Scalable Preparation of Polyketones

Abstract

The present invention particularly discloses a method for preparing polyketide compounds.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 883,491, filed on August 6, 2019, which is incorporated herein by reference in its entirety and for all purposes. Background Art

[0003] Although initial efforts proposed to rapidly translate small - molecule splicing regulators into the clinic for cancer patients, the inability to actually obtain gram - scale lead molecules with viable pharmacological properties has still hindered their clinical application. Here, we report a gram - scale method for the preparation of 17S - FD - 895, a highly potent and pharmacologically stable splicing regulator, which is supported by parallel, comprehensive structure - activity relationship (SAR) validation work. Summary of the Invention

[0004] In one aspect, a compound having the following formula is provided: wherein the compound is at least 95% enantiomerically pure.

[0005] In one aspect, a compound having the following formula is provided: wherein R 1 is a silyl protecting group and wherein the compound is at least 95% enantiomerically pure.

[0006] In one aspect, a compound having the following formula is provided: wherein the compound is at least 95% enantiomerically pure.

[0007] In one aspect, a compound having the following formula is provided: wherein the compound is at least 95% enantiomerically pure.

[0008] In one aspect, a compound having the following formula is provided: wherein R 1 is a silyl protecting group and wherein the compound is at least 95% enantiomerically pure.

[0009] In one aspect, a compound having the following formula is provided: wherein R 1 is a silyl protecting group and wherein the compound is at least 95% enantiomerically pure.

[0010] In one aspect, a compound having the following formula is provided: wherein the compound is at least 95% enantiomerically pure.

[0011] In one aspect, a compound having the following formula is provided: wherein the compound is at least 95% enantiomerically pure.

[0012] In one aspect, there is provided a compound having the following formula: wherein the compound is at least 95% enantiomerically pure.

[0013] In one aspect, there is provided a pharmaceutical composition comprising a compound having the following formula: and a pharmaceutically acceptable excipient, wherein the compound is at least 95% enantiomerically pure.

[0014] In one aspect, there is provided a method for preparing a compound having the following formula: comprising reacting a compound having the following formula: with 1-(dimethoxymethyl)-4-methoxybenzene in the presence of CBr4, an alcohol, a base, and one or more organic solvents.

[0015] In one aspect, there is provided a method for preparing a compound having the following formula: comprising reacting a compound having the following formula: with a transition metal catalyst for olefin metathesis in the presence of one or more organic solvents.

[0016] In one aspect, there is provided a method for preparing a compound having the following formula: comprising reacting a compound having the following formula: with the second-generation Hoveyda-Grubbs catalyst in the presence of toluene.

[0017] In one aspect, there is provided a method for preparing a compound having the following formula: comprising reacting a compound having the following formula: with a strong acid in the presence of an alcohol and one or more organic solvents.

[0018] In one aspect, there is provided a method for preparing a compound having the following formula: comprising reacting a compound having the following formula: with an acetylating agent in the presence of a strong acid and one or more organic solvents.

[0019] In an embodiment, there is provided a method for preparing a compound having the following formula: comprising reacting a compound having the following formula: with acetic anhydride in the presence of 4-dimethylaminopyridine and pyridine.

[0020] In one aspect, there is provided a method for preparing a linear polyketide compound.

[0021] On the one hand, a method for treating cancer is provided, which comprises administering to a subject in need an effective amount of a polyketide compound prepared by the method as described herein.

[0022] On the one hand, a method for preparing 17S-FD-895 is provided, which comprises using Compounds 6a, 6b, 6c, 6d and 6e as described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 . Synthetic design. The synthesis of 17S-FD-895 is generated by the coupling of two fragments given by side chain 3 and its related components 6e and 6d, and nucleus 2 and its three related components 6a-6c. The 11 sp 3 Stereocenters and stereochemical distributions of the three olefins of 1 are between the following components: 6a (containing the C6 and C7 stereodimers), 6b (inducing the C3 stereocenter and affecting the C8-C9 olefin), 6c (C10, C11 stereocenters, containing the C12-C13 olefin and inducing the C13-C14 stereochemistry and the C8-C9 olefin), 6d (containing the C20-C21 stereodimer, containing the functional group for installing the C18-C19 epoxide), and 6e (inducing the C16-C17 stereodimer). A list of the number of preparation steps (st), the number of chromatographic purifications (ch), the yield % (%y), and the amount of substance prepared so far (in g). Color-coded shading is used to highlight the assembly process.

[0024] Figures 2A - 2F . Synthetic problems. A list of the most prominent problems identified and remedied in the development of the gram-scale synthesis of 1. ( Figure 2A ) The conversion of 6a to 7 requires significant reaction adjustment. The solution stems from a method capable of in-situ converting the corresponding triol to the selectively protected pre-C6-C7 acetal 7. ( Figure 2B ) A two-day, 5-step method for converting 7 to 11 using single chromatographic purification was developed. This streamlined process can be carried out at an effective yield of ten grams of 11 obtained as a single stereoisomer substance. ( Figure 2C ) A problem in converting 7 to b is the formation of iso-11 due to the lack of enantiomeric purity of component 6c. 6c was resolved by esterification with (S)-mandelic acid to give 6c6 and 6c7, which can be separated by chromatography and subsequently hydrolyzed to give enantiopure 6a. ( Figure 2D ) Although it can be operated at the milligram level, the RCM on 16 provides a mixture of the desired product 18 and the related rearrangement product 17. ( Figure 2E ) Although the oxidation of 17 to remove the C7 alcohol enables the RCM to generate enone 19, reduction results in a 4:1 mixture of 20 and the desired 18. ( Figure 2F ) Impurities were observed at the compound 14 stage.

[0025] Figures 3A - 3B . Synthetic design.( Figure 3A ) The synthesis of 17S-FD-895 (1) is achieved by coupling of side chain 2 and core 3. The 11 sp 3 stereocenters and stereochemistry of 1 come from 12 precursors available in kilogram scale (inset). Key steps for preparation of each component are documented.( Figure 3B ) As by retrosynthetic analysis of related macrolide pladienolide B developed by Ghosh (25) from core 5a and Kotake (27) from core 5b. Color highlighting indicates source components as shown in the gray inset.

[0026] Figures 4A - 4F . Synthesis of 17S-FD-895 (1), single-carbon isotope-labeled materials, and stereoisomeric analogues.( Figure 4A ) Stille coupling of side chain 2 and core 3 gives 1 with an effective mass balance.( Figure 4B ) 13 C1-17S-FD-895 (Scheme AS1( Figure 10 )) and 13 C30-17S-FD-895 (Scheme AS2) are synthesized by installing the 13 C-containing precursors into the pathways of Scheme A1-A2. 13 C-NMR returns a singlet, indicating the presence of a single isomeric substance in these batches.( Figure 4C ) SAR determined by analogue development. Red spheres represent unexplored stereoisomers.( Figures 4D - 4F ) Synthetic analogues 1a-1c are made and their GI 50 values are evaluated in HCT-116 cells. Selected regions of the 1 H NMR spectra are provided to illustrate chemical shift alterations.( Figure 4D ) Inversion of the C3 stereocenter in 1a is achieved by replacing dichlorophenylborane and (-)-sparteine in the Sammakia aldol addition with TiCl4 and diisopropylamine (Scheme AS3).( Figure 4E ) The C7 nuclear isomer 1b is synthesized from 34 in 6 steps (Scheme AS4).( Figure 4F ) The C18-C19 epoxide isomer 1d is prepared by isolating the minor Sharpless epoxide during the preparation of 2.

[0027] Figures 5A - 5I . X-ray crystal structures depict the binding of pladienolide B (PDB ID 6EN4), FD-895 (18), and CYP (18) within the SF3B core. Observed The side chains of the residues within Figure 5A : Pradienolide B, Figure 5B : FD-895, and Figure 5C : CYPB, corresponding to SF3B1 and PHF5A. The van der Waals surface is drawn to depict the following nuclei: Figure 5D : Pradienolide B, Figure 5E : FD-895, and Figure 5F : CYPB and shows the following side chains: Figure 5G : Pradienolide B, Figure 5H : FD-895, and Figure 5I : CYPB. Surface plots depicting Pradienolide B, FD-895, or CYPB. The structure of Pradienolide B bound to the SF3B nucleus is described in (14). Discussions of the structures of FD-895 and the cyclopropane analogue CYPB are provided in (18).

[0028] Figure 6 . LC-MS traces. Using positive-ion mode electrospray ionization (ESI) as the ion source, 20 - 40 μL samples prepared in EtOH or DMSO were injected into an Agilent 1260 liquid chromatography (LC) system coupled with a Thermo LCQdeca mass spectrometer (MS). LC separation was carried out using a Phenomenex Kinetex EVO C18 (ID 2.1 mm × length 50 mm, particle size 5.0 μm), with water containing 0.1% formic acid as mobile phase A and acetonitrile containing 0.1% formic acid as mobile phase B. The LC flow rate was set at 0.30 mL / min. The LC gradient was set as follows: 0 min: 5% mobile phase B; 10 min: 95% mobile phase B; 12 min: 95% mobile phase B; 13 min: 5% mobile phase B; and 18 min: 5% mobile phase B. The total run time was 18 min. The UV detection wavelength was set at 254 nm (17S-FD-895 can be observed using 254 nm detection). MS and HRMS were generally observed in the positive mode of sodium ions (C 31 H 50 O9Na[M+Na] + Calculated HR-ESI-MS m / z value for

[0029] Figures 7A - 7C . NMR comparison. The bar chart depicts the 1 H (left) and 13 C (right) chemical shift differences between FD-895 (gray inset, upper right) and Figure 7A : 17S-FD-895 (1), Figure 7B: 3S, 17S-FD-895(1a) or Figure 7C : 7R, 17S-FD-895(1b).

[0030] Figure 8 . Protocol A1. The side chain 2 is synthesized in 11 steps starting from Crimmins auxiliary 6. The yields and stereoselectivities shown reflect improvements for gram-scale production. Compounds 6 and 7 are purified by recrystallization. The colored highlights indicate carbon ( Figure 1 ) from the source precursor. Abbreviations: DMAP, dimethylaminopyridine; DIBAL-H, diisobutylaluminum hydride; DET, diethyl tartrate; i-Pr, isopropyl; t-Bu, tert-butyl; TEMPO, 2,2,6,6-tetramethylpiperidine-N-oxide; n-Bu, butyl.

[0031] Figure 9 . Protocol A2. Synthesis of the core 3 from the monoprotected 1,4-butanediol 18. The yields and stereoselectivities shown reflect improvements for gram-scale production. Abbreviations: Ipc, isopinocampheyl; Ph, phenyl; TBSOTf; tert-butyldimethylsilyl trifluoromethanesulfonate; HGII, second-generation Hoveyda-Grubbs catalyst; CSA, (1S)-(+)-10-camphorsulfonic acid.

[0032] Figure 10 . Protocol AS1. The black spheres indicate 13 the positions labeled with

[0033] Figure 11 . Protocol AS3. The carbon attached to the -OTBS group and the carbon atoms on either side of this carbon include the isomer installation region.

[0034] Figure 12 . Protocol AS4. Carbon 7 and the two adjacent carbons include the isomer installation region. Detailed Description

[0035] I. Definitions

[0036] The abbreviations used herein have their conventional meanings in the fields of chemistry and biology. The chemical structures and chemical formulas described herein are constructed according to the standard rules of chemical valence known in the field of chemistry.

[0037] When substituents are described using a conventional chemical formula written from left to right, the substituents equally encompass the chemically identical substituents obtained from writing the structure from right to left, e.g., -CH2O- is equivalent to -OCH2-.

[0038] Unless otherwise specified, the term "alkyl" by itself or as part of another substituent means a straight-chain (i.e., unbranched) or branched carbon chain (or carbon) or a combination thereof, which may be fully saturated, mono-unsaturated, and / or poly-unsaturated, and may contain monovalent, divalent, and polyvalent groups. An alkyl may contain a specified number of carbons (e.g., C1-C 10 means from one carbon to ten carbons). In an embodiment, the alkyl is fully saturated. In an embodiment, the alkyl is mono-unsaturated. In an embodiment, the alkyl is poly-unsaturated. An alkyl is an uncyclized chain. Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, methyl, homologs and isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. An unsaturated alkyl group is an alkyl group having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. An alkoxy is an alkyl attached to the remainder of the molecule through an oxygen linker (-O-). The alkyl moiety may be an alkenyl moiety. The alkyl moiety may be an alkynyl moiety. An alkenyl contains one or more double bonds. An alkynyl contains one or more triple bonds. The alkyl moiety may be fully saturated. An alkenyl may contain more than one double bond and / or one or more triple bonds in addition to one or more double bonds. An alkynyl may contain more than one triple bond and / or one or more double bonds in addition to one or more triple bonds.

[0039] Unless otherwise specified, the term "alkylene" by itself or as part of another substituent means a divalent group derived from an alkyl, such as (but not limited to) -CH2CH2CH2CH2-. Generally, an alkyl (or alkylene) will have from 1 to 24 carbon atoms, and those groups having 10 or fewer carbon atoms are preferred herein. "Lower alkyl" or "lower alkylene" is a short-chain alkyl or alkylene group that generally has eight or fewer carbon atoms. Unless otherwise specified, the term "alkenylene" by itself or as part of another substituent means a divalent group derived from an alkene. Unless otherwise specified, the term "alkynylene" by itself or as part of another substituent means a divalent radical derived from an alkyne. In an embodiment, the alkylene is fully saturated. In an embodiment, the alkylene is mono-unsaturated. In an embodiment, the alkylene is poly-unsaturated. An alkenylene contains one or more double bonds. An alkynylene contains one or more triple bonds.

[0040] Unless otherwise specified, whether alone or in combination with another term, the term "heteroalkyl" means a stable straight-chain or branched-chain containing at least one carbon atom and at least one heteroatom (e.g., O, N, P, Si, and S, and wherein nitrogen and sulfur atoms may optionally be oxidized, and nitrogen heteroatoms may optionally be quaternized), or a combination thereof. The heteroatom (e.g., O, N, S, Si, or P) can be placed at any internal position of the heteroalkyl group or at the position where the alkyl group is attached to the remainder of the molecule. Heteroalkyl is an uncyclized chain. Examples include, but are not limited to: -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-S-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, -O-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms can be consecutive, such as -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. The heteroalkyl moiety can contain one heteroatom (e.g., O, N, S, Si, or P). The heteroalkyl moiety can contain two optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety can contain three optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety can contain four optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety can contain five optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety can contain up to 8 optionally different heteroatoms (e.g., O, N, S, Si, or P). Unless otherwise specified, the term "heteroalkenyl" by itself or in combination with another term means a heteroalkyl containing at least one double bond. In addition to one or more double bonds, heteroalkenyl can optionally contain more than one double bond and / or one or more triple bonds. Unless otherwise specified, the term "heteroalkynyl" by itself or in combination with another term means a heteroalkyl containing at least one triple bond. Heteroalkynyl can optionally contain more than one triple bond and / or one or more double bonds in addition to one or more triple bonds. In an embodiment, the heteroalkyl is fully saturated. In an embodiment, the heteroalkyl is monounsaturated. In an embodiment, the heteroalkyl is polyunsaturated.

[0041] Similarly, unless otherwise specified, whether alone or as part of another substituent, the term "heteroalkylene" means a divalent group derived from a heteroalkyl group, such as (but not limited to) -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. For heteroalkylene, the heteroatom(s) can also occupy any one or two of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkylamino, alkylenediamino, etc.). Still further, for alkylene and heteroalkylene linking groups, the direction in which the formula of the linking group is written does not imply the orientation of the linking group. For example, the formula -C(O)2R'- represents both -C(O)2R'- and -R'C(O)2-. As described above, the heteroalkyl groups used herein include those groups attached to the remainder of the molecule through a heteroatom, such as -C(O)R', -C(O)NR', -NR'R", -OR', -SR', and / or -SO2R'. It should be understood that in cases where "heteroalkyl" is recited after a specific heteroalkyl group such as -NR'R", the terms heteroalkyl and -NR'R" are not redundant or mutually exclusive. Instead, the specific heteroalkyl group is recited for increased clarity. Thus, the term "heteroalkyl" should not be construed herein to exclude specific heteroalkyl groups such as -NR'R". Unless otherwise specified, the term "heteroalkenylene" alone or as part of another substituent means a divalent group derived from a heteroalkene. Unless otherwise specified, the term "heteroalkynylene" alone or as part of another substituent means a divalent group derived from a heteroalkyne. In embodiments, the heteroalkylene is fully saturated. In embodiments, the heteroalkylene is mono-unsaturated. In embodiments, the heteroalkylene is poly-unsaturated. The heteroalkenylene contains one or more double bonds. The heteroalkynylene contains one or more triple bonds.

[0042] Unless otherwise specified, whether alone or in combination with other terms, the terms "cycloalkyl" and "heterocycloalkyl" respectively refer to the cyclic forms of "alkyl" and "heteroalkyl". Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, the heteroatom can occupy the position where the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidyl, 2-piperidyl, 3-piperidyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. "Cycloalkylene" and "heterocycloalkylene", whether alone or as part of another substituent, refer to divalent groups respectively derived from cycloalkyl and heterocycloalkyl. In an embodiment, the cycloalkyl is fully saturated. In an embodiment, the cycloalkyl is monounsaturated. In an embodiment, the cycloalkyl is polyunsaturated. In an embodiment, the heterocycloalkyl is fully saturated. In an embodiment, the heterocycloalkyl is monounsaturated. In an embodiment, the heterocycloalkyl is polyunsaturated.

[0043] In an embodiment, the term "cycloalkyl" refers to a monocyclic, bicyclic, or polycyclic cycloalkyl ring system. In an embodiment, the monocyclic ring system is a cyclic hydrocarbon group containing 3 to 8 carbon atoms, where such a group can be saturated or unsaturated, but not aromatic. In an embodiment, the cycloalkyl group is fully saturated. Examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. The bicyclic cycloalkyl ring system is a bridged monocyclic or fused bicyclic. In an embodiment, the bridged monocyclic contains a monocyclic cycloalkyl ring, where two non-adjacent carbon atoms of the monocyclic are connected by an alkylene bridge between one carbon atom and three other carbon atoms (i.e., of the form (CH2) wa bridging group, where w is 1, 2, or 3). Representative examples of bicyclic ring systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and bicyclo[4.2.1]nonane. In an embodiment, the fused bicycloalkyl ring system contains a monocycloalkyl ring fused to a phenyl, monocycloalkyl, monocycloalkenyl, monocyclic heterocyclic, or monocyclic heteroaryl group. In an embodiment, the bridged or fused bicycloalkyl is attached to the parent molecular moiety through any carbon atom contained within the monocycloalkyl ring. In an embodiment, the cycloalkyl group is optionally substituted with one or two groups independently selected from oxo or thio. In an embodiment, the fused bicycloalkyl is a 5- or 6-membered monocycloalkyl ring fused to a benzene ring, 5- or 6-membered monocycloalkyl, 5- or 6-membered monocycloalkenyl, 5- or 6-membered monocyclic heterocyclic, or 5- or 6-membered monocyclic heteroaryl group, where the fused bicycloalkyl is optionally substituted with one or two groups independently selected from oxo or thio. In an embodiment, the polycyclic cycloalkyl ring system is a monocycloalkyl ring (base ring) fused to any one of the following: (i) a ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicycloalkyl, bicycloalkenyl, and bicyclic heterocyclic; or (ii) two other ring systems independently selected from the group consisting of phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl, and monocyclic or bicyclic heterocyclic. In an embodiment, the polycyclic cycloalkyl is attached to the parent molecular moiety through any carbon atom contained within the base ring. In an embodiment, the polycyclic cycloalkyl ring system is a monocycloalkyl ring (base ring) fused to any one of the following: (i) a ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicycloalkyl, bicycloalkenyl, and bicyclic heterocyclic; or (ii) two other ring systems independently selected from the group consisting of phenyl, monocyclic heteroaryl, monocycloalkyl, monocycloalkenyl, and monocyclic heterocyclic. Examples of polycyclic cycloalkyl groups include, but are not limited to, tetradecahydrophenanthrenyl, perhydro-10H-phenothiazin-1-yl, and perhydro-10H-phenoxazin-1-yl. A bicyclic or polycyclic cycloalkyl ring system refers to multiple rings fused together, where at least one fused ring is a cycloalkyl ring, and where the multiple rings are attached to the parent molecular moiety through any carbon atom contained within the cycloalkyl rings of the multiple rings.

[0044] In an embodiment, the cycloalkyl group is a cycloalkenyl group. The term "cycloalkenyl" is used according to its ordinary and general meaning. In an embodiment, the cycloalkenyl group is a monocyclic, bicyclic or polycyclic cycloalkenyl ring system. In an embodiment, the monocyclic cycloalkenyl ring system is a cyclic hydrocarbon group containing 3 to 8 carbon atoms, wherein such a group is unsaturated (i.e., contains at least one cyclic carbon-carbon double bond), but not aromatic. Examples of the monocyclic cycloalkenyl ring system include cyclopentenyl and cyclohexenyl. In an embodiment, the bicyclic cycloalkenyl ring is a bridged monocyclic or fused bicyclic. In an embodiment, the bridged monocyclic contains a monocyclic cycloalkenyl ring, wherein two non-adjacent carbon atoms of the monocyclic are connected by an alkylene bridge between one carbon atom and three other carbon atoms (i.e., a bridging group of the form (CH2) w where w is 1, 2 or 3). Representative examples of the bicyclic cycloalkenyl include but are not limited to norbornyl and bicyclo[2.2.2]oct-2-enyl. In an embodiment, the fused bicyclic cycloalkenyl ring system contains a monocyclic cycloalkenyl ring fused to any one of phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclic or monocyclic heteroaryl. In an embodiment, the bridged or fused bicyclic cycloalkenyl is attached to the parent molecular moiety through any carbon atom contained within the monocyclic cycloalkenyl ring. In an embodiment, the cycloalkenyl group is optionally substituted with one or two groups independently selected from oxo or thio. In an embodiment, the polycyclic cycloalkenyl ring system contains a monocyclic cycloalkenyl ring (base ring) fused to any one of: (i) a ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl and bicyclic heterocyclic; or (ii) two ring systems independently selected from the group consisting of phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl and monocyclic or bicyclic heterocyclic. In an embodiment, the polycyclic cycloalkenyl is attached to the parent molecular moiety through any carbon atom contained within the base ring. In an embodiment, the polycyclic cycloalkenyl ring system contains a monocyclic cycloalkenyl ring (base ring) fused to any one of: (i) a ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl and bicyclic heterocyclic; or (ii) two ring systems independently selected from the group consisting of phenyl, monocyclic heteroaryl, monocyclic cycloalkyl, monocyclic cycloalkenyl and monocyclic heterocyclic. The bicyclic or polycyclic cycloalkenyl ring system refers to a plurality of rings fused together, wherein at least one of the fused rings is a cycloalkenyl ring, and wherein the plurality of rings are attached to the parent molecular moiety through any carbon atom contained within the cycloalkenyl ring of the plurality of rings.

