.

[0219] The aqueous phase was drained into a glass carboy. A solution of K2HPO4 (2.24 kg) in water (20.33 kg, 20.33 L) was prepared. A portion of the previous K2HPO4 aqueous solution (22.55 kg) was transferred to reactor 2. The contents of reactor 2 were stirred for a minimum of 10 minutes. The stirring was stopped and the phases were allowed to settle for 10 minutes. The aqueous phase was drained into a glass carboy. A solution of NaCl (1.19 kg) in water (21.46 kg, 21.45 L) was charged to reactor 2. The contents of reactor 2 were stirred for 10 minutes. The stirring was stopped and the phases were allowed to settle for 30 minutes. The aqueous phase was drained into a glass carboy.

[0220] The contents of Reactor 2 were passed through a carbon cartridge in a Graver Filter Housing and polish filtered into a glass carboy over 2.0±1.5 hours (actual time: 2 hours). Reactor 2 was charged with THF (32.55 kg, 36.61 L, 8.1 vol) and the contents of Reactor 2 were pumped through a Graver C941 carbon cartridge in a Graver Filter Housing and polish filtered into a glass carboy over 60±55 minutes (actual time: 1 hour, reported (HPLC): cpd. A17%wt / wt: 11.46%).

[0221] The contents of the glass carboy were charged to reactor 1 and distilled under reduced pressure to a target volume of 17 L (3.8 vol). EtOH (17.05 kg, 21.61 L, 4.8 vol) was passed through a polishing filter and charged to reactor 1. The contents of reactor 1 were distilled under reduced pressure to a target volume of approximately 20 L (4.4 vol). EtOH (17.05 kg, 21.61 L, 4.8 vol) was passed through a polishing filter and charged to reactor 1. The contents of reactor 1 were distilled under reduced pressure to a target volume of approximately 21 L (4.6 vol).

[0222] Water (4.00 kg, 4.00 L, 0.9 vol) was passed through a polishing filter and charged to reactor 1. Reactor 1 was adjusted to a temperature of 65±5° C. over 20 minutes. A slurry of Compound A17 seeds (19.9 g, 0.073 mmol, 0.4 wt%) in polishing filtered water (201 g, 201 mL) and polishing filtered EtOH (156 g, 198 mL) was charged to reactor 1. The contents of reactor 1 began to form a suspension. Reactor 1 was stirred at 60±5° C. for 20 minutes. Water (15.65 kg, 15.65 L, 3.5 vol) was passed through a polishing filter and charged to reactor 1 while maintaining the temperature above 55° C. Reactor 1 was stirred at 60±5° C. for 30 minutes. The temperature was adjusted to 0±5° C. over 4.5 hours and the contents of reactor 1 were stirred at 0±5° C. for 8.5 hours.

[0223] The IKA Magic Lab Mill was equipped with a Dispax Reactor DR Module and a 2G4M6F Stator / Rotor. The IKA Magic Lab Mill was cooled to 0±5° C. and the grinding speed was set to 26000 RPM. The contents of Reactor 1 were passed through the IKA Magic Lab Mill into the filter dryer and the filtrate was collected in Reactor 3. Grind filtered EtOH (12.65 kg, 16.03 L, 3.5 vol) and grind filtered water (15.85 kg, 15.85 L, 3.5 vol) were charged to a glass carboy to prepare a cake wash. The cake wash was charged to Reactor 1 and the temperature of Reactor 1 was adjusted to 5° C. The contents were stirred for 30 minutes and the remaining contents of Reactor 1 were passed through the IKA Magic Lab Mill into the filter dryer and the filtrate was collected in Reactor 3. The contents of the filter dryer were dried under vacuum with a nitrogen sweep at ambient temperature for 19 hours. The filter dryer was adjusted to 40±5° C. and the contents were dried for 4 days.

[0224] The filter dryer was adjusted to a temperature of 20±10° C. and the light brown orange solid product cyclopropyl-[(5S,7S)-7-fluoro-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-2-yl]methanone A17 (3.54 kg, 13.17 mol, 85% yield) was discharged into a sealed bag.