[0045] In an embodiment, the term "heteroalkyl" means a monocyclic, bicyclic, or polycyclic heteroalkyl ring system. In an embodiment, the heteroalkyl group is fully saturated. A bicyclic or polycyclic heteroalkyl ring system refers to multiple rings fused together, where at least one fused ring is a heteroalkyl ring, and where the multiple rings are attached to the parent molecular moiety through any carbon atoms contained within the heteroalkyl ring of the multiple rings.

[0046] In an embodiment, a heterocycloalkyl is a heterocyclic group. As used herein, the term "heterocyclic group" means a monocyclic, bicyclic, or polycyclic heterocycle. A heterocyclic group monocyclic heterocycle is a 3-, 4-, 5-, 6-, or 7-membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S, wherein the ring is saturated or unsaturated but not aromatic. A 3- or 4-membered ring contains 1 heteroatom selected from the group consisting of O, N, and S. A 5-membered ring may contain zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. A 6- or 7-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. The heterocyclic group monocyclic heterocycle is attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the heterocyclic group monocyclic heterocycle. Representative examples of heterocyclic group monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, thiazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofurfuryl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, tetrahydrothiazolyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. A heterocyclic group bicyclic heterocycle is a monocyclic heterocycle fused to any one of phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocycle, or monocyclic heteroaryl. The heterocyclic group bicyclic heterocycle is attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the monocyclic heterocycle portion of the bicyclic ring system. Representative examples of bicyclic heterocyclic groups include, but are not limited to, 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuran-3-yl, indol-1-yl, indol-2-yl, indol-3-yl, 2,3-dihydrobenzothiophen-2-yl, decahydroquinolinyl, decahydroisoquinolinyl, octahydro-1H-indolyl, and octahydrobenzofuranyl. In an embodiment, the heterocyclic group is optionally substituted with one or two groups independently being oxo or thio. In certain embodiments, the bicyclic heterocyclic group is a 5- or 6-membered monocyclic heterocyclic group ring fused to a benzene ring, 5- or 6-membered monocyclic cycloalkyl, 5- or 6-membered monocyclic cycloalkenyl, 5- or 6-membered monocyclic heterocyclic group, or 5- or 6-membered monocyclic heteroaryl, wherein the bicyclic heterocyclic group is optionally substituted with one or two groups independently being oxo or thio.The polycyclic heterocyclic ring system is a monocyclic heterocyclic ring (base ring) fused to any one of the following: (i) a ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclic; or (ii) two other ring systems independently selected from the group consisting of phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl, and monocyclic or bicyclic heterocyclic. The polycyclic heterocyclic is attached to the parent molecular moiety through any carbon atom or nitrogen atom contained in the base ring. In an embodiment, the polycyclic heterocyclic ring system is a monocyclic heterocyclic ring (base ring) fused to any one of the following: (i) a ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclic; or (ii) two other ring systems independently selected from the group consisting of phenyl, monocyclic heteroaryl, monocyclic cycloalkyl, monocyclic cycloalkenyl, and monocyclic heterocyclic. Examples of polycyclic heterocyclic groups include but are not limited to 10H-phenothiazin-10-yl, 9,10-dihydroacridin-9-yl, 9,10-dihydroacridin-10-yl, 10H-phenoxazin-10-yl, 10,11-dihydro-5H-dibenzo[b,t]azepin-5-yl, 1,2,3,4-tetrahydropyrido[4,3-g]isoquinolin-2-yl, 12H-benzo[b]phenoxazin-12-yl, and dodecahydro-1H-carbazol-9-yl.

[0047] Unless otherwise specified, the term "halo" or "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" mean including mono-haloalkyl and poly-haloalkyl. For example, the term "halo(C1-C4)alkyl" includes but is not limited to fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0048] Unless otherwise indicated, the term "acyl" means -C(O)R, where R is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0049] Unless otherwise specified, the term "aryl" means a polyunsaturated aromatic hydrocarbon substituent, which may be a monocyclic or fused together (i.e., fused-ring aryl) or covalently linked multiple rings (preferably, 1 to 3 rings). Fused-ring aryl means multiple rings fused together, where at least one fused ring is an aromatic ring, and where the multiple rings are attached to the parent molecular moiety through any carbon atoms contained in the aromatic rings of the multiple rings. The term "heteroaryl" means an aryl (or ring) containing at least one heteroatom such as N, O, or S, where the nitrogen and sulfur atoms are optionally oxidized, and one or more nitrogen atoms are optionally quaternized. Thus, the term "heteroaryl" includes fused-ring heteroaryl groups (i.e., multiple rings fused together, where at least one fused ring is a heteroaromatic ring, and where the multiple rings are attached to the parent molecular moiety through any atoms contained in the heteroaromatic rings of the multiple rings). 5,6-fused-ring heteroarylene means two rings fused together, where one ring has 5 members and the other ring has 6 members, and where at least one ring is a heteroaryl ring. Similarly, 6,6-fused-ring heteroarylene means two rings fused together, where one ring has 6 members and the other ring has 6 members, and where at least one ring is a heteroaryl ring. And 6,5-fused-ring heteroarylene means two rings fused together, where one ring has 6 members and the other ring has 5 members, and where at least one ring is a heteroaryl ring. Heteroaryl groups can be attached to the rest of the molecule through carbon or heteroatoms. Non-limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidinyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothienyl, isoquinolinyl, quinoxalinyl, quinolinyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolinyl, 5-isoquinolinyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolinyl, and 6-quinolinyl. Substituents of each of the above-indicated aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. Alone or as part of another substituent, "arylene" and "heteroarylene" mean divalent groups derived from aryl and heteroaryl, respectively. A heteroaryl substituent can be -O- bonded to the ring heteroatom nitrogen.

[0050] A fused polyheterocycloalkyl-aryl is an aryl fused to a heterocycloalkyl. A fused polyheterocycloalkyl-heteroaryl is a heteroaryl fused to a heterocycloalkyl. A fused polyheterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl. A fused polyheterocycloalkyl-heterocycloalkyl is a heterocycloalkyl fused to another heterocycloalkyl. A fused polyheterocycloalkyl-aryl, a fused polyheterocycloalkyl-heteroaryl, a fused polyheterocycloalkyl-cycloalkyl, or a fused polyheterocycloalkyl-heterocycloalkyl may each independently be unsubstituted or substituted with one or more substituents described herein.

[0051] A spiro ring is two or more rings in which adjacent rings are joined by a single atom. The individual rings within a spiro ring may be the same or different. The individual rings in a spiro ring may be substituted or unsubstituted and may have substituents different from those of other individual rings in the spiro ring collection. The possible substituents of an individual ring within a spiro ring are the possible substituents of the same ring when not part of a spiro ring (e.g., substituents of a cycloalkyl ring or a heterocycloalkyl ring). A spiro ring may be a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted cycloalkylene, a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted heterocycloalkylene, and the individual rings within a spiro ring group may be any of the rings in the immediately preceding list, including all rings of one type (e.g., all rings of a substituted heterocycloalkylene, where each ring may be the same or different substituted heterocycloalkylene). When referring to a spiro ring system, a heterospiro ring means a spiro ring in which at least one ring is a heterocycle and in which each ring may be a different ring. When referring to a spiro ring system, a substituted spiro ring means that at least one ring is substituted and each substituent may optionally be different.

[0052] The symbol represents the point of attachment of a chemical moiety to the remainder of a molecule or chemical formula.

[0053] As used herein, the term "oxo" means an oxygen double-bonded to a carbon atom.

[0054] As used herein, the term "alkylsulfonyl" refers to a moiety having the formula -S(O2)-R′, where R′ is a substituted or unsubstituted alkyl as defined above. R′ may have a specified number of carbons (e.g., "C1-C4 alkylsulfonyl").

[0055] The term "alkylene aryl" is an arylene moiety covalently bonded to an alkylene moiety (also referred to herein as an alkylene linker). In an embodiment, an alkylene aryl group has the following formula:

[0056]

[0057] The alkylarylene moiety may be substituted (e.g., by substituents) on the alkylene moiety or the arylene linker (e.g., at carbon 2, 3, 4, or 6) with: halogen, oxo, -N3, -CF3, -CCl3, -CBr3, -CI3, -CN, -CHO, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO2CH3, -SO3H, -OSO3H, -SO2NH2, NHNH2, ONH2, NHC(O)NHNH2, substituted or unsubstituted C1-C5 alkyl, or substituted or unsubstituted 2- to 5-membered heteroalkyl). In an embodiment, the alkylarylene is unsubstituted.

[0058] Each of the above terms (e.g., "alkyl", "heteroalkyl", "cycloalkyl", "heterocycloalkyl", "aryl", and "heteroaryl") includes both substituted and unsubstituted forms of the indicated group. Preferred substituents for each type of group are provided below.

[0059] Substituents of alkyl and heteroalkyl (including those commonly referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl and heterocycloalkenyl) can be selected from one or more of the following various groups: -OR′, =O, =NR′, =N-OR′, -NR′R″, -SR′, -halogen, -SiR′R″R″′, -OC(O)R′, -C(O)R′, -CO2R′, -CONR′R″, -OC(O)NR′R″, -NR″C(O)R′, -NR′-C(O)NR″R″′, -NR″C(O)2R′, -NR-C(NR′R″R″′)=NR″″, -NR-C(NR′R″)=NR″′, -S(O)R′, -S(O)2R′, -S(O)2NR′R″, -NRSO2R′, NR′NR″R″′, ONR′R″, NR′C(O)NR″NR″′R″″, -CN, -NO2, -NR′SO2R″, -NR′C(O)R″, -NR′C(O)-OR″, -NR′OR″, in the range of zero to (2m′ + 1), where m′ is the total number of carbon atoms in such groups. R, R′, R″, R″′ and R″″ each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1 - 3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy or thioalkoxy or aralkyl. When the compounds described herein contain more than one R group, for example, when there are more than one of these groups, each R group is independently selected as the R′ group, R″ group, R″′ group and R″″ group are independently selected. When R′ and R″ are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 4-, 5-, 6- or 7-membered ring. For example, -NR′R″ includes but is not limited to 1-pyrrolidinyl and 4-morpholinyl. Based on the above discussion of substituents, those skilled in the art will understand that the term “alkyl” is intended to include groups containing carbon atoms bonded to groups other than hydrogen groups, such as haloalkyl (e.g., -CF3 and -CH2CF3) and acyl (e.g., -C(O)CH3, -C(O)CF3, -C(O)CH2OCH3, etc.).

[0060] Substituents of aryl and heteroaryl are different from those described for alkyl and are selected from, for example, the following: -OR′, -NR′R″, -SR′, -halogen, -SiR′R″R″′, -OC(O)R′, -C(O)R′, -CO2R′, -CONR′R″, -OC(O)NR′R″, -NR″C(O)R′, -NR′-C(O)NR″R″′, -NR″C(O)2R′, -NR-C(NR′R″R″′)═NR″″, -NR-C(NR′R″)═NR″′, -S(O)R′, -S(O)2R′, -S(O)2NR′R″, -NRSO2R′, NR′NR″R″′, ONR′R″, NR′C(O)NR″NR″′R″″, -CN, -NO2, -R′, -N3, -CH(Ph)2, fluoro(C1-C4)alkoxy and fluoro(C1-C4)alkyl, -NR′SO2R″, -NR′C(O)R″, -NR′C(O)-OR″, -NR′OR″, in a number ranging from zero to the total number of open valences on the aromatic ring system; and wherein R′, R″, R″′ and R″″ are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl. When the compounds described herein contain more than one R group, each R group is independently selected, for example, as each R′ group, R″ group, R″′ group and R″″ group is independently selected when more than one of these groups are present.

[0061] Substituents of a ring (e.g., cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene) may be depicted as substituents on the ring rather than on a particular atom of the ring (commonly referred to as floating substituents). In such cases, the substituent may be attached to any of the ring atoms (subject to valence rules), and in the case of fused or spiro rings, a substituent depicted as associated with a member of the fused or spiro ring (a floating substituent on a monocyclic ring) may be a substituent on any of the fused or spiro rings (a floating substituent on a polycyclic ring). When a substituent is attached to a ring rather than a particular atom (a floating substituent) and the subscript of the substituent is an integer greater than one, multiple substituents may be on the same atom, the same ring, different atoms, different fused rings, different spiro rings, and each substituent may optionally be different. In cases where the point of attachment of the ring to the remainder of the molecule is not limited to a single atom (a floating substituent), the point of attachment may be any atom of the ring, and in the case of fused or spiro rings, may be any atom of any of the fused or spiro rings (subject to valence rules). In cases where the ring, fused ring, or spiro ring contains one or more ring heteroatoms and the ring, fused ring, or spiro ring is shown to have yet another floating substituent (including but not limited to the point of attachment to the remainder of the molecule), the floating substituent may be bonded to the heteroatom. In cases where a ring heteroatom is shown to be hydrogen-bonded to one or more in a structure or formula having a floating substituent (e.g., a ring nitrogen having two bonds to ring atoms and a third bond to hydrogen), when the heteroatom is bonded to the floating substituent, the substituent will be understood to replace hydrogen while following valence rules.

[0062] Two or more substituents may optionally be joined to form an aryl, heteroaryl, cycloalkyl, or heterocycloalkyl. Such so-called ring-forming substituents are generally (although not necessarily) attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring-forming substituents attached to adjacent members of a cyclic base structure produce a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring-forming substituents attached to a single member of a cyclic base structure produce a spiro ring structure. In another embodiment, the ring-forming substituents are attached to non-adjacent members of the base structure.

[0063] Two of the substituents on adjacent atoms of an aryl or heteroaryl ring may optionally form a ring of the formula -T-C(O)-(CRR′) q -U-, where T and U are independently -NR-, -O-, -CRR′-, or a single bond, and q is an integer from 0 to 3. Alternatively, two of the substituents on adjacent atoms of an aryl or heteroaryl ring may optionally be of the formula -A-(CH2) rSubstitution of the substituents of -B-, wherein A and B are independently -CRR′-, -O-, -NR-, -S-, -S(O)-, -S(O)2-, -S(O)2NR′- or a single bond, and r is an integer from 1 to 4. One of the single bonds of the new ring thus formed may optionally be replaced by a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced by substituents of the formula -(CRR′) s -x′-(C″R″R″′) d -, wherein s and d are independently integers from 0 to 3, and x′ is -O-, -NR′-, -S-, -S(O)-, -S(O)2- or -S(O)2NR′-. The substituents R, R′, R″ and R″′ are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl.

[0064] As used herein, the term "heteroatom" or "ring heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P) and silicon (Si).

[0065] As used herein, "substituent" means a group selected from the following moieties:

[0066] (A) oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CHCl2, -CHBr2, -CHF2, -CHI2, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CN, -OH, -NH2, -C(O)OH, -C(O)NH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI2, -OCHF2, -OCH2Cl, -OCH2Br, -OCH2I, -OCH2F, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2- to 8-membered heteroalkyl, 2- to 6-membered heteroalkyl or 2- to 4-membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C 10 aryl, C 10an aryl, or phenyl), or an unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl or a 5- to 6-membered heteroaryl), and

[0067] (B) an alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl substituted with at least one substituent selected from:

[0068] (i) oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CHCl2, -CHBr2, -CHF2, -CHI2, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CN, -OH, -NH2, -C(O)OH, -C(O)NH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI2, -OCHF2, -OCH2Cl, -OCH2Br, -OCH2I, -OCH2F, -N3, an unsubstituted alkyl (e.g., a C1-C8 alkyl, a C1-C6 alkyl or a C1-C4 alkyl), an unsubstituted heteroalkyl (e.g., a 2- to 8-membered heteroalkyl, a 2- to 6-membered heteroalkyl or a 2- to 4-membered heteroalkyl), an unsubstituted cycloalkyl (e.g., a C3-C8 cycloalkyl, a C3-C6 cycloalkyl or a C5-C6 cycloalkyl), an unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl or a 5- to 6-membered heterocycloalkyl), an unsubstituted aryl (e.g., a C6-C 10 aryl, C 10 aryl, or phenyl), or an unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl or a 5- to 6-membered heteroaryl), and

[0069] (ii) an alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl substituted with at least one substituent selected from:

[0070] (a) Oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CHCl2, -CHBr2, -CHF2, -CHI2, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CN, -OH, -NH2, -C(O)OH, -C(O)NH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI2, -OCHF2, -OCH2Cl, -OCH2Br, -OCH2I, -OCH2F, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2- to 8-membered heteroalkyl, 2- to 6-membered heteroalkyl or 2- to 4-membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C 10 aryl, C 10 aryl, or phenyl) or unsubstituted heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl or 5- to 6-membered heteroaryl), and

[0071] (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl substituted by at least one substituent selected from the following: oxo group, halogen, -CCl3, -CBr3, -CF3, -CI3, CHCl2, -CHBr2, -CHF2, -CHI2, -CH2C1, -CH2Br, -CH2F, -CH2I, -CN, -OH, -NH2, -C(O)OH, -C(O)NH2, -NO2, -SH, -SO3H, -SO4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI2, -OCHF2, -OCH2Cl, -OCH2Br, -OCH2I, -OCH2F, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2- to 8-membered heteroalkyl, 2- to 6-membered heteroalkyl or 2- to 4-membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C 10 aryl, C 10 aryl, or phenyl) or unsubstituted heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl or 5- to 6-membered heteroaryl).

[0072] As used herein, "size-limited substituent / size-limited substituent group" means a group selected from all the substituents described above for "substituent", wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C 20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2- to 20-membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C 10 aryl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5- to 10-membered heteroaryl.

[0073] As used herein, "lower substituent / lower substituent group" means a group selected from all the substituents described above for "substituent", wherein each substituted or unsubstituted alkyl group is a substituted or unsubstituted C1-C8 alkyl group, each substituted or unsubstituted heteroalkyl group is a substituted or unsubstituted 2- to 8-membered heteroalkyl group, each substituted or unsubstituted cycloalkyl group is a substituted or unsubstituted C3-C7 cycloalkyl group, each substituted or unsubstituted heterocycloalkyl group is a substituted or unsubstituted 3- to 7-membered heterocycloalkyl group, each substituted or unsubstituted aryl group is a substituted or unsubstituted C6-C 10 aryl group, and each substituted or unsubstituted heteroaryl group is a substituted or unsubstituted 5- to 9-membered heteroaryl group.

[0074] In some embodiments, each substituted group in the compounds of the present invention is substituted with at least one substituent. More specifically, in some embodiments, each substituted alkyl group, substituted heteroalkyl group, substituted cycloalkyl group, substituted heterocycloalkyl group, substituted aryl group, substituted heteroaryl group, substituted alkylene group, substituted heteroalkylene group, substituted cycloalkylene group, substituted heterocycloalkylene group, substituted arylene group, and / or substituted heteroarylene group in the compounds of the present invention is substituted with at least one substituent group. In other embodiments, at least one or all of these groups are substituted with at least one substituent group having a limited size. In other embodiments, at least one or all of these groups are substituted with at least one lower substituent group.

[0075] In other embodiments of the compounds herein, each substituted or unsubstituted alkyl group can be a substituted or unsubstituted C1-C 20 alkyl group, each substituted or unsubstituted heteroalkyl group is a substituted or unsubstituted 2- to 20-membered heteroalkyl group, each substituted or unsubstituted cycloalkyl group is a substituted or unsubstituted C3-C8 cycloalkyl group, each substituted or unsubstituted heterocycloalkyl group is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group, each substituted or unsubstituted aryl group is a substituted or unsubstituted C6-C 10 aryl group, and each substituted or unsubstituted heteroaryl group is a substituted or unsubstituted 5- to 10-membered heteroaryl group. In some embodiments of the compounds of the present invention, each substituted or unsubstituted alkylene group is a substituted or unsubstituted C1-C 20 alkylene group, each substituted or unsubstituted heteroalkylene group is a substituted or unsubstituted 2- to 20-membered heteroalkylene group, each substituted or unsubstituted cycloalkylene group is a substituted or unsubstituted C3-C8 cycloalkylene group, each substituted or unsubstituted heterocycloalkylene group is a substituted or unsubstituted 3- to 8-membered heterocycloalkylene group, each substituted or unsubstituted arylene group is a substituted or unsubstituted C6-C 10The arylene, and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5- to 10-membered heteroarylene.

[0076] In some embodiments, each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2- to 8-membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 7-membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C 10 The aryl, and / or each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5- to 9-membered heteroaryl. In some embodiments, each substituted or unsubstituted alkylene is a substituted or unsubstituted C1-C8 alkylene, each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2- to 8-membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C7 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3- to 7-membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted or unsubstituted C6-C 10 The arylene, and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5- to 9-membered heteroarylene. In some embodiments, the compound is a chemical species described in the Examples section, the figures, or the tables below.

[0077] In embodiments, a substituted or unsubstituted moiety (e.g., a substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and / or substituted or unsubstituted heteroarylene) is unsubstituted (e.g., an unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, unsubstituted alkylene, unsubstituted heteroalkylene, unsubstituted cycloalkylene, unsubstituted heterocycloalkylene, unsubstituted arylene, and / or unsubstituted heteroarylene, respectively). In embodiments, a substituted or unsubstituted moiety (e.g., a substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and / or substituted or unsubstituted heteroarylene) is substituted (e.g., a substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene, respectively).

[0078] In embodiments, a substituted moiety (e.g., a substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene) is substituted with at least one substituent, where if the substituted moiety is substituted with multiple substituents, each substituent can optionally be different. In embodiments, if the substituted moiety is substituted with multiple substituents, each substituent is different.

[0079] In embodiments, a substituted moiety (e.g., a substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene) is substituted with at least one size-limited substituent, where if the substituted moiety is substituted with multiple size-limited substituents, each size-limited substituent can optionally be different. In embodiments, if the substituted moiety is substituted with multiple size-limited substituent groups, each size-limited substituent group is different.

[0080] In embodiments, the substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene) is substituted with at least one lower substituent, where if the substituted moiety is substituted with a plurality of lower substituents, each lower substituent may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of lower substituent groups, each lower substituent group is different.

[0081] In embodiments, the substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene) is substituted with at least one substituent, size-limited substituent, or lower substituent; where if the substituted moiety is substituted with a plurality of groups selected from substituents, size-limited substituents, and lower substituents, each substituent, size-limited substituent, and lower substituent may optionally be different. In embodiments, if the substituted moiety is substituted with a plurality of groups selected from substituents, size-limited substituents, and lower-carbon-number substituents, then each substituent, size-limited substituent, and / or lower-carbon-number substituent is different.

[0082] Certain compounds of the present disclosure have asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms, and individual isomers that can be defined as (R)- or (S)- or (D)- or (L)- of an amino acid according to absolute stereochemistry are all encompassed within the scope of the present disclosure. Compounds of the present disclosure do not include compounds that are known in the art to be too unstable to be synthesized and / or isolated. The present disclosure is intended to include compounds in racemic and optically pure forms. Optically active (R)- and (S)- or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic bonds or other geometrically asymmetric centers, and unless otherwise specified, it is desired that the compounds include both E geometric isomers and Z geometric isomers.

[0083] As used herein, the term "isomer" refers to a compound having the same number and type of atoms, and thus the same molecular weight, but having a different structural arrangement or atomic configuration.

[0084] As used herein, the term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily convertible from one isomeric form to another.

[0085] It will be apparent to those skilled in the art that certain compounds of the present disclosure may exist in tautomeric forms, and all such tautomeric forms of the compounds are within the scope of the present disclosure.

[0086] Unless otherwise indicated, the structures depicted herein also mean to include all stereochemical forms of the structures; that is, the R configuration and the S configuration of each asymmetric center. Accordingly, single stereoisomers of the compounds of the invention, as well as enantiomeric and diastereomeric mixtures, are within the scope of the present disclosure.

[0087] Unless otherwise stated, the structures depicted herein also mean to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structures of the invention except for the replacement of hydrogen by deuterium or tritium or the replacement of carbon by 13 C- or 14 C-enriched carbon are within the scope of the present disclosure.

[0088] The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes at one or more of the atoms that make up such compounds. For example, the compounds may be radiolabeled with radioactive isotopes such as, for example, tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C). All isotopic variations of the compounds of the present disclosure, whether radioactive or non-radioactive, are encompassed within the scope of the present disclosure.

[0089] It should be noted that throughout the application, alternatives are written in Markush groups, for example, each amino acid position containing more than one possible amino acid. It is specifically contemplated that each member of the Markush group should be considered separately, so as to include another embodiment, and the Markush group should not be understood as a single unit.

[0090] As used herein, the terms "bioconjugate" and "bioconjugate linker" refer to an association that occurs between atoms or molecules of a bioconjugate reactive group or bioconjugate moiety. The association can be direct or indirect. For example, the conjugation between a first bioconjugate reactive group provided herein (e.g., -NH2, -C(O)OH, N-hydroxysuccinimide, or -maleimide) and a second bioconjugate reactive group (e.g., thiol, sulfur-containing amino acid, amine, amine side chain containing an amino acid, or carboxylate) can occur directly, for example, via a covalent bond or a linker (e.g., a first linker of a second linker), or indirectly, for example, via non-covalent bonds (e.g., electrostatic interactions (e.g., ionic bond, hydrogen bond, halogen bond), van der Waals interactions (e.g., dipole-dipole, dipole-induced dipole, London dispersion), ring stacking (π effect), hydrophobic interactions, etc.). In embodiments, the bioconjugate or bioconjugate linker is formed using bioconjugate chemistry (i.e., the association of two bioconjugate reactive groups), which includes but is not limited to nucleophilic substitution (e.g., the reaction of amines and alcohols with acyl halides, active esters), electrophilic substitution (e.g., enamine reaction), and addition to carbon-carbon and carbon-heteroatom multiple bonds (e.g., Michael reaction, Diels-Alder addition). These and other useful reactions are discussed in the following references: e.g., March, ADVANCED ORGANIC CHEMISTRY, 3rd Edition, John Wiley & Sons, New York, 1985; Hermanson, BIOCONJUGATE TECHNIQUES, Academic Press, San Diego, 1996; and Feeney et al., MODIFICATION OF PROTEINS; Advances in Chemistry Series, Vol. 198, American Chemical Society, Washington, D.C., 1982. In embodiments, a first bioconjugate reactive group (e.g., a maleimide moiety) is covalently attached to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a haloacetyl moiety) is covalently attached to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a pyridyl moiety) is covalently attached to a second bioconjugate reactive group (e.g., a thiol).In embodiments, a first bioconjugate reactive group (e.g., an -N-hydroxysuccinimide moiety) is covalently attached to a second bioconjugate reactive group (e.g., an amine). In embodiments, a first bioconjugate reactive group (e.g., a maleimide moiety) is covalently attached to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a sulfo-N-hydroxysuccinimide moiety) is covalently attached to a second bioconjugate reactive group (e.g., an amine).

[0091] Useful bioconjugate reactive moieties for the bioconjugate chemistry herein include, for example:

[0092] (a) Carboxyl groups and their various derivatives, including but not limited to N-hydroxysuccinimide esters, N-hydroxybenzotriazole esters, acid halides, acyl imidazoles, thioesters, p-nitrophenyl esters, alkyl, alkenyl, alkynyl, and aromatic esters;

[0093] (b) Hydroxyl groups, which can be converted to esters, ethers, aldehydes, etc.

[0094] (c) Haloalkyl groups, where the halide can be subsequently displaced by a nucleophilic group such as an amine, carboxylate anion, thiolate anion, carbanion, or alkoxide ion, resulting in a new group covalently linked at the site of the halogen atom;

[0095] (d) Dienophiles, which are capable of participating in Diels-Alder reactions, such as maleimido or maleimide groups;

[0096] (e) Aldehyde or ketone groups, which allow for subsequent derivatization through the formation of carbonyl derivatives (e.g., imines, hydrazones, semicarbazones, or oximes) or through mechanisms such as Grignard addition or alkyllithium addition;

[0097] (f) Sulfonyl halide groups, which are used for subsequent reaction with amines, for example, to form sulfonamides;

[0098] (g) Thiol groups, which can be converted to disulfides, react with acyl halides, or bond to metals such as gold, or react with maleimides;

[0099] (h) Amine or thiol groups (e.g., present in cysteine), which can be, for example, acylated, alkylated, or oxidized;

[0100] (i) Alkenes, which can undergo, for example, cycloadditions, acylations, Michael additions, etc.;

[0101] (j) Epoxides, which can react with, for example, amines and hydroxy compounds;

[0102] (k) Phosphoramidites and other standard functional groups useful for nucleic acid synthesis;

[0103] (l) Metal-oxide silicon bonding; and

[0104] (m) Metal bonding to a reactive phosphorus group (e.g., phosphine) to form, for example, a phosphodiester bond.

[0105] (n) Azides coupled to alkynes using copper-catalyzed cycloaddition click chemistry.

[0106] (o) Biotin conjugates can react with avidin or streptavidin to form an avidin-biotin complex or a streptavidin-biotin complex.

[0107] The bioconjugate reactive group can be selected such that it does not participate in or interfere with the chemical stability of the conjugates described herein. Alternatively, the reactive functional group can be protected from participating in the crosslinking reaction by the presence of a protecting group. In embodiments, the bioconjugate includes a molecular entity derived from the reaction of an unsaturated bond such as maleimide with a thiol group.

[0108] "Analog" is used according to its ordinary and general meaning within chemistry and biology and refers to a compound that is structurally similar but compositionally different from another compound (i.e., the so-called "reference" compound), for example, in the replacement of an atom with an atom of a different element, or in the presence of a particular functional group, or in the replacement of one functional group with another functional group, or in the absolute stereochemistry of one or more chiral centers of the reference compound. Thus, an analog is a compound that is similar or equivalent to the reference compound in function and appearance but not in structure or origin.

[0109] As used herein, the term "a / an" refers to one or more. Additionally, as used herein, the phrase "substituted with [n]..." means that the specified group can be substituted with one or more of any or all of the named substituents. For example, when a group such as an alkyl or heteroaryl group is "substituted with an unsubstituted C1-C 20 alkyl or an unsubstituted 2- to 20-membered heteroalkyl", the group can contain one or more unsubstituted C1-C 20 alkyl, and / or one or more unsubstituted 2- to 20-membered heteroalkyls.

[0110] In addition, when a moiety is substituted with an R substituent, the moiety may be referred to as "R-substituted". When a moiety is R-substituted, the moiety is substituted with at least one R substituent, and each R substituent is optionally different. When a particular R group is present in the description of a chemical species (such as formula (I)), Roman letter symbols may be used to distinguish each occurrence of that particular R group. For example, in the case where there are multiple R 13 substituents, each R 13 substituent can be distinguished as R 13.A , R 13.B , R 13.C , R 13.D , etc., where each of R 13.A , R 13.B , R 13.C , R 13.D , etc. is defined within the definition of R 13 and is optionally different.

[0111] "Oxidant" is used in accordance with its ordinary general chemical meaning in chemistry and biology and refers to a substance having the ability to oxidize other substances (i.e., remove electrons from substances). The term "oxidant" is a substance that removes one or more electrons from a substance (e.g., a reactant) during a chemical redox reaction, where the oxidant obtains one or more electrons from the substrate. In an embodiment, the oxidant is a chemical substance that transfers a negatively charged atom to another substrate (e.g., a reactant). In an embodiment, the oxidant is analogous to the term "electron acceptor" and may be used interchangeably herein. Non-limiting examples of oxidants include oxygen (O2), ozone (O3), hydrogen peroxide (H2O2), nitric acid (HNO3), sulfuric acid (H2SO4), hexavalent chromium, pyridinium chlorochromate (PCC), N-methylmorpholine-N-oxide (NMO), chromium trioxide (CrO3, Jones reagent), potassium permanganate (K2MnO4), potassium nitrate (KNO3), Dess-Martin periodinane (DMP), 2-iodoxybenzoic acid (IBX), 2,2,6,6-tetramethylpiperidinyloxy (TEMPO), and (F-TEDA-BF4, chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane bis(tetrafluoroborate)), potassium perchlorate, or ammonium persulfate.

[0112] The term "halogenating agent" is used according to its ordinary general chemical meaning in chemistry and refers to a substance (e.g., a compound or composition) capable of incorporating one or more halogen atoms (such as bromination, dibromination, tribromination, chlorination, dichlorination, trichlorination, iodination, diiodination, triiodination, fluorination, difluorination, trifluorination, etc.) into another substance (e.g., a compound or composition). Halogenating agents include chlorinating agents, brominating agents, iodinating agents, and fluorinating agents, where chlorinating agents incorporate chlorine atoms, brominating agents incorporate bromine atoms, iodinating agents incorporate iodine atoms, or fluorinating agents incorporate fluorine atoms. Brominating agents include, but are not limited to, N-bromosuccinimide (NBS), dibromo isocyanuric acid (DBI), bromine, bromotrichloromethane, 1,2-dibromo-1,1,2,2-tetrachloroethane, carbon tetrabromide, tetrabutylammonium tribromide, trimethylbenzylammonium tribromide, benzyltrimethylammonium tribromide, pyridinium perbromide, 4-dimethylaminopyridinium perbromide, 1-butyl-3-methylimidazolium tribromide, 1,8-diazabicyclo[5.4.0]-7-undecene, hydrogen tribromide, N-bromophthalimide, N-bromosaccharin, N-bromoacetamide, 2-bromo-2-cyano-N,N-dimethylacetamide, 1,3-dibromo-5,5-dimethylhydantoin, sodium monohydrate bromoisocyanurate, boron tribromide, phosphorus tribromide, dimethylsulfonium bromide bromide, 5,5-dibromomesoxalic acid, 2,4,4,6-tetrabromo-2,5-cyclohexadienone, or bis(2,4,6-trimethylpyridine)-iodonium hexafluorophosphate. Chlorinating agents include, but are not limited to, N-chlorosuccinimide (NCS), thionyl chloride, methanesulfonyl chloride, trichloromethanesulfonyl chloride, tert-butyl hypochlorite, chloromethyl methyl ether, dichloromethyl methyl ether, methoxyacetyl chloride, oxalyl chloride, cyanuric chloride, N-chlorophthalimide, sodium dichloroisocyanurate, trichloroisocyanuric acid, chloramine B hydrate, o-chloramine T dihydrate, chloramine T trihydrate, dichloramine B, dichloramine T, benzyltrimethylammonium, tetraiodochlorate. Iodinating agents include, but are not limited to, N-iodosuccinimide (NIS), 1,3-diiodo-5,5'-dimethylhydantoin (DIH), iodine, hydroiodic acid, diiodomethane, 1-chloro-2-iodoethane, carbon tetraiodide, tetramethylammonium dichloroiodate, benzyltrimethylammonium dichloroiodate, pyridinium monochloroiodide, N,N-dimethyl-N-(methylsulfanylmethylene)-ammonium iodide, N-iodosaccharin, trimethylsilyl iodide, bis(pyridine)iodonium tetrafluoroborate bis(2,4,6-trimethylpyridine)iodonium hexafluorophosphate. In an embodiment, the halogenating agent is not a fluorinating agent.

[0113] "Metal source" is used according to its ordinary and general chemical meaning in chemistry and biology and refers to a compound, salt, or complex that includes a transition metal (e.g., as seen in the periodic table of elements). In an embodiment, the metal source is a transition metal element (i.e., an element whose atoms have a partially filled d subshell, or an element that can produce a cation with an incomplete d subshell). The metal source can be a compound, salt, or complex and can contain one or more transition metals. In one embodiment, the metal source can be a "silver source", where the transition metal is silver. Non-limiting examples of silver sources include silver(I) tetrafluoroborate (AgBF4), silver(I) nitrate (AgNO3), silver(II) fluoride (AgF2), silver(I) fluoride (AgF), silver trifluoromethanesulfonate (AgOTf), silver bis(trifluoromethanesulfonyl)imide (AgNTf2), silver carbonate (Ag2CO3), silver(I) oxide (Ag2O), silver(I) acetate (AgOAc), silver(I) sulfate (Ag2SO4), silver methanesulfonate (AgOMs), silver(V) hexafluoroantimonate (AgSbF6), silver p-toluenesulfonate (AgOTs), silver(I) trifluoromethanethiolate (AgSCF3), and silver(I) bromide (AgBr). In one embodiment, the metal source can be a "copper source", where the transition metal is copper. Non-limiting examples of copper sources include copper(II) sulfate (CuSO4). In one embodiment, the metal source can be an "iron source", where the transition metal is iron. Non-limiting examples of iron sources include iron(III) chloride (FeCl3) and iron(I) nitrate (FeNO3). In one embodiment, the metal source can be a "manganese source", where the transition metal is manganese. Non-limiting examples of manganese sources include manganese(II) chloride (MnCl2), manganese(III) acetate (Mn(OAc)3), manganese(III) acetylacetonate (Mn(acac)3), and manganese(III) 2-pyridinecarboxylate (Mn(pic)3). See, Chem. Lett. 2017, 46, 1692, which is incorporated herein by reference in its entirety.

[0114] A "detectable agent" or "detectable moiety" is a composition, substance, element, or compound or a portion thereof that is detectable by suitable means such as spectroscopic, photochemical, biochemical, immunochemical, chemical, magnetic resonance imaging, or other physical means. For example, useful detectable agents include 18 F, 32 P, 33 P, 45 Ti, 47 Sc, 52 Fe, 59 Fe, 62 Cu, 64 Cu, 67 Cu, 67 Ga, 68 Ga,77 As 86 Y 90 Y 89 Sr 89 Zr 94 Tc 94 Tc 99 mTc 99 Mo 105 Pd 105 Rh 111 Ag 111 In 123 I 124 I 125 1 131 I 142 Pr 143 Pr 149 Pm 153 Sm 154-1581 Gd 161 Tb 166 Dy 166 Ho 169 Er 175 Lu 177 Lu 186 Re 188 Re 189 Re 194 Ir 198 Au 199 Au 211 At 211 Pb 212 Bi 212 Pb 213 Bi 223 Ra 225 Ac, Cr, V, Mn, Fe, Co, Ni, Cu, La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu 32P, fluorophores (e.g., fluorescent dyes), electron-dense reagents, enzymes (e.g., enzymes commonly used in ELISA), biotin, digoxigenin, paramagnetic molecules, paramagnetic nanoparticles, ultrasmall superparamagnetic iron oxide (“USPIO”) nanoparticles, USPIO nanoparticle aggregates, superparamagnetic iron oxide (“SPIO”) nanoparticles, SPIO nanoparticle aggregates, single-crystal iron oxide nanoparticles, single-crystal iron oxide, nanoparticle contrast agents, liposomes or other delivery vehicles containing gadolinium chelate (“Gd-chelate”) molecules, gadolinium, radioisotopes, radionuclides (e.g., carbon-11, nitrogen-13, oxygen-15, fluorine-18, rubidium-82), fluorodeoxyglucose (e.g., fluorine-18 labeled), any radioisotope emitting gamma rays, positron-emitting radioisotopes, radioactively labeled glucose, radioactively labeled water, radioactively labeled ammonia, biocolloids, microbubbles (e.g., containing a microbubble shell, containing albumin, galactose, lipid, and / or polymer; a microbubble gas core, containing air, one or more heavy gases, perfluorocarbon, nitrogen, octafluoropropane, perfluoroalkane lipid microspheres, perfluoroether, etc.), iodinated contrast agents (e.g., iohexol, iodixanol, iopamidol, iopromide, ioxilan, iopromide, diatrizoate, metrizoate, ioxaglic acid), barium sulfate, thorium dioxide, gold, gold nanoparticles, gold nanoparticle aggregates, fluorophores, two-photon fluorophores, or haptens and proteins or other entities, where the other entities can be detected, for example, by incorporating a radioactive label into a peptide or antibody that specifically reacts with a target peptide. The detectable moiety is a monovalent detectable agent or a detectable agent capable of forming a bond with another composition.

[0115] Radioactive substances (e.g., radioisotopes) that can be used as imaging agents and / or labeling agents according to embodiments of the present disclosure include, but are not limited to 18 F, 32 P, 33 P, 45 Ti, 47 Sc, 52 Fe, 59 Fe, 62 Cu, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 77 As, 86 Y, 90 Y. 89 Sr, 89 Zr, 94 Tc, 94 Tc, 99m Tc, 99 Mo, 105 Pd, 105 Rh,111 Ag, 111 In, 123 I, 124 I, 125 I, 131 I, 142 Pr, 143 Pr, 149 Pm, 153 Sm, 154-1581 Gd, 161 Tb, 166 Dy, 166 Ho, 169 Er, 175 Lu, 177 Lu, 186 Re, 188 Re, 189 Re, 194 Ir, 198 Au, 199 Au, 211 At, 211 Pb, 212 Bi, 212 Pb, 213 Bi, 223 Ra and 225 Ac. Paramagnetic ions that can be used as additional imaging agents according to embodiments of the present disclosure include, but are not limited to, ions of transition metals and lanthanide metals (e.g., metals having atomic numbers 21 - 29, 42, 43, 44, or 57 - 71). These metals include ions of Cr, V, Mn, Fe, Co, Ni, Cu, La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, and Lu.

[0116] The description of the compounds of the present disclosure is limited by the principles of chemical bonding known to those skilled in the art. Thus, in cases where a group can be substituted by one or more of a plurality of substituents, such substitutions are selected so as to conform to the principles of chemical bonding and to result in compounds that are not inherently unstable and / or that would not be likely to be unstable under environmental conditions (such as aqueous, neutral, and several known physiological conditions) as known to those of ordinary skill in the art. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule through a ring heteroatom in accordance with the principles of chemical bonding known to those skilled in the art, thus avoiding inherently unstable compounds.

[0117] The term "leaving group" is used according to its ordinary meaning in chemistry and refers to the portion (e.g., atom, functional group, molecule) that separates from a molecule after a chemical reaction (e.g., bond formation, reductive elimination, condensation, cross-coupling reaction), where the chemical reaction involves the atom or chemical moiety to which the leaving group is attached (also referred to herein as the "leaving group reactive moiety") and a complementary reactive moiety (i.e., the chemical moiety that reacts with the leaving group reactive moiety) to form a new bond between the remaining portion of the leaving group reactive moiety and the complementary reactive moiety. Thus, the leaving group reactive moiety and the complementary reactive moiety form a complementary reactive group pair. Non-limiting examples of leaving groups include hydrogen, hydroxide, organotin moieties (e.g., organotin heteroalkyls), halogens (e.g., Br), perfluoroalkyl sulfonates (e.g., trifluoromethanesulfonate), tosylates, mesylates, water, alcohols, nitrates, phosphates, thioethers, amines, ammonia, fluorides, carboxylates, phenolates, boronic acids, boronic esters, and alkoxides. In embodiments, two molecules having leaving groups are contacted, and upon reaction and / or bond formation (e.g., ketol condensation, aldol condensation, Claisen condensation, Stille reaction), the leaving group separates from the respective molecule. In embodiments, the leaving group is a bioconjugate reactive moiety. In embodiments, at least two leaving groups (e.g., R 1 and R 13 ) are contacted such that the leaving groups are close enough to react, interact, or physically contact. In embodiments, the leaving groups are designed to facilitate the reaction.

[0118] The term "protecting group" is used according to its ordinary meaning in organic chemistry and refers to a moiety that is covalently attached to a heteroatom, heterocycloalkyl, or heteroaryl to block the reactivity of the heteroatom, heterocycloalkyl, or heteroaryl during one or more chemical reaction processes prior to removal of the protecting group. In an embodiment, the protecting group is covalently attached to a heteroatom that is part of a heteroalkyl, heterocycloalkyl, or heteroaryl moiety. Typically, the protecting group is attached to a heteroatom (e.g., O) during a portion of a multi-step synthesis where reaction of the heteroatom with a reagent (e.g., chemical reduction) is not desired. After protection, the protecting group can be removed (e.g., by adjusting the pH). In an embodiment, the protecting group is an alcohol protecting group. Non-limiting examples of alcohol protecting groups include acetyl, benzoyl, benzyl, methoxymethyl ether (MOM), tetrahydropyranyl (THP), and silyl ethers (e.g., trimethylsilyl (TMS), tert-butyldimethylsilyl (TBS)). In an embodiment, the protecting group is an amine protecting group. Non-limiting examples of amine protecting groups include benzyloxycarbonyl (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), tert-butoxycarbonyl (BOC), 9-fluorenylmethoxycarbonyl (FMOC), acetyl (Ac), benzoyl (Bz), benzyl (Bn), carbamate, p-methoxybenzyl ether (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), pivaloyl (Piv), tosyl (Ts), and phthalimide.

[0119] The term "silyl protecting group" is used according to its ordinary meaning in organic chemistry and refers to a protecting group that contains a silicon atom covalently bonded to a heteroatom to prevent the reactivity of the heteroatom. In an embodiment, the silyl protecting group is covalently attached to an alkoxy group to form a silyl ether. Non-limiting examples of silyl protecting groups include trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBS / TBDMS), tert-butyldiphenylsilyl (TBDPS), and triisopropylsilyl (TIPS).