[0225] HPLC 99.1 A%, >99.95:0.05 dr. Chiral HPLC >99.9% ee. 1H NMR (600 MHz, DMSO-d6) δ 7.48 - 7.35 (m, 3H), 7.33 - 7.21 (m, 2H), 6.24 (ddd, J = 56.4, 7.2, 2.0 Hz, 1H), 5.73 (ddd, J = 8.5, 6.5, 3.1 Hz, 1H), 3.75 (dddd, J = 25.7, 15.6, 8.6, 7.2 Hz, 1H), 3.04 - 2.95 (m, 1H), 2.72 (dddd, J = 26.8, 15.2, 3.2, 2.0 Hz, 1H), 1.14 - 0.99 (m, 4H). 13 C NMR (150 MHz, DMSO-d6) δ 192.7, 165.6, 159.1, 138.3, 129.1, 128.7, 126.7, 83.2, 60.0, 43.0, 17.9, 11.9, 11.8. 19 F NMR (565 MHz, DMSO-d6) δ -167.5.C 15 H 15 FN3O [M+H] + The calculated HRMS(ESI) for is 272.1197, the measured value is 272.1202.

[0226] Example 4: Preparation of (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one A12 TIFF2024513394000094.tif29170 Scheme 8 TIFF2024513394000095.tif28170

[0227] Scheme 9 The synthesis of (3R,5S)-1-amino-3-hydroxy-5-phenylpyrrolidin-2-one A12 is shown in Schemes 8 and 9.

[0228] Ethyl 2,4-dioxo-4-phenyl-butanoate (A20) : TIFF2024513394000096.tif29170A glass reactor was charged with toluene (80.1 kg, 10 wt. / wt., 11.9 vol.) and 20% EtONa / EtOH solution (31.5 kg, 92.6 mol, 1.4 eq.) at 20-30 °C. The contents were cycled from vacuum to nitrogen three times and then cooled to -5 to 5 °C. To the mixture was added A18 (10.1 kg, 69.1 mol, 1.1 eq.) dropwise at 0-10 °C. After the addition was complete, the mixture was stirred for 10 min, followed by the addition of a solution of A19 (7.7 kg, 64.1 mol, 1.0 eq.) in toluene (24.1 kg, 3.1 wt. / wt., 3.56 vol.). After the addition was complete, the mixture was stirred at -5 to 5 °C for 2 h. The pH of the quench mixture was adjusted to 2-4 with 2 M HCl (48.2 kg, 6.3 wt. / wt., 6.2 vol.) and the pH of the mixture was adjusted to 4-5. Separate the two phases. Wash the organic layer twice with 10% NaCl aq. (24.0 kg, 3.1 wt. / wt., 4.0 vol. each). Concentrate the organic layer to 1-2 vol. (7.7-15.4 L). Charge the concentrated mixture with EtOH (32.0 kg, 4.15 wt. / wt., 5 vol.) and concentrate to 1-2 vol. (7.7-15.4 L). Charge the mixture with EtOH (15.2 kg, 2.4 vol., 2.0 wt / wt.). Stir the mixture at 5-15 °C. Add seed A18 (0.16 kg, 0.01 wt. / wt.). Add water (12.8 kg, 1.7 wt. / wt., 1.7 vol.) over 6 h. After addition, heat the mixture to 35-45 °C. Stir the mixture at 35-45 °C for 0.5 h, then cool the mixture to 0-5 °C over 5 h. Stir the mixture at 0-5 °C for 5 h. Filter and wash with EtOH / H2O (1 / 5) (15.0 kg, 2.0 wt. / wt., 2.4 vol.). Dry the wet cake at 25-30 °C for 10-30 h to obtain a yellow solid (12.6 kg, 1.6 wt. / wt., 57.3 mol, 89.4% yield, 99.5% HPLC purity, 98.3% HPLC assay).