[0120] The term "transition metal catalyst for olefin metathesis" is used according to its ordinary meaning in organic chemistry and refers to a transition metal catalyst that catalyzes the reaction of redistributing fragments of olefins (such as olefins) through the cleavage and regeneration of carbon-carbon double bonds. In an embodiment, olefin metathesis is cross-metathesis. In an embodiment, olefin metathesis involves a ring closure between two terminal vinyl groups (ring-closing metathesis). In an embodiment, the transition metal catalyst is a heterogeneous catalyst. In an embodiment, the transition metal catalyst is a molybdenum-based catalyst. In an embodiment, the transition metal catalyst is a molybdenum(VI)-based catalyst. In an embodiment, the transition metal catalyst is a tungsten-based catalyst. In an embodiment, the transition metal catalyst is a tungsten(VI)-based catalyst. In an embodiment, the transition metal catalyst is a ruthenium-based catalyst. In an embodiment, the transition metal catalyst is a ruthenium(II)-based catalyst. In an embodiment, the transition metal catalyst is a Grubbs catalyst. In an embodiment, the transition metal catalyst is a Schrock catalyst. In an embodiment, the transition metal catalyst is a Hoveyda-Grubbs catalyst.Non-limiting examples of transition metal catalysts for olefin metathesis include: Grubbs 1st generation catalysts [benzylidene-bis(tricyclohexylphosphine) ruthenium dichloride, bis(tricyclohexylphosphine)benzylidene ruthenium(IV) dichloride, or dichloro(benzylidene)bis(tricyclohexylphosphine)ruthenium(II)]; Grubbs 2nd generation catalysts [(1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinyl)dichloro(phenylmethylene)(tricyclohexylphosphine)ruthenium, benzyl[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinyl)dichloro(phenylmethylene)(tricyclohexylphosphine)ruthenium, bis(3-bromopyridine)ruthenium(II)]; [1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(tricyclohexylphosphine)ruthenium or dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](benzylidene)(tricyclohexylphosphine)ruthenium(II)]; Grubbs third generation catalyst [dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](benzylidene)bis(3-bromopyridine)ruthenium(II), [1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)bis(3- bromopyridine) ruthenium (II), or [1,3-dimethyl-2-imidazolidinyl] dichloro (phenylmethylene) bis (3-bromopyridine) ruthenium (II)]; Hoveyda-Grubbs first generation catalyst [dichloro (2-isopropoxyphenylmethylene) (tricyclohexylphosphine) ruthenium (II) or dichloro (o-isopropoxyphenylmethylene) (tricyclohexylphosphine) ruthenium (II)]; Hoveyda-Grubbs second generation catalyst [(1,3-bis-(2,4,6-trimethylphenyl)-2-imidazole dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](2-isopropoxyphenylmethylene)ruthenium(II)]; NitroGrela[(1,3-dimesityl imidazolin-2-ylidene)dichloro(2-isopropoxy-5-nitrobenzylidene)ruthenium(II)]; dichloro[1,3-bis(2-methylphenyl)-2-imidazolidinylidene](benzylidene)(tricyclohexylphosphine)ruthenium(II)[Grubbs. M2a SI (o-Tol) (C793)]; dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](3-methyl-2-butenylidene)(tricyclohexylphosphine)ruthenium(II) [Grubbs M2b(C827)]; dichloro[1,3-bis(2-methylphenyl)-2-imidazolidinylidene](2-isopropoxyphenylmethylene)ruthenium(II)[Hoveyda-Grubbs M72SI (o-Tol) (C571) or Stewart-Grubbs catalyst]; and dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][3-(2-pyridyl)propylene]ruthenium(II) (Grubbs C598).

[0121] The term "alcohol" is used according to its ordinary meaning in organic chemistry and refers to an organic compound carrying at least one hydroxyl functional group (-OH) bonded to a saturated carbon atom. In an embodiment, the alcohol is a primary alcohol. In an embodiment, the alcohol is a secondary alcohol. In an embodiment, the alcohol is a tertiary alcohol. Non-limiting examples of alcohols include: methanol, ethanol, n-propanol (propan-1-ol or 1-propanol), isopropanol (propan-2-ol or 2-propanol), cyclohexanol, isobutanol (2-methylpropan-1-ol or 2-methyl-1-propanol), or tert-amyl alcohol (2-methylbutan-2-ol or 2-methyl-2-butanol).

[0122] The term "base" is used according to its ordinary meaning in organic chemistry and refers to a substance that accepts a proton from any proton donor or contains fully or partially displaceable OH- ions. In an embodiment, the base is an inorganic base. In an embodiment, the base is an organic base. Non-limiting examples of inorganic bases include: NaOH, LiOH, Ca(OH)2, magnesium hydroxide, sodium carbonate, sodium bicarbonate, sodium hydrogen carbonate, or ammonium hydroxide. Non-limiting examples of organic bases include: pyridine, alkylamines (such as methylamine), imidazole, benzimidazole, histidine, guanidine, or phosphazene bases.

[0123] The compound "17S-FD-895" corresponds to the following structure:

[0124] One of ordinary skill in the art will understand that when variables (e.g., moieties or linkers) of a compound or a genus of compounds (e.g., the genus described herein) are described by the name or molecular formula of an independent compound having all filled valences, one or more unfilled valences of the variable will be determined by the context in which the variable is used. For example, when a variable of a compound described herein is connected (e.g., bonded) to the rest of the compound by a single bond, the variable should be understood to represent the monovalent form of the independent compound (i.e., capable of forming a single bond due to the unfilled valence) (e.g., in one embodiment, if the variable is named "methane" but it is known that the variable is connected to the rest of the compound by a single bond, one of ordinary skill in the art will understand that the variable is actually the monovalent form of methane, i.e., methyl or -CH3). Similarly, for linker variables (e.g., L 1 、L 2 or L 3), one of ordinary skill in the art will understand that the variable is the divalent form of an independent compound (e.g., in one embodiment, if the variable is designated as "PEG" or "polyethylene glycol" but the variable is attached to the rest of the compound by two separate bonds, one of ordinary skill in the art will understand that the variable is the divalent form of PEG (i.e., capable of forming two bonds through two unfilled valences), rather than the independent compound PEG).

[0125] The term "exogenous" refers to a molecule or substance (e.g., a compound, nucleic acid, or protein) that is derived from outside a given cell or organism. For example, an "exogenous promoter" as referred to herein is a promoter that is not derived from the plant in which it is expressed. In contrast, the term "endogenous" or "endogenous promoter" refers to a molecule or substance that is native to or derived from within a given cell or organism.

[0126] The term "lipid moiety" is used in its ordinary sense in chemistry and refers to a hydrophobic molecule that is typically characterized by an aliphatic hydrocarbon chain. In embodiments, the lipid moiety includes a carbon chain having 3 to 100 carbons. In embodiments, the lipid moiety includes a carbon chain having 5 to 50 carbons. In embodiments, the lipid moiety includes a carbon chain having 5 to 25 carbons. In embodiments, the lipid moiety includes a carbon chain having 8 to 25 carbons. The lipid moiety may include a saturated or unsaturated carbon chain and may optionally be substituted. In embodiments, the lipid moiety is optionally substituted at the terminus with a charged moiety. In embodiments, the lipid moiety is an alkyl or heteroalkyl optionally substituted at the terminus with a carboxylic acid moiety.

[0127] A charged moiety refers to a functional group having a rich electron density (i.e., negative electronegativity) or a lack of electron density (i.e., positive electronegativity). Non-limiting examples of charged moieties include carboxylic acid, alcohol, phosphate, aldehyde, and sulfonamide. In embodiments, the charged moiety is capable of forming a hydrogen bond.

[0128] The term "coupling reagent" is used according to its ordinary and general meaning in the art and refers to a substance (e.g., a compound or solution) that participates in a chemical reaction and results in the formation of a covalent bond (e.g., between bioconjugate reactive moieties, between a bioconjugate reactive moiety and a coupling reagent). In an embodiment, the reagent level is depleted during the chemical reaction. This is in contrast to a solvent, which is generally not consumed during a chemical reaction. Non-limiting examples of coupling reagents include benzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (PyBOP), 7-azabenzotriazol-1-yloxy)tris(pyrrolidino)phosphonium hexafluorophosphate (PyAOP), 6-chloro-benzotriazol-1-yloxy-tris-pyrrolidinophosphonium hexafluorophosphate (PyClock), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), or 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU).

[0129] The term "solution" is used according to its ordinary meaning and refers to a liquid mixture in which a minor component (e.g., a solute or compound) is uniformly distributed within a major component (e.g., a solvent).

[0130] As used herein, the term "organic solvent" is used according to its ordinary meaning in chemistry and refers to a solvent that contains carbon. Non-limiting examples of organic solvents include acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 2-butanone, tert-butanol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, 1,2-dichloroethane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether (diethylene glycol, dimethyl ether), 1,2-dimethoxyethane (glycol dimethyl ether, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4-dioxane, ethanol, ethyl acetate, ethylene glycol, glycerol, heptane, hexamethylphosphoramide (HMPA), hexamethylphosphorous triamide (HMPT), hexane, methanol, methyl tert-butyl ether (MTBE), dichloromethane, N-methyl-2-pyrrolidone (NMP), nitromethane, pentane, petroleum ether (ligroine), 1-propanol, 2-propanol, pyridine, tetrahydrofuran (THF), toluene, triethylamine, o-xylene, m-xylene, or p-xylene. In an embodiment, the organic solvent is or includes chloroform, dichloromethane, methanol, ethanol, tetrahydrofuran, or dioxane.

[0131] As used herein, the term "enantiomerically pure" is used according to its ordinary meaning in organic chemistry and refers to a molecule of a specified chirality having a specified purity. For example, a 99% enantiomerically pure sample has a molar ratio of 99:1 of the specified enantiomer relative to one or more alternative enantiomeric configurations. In an embodiment, enantiomeric purity can be measured using NMR, LC-MS, or chiral-HPLC.

[0132] As used herein, the term "salt" refers to an acid or base salt of a compound used in the methods provided herein. Exemplary examples of acceptable salts are salts of mineral acids (hydrochloric acid, hydrobromic acid, phosphoric acid, etc.), salts of organic acids (acetic acid, propionic acid, glutamic acid, citric acid, etc.), quaternary ammonium (methyl iodide, ethyl iodide, etc.) salts.

[0133] As used herein, the terms "bind" and "bound" are used according to their ordinary and general meanings and refer to the association between atoms or molecules. The association can be direct or indirect. For example, the bound atoms or molecules can be bound by, for example, covalent bonds, linkers (such as a first linker or a second linker), or non-covalent bonds (such as electrostatic interactions (such as ionic bonds, hydrogen bonds, halogen bonds), van der Waals interactions (such as dipole-dipole, dipole-induced dipole, London dispersion), ring stacking (π effects), hydrophobic interactions, etc.).

[0134] As used herein, the term "capable of binding" refers to a moiety (such as a compound as described herein) that is capable of measurably binding to a target (e.g., NF-κB, Toll-like receptor protein). In embodiments, when the moiety is capable of binding to the target, the moiety is capable of binding with a Kd of less than about 10 μM, 5 μM, 1 μM, 500 nM, 250 nM, 100 nM, 75 nM, 50 nM, 25 nM, 15 nM, 10 nM, 5 nM, 1 nM, or about 0.1 nM.

[0135] As used herein, when referring to two moieties, the term "conjugated" means that the two moieties are bonded, and one or more bonds connecting the two moieties can be covalent or non-covalent. In embodiments, the two moieties are covalently bonded to each other (e.g., directly or through a covalently bonded intermediate). In embodiments, the two moieties are non-covalently bonded (e.g., by ionic bonds, van der Waals bonds / interactions, hydrogen bonds, polar bonds, or a combination or mixture thereof).

[0136] As used herein, the term "non-nucleophilic base" refers to any sterically hindered base that is a poor nucleophile.

[0137] As used herein, the term "nucleophile" refers to a chemical species that donates an electron pair to an electrophile to form a chemical bond relevant to a reaction. All molecules or ions having a pair of free electrons or at least one π bond can act as nucleophiles.

[0138] The term "strong acid" is used according to its ordinary and general meaning in the art and includes acids that are completely dissociated or ionized in aqueous solution. Examples of common strong acids include hydrochloric acid (HCl), nitric acid (HNO3), sulfuric acid (H2SO4), hydrobromic acid (HBr), hydroiodic acid (HI), perchloric acid (HClO4), or chloric acid (HClO3). In an embodiment, the strong acid is a sulfonic acid, such as p-toluenesulfonic acid (TsOH), pyridinium p-toluenesulfonate, or camphorsulfonic acid (CSA).

[0139] As used herein, the term "carbocation-stabilizing solvent" refers to any polar protic solvent that is capable of forming dipole-dipole interactions with a carbocation to stabilize the carbocation.

[0140] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that act in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code and those that are subsequently modified, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an α-carbon bonded to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have a modified R group (e.g., norleucine) or a modified peptide backbone, but retain the same basic chemical structure as the naturally occurring amino acid. Amino acid mimetics are compounds that have a structure different from the general chemical structure of an amino acid, but act in a manner similar to a naturally occurring amino acid. The terms "non-naturally occurring amino acid" and "unnatural amino acid" refer to amino acid analogs, synthetic amino acids, and amino acid mimetics that are not found in nature.

[0141] Amino acids are represented herein by their commonly known three-letter symbols or by the single-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Similarly, nucleotides can be represented by their commonly accepted single-letter codes.

[0142] The terms "polypeptide", "peptide", and "protein" are used interchangeably herein to refer to a polymer of amino acid residues, where the polymer can, in an embodiment, be conjugated to moieties that are not composed of amino acids. The terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. A "fusion protein" refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed as a single moiety.

[0143] As used herein, the terms "nucleic acid", "nucleic acid molecule", "nucleic acid oligomer", "oligonucleotide", "nucleic acid sequence", "nucleic acid fragment", and "polynucleotide" are used interchangeably and are intended to include, without limitation, polymeric forms of nucleotides that are covalently linked together and can have various lengths, where the nucleotides are deoxyribonucleotides or ribonucleotides, or analogs, derivatives, or modifications thereof. Different polynucleotides can have different three-dimensional structures and can perform various known or unknown functions. Non-limiting examples of polynucleotides include genes, gene fragments, exons, introns, intergenic DNA (including, but not limited to, heterochromatic DNA), messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of a sequence, isolated RNA of a sequence, nucleic acid probes, and primers. Polynucleotides that can be used in the methods of the present disclosure can include natural nucleic acid sequences and variants thereof, artificial nucleic acid sequences, or combinations of such sequences.

[0144] Polynucleotides typically consist of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (when the polynucleotide is RNA, uracil (U) replaces thymine (T)). Thus, the term "polynucleotide sequence" is the alphabetical representation of a polynucleotide molecule; alternatively, the term can apply to the polynucleotide molecule itself. This alphabetical representation can be entered into a database in a computer with a central processing unit and used in bioinformatics applications such as functional genomics and homology searching. Polynucleotides can optionally include one or more non-standard nucleotides, nucleotide analogs, and / or modified nucleotides.

[0145] "Contact" is used in accordance with its ordinary and general meaning and is the process of bringing at least two different species (such as compounds including biomolecules, or cells) close enough to react, interact, or physically touch. However, it should be understood that the resulting reaction product can be directly produced by the reaction between the added reagents or by intermediates from one or more of the added reagents, which can be generated in the reaction mixture. The term "contact" can encompass allowing two species to react, interact, or physically touch, where the two species can be a compound and a protein or enzyme as described herein. In some embodiments, contact includes interacting a compound described herein with a protein or enzyme involved in a signal transduction pathway.

[0146] As used herein, a "therapeutic agent" or "pharmaceutical agent" refers to an agent (e.g., a compound or composition) that will have an intended prophylactic effect or an intended therapeutic effect when administered to a subject, where the prophylactic effect is, for example, preventing or delaying the onset (or recurrence) of an injury, disease, pathology, or disorder, or reducing the likelihood of the onset (or recurrence) of an injury, disease, pathology, or disorder or their symptoms, and the therapeutic effect is, for example, treating or ameliorating an injury, disease, pathology, or disorder or their symptoms, including any objective or subjective parameter of the treatment, such as elimination; alleviation; reduction of symptoms or making a patient more tolerable of an injury, pathology, or disorder; slowing the rate of degeneration or decline; making the end point of degeneration less debilitating; or improving the physical or mental health of the patient. A pharmaceutical moiety is a monovalent pharmaceutical. A therapeutic moiety is a monovalent therapeutic agent.

[0147] As used herein, the term "nucleophilic reaction product" is the reaction product between a haloalkylamine and a nucleophile (e.g., a monovalent nucleophile).

[0148] The term "nucleophile" is used according to its ordinary general chemical meaning and refers to a nucleophilic chemical group (e.g., a monovalent chemical group). A nucleophile can be an ion. A nucleophile can be monovalent. A nucleophile can be a moiety attached to the remainder of a compound (e.g., -OH) (e.g., a compound such as methanol where the remainder is -CH3). A nucleophile donates an electron pair to a substance (e.g., an electrophile), which results in the formation of a covalent bond between the nucleophile and the electrophile. Compounds or ions having a pair of free electrons or at least one π bond can act as nucleophiles. Quantification of relative nucleophilic strength, called nucleophilicity, has been devised by various methods (e.g., the Swain-Scott equation, the Ritchie equation, the Mayr-Patz equation, or the unified equation). In embodiments where there are multiple nucleophiles in a reaction (e.g., -OH or -SH), the nucleophile participating in the reaction (i.e., the reaction between a haloalkylamine and a nucleophile) is the stronger nucleophile as determined by one of the methods known in the art (e.g., the Swain-Scott equation, the Ritchie equation, the Mayr-Patz equation, or the unified equation). In an embodiment, the nucleophile includes an enol. In an embodiment, the nucleophile is -OH, an alcohol, an alkoxide anion, hydrogen peroxide, or a carboxylate anion. In an embodiment, the nucleophile is hydrogen sulfide, a thiol (-SH), a thiolate anion, a thiolcarboxylate anion (-C(O)-S-), a dithiocarbonate anion (-O-C(S)-S-), or a dithiocarbamate anion (-N-C(S)-S-). In an embodiment, the nucleophile is ammonia, an azide, an amine, a nitrite, hydroxylamine, hydrazine, carbohydrazide, phenylhydrazine, semicarbazide, or an amide. In an embodiment, the nucleophile includes ammonia, an azide, an amine, a nitrite, hydroxylamine, hydrazine, carbohydrazide, phenylhydrazine, semicarbazide, or an amide. In an embodiment, the nucleophile includes -OH, an alcohol, an alkoxide anion, hydrogen peroxide, or a carboxylate anion. In an embodiment, the nucleophile includes hydrogen sulfide, a thiol (-SH), a thiolate anion, a thiolcarboxylate anion (-C(O)-S-), a dithiocarbonate anion (-O-C(S)-S-), or a dithiocarbamate anion (-N-C(S)-S-). In an embodiment, the nucleophile is a haloester.

[0149] The terms "disease" or "disorder" refer to the state or health condition of a patient or subject that can be treated with the compounds or methods provided herein. The disease can be cancer. The disease can be an autoimmune disease. The disease can be an inflammatory disease. The disease may be an infectious disease. In some additional cases, "cancer" refers to human cancers and carcinomas, sarcomas, adenocarcinomas, lymphomas, leukemias, etc., including solid cancers and lymphomas, kidney cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, gastric cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, glioma, esophageal cancer, and liver cancer (including hepatoma), lymphoma (including acute B-lymphoblastic lymphoma), non-Hodgkin's lymphoma (e.g., Burkitt's lymphoma, small cell lymphoma, and large cell lymphoma), Hodgkin's lymphoma, leukemia (including AML, ALL, and CML), or multiple myeloma.

[0150] The terms "lung disease", "pulmonary disease", "pulmonary disorder", etc. are used interchangeably herein. This term is used to generally refer to pulmonary conditions characterized by difficulty breathing, coughing, airway discomfort and inflammation, increased mucus, and / or pulmonary fibrosis. Examples of lung diseases include lung cancer, cystic fibrosis, asthma, chronic obstructive pulmonary disease (COPD), bronchitis, emphysema, bronchiectasis, pulmonary edema, pulmonary fibrosis, sarcoidosis, pulmonary hypertension, pneumonia, tuberculosis, interstitial pulmonary fibrosis (IPF), interstitial lung disease (ILD), acute interstitial pneumonia (AlP), respiratory bronchiolitis-associated interstitial lung disease (RBILD), desquamative interstitial pneumonia (DIP), nonspecific interstitial pneumonia (NSIP), idiopathic interstitial pneumonia (IIP), bronchiolitis obliterans, organizing pneumonia (BOOP), restrictive lung disease, or pleurisy.

[0151] As used herein, the term "inflammatory disease" refers to a disease or condition characterized by abnormal inflammation (e.g., an increased level of inflammation compared to a control, such as a healthy individual without the disease). Examples of inflammatory diseases include autoimmune diseases, arthritis, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), myasthenia gravis, juvenile diabetes, type 1 diabetes, graft-versus-host disease (GvHD), Guillain-Barre syndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, ankylosing spondylitis, psoriasis, Sjogren's syndrome, vasculitis, glomerulonephritis, autoimmune thyroiditis, Behcet's disease, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, Graves ophthalmopathy, inflammatory bowel disease, Addison's disease, vitiligo, asthma, allergic asthma, acne vulgaris, celiac disease, chronic prostatitis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, ischemic reperfusion injury, stroke, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, scleroderma, and atopic dermatitis.

[0152] As used herein, the term "cancer" refers to all types of cancer, neoplasm, or malignancy found in mammals (e.g., humans), including leukemia, lymphoma, carcinoma, and sarcoma. Exemplary cancers that can be treated with the compounds or methods provided herein include brain cancer, glioma, glioblastoma, neuroblastoma, prostate cancer, colorectal cancer, pancreatic cancer, medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, head cancer, Hodgkin's disease, and non-Hodgkin's lymphoma. Exemplary cancers that can be treated with the compounds or methods provided herein include thyroid cancer, endocrine system cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, ovarian cancer, pancreatic cancer, rectal cancer, gastric cancer, and uterine cancer. Additional examples include thyroid cancer, bile duct cancer, pancreatic adenocarcinoma, cutaneous melanoma, colon adenocarcinoma, rectal adenocarcinoma, gastric adenocarcinoma, esophageal adenocarcinoma, head and neck squamous cell carcinoma, breast invasive carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung cancer, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, essential thrombocythemia, Waldenström's macroglobulinemia, primary brain tumor, malignant pancreatic insulinoma, malignant carcinoid, bladder cancer, precancerous skin lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary cancer, malignant hypercalcemia, endometrial cancer, adrenocortical cancer, endocrine or exocrine pancreatic tumors, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, or prostate cancer.

[0153] The term "leukemia" generally refers to a progressive malignant disease of the blood-forming organs and is generally characterized by the abnormal proliferation and development of white blood cells and their precursors in the blood and bone marrow. Leukemias are typically clinically classified based on: (1) the duration and nature of the acute or chronic disease; (2) the type of cells involved; myeloid (myelogenous), lymphoid (lymphocytic), or monocytic; and (3) the increase or non-increase in the number of abnormal cells in the blood - leukemic or non-leukemic (sub-leukemic). Exemplary leukemias that can be treated with the compounds or methods provided herein include, for example, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, leukemic leukemia, basophilic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, cutaneous leukemia, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy cell leukemia, hemoblastic leukemia, hemocytoblast leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, microgranulocytic leukemia, monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myelogranulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, multiple myeloma, plasmacytic leukemia, promyelocytic leukemia, Rieder cell leukemia, Schilling's leukemia, stem cell leukemia, sub-leukemic leukemia, or undifferentiated cell leukemia.

[0154] As used herein, the term "lymphoma" refers to a group of cancers that affect hematopoietic and lymphoid tissues. It begins in lymphocytes, which are blood cells found mainly in lymph nodes, spleen, thymus, and bone marrow. The two main types of lymphoma are non-Hodgkin lymphoma and Hodgkin disease. Hodgkin disease accounts for approximately 15% of all diagnosed lymphomas. It is a cancer associated with Reed-Sternberg malignant B lymphocytes. Non-Hodgkin lymphoma (NHL) can be classified based on the growth rate of the cancer and the cell types involved. There are aggressive (high-grade) and indolent (low-grade) types of NHL. Based on the cell types involved, there are B-cell and T-cell NHL. Exemplary B-cell lymphomas that can be treated with the compounds or methods provided herein include, but are not limited to, small lymphocytic lymphoma, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, extranodal (MALT) lymphoma, nodular (monocytoid B-cell) lymphoma, splenic lymphoma, diffuse large cell B lymphoma, Burkitt lymphoma, lymphoblastic lymphoma, immunoblastic large cell lymphoma, or precursor B lymphoblastic lymphoma. Exemplary T-cell lymphomas that can be treated with the compounds or methods provided herein include, but are not limited to, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, anaplastic large cell lymphoma, mycosis fungoides, and precursor T lymphoblastic lymphoma.

[0155] The term "sarcoma" generally refers to a tumor composed of substances such as embryonic connective tissue and generally consisting of closely packed cells embedded in a fibrous or homogeneous substance. Sarcomas that can be treated with the compounds or methods provided herein include chondrosarcoma, fibrosarcoma, lymphosarcoma, melanocarcinoma, myxosarcoma, osteosarcoma, Abemethy's sarcoma, liposarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma, choriocarcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcoma of B cells, lymphoma, immunoblastic sarcoma of T cells, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukosarcoma, malignant mesothelial sarcoma, extraskeletal sarcoma, reticulosarcoma, Rous sarcoma, serocystic sarcoma, synovial sarcoma or telangiectaltic sarcoma.

[0156] The term "melanoma" refers to a tumor originating from the melanocyte system of the skin and other organs. Melanomas that can be treated with the compounds or methods provided herein include, for example, acral lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna, malignant melanoma, nodular melanoma, subungual melanoma or superficial spreading melanoma.

[0157] The term "carcinoma" refers to a malignant new growth composed of epithelial cells, which tend to infiltrate surrounding tissues and cause metastasis. Exemplary carcinomas that can be treated with the compounds or methods provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, acinar carcinoma, acinar cell carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, adenocarcinoma, adrenocortical carcinoma, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, basaloid cell tumor, basal cell-like carcinoma, basal squamous cell carcinoma, bronchioloalveolar carcinoma, bronchiolar carcinoma, bronchial carcinoma, cerebriform carcinoma, cholangiocarcinoma, choriocarcinoma, colloid carcinoma, comedocarcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, cutaneous carcinoma, columnar carcinoma, columnar cell carcinoma, ductal carcinoma, scirrhous carcinoma, embryonal carcinoma, medullary carcinoma, epidermoid carcinoma, adenoid epithelioma, explanted carcinoma, carcinoma ex ulcere, fibroid carcinoma, gelatiniform carcinoma, colloid carcinoma, giant cell carcinoma, megalocytic carcinoma, adenocarcinoma, granulosa cell carcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, adrenoid carcinoma, infantile embryonal carcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma, Krompecher's carcinoma, Kulchitzky-cell carcinoma, large cell carcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma, lymphoepithelial carcinoma, medullary carcinoma, myeloid carcinoma, melanoma, encephaloid carcinoma, mucinous carcinoma, mucinous adenocarcinoma, mucinous cell carcinoma, mucoepidermoid carcinoma, myxoid carcinoma, myxomatous carcinoma, nasopharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans, osteoid carcinoma, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, spinous cell carcinoma, pultaceous carcinoma, renal cell carcinoma of the kidney, reserve cell carcinoma, sarcomatoid carcinoma, schneiderian carcinoma, scirrhous carcinoma, scrotal carcinoma, signet ring cell carcinoma, simplex carcinoma, small cell carcinoma, tuberosum carcinoma, globular cell carcinoma, spindle cell carcinoma, carcinoma spongiosum, squamous carcinoma, squamous cell carcinoma, string carcinoma, carcinomatelangiectaticum), carcinomatelangiectodes, transitional cell carcinoma, carcinoma tuberosum, tuberous carcinoma, verrucous carcinoma or villous carcinoma.

[0158] As used herein, the terms "cancer metastasis", "metastatic", and "metastatic cancer" are used interchangeably and refer to the spread of a proliferative disease or disorder (such as cancer) from one organ to another non-adjacent organ or body part. "Metastatic cancer" is also referred to as "stage IV cancer". Cancer occurs at the site of origin, such as the breast, and this site is called the primary tumor, such as primary breast cancer. Some of the cancer cells in the primary tumor or site of origin acquire the ability to penetrate and invade the normal tissue surrounding the local area, and / or the ability to penetrate the lymphatic system or the walls of the vascular system of the systemic circulation to other parts and tissues within the body. The second clinically detectable tumor formed by the cancer cells of the primary tumor is called a metastatic or secondary tumor. When cancer cells metastasize, it is presumed that the metastatic tumor and its cells are similar to the primary tumor. Thus, if lung cancer metastasizes to the breast, the secondary tumor in the breast is composed of abnormal lung cells rather than abnormal breast cells. The secondary tumor in the breast is a metastatic lung cancer. Thus, the phrase metastatic cancer refers to a disease in which a subject has or has had a primary tumor and has one or more secondary tumors. The phrase non-metastatic cancer or an individual with non-metastatic cancer refers to a disease in which an individual has a primary tumor but does not have one or more secondary tumors. For example, metastatic lung cancer refers to a disease in an individual who has a primary lung tumor or a history of a primary lung tumor and has one or more secondary tumors in a second location or multiple locations (such as in the breast).

[0159] The term "cutaneous metastasis" or "skin metastasis" refers to the growth of secondary malignant cells in the skin, where the malignant cells originate from the primary cancer site (such as the breast). In cutaneous metastasis, cancer cells from the primary cancer site can migrate to the skin, where these cells divide and cause lesions. Cutaneous metastasis can be caused by the migration of cancer cells from a breast cancer tumor to the skin.

[0160] The term "visceral metastasis" refers to the growth of secondary malignant cells in visceral organs (such as the heart, lungs, liver, pancreas, intestines) or body cavities (such as the pleura, peritoneum), where the malignant cells originate from a primary cancer site (such as the head and neck, liver, breast). In visceral metastasis, cancer cells from the primary cancer site can migrate to visceral organs, where these cells divide and cause lesions. Visceral metastasis can be caused by the migration of cancer cells from a liver cancer tumor or a head and neck tumor to visceral organs.

[0161] The term "treating" or "treatment" refers to any indication of successful treatment or improvement of an injury, disease, pathology, or condition, including any objective or subjective parameter, such as elimination; alleviation; reduction of symptoms or making the injury, pathology, or condition more tolerable to the patient; slowing the rate of degeneration or decline; or making the endpoint of degeneration less regressive; improving the physical or mental health of the patient. Treatment or improvement of symptoms can be based on objective or subjective parameters; including the results of physical examination, neuropsychiatric examination, and / or psychiatric evaluation. The term "treatment" and its conjugates can include prevention of an injury, pathology, condition, or disease. In an embodiment, treatment is prevention. In an embodiment, treatment does not include prevention.

[0162] As used herein (and as is well understood in the art), "treating" or "treatment" also broadly includes any method for obtaining a beneficial or desired result (including a clinical result) with respect to the condition of a subject. Beneficial or desired clinical results can include, but are not limited to, alleviating or improving one or more symptoms or conditions, reducing the severity of the disease, stabilizing the disease state (i.e., not worsening), preventing the transmission or spread of the disease, delaying or slowing the progression of the disease, improving or alleviating the disease state, reducing disease recurrence, and remission, whether partial or complete, and whether detectable or not. In other words, "treatment" as used herein includes any cure, improvement, or prevention of a disease. Treatment can prevent the occurrence of a disease; inhibit the spread of a disease; alleviate the symptoms of a disease; completely or partially remove the underlying cause of a disease; shorten the duration of a disease; or a combination of these things.

[0163] As used herein, "treating" and "treatment" include prophylactic treatment. A method of treatment comprises administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may comprise a series of administrations. The length of the treatment period depends on a variety of factors such as the severity of the condition, the age of the patient, the concentration of the active agent, the activity of the composition used in the treatment, or a combination thereof. It should also be understood that the effective dose of a reagent for treatment or prophylaxis may be increased or decreased during the course of a particular treatment or prophylaxis regimen. By standard diagnostic assays known in the art, changes in dose can be made and become apparent. In some cases, long-term administration may be required. For example, a composition is administered to a subject in an amount sufficient to treat the patient and for a sufficient duration. In an embodiment, "treating" or "treatment" is not prophylactic treatment.

[0164] The term "prophylaxis" refers to reducing the occurrence of disease symptoms in a patient. As noted above, prophylaxis can be complete (no detectable symptoms) or partial, such that fewer symptoms are observed than would occur in the absence of treatment.

[0165] A "patient" or "subject in need" refers to a living organism that is suffering from or is susceptible to a disease or condition that can be treated by administering a pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient is a human.

[0166] "Effective amount" means an amount that, relative to the absence of the compound, is sufficient for the compound to achieve the stated purpose (e.g., achieve its administration effect, treat a disease, reduce enzyme activity, increase enzyme activity, reduce a signal transduction pathway, or alleviate one or more symptoms of a disease or condition). An example of an "effective amount" is an amount sufficient to effect treatment, prevention, or reduction of one or more symptoms of a disease, which amount may also be referred to as a "therapeutically effective amount". "Reduction" of one or more symptoms (and grammatical equivalents of this phrase) means a decrease in the severity or frequency of one or more symptoms, or the elimination of one or more symptoms. A "prophylactically effective amount" of a drug is an amount of the drug that, when administered to a subject, will have the desired prophylactic effect, e.g., prevent or delay the onset (or recurrence) of an injury, disease, pathology, or condition, or reduce the likelihood of the onset (or recurrence) of an injury, disease, pathology, or condition or its symptoms. A complete prophylactic effect does not necessarily occur upon administration of a single dose and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in a single or multiple administrations. As used herein, "amount reducing activity" means the amount of an antagonist required to reduce the activity of an enzyme relative to the absence of the antagonist. As used herein, "amount disrupting function" means the amount of an antagonist required to disrupt the function of an enzyme or protein relative to the absence of the antagonist. The exact amount will depend on the purpose of the treatment and will be determinable by those of skill in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th ed., 2003, edited by Gennaro, Lippincott, Williams & Wilkins).

[0167] For any compound described herein, a therapeutically effective amount can initially be determined according to cell culture assays. The target concentration will be the concentration of one or more active compounds that is capable of achieving the methods described herein, as measured using the methods described herein or known in the art.

[0168] As is well known in the art, a therapeutically effective amount for humans can also be determined by animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dose in humans can be adjusted by monitoring the effectiveness of the compound and upregulating or downregulating the dose as described above. Adjusting the dose based on the methods described above and other methods to achieve maximum efficacy in humans is well within the capabilities of a person of ordinary skill in the art.

[0169] As used herein, the term "therapeutically effective amount" refers to the amount of a therapeutic agent sufficient to ameliorate a disorder as described above. For example, for a given parameter, a therapeutically effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90% or at least 100%. Therapeutic efficacy can also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount can have at least 1.2-fold, 1.5-fold, 2-fold, 5-fold or more effect relative to a control.

[0170] The dose can vary depending on the needs of the patient and the compound employed. In the context of the present disclosure, the dose administered to a patient should be sufficient to produce a beneficial therapeutic response in the patient over time. The magnitude of the dose will also be determined by the presence, nature and extent of any adverse side effects. Determining the appropriate dose for a particular situation is within the skill of the practitioner. Generally, treatment is initiated at a smaller dose that is less than the optimal dose of the compound. Thereafter, the dose is increased in small increments until the optimal effect is achieved in a number of cases. The dose and interval can be adjusted individually to provide an effective level of the administered compound for the particular clinical indication being treated. This will provide a treatment regimen commensurate with the severity of the individual disease state.

[0171] As used herein, the term "administer" means to administer orally to a subject, administer in the form of a suppository, surface contact, intravenously, parenterally, intraperitoneally, intramuscularly, intralesionally, intrathecally, intranasally or subcutaneously, or implant a slow release device, such as a mini-osmotic pump. Administration is effected by any route, including parenterally and transmucosally (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular and intracranial administration. Other delivery modes include, but are not limited to, the use of liposomal formulations, intravenous infusions, transdermal patches, etc. In an embodiment, the administration does not include the administration of any active agent other than the recited active agent.

[0172] "Co-administered" means that the compositions described herein are administered at the same time, immediately before, or immediately after the administration of one or more additional therapies. The compounds provided herein can be administered to a patient alone or can be co-administered. Co-administration is intended to include the compounds being administered simultaneously or sequentially, either alone or in combination (more than one compound). Thus, when desired, the formulations can also be combined with other active substances (e.g., to reduce metabolic degradation). The compositions of the present disclosure can be transdermal, delivered via a surface route, or formulated in the form of applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.

[0173] As used herein, a cancer model organism is an organism that exhibits a phenotype suggestive of cancer, or the activity of a component that causes cancer, within the organism. The term cancer is defined above. A variety of organisms can be used as cancer model organisms and include, for example, cancer cells and mammalian organisms such as rodents (e.g., mice or rats) and primates (such as humans). Those skilled in the art widely understand cancer cell lines to be cells that exhibit a phenotype or genotype similar to cancer in vivo. As used herein, cancer cell lines include cell lines from animals (e.g., mice) and from humans.

[0174] The term "pharmaceutically acceptable salts" is intended to include salts of the active compounds prepared with relatively non-toxic acids or bases on the basis of specific substituents found on the compounds described herein. When the compounds of the present disclosure 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 pure or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include sodium salts, potassium salts, calcium salts, ammonium salts, organic amine salts, or magnesium salts or similar salts. When the compounds of the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either pure or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate, phosphoric acid, hydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfide, hydroiodic acid, or phosphorous acid, etc.; and salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, oxalic acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginine salts, and salts of organic acids such as glucuronic acid or galacturonic acid (see, e.g., Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure contain both basic and acidic functional groups that permit the conversion of these compounds into base addition salts or acid addition salts.

[0175] Accordingly, the compounds of the present disclosure can exist, for example, in the form of salts with pharmaceutically acceptable acids. The present disclosure encompasses such salts. Non-limiting examples of such salts include hydrochlorides, hydrobromides, phosphates, sulfates, mesylates, nitrates, maleates, acetates, citrates, fumarates, propionates, tartrates (e.g., (+)-tartrates, (-)-tartrates, or mixtures thereof including racemic mixtures), succinates, benzoates, and salts with amino acids such as glutamic acid, and quaternary ammonium salts (e.g., methyl iodide, ethyl iodide, etc.). These salts can be prepared by methods known to those skilled in the art.

[0176] The neutral form of the compounds is preferably regenerated by contacting the salt with a base or an acid and isolating the parent compound in a conventional manner. The parent form of the compound may differ in certain physical properties from the various salt forms, such as solubility in polar solvents.

[0177] In addition to the de-salted forms, the present disclosure also provides prodrug forms of the compounds. Prodrugs of the compounds described herein are those compounds that are readily subjected to chemical changes under physiological conditions to provide the compounds of the present disclosure. Prodrugs of the compounds described herein can be converted in vivo after administration. Additionally, prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical means in an ex vivo environment (such as when contacted with a suitable enzyme or chemical reagent).

[0178] Certain compounds of the present disclosure can exist in unsolvated forms as well as solvated forms (including hydrated forms). Generally speaking, the solvated forms are equivalent to the unsolvated forms and are encompassed within the scope of the present disclosure. Certain compounds of the present disclosure can exist in multiple crystalline or amorphous forms. Generally speaking, all physical forms are equivalent for the uses contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0179] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that assist in the administration of an active agent to a subject and facilitate the absorption of the subject, and such substances can be included in the compositions of the present disclosure without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, saline solution, lactated Ringer's solution, common sucrose, common glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents, salt solutions (such as Ringer's solution), alcohols, oils, gelatin, carbohydrates (such as lactose, amylose or starch), fatty acid esters, carboxymethyl cellulose, polyvinylpyrrolidone and pigments, etc. Such preparations can be sterilized and, if necessary, can be mixed with adjuvants (such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts for influencing osmotic pressure, buffers, coloring substances and / or aromatic substances, etc.), which do not react harmfully with the compounds of the present disclosure. Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present disclosure.

[0180] The term "formulation" is intended to include a formulation of an active compound with an encapsulating material in the form of a carrier that provides a capsule, in which the active component, with or without other carriers, is surrounded by the carrier and the active component is thus associated with the carrier. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets and lozenges can be used as solid dosage forms suitable for oral administration.

[0181] As used herein, the term "about" means a range of values including the specified value, which would be reasonably similar to the specified value by those of ordinary skill in the art. In an embodiment, about means within the standard deviation using measurements generally acceptable in the art. In an embodiment, about means extending to a range of + / - 10% of the specified value. In an embodiment, about includes the specified value.

[0182] II. Compound

[0183] On the one hand, a compound having the following formula is provided: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0184] On the one hand, a compound having the following formula is provided: R 1 is a silyl protecting group. In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure. In an embodiment, R 1 is trimethylsilyl (TMS). In an embodiment, R 1 is triethylsilyl (TES). In an embodiment, R 1 is tert-butyldimethylsilyl (TBS / TBDMS). In an embodiment, R 1 is tert-butyldiphenylsilyl (TBDPS). In an embodiment, R 1 is triisopropylsilyl (TIPS).

[0185] In an embodiment, a compound having the following formula is provided: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0186] In one aspect, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0187] In one aspect, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0188] In one aspect, there is provided a compound having the following formula: R 1is a silyl protecting group. In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure. In an embodiment, R 1 is trimethylsilyl (TMS). In an embodiment, R 1 is triethylsilyl (TES). In an embodiment, R 1 is tert-butyldimethylsilyl (TBS / TBDMS). In an embodiment, R 1 is tert-butyldiphenylsilyl (TBDPS). In an embodiment, R 1 is triisopropylsilyl (TIPS).

[0189] In an embodiment, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0190] On the one hand, there is provided a compound having the following formula: In an embodiment, R 1is a silyl protecting group and wherein the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure. In an embodiment, R 1 is trimethylsilyl (TMS). In an embodiment, R 1 is triethylsilyl (TES). In an embodiment, R 1 is tert-butyldimethylsilyl (TBS / TBDMS). In an embodiment, R 1 is tert-butyldiphenylsilyl (TBDPS). In an embodiment, R 1 is triisopropylsilyl (TIPS).

[0191] In an embodiment, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0192] In one aspect, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0193] In one aspect, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0194] In one aspect, there is provided a compound having the following formula: In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0195] In an embodiment, a compound as described herein comprises at least 5 grams of the compound, with or without a pharmaceutically acceptable excipient.

[0196] In an embodiment, the compound is R 1 is a silyl protecting group. In an embodiment, the compound is

[0197]

[0198] In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound is In an embodiment, the compound comprises at least 10 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 25 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 50 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 100 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 250 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 500 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 1000 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 2000 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 3000 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 4000 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 5000 grams of the compound with or without a pharmaceutically acceptable excipient. In an embodiment, the compound comprises at least 10,000 grams of the compound with or without a pharmaceutically acceptable excipient.

[0199] III. Pharmaceutical Composition

[0200] On the one hand, a pharmaceutical composition is provided, which comprises a compound having the following formula: and a pharmaceutically acceptable excipient. In an embodiment, the compound is at least 99% enantiomerically pure. In an embodiment, the compound is at least 98% enantiomerically pure. In an embodiment, the compound is at least 97% enantiomerically pure. In an embodiment, the compound is at least 96% enantiomerically pure. In an embodiment, the compound is at least 95% enantiomerically pure. In an embodiment, the compound is at least 94% enantiomerically pure. In an embodiment, the compound is at least 93% enantiomerically pure. In an embodiment, the compound is at least 92% enantiomerically pure. In an embodiment, the compound is at least 91% enantiomerically pure. In an embodiment, the compound is at least 90% enantiomerically pure.

[0201] In an embodiment, the pharmaceutically acceptable excipient is Kolliphor HS15, Kolliphor EL, Cremaphor RH40, Kolliphor P188 or Kolliphor P407. In an embodiment, the pharmaceutically acceptable excipient is Kolliphor HS15. In an embodiment, the pharmaceutically acceptable excipient is Kolliphor EL. In an embodiment, the pharmaceutically acceptable excipient is Cremaphor RH40, Kolliphor P188. In an embodiment, the pharmaceutically acceptable excipient is Kolliphor P407.

[0202] IV. Method for preparing the compound

[0203] On the one hand, a method for preparing a compound having the following formula is provided: The method comprises reacting a compound having the following formula: with 1-(dimethoxymethyl)-4-methoxybenzene in the presence of CBr4, an alcohol, a base and one or more organic solvents. In an embodiment, the method comprises reacting a compound having the following formula: with 1-(dimethoxymethyl)-4-methoxybenzene in the presence of CBr4, isopropanol, imidazole and dichloromethane.

[0204] In an embodiment, the alcohol is methanol, ethanol or isopropanol. In an embodiment, the alcohol is methanol. In an embodiment, the alcohol is ethanol. In an embodiment, the alcohol is isopropanol. In an embodiment, the base is imidazole. In an embodiment, the organic solvent is dichloromethane or chloroform. In an embodiment, the organic solvent is dichloromethane. In an embodiment, the organic solvent is chloroform.

[0205] On the one hand, a method for preparing a compound having the following formula is provided: The method comprises reacting a compound having the following formula: React with a transition metal catalyst for olefin metathesis in the presence of one or more organic solvents. In an embodiment, the method includes reacting a compound having the following formula: React with a Hoveyda-Grubbs 2nd generation catalyst in the presence of toluene. In an embodiment, the method includes reacting a compound having the following formula: React with a Hoveyda-Grubbs 2nd generation catalyst at 120 °C in the presence of toluene.

[0206] In an embodiment, the transition metal catalyst is a ruthenium-based catalyst. In an embodiment, the transition metal catalyst is a Grubbs catalyst. In an embodiment, the transition metal catalyst is a Hoveyda-Grubbs catalyst.

[0207] In an embodiment, the transition metal catalyst is a Grubbs 1st generation catalyst, a Grubbs 2nd generation catalyst, a Grubbs 3rd generation catalyst, a Hoveyda-Grubbs 1st generation catalyst, a Hoveyda-Grubbs 2nd generation catalyst, NitroGrela, dichloro[1,3-bis(2-methylphenyl)-2-imidazolidinylidene](benzylidene)(tricyclohexylphosphine)ruthenium(II) [Grubbs M2a SI(o-Tol)(C793)], dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](3-methyl-2-butenylidene)(tricyclohexylphosphine)ruthenium(II) [Grubbs M2b(C827)], dichloro[1,3-bis(2-methylphenyl)-2-imidazolidinylidene](2-isopropoxybenzylidene)ruthenium(II) [Hoveyda-Grubbs M72 SI(o-Tol)(C571) or Stewart-Grubbs catalyst] or dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][3-(2-pyridyl)propylidene]ruthenium(II) (Grubbs C598). In an embodiment, the transition metal catalyst is a Grubbs 1st generation catalyst, a Grubbs 2nd generation catalyst, a Hoveyda-Grubbs 1st generation catalyst, a Hoveyda-Grubbs 2nd generation catalyst or NitroGrela.