[0229] (-)-Ethyl (R)-2-hydroxy-4-oxo-4-phenylbutyrate (A7) : Alternative 1 : TIFF2024513394000097.tif40170A 300 L stirred tank is charged with Na2CO3 (6.8 kg, 0.54 w / w, 1.12 equiv.) and H2O (68.0 kg, 5.4 w / w, 5.4 vol.) and stirred at room temperature to dissolve. Place in barrel and label as 9% Na2CO3 solution.A 300 L stirred tank is charged with Na2CO3 (10.1 kg, 0.80 w / w, 1.66 equiv.) and H2O (50.4 kg, 4.0 w / w, 4.0 vol.) and stirred at room temperature to dissolve. Place in barrel and label as 16.7% Na2CO3 solution.The 300 L stirred tank is washed to neutral. A cleaned 300 L stirred tank was charged with HCl (18.0 kg, 1.43 w / w, 3.18 equiv.) and HO (15.4 kg, 1.22 w / w, 1.22 vol.) and allowed to dissolve with stirring at room temperature. Place in a barrel and label as 6M HCl solution. A 500 L stirred tank was charged with A18 (12.6 kg, 1 w / w, 57.3 mol, 1.00 equiv.) and ACN (2.6 kg, 0.21 w / w, 0.26 vol.) and allowed to dissolve with stirring at room temperature.

[0230] Charge a 50 L preparation vessel with K2HPO4 (0.4 kg, 0.032 w / w, 0.04 equiv.), KH2PO4 (0.6 kg, 0.048 w / w, 0.077 equiv.), glucose (16.8 kg, 1.33 w / w, 1.63 equiv.), and HO (50.5 kg, 4.0 w / w, 4 vol.) and dissolve by stirring at room temperature. Place in a barrel and label as Glycosylation Buffer Solution. Transfer to a 500 L stirred tank at 25-30 °C (recommended: 28 °C). Rinse the vessel with HO (6.2 kg, 0.49 w / w, 0.49 vol.), then transfer to a 500 L stirred tank. Stir at 25-30 °C and adjust pH to 6.0-6.5 by titrating with 9% Na2CO3. A 50 L preparation vessel is charged with NADP (0.069 kg, 0.0054 w / w, 0.0015 equiv.), KRED 1-200-0-16 (0.284 kg, 0.023 w / w), coenzyme 1-030-0-05 (0.035 kg, 0.0028 w / w), and HO (3.33 kg, 0.26 w / w, 0.26 vol.) and dissolved by stirring at room temperature. Then it is transferred to a 500 L stirred tank. The vessel is washed with HO (5.0 kg, 0.40 w / w, 0.4 vol.) and then it is transferred to a 300 L stirred tank. Stir at 25-30 °C (recommended: 28 °C) and titrate with 9% Na2CO3 (37.5 kg, approximately 2.97 vol%) for 7 h to maintain the pH between 6.0-6.5 (recommended: pH = 6.3).

[0231] Adjust the pH to 1.5-2.0 (recommended: pH=1.7) with 6M HCl solution (12.2 kg, approx. 0.97 vol.). Stir for 1 hour. Then, adjust the pH back to 6.0-6.5 with 16.7% Na2CO3 solution (22.6 kg, 1.79 w / w, approx. 1.8 vol.). Fill a 100L filter tank with Celite (2.1 kg, 0.17 w / w). Filter the reaction slurry to obtain filtrate 1 and cake 1. Transfer filtrate 1 to a 500L stirred tank and cake 1 to a 500L stirred tank. Fill the 500L stirred tank with MTBE (93.6 kg, 7.43 w / w) and stir at 25-30°C for 25-35 minutes (recommended: 30 minutes). Filter the slurry to obtain filtrate 2 and cake 2. Transfer filtrate 2 to a 500L stirred tank and extract filtrate 1. After phase separation, transfer the upper organic phase 1 to a transit drum and the lower aqueous phase 1 to a 500L stirred tank. Transfer cake 2 to a 300L stirred tank. Charge MTBE (93.2 kg, 7.40 w / w, 10 vol.) into the 500L stirred tank and stir for 25-35 minutes (recommended: 30 minutes). Filter the slurry to obtain filtrate 3 and cake 3. Discard cake 3. Transfer filtrate 3 to a 500L stirred tank and extract aqueous phase 1. After phase separation, transfer the upper organic phase 2 to a transit drum and the lower aqueous phase 2 to a 500L stirred tank.