[0208] In an embodiment, the transition metal catalyst is the 1st generation Grubbs catalyst. In an embodiment, the transition metal catalyst is the 2nd generation Grubbs catalyst. In an embodiment, the transition metal catalyst is the 3rd generation Grubbs catalyst. In an embodiment, the transition metal catalyst is the 1st generation Hoveyda-Grubbs catalyst. In an embodiment, the transition metal catalyst is the 2nd generation Hoveyda-Grubbs catalyst. In an embodiment, the transition metal catalyst is NitroGrela. In an embodiment, the transition metal catalyst is dichloro[1,3-bis(2-methylphenyl)-2-imidazolidinylidene](benzylidene)(tricyclohexylphosphine)ruthenium(II) [Grubbs M2a SI(o-Tol)(C793)]. In an embodiment, the transition metal catalyst is dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](3-methyl-2-butenylidene)(tricyclohexylphosphine)ruthenium(II) [Grubbs M2b(C827)]. In an embodiment, the transition metal catalyst is dichloro[1,3-bis(2-methylphenyl)-2-imidazolidinylidene](2-isopropoxybenzylidene)ruthenium(II) [Hoveyda-Grubbs M72 SI(o-Tol)(C571) or Stewart-Grubbs catalyst]. In an embodiment, the transition metal catalyst is dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][3-(2-pyridyl)propylidene]ruthenium(II) (Grubbs C598).

[0209] In an embodiment, the organic solvent is toluene.

[0210] On the one hand, a method for preparing a compound having the following formula is provided: The method includes reacting a compound having the following formula: with the 2nd generation Hoveyda-Grubbs catalyst in the presence of toluene.

[0211] On the one hand, a method for preparing a compound having the following formula is provided: The method includes reacting a compound having the following formula: with a strong acid in the presence of an alcohol and one or more organic solvents. In an embodiment, the method includes reacting a compound having the following formula: with camphorsulfonic acid in the presence of methanol and dichloromethane. In an embodiment, the strong acid is camphorsulfonic acid, pyridinium p-toluenesulfonate or p-toluenesulfonic acid. In an embodiment, the strong acid is camphorsulfonic acid. In an embodiment, the strong acid is pyridinium p-toluenesulfonate In an embodiment, the strong acid is p-toluenesulfonic acid. In an embodiment, the organic solvent is dichloromethane or chloroform. In an embodiment, the strong acid is camphorsulfonic acid and the solvent is dichloromethane.

[0212] On the one hand, a method for preparing a compound having the following formula is provided: This method includes reacting a compound having the following formula: with an acetylating agent in the presence of a strong acid and one or more organic solvents. This method includes reacting a compound having the following formula: with 1,1,1-trimethoxyethane in the presence of camphorsulfonic acid and dichloromethane. In an embodiment, the acetylating agent is acetic anhydride or 1,1,1-trimethoxyethane. In an embodiment, the acetylating agent is acetic anhydride. In an embodiment, the acetylating agent is 1,1,1-trimethoxyethane. In an embodiment, the strong acid is camphorsulfonic acid, pyridinium p-toluenesulfonate or p-toluenesulfonic acid. In an embodiment, the strong acid is camphorsulfonic acid. In an embodiment, the strong acid is pyridinium p-toluenesulfonate In an embodiment, the strong acid is p-toluenesulfonic acid. In an embodiment, the organic solvent is dichloromethane or chloroform.

[0213] In an embodiment, a method for preparing a compound having the following formula is provided: In an embodiment, this method includes reacting a compound having the following formula: with acetic anhydride in the presence of 4-dimethylaminopyridine and pyridine. In an embodiment, a method for preparing a compound having the following formula is provided: In an embodiment, this method includes reacting a compound having the following formula: with acetic anhydride in the presence of acetylimidazole, 4-dimethylaminopyridine and tetrahydrofuran. In an embodiment, a method for preparing a compound having the following formula is provided: In an embodiment, this method includes reacting a compound having the following formula: with acetyl chloride in the presence of triethylamine and dichloromethane at a reduced temperature. In an embodiment, a method for preparing a compound having the following formula is provided: In an embodiment, this method includes reacting a compound having the following formula: with methyl S-thioacetate in the presence of dichloromethane.

[0214] On the one hand, a method for preparing a linear polyketone compound is provided.

[0215] In an embodiment, the polyketone compound is a splicing regulator.

[0216] In an embodiment, the polyketone compound is 17S-FD-895.

[0217] On the one hand, a method for preparing 17S-FD-895 is provided, which method comprises using compounds 6a, 6b, 6c, 6d and 6e as described herein.

[0218] V. Method of treatment

[0219] On the one hand, a method for treating cancer is provided, which method comprises administering to a subject in need thereof an effective amount of a polyketide compound prepared by the method as described herein.

[0220] In an embodiment, the cancer is a blood cancer.

[0221] VI. Embodiments

[0222] Embodiment P1. A method for preparing a linear polyketide compound.

[0223] Embodiment P2. The method according to Embodiment P1, wherein the polyketide compound is a splicing regulator.

[0224] Embodiment P3. The method according to Embodiment P1, wherein the polyketide compound is 17S-FD-895.

[0225] Embodiment P4. A method for treating cancer, which method comprises administering to a subject in need thereof an effective amount of the polyketide compound prepared by the method according to any one of Embodiments P1 to P3.

[0226] Embodiment P5. The method according to Embodiment P4, wherein the cancer is a blood cancer.

[0227] Embodiment P6. A method for preparing 17S-FD-895, which method comprises using compounds 6a, 6b, 6c, 6d and 6e as shown in Scheme 1.

[0228] VII. Additional embodiments

[0229] Embodiment 1. A compound having the following formula:

[0230] wherein the compound is at least 95% enantiomerically pure.

[0231] Embodiment 2. The compound according to Embodiment 1, wherein the compound is at least 98% enantiomerically pure.

[0232] Embodiment 3. A compound having the following formula:

[0233] wherein the compound is at least 95% enantiomerically pure.

[0234] Embodiment 4. The compound according to Embodiment 3, wherein the compound is at least 98% enantiomerically pure.

[0235] Embodiment 5. A compound having the following formula:

[0236] wherein the compound is at least 95% enantiomerically pure.

[0237] Embodiment 6. The compound according to Embodiment 5, wherein the compound is at least 98% enantiomerically pure.

[0238] Embodiment 7. A compound having the following formula:

[0239] wherein the compound is at least 95% enantiomerically pure.

[0240] Embodiment 8. The compound according to Embodiment 7, wherein the compound is at least 98% enantiomerically pure.

[0241] Embodiment 9. A compound having the following formula:

[0242] wherein the compound is at least 95% enantiomerically pure.

[0243] Embodiment 10. The compound according to Embodiment 9, wherein the compound is at least 98% enantiomerically pure.

[0244] Embodiment 11. A compound having the following formula:

[0245] wherein the compound is at least 95% enantiomerically pure.

[0246] Embodiment 12. The compound according to Embodiment 11, wherein the compound is at least 98% enantiomerically pure.

[0247] Embodiment 13. A compound having the following formula:

[0248] wherein the compound is at least 95% enantiomerically pure.

[0249] Embodiment 14. The compound according to Embodiment 13, wherein the compound is at least 98% enantiomerically pure.

[0250] Embodiment 15. A compound having the following formula:

[0251] wherein the compound is at least 95% enantiomerically pure.

[0252] Embodiment 16. The compound according to embodiment 15, wherein the compound is at least 98% enantiomerically pure.

[0253] Embodiment 17. A compound having the following formula:

[0254] wherein the compound is at least 95% enantiomerically pure.

[0255] Embodiment 18. The compound according to embodiment 17, wherein the compound is at least 98% enantiomerically pure.

[0256] Embodiment 19. The compound according to embodiments 1 to 18, the compound comprising at least 5 grams of the compound with or without a pharmaceutically acceptable excipient.

[0257] Embodiment 20. A pharmaceutical composition, the pharmaceutical composition comprising a compound having: the formula and a pharmaceutically acceptable excipient, wherein the compound is at least 95% enantiomerically pure.

[0258] Embodiment 21. The pharmaceutical composition according to embodiment 20, wherein the compound is at least 98% enantiomerically pure.

[0259] Embodiment 22. A method for preparing a compound having: the formula; the method comprising reacting a compound having the formula: with 1-(dimethoxymethyl)-4-methoxybenzene in the presence of CBr4, an alcohol, a base and one or more organic solvents.

[0260] Embodiment 23. The method according to embodiment 22, wherein the alcohol is methanol, ethanol or isopropanol.

[0261] Embodiment 24. The method according to embodiment 22, wherein the alcohol is isopropanol.

[0262] Embodiment 25. The method according to embodiment 22, wherein the base is imidazole.

[0263] Embodiment 26. The method according to embodiment 22, wherein the organic solvent is dichloromethane or chloroform.

[0264] Embodiment 27. A method for preparing a compound having the formula: the method comprising reacting a compound having the formula: with a transition metal catalyst for olefin metathesis in the presence of one or more organic solvents.

[0265] Embodiment 28. The method according to embodiment 27, wherein the transition metal catalyst is a ruthenium-based catalyst.

[0266] Embodiment 29. The method according to embodiment 27, wherein the transition metal catalyst is a first-generation Grubbs catalyst, a second-generation Grubbs catalyst, a first-generation Hoveyda-Grubbs catalyst, a second-generation Hoveyda-Grubbs catalyst, or NitroGrela.

[0267] Embodiment 30. The method according to embodiment 27, wherein the transition metal catalyst is a second-generation Hoveyda-Grubbs catalyst.

[0268] Embodiment 31. The method according to embodiment 27, wherein the organic solvent is toluene.

[0269] Embodiment 32. A method for preparing a compound having the following formula: The method comprises reacting a compound having the formula: with a strong acid in the presence of an alcohol and one or more organic solvents.

[0270] Embodiment 33. The method according to embodiment 32, wherein the strong acid is camphorsulfonic acid.

[0271] Embodiment 34. The method according to embodiment 32, wherein the organic solvent is dichloromethane or chloroform.

[0272] Embodiment 35. A method for preparing a compound having the formula: The method comprises reacting a compound having the formula: with an acetylating agent in the presence of a strong acid and one or more organic solvents.

[0273] Embodiment 36. The method according to embodiment 35, wherein the acetylating agent is acetic anhydride or 1,1,1-trimethoxyethane.

[0274] Embodiment 37. The method according to embodiment 35, wherein the strong acid is camphorsulfonic acid.

[0275] Embodiment 38. The method according to embodiment 37, wherein the organic solvent is dichloromethane or chloroform.

[0276] Examples

[0277] Example 1. Initial synthesis work

[0278] The compound numbers used in Examples 1 and 2 correspond to the compounds described in these examples, andFigure 1 , Figures 2A - 2F , the compounds described in Scheme 1, Schemes S1 - S6, and the embodiments.

[0279] A practical 14 - step assembly method has been developed, which can economically and efficiently produce gram - scale amounts of the highly potent splicing regulator 17S - FD - 895, the result of installing 11 stereocenters, a 12 - membered macrolide, and a complex and dense linear polyketide tail.

[0280] Since their first discovery in the mid - 1990s, studies on the mode of action (MOA) a decade later revealed that a family of polyketides, including FD - 895, pladienolide, spliceostatin, herboxadiene, GEX1, FR901464, and thailanstatin, share a similar ability to regulate splicing through interactions within the SF3b component of the spliceosome. First thought to be a common motif and subsequently confirmed by structural biology analysis, these small molecules uniquely position themselves at the interface between SF3B1, PHF5A, and SF3B3. Here, the importance and positioning of the stereochemical centers within these molecules clearly demonstrate the unique geometric requirements for functional binding.

[0281] Although many natural products, homologs, and semi - synthetic analogs show the necessary functional space display to be able to bind easily to the SF3B pocket and thus exhibit highly potent splicing regulation, the high density of their functional groups leads to reduced material stability. Notably, many of these natural products are very unstable in aqueous media, with half - lives typically less than 30 minutes. Recent studies have shown that synthetic modifications at C16 - C17 are not only tolerated but also allow for a three - dimensional arrangement, which greatly reduces the degradation rate while meeting the requirements for active binding to the binding pocket in SF3B, ultimately leading to the identification of 17S - FD - 895 as a potential therapeutic lead.

[0282] The severity of this problem was partially manifested in the first clinical trial of the splicing regulator E7107. Although developed with a significant level of diligence and optimization, the instability was partly a cause for concern regarding the results of the first clinical trials of this new class of agents. In the development of the problems observed with E7107, subsequent procedures led to H3B-8800 entering a Phase 1 clinical trial to evaluate the safety, pharmacokinetics, and pharmacodynamics of H3B-8800 in subjects with myelodysplastic syndromes, acute myeloid leukemia (AML), and chronic myelomonocytic leukemia. Although access to E7107 and H3B-8800 was achieved through synthesis, the final preclinical and clinical deliveries were generated through semi-synthetic preparations, partly due to the complexity associated with converting many of the published milligram-scale routes to gram-scale methods to date. To date, a gram-scale synthetic method has not been achieved to access all but the simpler hopene family of substances. Here, we describe the development of a practical gram-scale route to 17S-FD-895 (1), which employs a highly convergent route enabling the preparation of potentially superior derivatives for clinical applications.

[0283] Developed in the context of a rich synthetic work forum, the synthetic challenge was to provide multi-gram preparations of a molecule containing 31 carbons, of which 11 carbons and 6 carbons respectively occupy sp 3 and sp 2 stereochemistry. Notably, only 5 carbons lack stereochemical requirements and are functionalized throughout the molecule. In addition to this high density of functionality, the molecule also contains a 12-membered lactone ring, which is a class of small polyketides, including mycolactone and other polyketides that exhibit this rare ring size.

[0284] Our method was developed from multiple synthetic activities that identified the importance of component assembly and operated through a three-pronged strategy. As Figure 1 shown, the first prong, component preparation, began with establishing practical methods to synthesize multi-hundred-gram quantities of six components 6a - 6e. Here, the aim of these studies was to perform the efficient preparation of 6a - 6e in a cost-effective manner using the overall yields and materials listed in Figure 1 . With these materials available, our focus then shifted to assembly. Here, the goal was to achieve a method that would enable the preparation of gram-scale quantities of 1 from five components in less than a month.

[0285] Furthermore, our design goal was to provide the bioactive substance only in the final step. Since these compounds exhibit very potent bioactivity and early clinical trials have shown a very low MTD in humans, we chose a route with two-sided assembly that begins with the preparation of two bio-inactive fragments, such as the core 2 (Figure 1 ) and side chain 3( Figure 1 ), followed by a final coupling to afford 17S-FD-895(1). Supported by structural studies, the lack of a link between the side chain and the nucleus should abrogate binding to SF3B, a finding confirmed by activity analysis of the side chain or nuclear intermediates, all of which failed to demonstrate the activity of a splicing modulator.

[0286] Having established a safe and convergent method( Figure 1 ), we turned our attention to the assembly of nucleus 2. Our route (Scheme 1) began with the development of a method to convert 6a to alcohol 7. After screening a wide range of acidic conditions, we modified the previously described protocol for in situ generation of HBr by slow reaction with iPrOH. Although our initial goal was to isolate the corresponding triol( Figure 2A ), the lack of stability of this species prompted the development of an in situ method to trap this species as a 5:1 mixture of the α∶β acetal isomers 7, (the straight line in 7, Scheme 1). After a series of logical reaction optimization steps, we established an overnight one-pot conversion of 6a to 7, which was carried out on a ten-gram scale with a single chromatographic purification operation. As described above, this process involves three operations, removal of the MEM and TBS ethers, followed by selective protection of the C6-C7 diol as its PMP acetal.

[0287] Having secured this conversion, our next goal was to develop a method that would facilitate the conversion of 7 to 11. By a detailed evaluation of each step, we were able to identify a method that allowed a five-step conversion of 7 to 11 over a 48-hour period (Scheme 1)( Figure 2B ). This began with the oxidation of alcohol 7 to aldehyde 8, which was readily accomplished using DMSO as a solvent. The resulting aldehyde was then subjected to an acetyl-Crimmins addition reaction using (−)-sparteine as a chiral additive (34). Mild hydrolysis after TBS protection gave acid 4, which was obtained in high yield and purity after flash dry-column vacuum chromatography (DCVC). Advantageously, this method allowed us to successfully recycle the auxiliary 6b1 (see Supporting Information) as well as (−)-sparteine. Overall, we were able to easily carry out ten grams of 7 to 4 per week.

[0288] The next esterification of 4 with 6c to afford 7 proved challenging. While many viable esterification conditions exist, many of these methods result in β-elimination of the TBS group at C3 in 4, unwanted ring-opening of the PMP acetal at C6–C7 in 4, and dehydration of the alcohol in 6c. After extensive screening, we found that treatment of equimolar amounts of 4 and 6c with 10 mol% of neat DMAP in pivalic anhydride at 70 °C gave 11 in near-quantitative yield, which could be used without purification. While effective, we soon realized that NMR studies of 11 (which was already a mixture of two species due to being a mixture of acetal isomers) indicated the presence of four compounds. Careful analysis revealed that the scale-up preparation of 6c provided one species. If care was not taken at this stage, introduction of this species into the synthetic pathway would result in sample 1 being contaminated with 5–10% of the wrong stereochemistry at C10–C11, which was observed in early runs of this pathway. In response, we developed a method to eliminate the possibility of forming iso-11 ( Figure 2C ), by resolution with (S)-mandelic acid. As detailed in the Supporting Information, a method was developed for the rapid preparation of 6c6 and 6c7 from large quantities of 6c, chromatographic purification of 6c6, and hydrolysis to afford enantiopure 6c.

[0289] At this point, we were able to obtain ten-gram quantities of 11 from 6a in approximately 5–6 days. Here, removal of the PMP ester followed by ring-closing metathesis ( Figure 2D ) of 16 provided a direct relay to 2. Unfortunately, this process was not reproducible due to unwanted competing ruthenium-catalyzed isomerization of the allylic alcohol in 16 to the ketone in 17. While 18 was accessible, its yield varied unpredictably between 25% ± 15%. A solution was effected by oxidation of 16 to 17, accomplished quantitatively using IBX. Here, elimination of the possibility of isomerization by oxidation at C7 provided an efficient RCM to enone 19. Unfortunately, reaction screening efforts using various reducing agents, chiral reduction methods, could not provide more than a 3:1 mixture that favored unwanted 20 over 18. Examination of the X-ray crystal structure of 19 explained this result, as addition of hydride to afford the desired 18 required a trajectory generated from within the macrocyclic lactone ring. With these options exhausted, we turned our attention back to 11 and conducted an exhaustive screen of catalysts (13 were tried), temperature, addition rate, and found that reverse addition (of the catalyst to 11) provided an efficient way to afford 11 with minimal byproduct formation. Here, we were able to consistently provide the product using slow addition of the Hoveyda–Grubbs II catalyst to 11 in refluxing toluene, a very simple solution to a decade-long problem in the synthesis of these and related 12-membered macrocyclic lactones.

[0290] At this stage, we are now able to convert ten grams of 6a to afford 12 (Scheme 1), a process that takes eight days to complete. At this point, the screening effort enabled us to identify a two-step method that involves complete deprotection to the stable triol 13 by mild acid hydrolysis followed by acetylation of 2 by treatment with trimethyl orthoacetate under acid catalysis by CSA. Although each of the last three steps (11 to 12, 12 to 13, and 13 to 2) requires flash chromatography, an optimal method was established that minimized the effort required for gram-scale preparation. To date, we have used this method to prepare nucleus 2. Stability studies of 2 indicate its stability for over six months under ambient conditions.

[0291] The preparation of side chain 3 is most efficiently carried out on a multigram scale by converting 6d to alkyne 5, which is an advantage point for purification and storage. First, Sharpless epoxidation of 6d is carried out, followed by oxidation of the corresponding alcohol 14 with IBX in DMSO, a two-step method that can be carried out without flash purification. Then, alkyne 15 is prepared on a 20 g scale from 6e and 15. Although stable upon storage at 0 °C under argon, the allenylic stannane is optimally prepared by distillation and used within 2–3 months of preparation. Here, the method developed by Marshall provided high selectivity in the installation of the C16–C17 center, affording a single isomer 5 from 6d. The process was carried out within four days of work. Similar to component 2, alkyne 15 is also stable for six months under ambient conditions.

[0292] At this stage, we are ready to perform the final coupling. The alkyne 5 is converted to the Z-stannane 3 by hydrostannylation using PdCl2(PPh3). The yield of this method was further optimized by using the Figueroa catalyst. The stannane 3 was then purified by flash chromatography and directly subjected to a Stille coupling with CuCl, KF in anhydrous tBuOH using the Buchwald-optimized XPhos G2 (38) catalyst. Given the high bioactivity of 1 and the potential toxicity risk, we carried out this process on a small scale with a handling criterion of not exceeding one gram. Through careful evaluation, we were able to complete this step with a minimal exposure time (2–4 h) of 1 by tandem use of DCVC and flash chromatography. We were able to recover 2 from this process, which can be recycled in the conversion of 2 to 1. Unfortunately, this process destroys the side chain 3, and the loss is limited since 2 is the limiting reagent in this process.

[0293] In our hands, each run of the process can be completed within a 16-day period, providing gram quantities of 1. To date, we have applied this route to prepare 1 from 6a (the most distal linear precursor). We have successfully accomplished the multigram synthesis of 17S-FD-895, using methods that couple components on the hectogram scale and couple them on the decagram scale in a 14-step, 2-route assembly. We have successfully been able to complete this entire process with two process chemists in three months, with eight weeks dedicated to component preparation and four weeks for assembly, meaning that 5 g of 1 can be prepared at a certain cost, which can be significantly reduced with future pilot work.

[0294] To further demonstrate the streamlined nature of this route, we examined the preparation of 17S-FD-895 analogues that are derived from isomeric by-products that are not required but can be collected in this route. We were able to prepare three untested analogues. This work was easily accomplished and lays the groundwork for completing a full SAR study on such substances.

[0295]

[0296] Scheme 1. Schematic of the assembly process developed for the multigram synthesis of 17S-FD-895. Here, five components 6a - 6e are processed in a stepwise manner to provide component, nucleus 2, and side chain 3. A 4-step sequence was developed to prepare 3 starting from 6d and later applying 6e to install the C16 - C17 stereodimer. The preparation of component 2 was accomplished in a 9-step sequence starting from component 6a. The process requires a series of operations that begin with the conversion of 6a to 7, the relay of 7 to 11 to install the C3 stereocenter using component 6b, ring-closing metathesis to 12, and appropriate adjustment of the functional groups in 2. Reagents and conditions as well as the observed yields are provided along each arrow.

[0297] Example 2. Experimental data for the initial synthesis

[0298] A. General Experimental Methods: Chemical reagents were purchased from Acros, Fluka, Sigma-Aldrich or TCI. Deuterated NMR solvents were purchased from Cambridge Isotope Laboratories. All reactions were carried out using strictly dried anhydrous solvents obtained by solvent columns consisting of activated Al alumina. Trace amounts of dimethylamine were removed by activated molecular sieves and subsequent NaOCN columns to obtain anhydrous N, N-dimethylformamide. Triethylamine (Et3N) was dried over Na and freshly distilled. Ethyl-N, N-diisopropylamine (EtNiPr2) was then distilled from potassium hydroxide from ninhydrin. Anhydrous CH3CN was obtained by distillation from CaH2. All reactions were carried out in oven-dried glassware sealed with septa under positive Ar pressure and stirred with a Teflon-coated stirring rod using an IKAMAG RCT-Basic mechanical stirrer (IKA GmbH). The solution was heated using a sand bath or silicone oil bath. Analytical thin layer chromatography (TLC) was performed on silica gel 60 F254 pre-coated glass plates (EM Sciences). Preparative TLC (pTLC) was performed on silica gel 60 plates (EMSciences). Visualization was achieved with UV light and / or appropriate staining agents (I on SiO2, KMnO4, bromocresol green, dinitrophenylhydrazine, ninhydrin and ceric ammonium molybdate). Flash chromatography was performed using Geduran silica gel 60 (40-63 mesh) from EMBiosciences. Yield and characterization data correspond to separated, chromatographic and spectrally homogeneous materials. Recorded on Varian Mercury 300, Varian Mercury 400 spectrometer, Varian MercuryPlus400, JEOL ECA500 or Varian VX500 spectrometer. 1 H NMR spectra. Most of the samples were recorded at 125 MHz on a Varian VX500 spectrometer equipped with an XsensCold probe. 13 C NMR spectra. The remaining spectra were collected at 125 MHz on a JEOL ECA 500, at 100 MHz on a Varian Mercury 400, or at 100 MHz on a Varian Mercury Plus 400 spectrometer. 1 H NMR and 13 The chemical shifts of C NMR analysis were referenced to the values reported by Gottlieb and were obtained using the signal from the residual solvent. 1 H spectra, or reference from deuterated solvents 13 C signal. 1 H and 13The chemical shift δ values of the C spectra are reported in parts per million (ppm) relative to these reference values, and the multiplicities are abbreviated as s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad. All 13 13C NMR spectra were recorded with complete proton decoupling. The FID files were processed using MestraNova 6.0.2. (MestreLab Research). Electrospray (ESI) mass spectrometry was performed using a ThermoFinnigan LCQDeca spectrometer, and high-resolution analysis was performed using a ThermoFinnigan MAT900XL mass spectrometer with electron impact (EI) ionization. High-resolution electrospray ionization mass spectrometry (HR-ESI-MS) was performed using a Thermo Scientific LTQ Orbitrap XL mass spectrometer. FTIR spectra were obtained as thin films on KBr or NaCl disks on a Nicolet magna 550 II series spectrometer, and the peaks were reported in wave numbers (cm -1 -1). The specific rotation [α] D was measured using a Perkin-Elmer Model 241 polarimeter with the specified solvent and concentration, and was reported in deg cm 2 2 g -1 -1. Spectral data and procedures for all new compounds were provided, and copies of selected spectra were provided.

[0299] B. Synthesis of Component 6a. A four-step sequence was developed to prepare Component 6a starting from commercially available 6a1, as shown in Scheme S1.

[0300]

[0301] Scheme S1. Synthesis of Side Chain Component 6a

[0302] 4-((tert-Butyldimethylsilyl)oxy)butanal (6a2). A solution of KBr (6.99 g, 58.7 mmol) in H2O (60 mL) was added to a solution of 4-((tert-butyldimethylsilyl)oxy)butan-1-ol (6a1) (100 g, 489 mmol) in CH2Cl2 (1.0 L), followed by the addition of saturated NaHCO3 (100 mL) and 2,2,6,6-tetramethylpiperidin-1-olate (2.29 g, 14.7 mmol). The reaction mixture was cooled to -3 °C and a mixture of NaOCl (0.33 L, 636 mmol) and saturated NaHCO3 (300 mL) was added in portions via a dropping funnel. The mixture was allowed to warm to room temperature. After stirring for 3 h at room temperature, the reaction mixture was extracted with CH2Cl2 (3 × 250 mL). The combined organic phases were washed with H2O (500 mL), saturated NaCl (500 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator to give 6a2 (100 g, quantitative yield).

[0303] Aldehyde 6a2: 1 H NMR (CDCl3, 300 MHz) δ 9.79 (t, J = 1.7 Hz, 1H), 3.65 (t, J = 6.0 Hz, 2H), 2.50 (td, J = 7.1, 1.7 Hz, 2H), 1.86 (tt, J = 7.1, 5.9 Hz, 2H), 0.94 - 0.84 (m, 9H), 0.04 (s, 6H); 13 C NMR (CDCl3, 75 MHz) δ 202.54, 62.06, 40.77, 25.87, 25.49, 18.24, -5.44.

[0304] (8S,9S)-14,14,15,15-Tetramethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadecan-9-ol (6a3). A solution of s-BuLi (1.4 M in cyclohexane, 353 mL, 494 mmol) was added dropwise over a 30-minute period to a solution of 3-((2-methoxyethoxy)methoxy)prop-1-ene (86.7 g, 593 mmol) in anhydrous THF (1 L), cooled to -78 °C under a N2 atmosphere. It was crucial to maintain the temperature below -70 °C during this addition. After stirring at -78 °C for 1 hour, a solution of methoxybis((1S,2R,3S,5S)-2,6,6-trimethylbicyclo[3.1.1]hept-3-yl)borane (156 g, 494 mmol) in anhydrous THF (500 mL) was added. The reaction mixture was stirred again at -78 °C for 1 hour. BF3·Et2O (79.3 mL, 642 mmol) was added, followed by a solution of 4-((tert-butyldimethylsilyl)oxy)butanal (6a2) (100 g, 494 mmol) in anhydrous THF (200 mL). The reaction mixture was stirred at -78 °C for 3 hours and then warmed to room temperature overnight. After cooling to between -4 °C and 0 °C, saturated NH4Cl (500 mL) was added to the mixture, and it was extracted with CH2Cl2 (3 × 250 mL). The combined organic phases were washed with H2O (500 mL), saturated NaCl (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Pure 6a3 (89 g, 52%) was obtained by flash chromatography, eluting with a gradient of heptane to EtOAc.

[0305] Alcohol 6a3: TLC (5∶1 hexane / EtOAc): R f = 0.25; 1 1H NMR (CDCl3, 300 MHz) δ 5.78 - 5.59 (m, 1H), 5.36 - 5.23 (m, 2H), 4.78 (d, J = 6.9 Hz, 1H), 4.69 (d, J = 6.9 Hz, 1H), 3.89 (dt, J = 8.2, 7.1 Hz, 1H), 3.85 - 3.75 (m, 1H), 3.71 - 3.61 (m, 3H), 3.61 - 3.47 (m, 4H), 3.38 (s, 3H), 2.94 (s, 1H), 1.79 - 1.54 (m, 4H), 1.50 - 1.31 (m, 1H), 0.87 (s, 10H), 0.10 (d, J = 0.6 Hz, 5H); 1313C NMR (CDCl3, 75 MHz) δ 134.81, 119.73, 92.99, 81.51, 73.15, 71.75, 67.37, 63.12, 58.96, 29.35, 28.81, 25.91, 18.30, -5.35.

[0306] (S)-14,14,15,15-Tetramethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadec-9-one (6a4). A solution of KBr (3.646 g, 30.64 mmol) in H2O (100 mL) was added to a solution of (8S,9S)-14,14,15,15-tetramethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadecan-9-ol (6a3) (89.00 g, 255.3 mmol) in DCM (400 mL), followed by the addition of saturated NaHCO3 (250 mL) and 2,2,6,6-tetramethylpiperidin-1-olate (3.990 g, 25.53 mmol). The reaction mixture was cooled to 0 °C and NaOCl (0.32 kg, 510.7 mmol) and saturated NaHCO3 (300 mL) were added dropwise (20 mL at a time) via a dropping funnel while maintaining the temperature below 1.5 °C. The reaction mixture was warmed to room temperature and stirred for 2 h. The phases were separated. The aqueous phase was extracted with CH2Cl2 (200 mL). The combined organic phases were washed with saturated NaCl (500 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator to give 6a4 (88.0 g, 99%).

[0307] Ketone 6a4: TLC (3:1 hexane / EtOAc): R f = 0.40; 1 1H NMR (CDCl3, 300 MHz) δ 5.75 (ddd, J = 17.0, 10.3, 6.6 Hz, 1H), 5.49 - 5.37 (m, 1H), 5.37 - 5.24 (m, 1H), 4.74 (q, J = 6.9 Hz, 2H), 4.59 (d, J = 6.6 Hz, 1H), 3.80 - 3.67 (m, 1H), 3.67 - 3.53 (m, 3H), 3.49 (t, J = 4.6 Hz, 2H), 3.34 (s, 3H), 2.60 (td, J = 7.2, 3.9 Hz, 2H), 1.74 (p, J = 6.7 Hz, 2H), 0.85 (s, 9H), 0.01 (s, 6H); 1313C NMR (CDCl3, 75 MHz) δ 208.01, 132.56, 119.85, 93.63, 82.58, 71.63, 67.40, 61.95, 58.94, 34.63, 26.25, 25.87, 18.23, -5.39.

[0308] (8S,9R)-9,14,14,15,15-Pentamethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadecan-9-ol (6a). MeMgBr (3 M solution in Et2O, 462 mL, 1385.1 mmol) was added dropwise to a solution of (S)-14,14,15,15-tetramethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexan-9-one (6a4) (160.0 g, 461.7 mmol) in anhydrous THF (1.5 L) at -85 °C. The reaction mixture was stirred at -85 °C for 2 h, warmed to room temperature, and then stirred for an additional 16 h. After cooling back to 0 °C, saturated NH4Cl (500 mL) was added dropwise to the mixture. The mixture was diluted with H2O (1 L) and extracted with TBME (2 × 500 mL). The combined organic phases were washed with H2O (500 mL) and saturated NaCl (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Pure 6a (78.9 g, 47%) was obtained by flash chromatography using a gradient elution of hexane to EtOAc.

[0309] Component 6a: TLC (5:1 hexane / EtOAc): Rf = 0.30; 1 1H NMR (CDCl3, 300 MHz) δ 5.74 (ddd, J = 17.1, 10.6, 8.0 Hz, 1H), 5.31 (dd, J = 1.9, 0.7 Hz, 1H), 5.32 - 5.19 (m, 1H), 4.75 (d, J = 6.9 Hz, 1H), 4.67 (d, J = 6.9 Hz, 1H), 3.94 - 3.71 (m, 2H), 3.68 - 3.57 (m, 1H), 3.63 - 3.51 (m, 2H), 3.57 - 3.44 (m, 2H), 3.36 (s, 3H), 2.66 (s, 1H), 1.76 - 1.51 (m, 3H), 1.56 - 1.32 (m, 1H), 1.14 (s, 3H), 0.87 (s, 9H), 0.02 (s, 6H); 1313C NMR (CDCl3, 75 MHz) δ 134.27, 120.02, 93.20, 84.42, 73.27, 71.74, 67.43, 63.77, 58.97, 33.77, 26.50, 25.92, 23.41, 18.30, -5.34.

[0310] C. Synthesis of Auxiliary Agent 6b. A two-step sequence was developed to prepare Component 6b starting from commercially available 6b1, as shown in Scheme S2.

[0311]

[0312] Scheme S2. Synthesis of Component 6b

[0313] (S)-4-(tert-Butyl)thiazolidine-2-thione (6b2). KOH (2.63 kg, 46.9 mol) was dissolved in H2O (9 L) and stirred in a 20 L reactor equipped with a mechanical stirrer and two reflux condensers. (S)-2-Amino-3,3-dimethylbutan-1-ol (6b1) (250 g, 2.13 mol) was added, followed by the dropwise addition of CS2 (1.03 L, 17.1 mol) under a N2 atmosphere. The reaction mixture was heated at 95 °C for 16 h. After the reaction mixture was cooled to 50 °C, an additional portion of CS2 (1 L) was added dropwise and the reaction mixture was heated at 70 °C for 16 h. The reaction mixture was cooled to 50 °C again, and an additional portion of CS2 (500 mL) was added dropwise. The mixture was heated at 65 °C and stirred over the weekend. After the reaction mixture was cooled to room temperature, the solid was collected by filtration and washed with H2O. The white solid was dried in air at room temperature. Pure 6b2 (175.7 g, 47%) was obtained by flash chromatography, eluting with DCM.

[0314] Auxiliary Agent 6b2: TLC (100% DCM): R f = 0.7; 1 1H NMR (CDCl3, 300 MHz) δ 7.58 (s, 1H), 4.01 (t, J = 9.6, 8.5, 1.2 Hz, 1H), 3.50 - 3.32 (m, 2H), 1.01 (s, 9H); 13 13C NMR (CDCl3, 75 MHz) δ 73.3, 34.5, 34.4, 25.9.

[0315] (S)-1-(4-(tert-Butyl)-2-thioxothiazolidin-3-yl)ethan-1-one (6b). Under a N2 atmosphere, n-Butyllithium (2.5 M solution in hexanes, 0.46 L, 1.1 mol) was added dropwise to a cooled (78 °C) solution of (S)-4-(tert-butyl)thiazolidine-2-thione (6b2) (181.83 g, 1.04 mol) in anhydrous THF (1.8 L). The mixture was stirred at -78 °C for 30 minutes, acetyl chloride (82 mL, 1.2 mol) was added dropwise, and the mixture was stirred for an additional 1.5 hours under the above conditions. Thereafter, the reaction mixture was warmed to room temperature, stirred for 1 hour, cooled to 0 °C and quenched with saturated NH4Cl (800 mL). The phases were separated. The aqueous phase was extracted with DCM (2 × 200 mL). The combined organic phases were dried over Na2SO4, filtered and concentrated on a rotary evaporator. Pure 6b3 (190.5 g, 85%) was obtained by flash chromatography using a gradient elution of heptane to DCM.

[0316] Auxiliary 6b: TLC (1:1 heptane / DCM): R f = 0.8; 1 1H NMR (CDCl3, 300 MHz) δ 5.30 (d, J = 8.4, 0.9 Hz, 1H), 3.60 - 3.44 (m, 1H), 3.09 (d, J = 11.8, 0.9 Hz, 1H), 2.77 (s, 3H), 1.03 (s, 9H); 13 13C NMR (CDCl3, 75 MHz) δ 205.3, 170.3, 72.0, 38.0, 30.4, 26.8, 26.8; LCMS (ES-API) [M+1] + : 218.0.

[0317] D. Synthesis of Component 6c. A seven-step sequence was developed to prepare Component 6c starting from commercially available 6c1 as shown in Schemes S3 - S4.

[0318]

[0319] Scheme S3. Synthesis of Component 6c

[0320] Dimethyl 2-(diiodomethyl)-2-methylmalonate (6c2). Under a N2 atmosphere, a solution of dimethyl 2-methylmalonate (6ca) (310 mL, 2.33 mol) in THF (800 mL) was added dropwise over a 20-minute period to a suspension of NaH (150 g, 3.8 mol) in THF (800 mL). The reaction was stirred under reflux for 1.5 hours. A solution of iodoform (801.9 g, 2.037 mol) in THF (2 L) was added dropwise over a 40-minute period. The reaction mixture was cooled to 50 °C and stirred under these conditions for 16 hours. After cooling to 0 °C, 2 M HCl (1.5 L) was added to the reaction mixture. The phases were separated. The aqueous phase was extracted with EtOAc (2 × 300 mL). The combined organic phases were dried over Na2SO4, filtered and concentrated on a rotary evaporator to give 6c2 (1008.8 g, quantitative yield).

[0321] Diester 6c2: 1 H NMR (CDCl3, 300 MHz) δ 3.77 (s, 6H), 3.22 (q, J = 6.7 Hz, 1H), 1.81 (s, 3H), 0.85 (t, J = 8.2 Hz, 7H); 13 C NMR (CDCl3, 75 MHz) δ 166.6, 53.6, 52.6, 20.42, 9.0.

[0322] (E)-3-Iodo-2-methylacrylic acid (6c3). Dimethyl 2-(diiodomethyl)-2-methylmalonate (6c2) (1008.8 g, 2.45 mol) was dissolved in a mixture of EtOH (2 L) and H2O (500 mL). KOH (300 g, 4.5 mol) was added in portions. Due to the large amount of heat released, the remaining KOH (400 g, 6.06 mol) was dissolved in H2O (300 mL) and added dropwise over a 1-hour period. The reaction mixture was heated to reflux and stirred for 16 hours. After cooling to room temperature, the mixture was concentrated on a rotary evaporator. The residue was acidified to pH 1 with concentrated HCl. The solid formed was collected by filtration and washed with CH2Cl2. The organic phase was washed with H2O (1 × 1 L), and the aqueous phase was extracted with CH2Cl2 (3 × 600 mL). The combined organic phases were dried over Na2SO4, filtered and concentrated on a rotary evaporator to give 6c3 (288.53 g, 65%).

[0323] Acid 6c3: 1 H NMR (CDCl3, 300 MHz) δ 9.65 (bs, 1H), 8.02 (s, 1H), 2.06 (s, 3H); 1313C NMR (CDCl3, 75 MHz) δ 168.9, 139.0, 101.8, 19.8.

[0324] (E)-3-Iodo-2-methylprop-2-en-1-ol (6c4). Under a N2 atmosphere, a solution of (E)-3-iodo-2-methylacrylic acid (6c3) (288.53 g, 1.3 mol) in Et2O (400 mL) was added dropwise over 20 minutes to a suspension of LiAlH4 (76.4 g, 2.01 mol) in Et2O (800 mL) cooled to 5 °C. The reaction mixture was stirred at -5 °C for 1 hour, warmed to room temperature and stirred for an additional 2 hours. After cooling the mixture to -78 °C, acetone (200 mL) was added dropwise over a 35-minute period, followed by 2 M HCl (750 mL) added dropwise over a 1-hour period. The resulting mixture was filtered through a Buchner funnel. The phases were separated, and the aqueous phase was extracted with TBME (3 × 1 L). The combined organic phases were washed with saturated NaCl (3 × 500 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator. Pure 6c4 (146.4 g, 56%) was obtained by flash chromatography using a gradient elution of heptane to CH2Cl2.

[0325] Alcohol 6c4: TLC (0:1 heptane / CH2Cl2): R f = 0.6; 1 1H NMR (CDCl3, 300 MHz) δ 6.24 (m, J = 1.3 Hz, 1H), 4.12 - 4.04 (d, 2H), 2.43 (t, J = 5.9 Hz, 1H), 1.82 (s, 3H). 13 13C NMR (CDCl3, 75 MHz) δ 147.2, 67.0, 21.4.

[0326] (E)-3-Iodo-2-methylacrolein (6c5). Under a N2 atmosphere, activated MnO2 (642.8 g, 7.394 mol) was added to a solution of (E)-3-iodo-2-methylprop-2-en-1-ol (6c4) (146.4 g, 739.4 mmol) in CH2Cl2 (1 L). The reaction mixture was stirred at room temperature for 16 hours. After filtration through Celite and concentration on a rotary evaporator, pure 6c5 (142.4 g, 84%) was obtained.

[0327] Aldehyde 6c5: 1 1H NMR (CDCl3, 300 MHz) δ 9.52 (s, 1H), 7.80 (d, J = 1.3 Hz, 1H), 5.29 (s, 1H), 1.92 (d, J = 1.2 Hz, 3H). 1313C NMR (CDCl3, 75 MHz) δ 189.4, 150.8, 109.4, 16.4.

[0328] (3S, 4S, E)-1-Iodo-2,4-dimethylhexa-1,5-dien-3-ol (6c). (E)-But-2-ene (200 mL, 2 mol) was condensed at -78 °C under a N2 atmosphere and added to THF (1.5 L). KOtBu (113.8 g, 1.014 mol) was added and the reaction mixture was stirred for 30 min under the above conditions. n-BuLi (2.5 M solution in hexane, 400 mL, 1.0 mol) was added dropwise over a 15 min period and the mixture was stirred at -78 °C for 30 min. A solution of methoxybis((1S,2R,3S,5S)-2,6,6-trimethylbicyclo[3.1.1]hept-3-yl)borane (253 g, 800 mmol) in THF (1 L) was added dropwise over a 15 min period. After the mixture was stirred for 30 min, BF3·Et2O (170 mL, 1.34 mol) was added dropwise over a 10 min period and the mixture was stirred for 10 min. After the reaction mixture was cooled to -94 °C, a solution of (E)-3-iodo-2-methylacrolein (6c5) (121 g, 617 mmol) in THF (750 mL) was added dropwise over a 45 min period. After addition was complete, the reaction mixture was warmed to room temperature and stirred for 16 h. H2O (2 L) was added and the mixture was concentrated on a rotary evaporator. Component 6c (78 g, 50%) was obtained by flash chromatography, eluting with CH2Cl2.

[0329] Intermediate 6c: 1 1H NMR (CDCl3, 300 MHz) δ 6.26 (s, 1H), 5.72 (ddd, J = 17.8, 9.9, 8.1 Hz, 1H), 5.24 - 4.94 (m, 2H), 3.87 (dd, J = 8.1, 2.3 Hz, 1H), 2.35 (q, J = 7.4 Hz, 1H), 1.88 - 1.55 (s, 3H), 0.92 (d, J = 6.8 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 148.0, 139.9, 117.2, 80.1, 79.7, 42.2, 19.3, 16.5. Chiral GC: 78.8% ee.

[0330]

[0331] Scheme S4. Resolution and provision of enantiopure 6c.

[0332] (3S, 4S, E)-1-Iodo-2,4-dimethylhexa-1,5-dien-3-yl (R)-2-methoxy-2-phenylacetate. A mixture of 6c (9.3 g, 36.8 mmol) was added to a 100 mL pear-shaped round-bottom flask and dried by azeotrope with toluene (2 × 25 mL). Solid (R)-2-methoxy-2-phenylacetic acid (6.74 g, 40.6 mmol) and DMAP (678.0 mg, 5.5 mmol) were added, followed by pivalic anhydride (15 mL). The mixture was heated to 70 °C in a 100 mL Heat-On attachment with a Hei-Tec magnetic stirrer. After 2 h, the reaction was cooled and dried by rotary evaporation and air stream. The resulting crude wax was flash chromatographed with a gradient from hexane to 20:1 hexane:Et2O to give pure major 6c6 (80% ± 3%) and minor 6c7 (6% ± 2%) fractions and a recyclable mixed fraction (4% ± 1%). The pure major isomer 6c6 was immediately subjected to the following step.

[0333] Ester 6c6: 1 H NMR (CDCl3, 300 MHz) δ 7.37 (m, 5H), 5.96 (s, 1H), 5.61 (ddd, J = 7.9, 10.2, 18.1 Hz, 1H), 5.15 (d, J = 7.9 Hz, 1H), 5.01 (dd, J = 1.4, 17.1 Hz, 1H), 4.99 (d, J = 9.7 Hz, 1H), 4.73 (s, 1H), 3.39 (s, 3H), 2.46 (dt, J = 6.8, 7.3 Hz, 1H), 1.51 (d, J = 1.5 Hz, 3H), 0.89 (d, J = 6.9 Hz, 1H); 13 C NMR (CDCl3, 75 MHz) δ 143.7, 138.9, 136.0, 129.0, 128.8, 127.4, 116.2, 82.4, 81.6, 81.0, 57.4, 40.0, 20.0, 16.5.