[0232] A 500L stirred tank was charged with MTBE (93.1 kg, 7.39 w / w, 10 vol.) and stirred for 25-35 min (recommended: 30 min). The slurry was filtered to obtain filtrate 4 and cake 4. The filtrate 4 was transferred to a 500L stirred tank to extract the aqueous phase 2. After phase separation, the upper organic phase 3 was transferred to a transit drum and the lower aqueous phase 3 was discarded. The organic phases 1, 2, and 3 were combined and charged into a 500L stirred tank. Concentrate to 3-4V (37.8-57.4L) at -0.06MPa to -0.10MPa and 30-40℃ (optimum 40℃). Ethanol (approximately 59 kg*3, approximately 4.7V*3) was charged into the 500L stirred tank, and then concentrated three times to 3-4V (37.8-50.4L) to switch the solvent. A7 was obtained with purity >95.0% and chiral purity >99.0% (HPLC 97 A%, chiral HPLC >99% ee) The resulting solution was used directly in the next step.

[0233] Alternative 2 : TIFF2024513394000098.tif43170A 30 L reactor equipped with overhead stirring, pH probe, peristaltic pump, and jacket temperature 20-25 °C was charged with 0.2 M aqueous sodium acetate pH 7 (3.76 kg) followed by D-(+)-glucose (1.00 kg, 5064 mmol, 1.10 equiv). The reactor was rinsed by adding 0.2 M aqueous sodium acetate (3.76 kg) followed by 0.1 M aqueous magnesium chloride (0.20 kg). The contents of reactor 1 were stirred until dissolution was achieved (approximately 10 min). Next, A20 (1.00 kg, 90.3 wt%, 4100 mmol, 1.0 equiv) was charged as a solid, followed by ethanol (0.4 kg) and a pH probe was set up to continuously adjust the pH to 7 with 1 N aqueous sodium hydroxide (final consumption of 0.19 kg). NAD (0.01 kg, 15.0 mmol, 1 wt%) was charged as a solid, followed by a solution of GDH-105 (0.01 kg, 1 wt%) in 0.2 M aqueous sodium acetate pH 7 (0.2 kg) and a solution of ADH-114 (0.01 kg, 1 wt%) in 0.2 M aqueous sodium acetate pH 7.0 (0.2 kg). The reaction was incubated at 4°C for 1 h at 37°C for 2 h at 37°C. i Stirred at 25° C. for 24 h (>95 A% A7 in 24 h).

[0234] After the reaction was complete, the pH control was stopped and 6N aqueous hydrochloric acid (1.03 kg) was added to the reaction mixture until the pH was <2 and stirred for 1 hour. Then MTBE (7.37 kg) was added and the reaction was stirred vigorously for an additional 30 minutes. The layers were separated and the aqueous layer was back extracted two more times with MTBE (7.44 kg and 3.83 kg). The organic layers were then combined in another 30 L reactor and concentrated to 8 V by distillation (pressure 260 mbar, T j 60~70℃, T i 40-45 °C). Ethanol (9.32 kg) was then added and the reaction was concentrated to 8 V. A7 was obtained in approximately 95% purity and >99.0% chiral purity (HPLC 95 A%, Chiral HPLC >99% ee). The resulting solution was used directly in the next step.

[0235] tert-Butyl (R,E)-2-(4-ethoxy-3-hydroxy-4-oxo-1-phenylbutylidene)hydrazine-1-carboxylate (A8) : Alternative 1: TIFF2024513394000099.tif37170A glass reactor was charged with EtOH (97.4 kg, 10 vol, 7.9 wt / wt) and A7 (12.3 kg, 1.0 equiv, 55.3 mol). Formic acid (630 g, 0.25 equiv, 13.8 mol) was charged to the reactor. NH2NHBoc (10.1 kg, 1.3 equiv, 76.5 mol) was charged to the reactor in EtOH solution. The mixture was heated at 40-50 °C for 14 h. The mixture was concentrated to 1.5-2.5 vol. IPAc (109.2 kg, 10 vol., 8.9 wt / wt) was charged to the reactor. The mixture was concentrated to 3.0-3.5 vol. A sample was taken to check for remaining EtOH (Spec: ≦1.0%). Stir the mixture at 50-60 °C for 10-20 min. Add n-heptane (67.3 kg, 8.0 vol. 5.4 wt / wt) to the reactor over 5 min. Stir the mixture at 50-60 °C for 1-3 h. Cool to 10-20 °C over 5 h. Stir the mixture at 10-20 °C for 13-20 h. Take a sample to check the ratio of n-heptane to IPAc. (Spec: 2.6-3.5 (n-heptane / IPAc is 3 / 1 to 5 / 1)). Filter and wash the wet cake with n-heptane (21.6 kg, 2.5 vol., 1.75 wt / wt) to get 17.8 kg (1.45 wt / wt) of wet A8. Dry 15.5 kg (1.26 wt / wt) of wet cake at 40-50 °C for 10-20 h. A solid is obtained (13.7 kg, 40.7 mol, 81.2% yield for two steps, 99.8% HPLC purity, >99.9% chiral purity, 98.8% qNMR assay, EtOH, n-heptane, and IPAc residues were 817 ppm, 834 ppm, and 491 ppm, respectively. Other solvents were ND, enzyme residue was 26 ppm. KF was 0.10%. ROI was 0.08%).

[0236] Alternative 2: TIFF2024513394000100.tif36170A7 is T iA 30 L reactor containing a solution of formic acid (0.05 kg, 1024.8 mmol, 0.25 equiv.), tert-butyl carbazate (0.90 kg, 6148.8 mmol, 1.5 equiv.), and ethanol (1.31 kg) at T = 25° C. was charged. The reaction mixture was stirred overnight at T i =60°C (A8 to A7:>95:5 (A%:A%)). The reaction mixture was concentrated in vacuum to 5V (~50°C internal, ~275mbar). n-Heptane (3.26kg) was added and the contents of reactor 2 were concentrated in vacuum to 5V by vacuum distillation. n-Heptane (3.26kg) was added and the contents of the 30L reactor were concentrated in vacuum to 5V by vacuum distillation. n-Heptane (3.40kg) was added to the suspension and stirred at 45-50°C. Then MTBE (3.00kg) was added. The mixture was cooled to 0-5°C over 1 hour. Additional n-heptane (5.11kg) was added over 1 hour. The slurry was aged overnight and transferred to a filter dryer for filtration. The cake was washed with n-heptane (5.11kg) and then dried over the weekend at 25°C under a stream of nitrogen. A fine off-white solid was obtained in 67% yield with HPLC purity of 96.2 A%, KF=0.69 wt%.

[0237] Additional product (15% yield) was recovered from the wall cake by isolation and recrystallization using the following procedure: A 5 L reactor was charged with a slurry of A8 in MTBE (1.6 kg). The reactor contents were heated to 50° C. and stirred until dissolved. The temperature was then brought to 45° C., seeds were added (1 wt%), and the contents were then aged at 40° C. for 30 min. n-heptane (2.41 kg) was added over 1.5 h, followed by cooling to 5° C. over 1 h. The final slurry was aged at 5° C. for 1 h. The solid was filtered using a 3 L glass filter. The cake was washed with n-heptane (0.602 kg) and then dried overnight at 25° C. under a nitrogen stream. A fine off-white solid of A8 was obtained in 85% recovery with a purity of 96.2 A% by HPLC (HPLC 96.2 A%, 98.5 : 1.5 dr, chiral HPLC > 99% ee).

[0238] Exemplary compounds prepared by the above process are: 1 The H NMR data are provided herein along with the chiral column retention time (min) for the particular stereoisomer. Unless otherwise noted, the stereochemistry shown in each structure represents the relative configuration of a single stereoisomer, and the absolute configuration (i.e., "R" and / or "S") has been arbitrarily assigned.

[0239] All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, international publications, and non-patent literature cited herein are hereby incorporated by reference in their entirety.

[0240] Although the above-described embodiments of the invention have been described in some detail to facilitate understanding, it will be apparent that certain changes and modifications can be practiced within the scope of the appended claims. Accordingly, the described embodiments are considered to be illustrative and not restrictive, and the invention is not to be limited to the details shown herein, but may be modified within the scope of the appended claims and their equivalents.

Claims

1. A chiral bicyclic ketone compound of formula (I), or a stereoisomer thereof: or a pharma- ceutically acceptable salt thereof, comprising the formula: R １ is C １ -C ６ Alkyl, C ３ -C ６ Cycloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkyl, C １ -C ６ Haloalkoxy, C １ -C ６ Alkyl-N(R Ｎ ) ２ , phenyl, benzyl, 4- to 8-membered heterocyclyl, and 5- to 6-membered heteroaryl, where R １ is attached to the adjacent carbonyl by a carbon atom, and R １ is F, Cl, Br, C １ -C ６ Alkyl, C ３ -C ６ Cycloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkyl, C １ -C ６ Haloalkoxy, C １ -C ６ Alkyl-N(R Ｎ ) ２ , hydroxyl, hydroxymethyl, cyano, cyanomethyl, cyanoethyl, C(O)C １ -C ６ Alkyl, phenyl, benzyl, CH ２ - (C ３ -C ６ cycloalkyl), 5-6 membered heteroaryl, and CH ２ -(5- to 6-membered heteroaryl); Each R Ｎ is H, C １ -C ６ Alkyl, C ３ -C ６ Cycloalkyl, C １ -C ６ Alkoxy, and C １ -C ６ haloalkyl; or two R Ｎ may form a 4- to 6-membered ring together with the adjacent N; R ２ is H, C １ -C ６ Alkyl, C １ -C ６ Haloalkyl, C ３ -C ６ Cycloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, C １ -C ６ Thioalkyl, phenyl, benzyl, CH ２ - (C ３ -C ６ Cycloalkyl), CH ２ CH ２ - (C ３ -C ６ Cycloalkyl), CH ２ -(4- to 6-membered heterocyclyl), CH ２ CH ２ -(4- to 6-membered heterocyclyl), 5- to 6-membered heteroaryl, and CH ２ -(5-6 membered heteroaryl); where the phenyl ring, if present, is selected from the group consisting of halogen, C １ -C ４ Alkyl, C １ -C ４ Haloalkyl, C １ -C ４ Alkoxy, C １ -C ４ optionally substituted with 1 to 3 substituents selected from the group consisting of haloalkoxy and cyano; R ３ is D, halogen, OH, CN, C １ -C ４ Alkyl, C １ -C ４ Haloalkyl, cyclopropyl, C １ -C ４ Alkoxy and C １ -C ４ haloalkoxy; n is 1, 2, or 3; (a) a chiral N-aminolactam compound of formula p, or a stereoisomer thereof: or a salt thereof, in the presence of an acid additive and an alcohol solvent, with an imidate compound of formula c: or a salt thereof to produce a chiral bicyclic triazole compound of formula x, or a stereoisomer thereof: or a salt thereof, Pg １ is an optional hydroxyl protecting group, which may be the same or different at each occurrence; The chiral bicyclic triazole compound of formula x, or a stereoisomer thereof, is an intermediate compound in the preparation of the chiral bicyclic ketone compound of formula (I), or a stereoisomer thereof: method.

2. 2. The method of claim 1, wherein the acid additive is acetic acid and the organic solvent is EtOH.

3. (b) deoxyhalogenating a chiral bicyclic triazole compound of formula x, or a stereoisomer thereof, in the presence of a halogenating agent to obtain a chiral halogenated bicyclic compound of formula y, or a stereoisomer thereof:

3. The method of claim 1 or 2, further comprising forming a compound represented by the formula (I) or a salt thereof, wherein X is a halogen.

4. The method of claim 3, wherein the halogenating agent is PBSF or 2-pyridinesulfonyl fluoride.

5. 4. The process of claim 3, wherein the deoxyhalogenation step (b) is carried out in the presence of an organic base and an organic solvent.

6. The process of claim 5, wherein the organic base is N,N-diisopropylethylamine and the organic solvent is acetonitrile.

7. 4. The method of claim 3, wherein the deoxyhalogenation step (b) is further carried out in the presence of an additive that acts as a fluoride source.

8. 8. The method of claim 7, wherein the additive is triethylamine trihydrofluoride.

9. (c) contacting a chiral halogenated bicyclic compound of formula y, or a stereoisomer thereof, with an acid in the presence of an ethereal solvent / water mixture to produce a halogenated bicyclic carboxylic acid compound of formula z, or a stereoisomer thereof:

4. The method of claim 3, further comprising forming the compound, or a salt thereof.

10. 10. The process of claim 9, wherein the ethereal solvent / water mixture is a THF / water mixture and the acid is HCl.

11. (d) reacting a halogenated bicyclic carboxylic acid compound of formula z, or a stereoisomer thereof, with a compound of formula aa: or a salt thereof in the presence of a coupling agent to produce a chiral bicyclic amide compound of formula bb, or a stereoisomer thereof: or a salt thereof, wherein each Pg ５ The method of claim 9 , wherein:

12. The method of claim 11 , wherein the coupling agent is EDCI.

13. (e) reacting a chiral bicyclic amide compound of formula bb, or a stereoisomer thereof, with a compound of formula cc: or a salt thereof to produce a chiral bicyclic ketone compound of formula dd, or a stereoisomer thereof:

12. The method of claim 11, further comprising forming the compound, or a salt thereof.

14. 14. The method of claim 13 further comprising adding a seed of a chiral bicyclic ketone compound of formula dd.

15. The chiral bicyclic ketone compound of formula (I) or a pharma- ceutically acceptable salt thereof, wherein Each R ４ is H, F, Cl, C １ -C ６ Alkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, and C １ -C ６ haloalkoxy; m is 0, 1, 2, or 3; The method of claim 1.

16. The chiral bicyclic ketone compound of formula (I) or a pharma- ceutically acceptable salt thereof.

17. The stereoisomers of the chiral bicyclic ketone compound of formula (I) are represented by the formula (II): or a pharma- ceutically acceptable salt thereof.

18. The compound of formula (II) or a pharma- ceutically acceptable salt thereof, wherein Each R ４ is H, F, Cl, C １ -C ６ Alkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, and C １ -C ６ haloalkoxy; m is 0, 1, 2, or 3; 20. The method of claim 17.

19. The compound of formula (II) or a pharma- ceutically acceptable salt thereof.

20. 18. The method of claim 15 or 17, wherein m is 0.

21. R １ 20. The method of any one of claims 1, 2, 15 and 17, wherein is cyclopropyl.

22. A chiral N-aminolactam compound of formula p, or a stereoisomer thereof: or a salt thereof, comprising the formula: R ２ is H, C １ -C ６ Alkyl, C １ -C ６ Haloalkyl, C ３ -C ６ Cycloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, C １ -C ６ Thioalkyl, phenyl, benzyl, CH ２ - (C ３ -C ６ Cycloalkyl), CH ２ CH ２ - (C ３ -C ６ Cycloalkyl), CH ２ -(4- to 6-membered heterocyclyl), CH ２ CH ２ -(4- to 6-membered heterocyclyl), 5- to 6-membered heteroaryl, and CH ２ -(5-6 membered heteroaryl), and the phenyl ring, if present, is selected from the group consisting of halogen, C １ -C ４ Alkyl, C １ -C ４ Haloalkyl, C １ -C ４ Alkoxy, C １ -C ４ and optionally substituted with 1 to 3 substituents selected from the group consisting of haloalkoxy and cyano; n is 1, 2, or 3; (a) a chiral hydroxydicarboxylic acid compound of formula d, or a stereoisomer thereof: or a salt thereof, with an organic acid chloride of formula e: or a salt thereof to obtain a chiral cyclic carboxylic acid anhydride compound of formula f, or a stereoisomer thereof: or a salt thereof; (b) a protected hydrazone compound of formula l, or a stereoisomer thereof: or a salt thereof in the presence of an acid additive to produce a chiral hydroxyester hydrazine compound of formula m, or a stereoisomer thereof: or a salt thereof, Pg ２ is C １ -C ６ Alkyl, C ３ -C ６ may be substituted with cycloalkyl or aryl; Pg ３ is an optional hydroxyl protecting group, which may be the same or different in each occurrence; Pg ４ is an optional amine protecting group, which may be the same or different in each occurrence; A chiral cyclic carboxylic acid anhydride compound of formula f, or a stereoisomer thereof, is an intermediate in the preparation of a protected hydrazone compound of formula l, or a stereoisomer thereof, Chiral hydroxyester hydrazine compounds of formula m, or stereoisomers thereof, are intermediates in the preparation of chiral N-aminolactam compounds of formula p, or stereoisomers thereof: method.

23. 23. The method of claim 22, wherein the acid additive is AcOH.

24. (c) contacting a chiral cyclic carboxylic acid anhydride compound of formula f, or a stereoisomer thereof, with a reactive arene compound to produce a compound of formula h, or a stereoisomer thereof:

24. The method of claim 22 or 23, further comprising forming the compound or a salt thereof.

25. 25. The method of claim 24, wherein the reactive arene compound is benzene.

26. 25. The method of claim 24, wherein the contacting step (c) is carried out in the presence of a Lewis acid in an organic solvent.

27. Lewis acid is AlCl ３ and the organic solvent is CH ２ C ２ 27. The method of claim 26, wherein:

28. (d) reacting a compound of formula h, or a stereoisomer thereof, with a compound of formula i: to obtain a compound of formula j, or a stereoisomer thereof:

25. The method of claim 24, further comprising forming the compound, or a salt thereof.

29. 29. The method of claim 28, wherein the alcohol solvent of formula i is EtOH.

30. 30. The method of claim 28, wherein the reacting step (d) is carried out in the presence of an acid.

31. Acid is H ２ SO ４ The method of claim 30, wherein

32. (e) reacting a compound of formula j, or a stereoisomer thereof, with a hydrazine compound of formula k: or a salt thereof to form a protected hydrazone compound of formula l, or a stereoisomer thereof, or a salt thereof.

33. 33. The method of claim 32, wherein the contacting step (e) is carried out in the presence of an acid additive.

34. 34. The method of claim 33, wherein the acid additive is formic acid.

35. (f) reacting a protected hydrazone compound of formula l, or a stereoisomer thereof, to obtain a chiral protected N-aminolactam compound of formula n, or a stereoisomer thereof: or a salt thereof; (g) deprotecting the chiral protected N-aminolactam compound of formula n, or a stereoisomer thereof, to obtain a salt compound of formula o, or a stereoisomer thereof: and forming (h) reacting a salt compound of formula o, or a stereoisomer thereof, in the presence of a base to form a chiral N-aminolactam compound of formula p, or a stereoisomer thereof; 33. The method of claim 32, further comprising:

36. 36. The method of claim 35, wherein the base is NaOH in an aqueous base solution.

37. Optional Amine Protecting Group Pg ４ The method of claim 22 or 23, wherein is Boc.

38. 24. The method of any one of claims 1, 2, 15, 17, 22 and 23, wherein n is 1.

39. Imidate salt compounds of formula b: A process for the preparation of (a) a cyanoformate compound of formula a reacting in the presence of an anhydrous acid source in an alcohol solvent to form an imidate salt compound of formula b, １ is an optional hydroxyl protecting group, which may be the same or different in each occurrence.

40. 40. The method of claim 39, wherein the alcohol solvent is EtOH in MTBE.

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