[0334] Enantiopure (3S, 4S, E)-1-iodo-2,4-dimethylhexa-1,5-dien-3-ol (6c). Pure 6d6 (12.2 g, 30.4 mmol) was dissolved in MeOH (400 mL) and H2O (∼80 mL) until the solution became slightly turbid. NaOH (1 M) was added in 50 mL portions until TLC analysis indicated complete hydrolysis (usually completed by adding 5 - 6 times within 1.5 h). Once the addition of H2O (100 mL) was complete, the resulting mixture was extracted with CH2Cl2 (3 × 300 mL), washed with brine (100 mL) and dried over Na2SO4. The resulting resolved 6c (7.4 g, 79%) was used as is.

[0335] Enantiopure 6c: 1 H NMR(CDCl3, 300 MHz) δ 6.26 (s, 1H), 5.72 (ddd, J = 17.8, 9.9, 8.1 Hz, 1H), 5.24 - 4.94 (m, 2H), 3.87 (dd, J = 8.1, 2.3 Hz, 1H), 2.35 (q, J = 7.4 Hz, 1H), 1.88 - 1.55 (s, 3H), 0.92 (d, J = 6.8 Hz, 3H); 13 C NMR(CDCl3, 75 MHz) δ 148.0, 139.9, 117.2, 80.1, 79.7, 42.2, 19.3, 16.5; Chiral GC: 99% ee.

[0336] Repeat this procedure to provide a total of >50 g of 6c in 5 batches.

[0337] E. Synthesis of Component 6d. A seven-step sequence was developed to prepare Component 6d starting from commercially available 6d1 as shown in Scheme S5.

[0338]

[0339] Scheme S5. Synthesis of Component 6d

[0340] (R)-1-(4-Benzyl-2-thioxothiazolidin-3-yl)propan-1-one (6d2). Triethylamine (0.7 L, 5.2 mol) and N,N-dimethylpyridin-4-amine (105.1 g, 0.86 mol) were added to a solution of (R)-4-benzylthiazolidine-2-thione (6d1) (891.8 g, 4.3 mol) in CH2Cl2 (9.0 L) at room temperature. The reaction mixture was cooled to 0 °C and a solution of propionyl chloride (490 mL, 5.61 mol) in CH2Cl2 (2.25 L) was added dropwise over a 1.5-hour period while maintaining the temperature below 5 °C. The reaction mixture was stirred at room temperature for 18 hours. Thereafter, the mixture was cooled to 0 °C and saturated NH4Cl (5.8 L) was added dropwise while maintaining the temperature below 5 °C. The mixture was extracted with DCM (3 × 2 L). The combined organic phases were washed with saturated NaHCO3 (4 L) and saturated NaCl (4 L), dried over Na2SO4, filtered and concentrated on a rotary evaporator. This batch was combined with a smaller batch of 6d2 (130 g). Pure 6d2 (950.1 g, 84%) was obtained by crystallization from MeCN.

[0341] Auxiliary 6d2: 11H NMR (CDCl3, 300 MHz) δ 7.40 - 7.21 (m, 5H), 5.38 (m, 1H), 3.52 - 3.40 (m, 1H), 3.40 - 3.32 (m, 1H), 3.28 - 2.96 (m, 3H), 2.88 (dd, J = 11.5, 0.7 Hz, 1H), 1.19 (t, J = 7.2 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 174.9, 136.6, 129.5, 128.9, 127.2, 68.7, 36.8, 32.3, 31.9, 8.82. LCMS (ES-API) [M+1] + : 266.40.

[0342] (2R, 3S)-1-((S)-4-Benzyl-2-thiazolidinyl)-3-hydroxy-2-methylpentan-1-one (6d3). Under mechanical stirring, (S)-1-(4-benzyl-2-thiazolidinyl)propan-1-one (6d2) (235.3 g, 887 mmol) was dissolved in CH2Cl2 (7.05 L). The reaction mixture was cooled to below 0 °C. TiCl4 (1 M solution in CH2Cl2, 922 mL, 922 mmol) was added dropwise over a 1-hour period while maintaining the temperature below 0 °C. EtN(iPr)2 (168 mL, 966 mmol) was added dropwise over a 30-minute period and the reaction mixture was stirred at 0 °C for 15 minutes. After cooling the reaction mixture to below -82 °C, a solution of propionaldehyde (71 mL, 984 mmol) in CH2Cl2 (350 mL) was added dropwise over a 6-hour period while maintaining the temperature below -82 °C. The reaction mixture was stirred under the above conditions for 30 minutes and slowly warmed to room temperature overnight. Saturated NaHCO3 (1.67 L) was added dropwise to the mixture. Note: A large exotherm was observed and the temperature was kept below 5 °C. The phases were separated. The aqueous phase was extracted with CH2Cl2 (3 × 1 L). The combined organic phases were washed with saturated NaCl (2 L), dried over Na2SO4, filtered and concentrated on a rotary evaporator. Pure 6d3 (249.5 g, 87%) was obtained by flash chromatography, eluting with a gradient of heptane to EtOAc.

[0343] Adduct 6d3: TLC (3∶1 heptane / EtOAc): R f = 0.63; 11H NMR (CDCl3, 300 MHz) δ 7.41 - 7.22 (m, 5H), 5.43 - 5.32 (m, 1H), 4.72 (dd, J = 7.1, 2.3 Hz, 1H), 3.97 (tt, J = 5.2, 2.6 Hz, 1H), 3.37 (ddd, J = 11.5, 7.1, 1.0 Hz, 1H), 3.24 (dd, J = 13.2, 4.1 Hz, 1H), 3.04 (dd, J = 13.2, 10.4 Hz, 1H), 2.89 (dd, J = 11.6, 0.8 Hz, 1H), 2.77 (dd, J = 2.9, 0.9 Hz, 1H), 1.70 - 1.35 (m, 3H), 1.18 (d, J = 7.1 Hz, 3H), 0.98 (t, J = 7.4 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 201.6, 178.5, 136.4, 129.5, 128.9, 127.3, 72.5, 68.9, 42.3, 36.9, 31.8, 26.7, 10.5, 10.5; LCMS (ES-API) [M + 1] + : 324.40.

[0344] (2R, 3S)-3-Hydroxy-N-methoxy-N,2-dimethylpentanamide (6d4). At room temperature, N,O-dimethylhydroxylamine hydrochloride (174.0 g, 1.78 mol) and imidazole (182.2 g, 2.68 mol) were successively added to a solution of (2R, 3S)-1-((S)-4-benzyl-2-thiazolidin-3-yl)-3-hydroxy-2-methylpentan-1-one (6d3) (288.5 g, 0.89 mol) in CH2Cl2 (12.5 L). The reaction mixture was stirred at room temperature for an additional 16 h. H2O (3.0 L) was added, and the aqueous phase (pH ~ 7) was extracted with CH2Cl2 (3 × 2.5 L). The combined organic phases were washed with saturated NaCl (5.0 L), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. A yellow oil (344.0 g) was obtained. Pure 6d4 (155.0 g, 99%) was obtained by flash chromatography eluting with a gradient of heptane to EtOAc.

[0345] Amide 6d4 TLC (3∶1 heptane / EtOAc): R f = 0.17; 11H NMR (CDCl3, 300 MHz) δ 3.73 (ddd, J = 8.1, 5.4, 2.9 Hz, 1H), 3.67 (s, 3H), 3.17 (s, 3H), 2.91 - 2.83 (br, 1H), 1.55 (dt, J = 13.5, 7.5 Hz, 1H), 1.37 (ddd, J = 11.8, 7.4, 5.4 Hz, 1H), 1.13 (d, J = 7.1 Hz, 3H), 0.93 (t, J = 7.4 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 178.5, 73.0, 61.5, 38.2, 31.8, 26.7, 10.4, 10.0; LCMS (ES-API) [M+1] + : 176.40.

[0346] (2R, 3S)-N,3-Dimethoxy-N,2-dimethylpentanamide (6d5). At room temperature, MeI (1.1 L, 18.0 mol) was added to a solution of (2R, 3S)-3-hydroxy-N-methoxy-N,2-dimethylpentanamide (6d4) (155.0 g, 0.89 mol) in a mixture of THF (6.1 L) and DMF (1.5 L). The reaction mixture was cooled to 0 °C and NaH (60% in mineral oil, 88.5 g, 2.21 mol) was added in portions. The reaction mixture was slowly warmed to room temperature and stirred for 16 h. After cooling the reaction mixture to 0 °C, an aqueous pH 7 phosphate buffer solution (1.5 L) was added dropwise. The volatiles were evaporated on a rotary evaporator. H2O (4.5 L) was added to the residue and the resulting mixture was extracted with TBME (3 × 3 L). The combined organic phases were washed with saturated NaCl (3 L), dried over Na2SO4, filtered and evaporated on a rotary evaporator. Pure 6d5 (152.3 g, 91%) was obtained by flash chromatography, eluting with a gradient of heptane to EtOAc.

[0347] Amide 6d5: TLC (3:1 heptane / EtOAc): R f = 0.27; 1 1H NMR (CDCl3, 300 MHz) δ 3.59 (s, 3H), 3.30 (s, 3H), 3.28 - 3.14 (m, 1H), 3.08 (s, J = 5.1 Hz, 3H), 2.98 - 2.87 (m, 1H), 1.49 (ddt, J = 14.5, 7.4, 3.7 Hz, 1H), 1.33 (dt, J = 14.2, 7.1 Hz, 1H), 1.11 (d, 3H), 0.83 (t, J = 7.4, 6.1 Hz, 3H); 1313C NMR (CDCl3, 75 MHz, ) δ 176.0, 171.0, 83.5, 61.1, 59.9, 58.0, 42.8, 39.1, 35.1, 26.1, 26.0, 24.7, 22.6, 20.6, 13.9, 9.2; LCMS (ES-API) [M+1] + : 190.40.

[0348] (4S, 5S, E)-Ethyl 5-methoxy-4-methylhept-2-enoate (6d7). (2R, 3S)-N,3-Dimethoxy-N,2-dimethylpentanamide (6d5) (107 g, 565 mmol) was dissolved in CH2Cl2 (2.14 L). The reaction mixture was cooled to below -78 °C. DIBAL-H (1.1 M solution in heptane, 0.8 L, 0.88 mol) was added dropwise over a 45-minute period while maintaining the temperature below -78 °C. The reaction mixture was stirred under the above conditions for 15 minutes. Acetone (64.1 mL, 0.88 mol) was added dropwise over a 10-minute period. The reaction mixture was warmed to 0 °C. Saturated Rochelle salt (1.75 L) was added over a 30-minute period and the mixture was stirred at room temperature for 1.5 hours. The phases were separated. The aqueous phase was extracted with a mixture of CH2Cl2 (520 mL) and heptane (52 mL). The combined organic phases were dried over Na2SO4, filtered and concentrated on a rotary evaporator. The residue was co-evaporated with toluene (460 mL) to afford aldehyde 6d6, which was used immediately after preparation.

[0349] Aldehyde 6d6: 1 1H NMR (CDCl3, 300 MHz) δ 9.77 (s, 1H), 3.56 - 3.48 (m, 1H), 3.35 (s, 3H), 2.58 - 2.48 (m, 1H), 1.73 - 1.45 (m, 2H), 1.10 (d, J = 7.1 Hz, 3H), 0.94 (t, J = 6.0, 3H).

[0350] A solution of ethyl 2-(diethoxyphosphoryl)acetate (572 mL, 2.88 mol) in anhydrous THF (400 mL) was added dropwise over a 30-minute period to a cooled suspension of NaH (60% in mineral oil, 97.4 g, 2.44 mol) in anhydrous THF (1.0 L) cooled to 0 °C. The reaction mixture was stirred at 0 °C for 15 minutes and a solution of 6d6 in anhydrous THF was added dropwise over a 30-minute period. The reaction mixture was stirred at room temperature for 16 hours, cooled to 0 °C and quenched with saturated NH4Cl (1.6 L). The volatiles were evaporated on a rotary evaporator and H2O (400 mL) was added. The mixture was extracted with EtOAc (2 × 1 L). The combined organic phases were dried over Na2SO4, filtered and concentrated on a rotary evaporator. The ester 6d7 was purified by flash chromatography, eluting with a gradient of CH2Cl2 to EtOAc. The separated 6d7 was further stirred in a mixture of saturated NaHSO3 (500 mL), EtOAc (450 mL) and heptane (50 mL) for 40 minutes. H2O (250 mL) was added. The phases were separated. The aqueous phase was extracted with a mixture of EtOAc and heptane (3 × 250 mL, 9:1). The combined organic phases were dried over Na2SO4, filtered and concentrated on a rotary evaporator to give 6d7 (57.9 g, 51%).

[0351] Ester 6d7: TLC (100% DCM): R f = 0.14; 1 1H NMR (CDCl3, 300 MHz) δ 6.95 (dd, J = 15.8, 7.7 Hz, 1H), 5.82 (dd, J = 15.8, 1.3 Hz, 1H), 4.18 (q, J = 7.1 Hz, 2H), 3.37 (s, 3H), 3.01 (m, 1H), 2.57 (m, 1H), 1.62 - 1.28 (m, 2H), 1.29 (t, J = 7.5 Hz, 3H), 1.07 (d, J = 6.8 Hz, 3H), 0.91 (t, J = 7.4 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 166.5, 151.1, 120.9, 85.4, 60.0, 57.7, 39.1, 23.7, 14.6, 14.2, 9.7; LCMS (ES-API) [M+NH4] + : 218.6.

[0352] (4S, 5S, E)-5-methoxy-4-methylhept-2-en-1-ol (6d). DIBAL-H (1.1 M solution in heptane, 0.77 L, 0.85 mol) was added dropwise over a 60-minute period to a solution of ethyl (4S, 5S, E)-5-methoxy-4-methylhept-2-enoate (6d7) (56.5 g, 282 mmol) in CH2Cl2 (1.5 L) cooled to -78 °C. The reaction mixture was stirred for 1 hour under the above conditions. Acetone (57 mL, 0.78 mol) was added dropwise over a 25-minute period. The reaction mixture was warmed to 0 °C and saturated Rochelle salt (1030 mL) was added over a 40-minute period. The mixture was stirred at room temperature for 1 hour 45 minutes. The phases were separated. The aqueous phase was extracted with CH2Cl2 (3 × 500 mL). The combined organic phases were washed with saturated NaCl (250 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator. Pure 6d (39.0 g, 87%) was obtained by flash chromatography, eluting with a gradient of heptane to EtOAc.

[0353] Intermediate 6d: TLC (3:1 heptane / EtOAc): R f = 0.26. 1 1H NMR (CDCl3, 300 MHz) δ 5.73 - 5.57 (m, 2H), 4.11 (m, 2H), 3.36 (s, 3H), 2.92 (ddd, J = 7.4, 5.7, 4.3 Hz, 1H), 2.44 (m, 1H), 1.57 - 1.34 (m, 2H), 1.02 (d, J = 6.8 Hz, 3H), 0.91 (t, J = 7.4 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 134.3, 129.2, 86.4, 63.2, 57.4, 38.8, 23.2, 15.8, 9.8; chiral GC: 98.4% e.e.

[0354] F. Synthesis of Component 6e. A two-step sequence was developed to prepare Component 6e starting from commercially available 6e1, as shown in Scheme S6.

[0355]

[0356] Scheme S6. Synthesis of Component 6e

[0357] (R)-But-3-yn-2-yl methanesulfonate (6e2). Et3N (198 mL, 1.43 mol) was added dropwise to a solution of (R)-but-3-yn-2-ol (6e1) (50.0 g, 713 mmol) in CH2Cl2 (750 mL) cooled to -78 °C over a 15-minute period. After 10 minutes, MsCl (83.4 mL, 1.07 mol) was added dropwise over a 2-hour period. The reaction mixture was stirred at the above conditions for 1 hour. Saturated NaHCO3 (750 mL) was added dropwise over a 4-hour period. The reaction mixture was warmed to room temperature. H2O (250 mL) was added and the phases were separated. The aqueous phase was extracted with CH2Cl2 (250 mL). The combined organic phases were washed with saturated NaCl (250 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator. The impure product was partitioned between DCM (750 mL) and saturated NaHCO3 (750 mL), and the mixture was stirred at room temperature for 2 hours. The phases were separated. The organic phase was dried over Na2SO4, filtered and concentrated on a rotary evaporator to give 6e2 (21.1 g, 20.0%).

[0358] Methanesulfonate 6e2: 1 1H NMR (CDCl3, 300 MHz) δ 5.29 (qd, J = 6.7, 2.1 Hz, 1H), 3.12 (s, 3H), 2.70 (d, J = 2.2 Hz, 1H), 1.66 (d, J = 6.7 Hz, 3H); 13 13C NMR (CDCl3, 75 MHz) δ 80.1, 76.4, 67.5, 39.1, 22.4.

[0359] (S)-Buta-1,2-dien-1-yltributylstannane (6e). n-BuLi (2.5 M solution in hexanes, 172 mL, 429 mmol) was added dropwise to a solution of diisopropylamine (60.7 mL, 429 mmol) in THF (800 mL) over a 10-minute period at 0 °C. After 15 minutes, nBu3SnH (135 mL, 501 mmol) was added dropwise over a 7-minute period and the reaction mixture was stirred at 0 °C for 2.5 h. After cooling the reaction mixture to -85 °C (less than -78 °C), CuBr·DMS (88.2 g, 429 mmol) was added in portions over a 40-minute period. The mixture was stirred at -85 °C (or less than -78 °C) for 30 minutes. (R)-But-3-yn-2-yl methanesulfonate (6e2) (53.0 g, 358 mmol) was added dropwise over a 2-minute period and the mixture was stirred for an additional 8 minutes. The reaction mixture was poured into a mixture of TBME (1.75 L), 25% aqueous NH3 (260 mL), and saturated NH4Cl (2.12 L) and stirred vigorously for 1 h. The phases were separated. The organic phase was dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Component 6e (77.2 g, 62.9%) was obtained by falling-film distillation.

[0360] Intermediate 6e: 1 1H NMR (CDCl3, 300 MHz) δ 5.08 - 4.88 (m, 1H), 4.56 (p, J = 7.0 Hz, 1H), 1.74 - 1.41 (m, 12H), 1.31 (h, J = 7.2 Hz, 6H), 0.92 (dt, J = 11.6, 7.7 Hz, 12H); 13 13C NMR (CDCl3, 75 MHz) δ 209.1, 75.2, 74.3, 30.6, 28.9, 13.7, 10.3; Chiral GC: 94.2% e.e.

[0361] Derivatization of 6e for determination of enantiomeric excess: A solution of isobutyraldehyde (40 μL, 0.44 mmol) in CH2Cl2 (4 mL) was added dropwise to a solution of (S)-buta-1,2-dien-1-yltributylstannane (6e) (200 mg, 583 μmol) and BF3·OEt2 (210 μL, 1.66 mmol) cooled to -78 °C. After stirring at -78 °C for 1 h, the reaction was quenched with saturated NaHCO3 (4 mL). The mixture was warmed to room temperature and the phases were separated. The organic phase was stirred with KF on Celite (50 wt%, 100 mg) and Na2SO4 (100 mg). The solids were removed by filtration and an aliquot of the filtrate was used for chiral GC analysis, indicating 96% ee.

[0362] Group G was assembled into 17S-FD-895(1). The following procedures and spectral data were developed for the assembly of components 6a-6c to 2 and 6d-6e to 3 and the coupling of 2 and 3 to afford 17S-FD-895(1), as shown in Scheme 1.

[0363] 3-((4R,5S)-2-(4-Methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)propan-1-ol (7). To a 3 L round-bottom flask equipped with a magnetic stir bar were sequentially added alcohol 6a (15.0 g, 42.5 mmol), wet iPrOH (1.5 L), CBr4 (19.9 g, 63.8 mmol), and imidazole (0.145 g, 2.1 mmol). The mixture was heated to reflux and stirred overnight, at which point the mixture became a clear light brown solution. Complete conversion of 6a to the intermediate was determined by NMR. The mixture was quenched with 4A molecular sieves (200 g) and cooled to room temperature. The mixture was filtered through an oven-dried vacuum funnel into a flame-dried 2 L flask and concentrated in vacuo to afford a dark brown oil. The crude was immediately taken up in anhydrous CH2Cl2 (300 mL) and purged with an Ar atmosphere. Anisaldehyde dimethyl acetal (14.5 mL, 85.1 mmol) was added in one aliquot, and the mixture turned purple after stirring for 10 minutes at room temperature. The reaction was stirred for a further 2 hours at room temperature. Saturated aqueous NaHCO3 (100 mL) was added and the mixture was extracted into CH2Cl2. The organics were combined and concentrated in vacuo to afford a brown oil. Pure 7 (7.7 g, 65%) was obtained as a 5:3 mixture of acetal diastereomers by flash chromatography using a hexanes to 35% EtOAc / hexanes gradient elution. Note 1: Formation of the intermediate is generally quantitative as determined by NMR and is sufficiently pure in practice to proceed. Note 2: The intermediate is somewhat unstable and best yields are obtained when anisaldehyde dimethyl acetal is added as quickly as possible.

[0364] Alcohol 8: TLC (1:1 hexanes / EtOAc): Rf = 0.37; CAM staining; single spot; 11H NMR (C6D6, 500 MHz) δ 7.55 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.7 Hz, 2H), 5.91 (s, 1H), 5.84 - 5.75 (m, 1H), 5.31 (dt, J = 17.1, 1.3 Hz, 1H), 5.07 (dt, J = 10.4, 1.2 Hz, 1H), 4.09 (dt, J = 7.1, 1 Hz, 1H), 4.24 (dt, J = 7.1, 1 Hz), 3.25 (s, 3H), 3.39 (dd, J = 9.7, 5.6 Hz, 1H), 3.64 - 3.58 (m, 1H), 1.83 - 1.63 (m, 4H), 1.38 (s, 3H); Minor: δ 7.50 (d, J = 8.7 Hz, 2H), 6.80 (d, J = 8.6 Hz, 2H), 6.16 (s, 1H), 5.84 - 5.75 (m, 1H), 5.31 (dt, J = 17.1, 1.3 Hz, 1H), 5.07 (dt, J = 10.4, 1.2 Hz, 1H), 4.09 (dt, J = 7.1, 1 Hz, 1H), 4.24 (dt, J = 7.1, 1 Hz), 3.25 (s, 3H), 3.39 (dd, J = 9.7, 5.6 Hz, 1H), 3.64 - 3.58 (m, 1H), 1.83 - 1.63 (m, 4H), 1.38 (s, 3H); 13 13C NMR (500 MHz) δ 160.4, 160.2, 133.5, 133.4, 132.5, 130.7, 128.2, 127.7, 117.6, 117.5, 113.6, 113.5, 102.2, 101.9, 87.6, 85.6, 83.2, 82.0, 62.7, 62.6, 54.4, 33.4, 32.3, 31.0, 29.5, 28.2, 27.1, 26.9, 26.7, 22.1, 21.7; FTIR (film) νmax 3421, 3080, 2938, 1718, 1614, 1516, 1932, 1303, 1249, 1170, 1032 cm -1 ; C 16 H 22 O4 Na [M+Na] + HR-ESI-MS m / z calcd for C15H17O4Na: 301.1410, found 301.1411.

[0365] 3-((4R,5S)-2-(4-Methoxyphenyl)-4-methyl-5-ethenyl-1,3-dioxolan-4-yl)propanal (8). Alcohol 7 (7.5 g, 26.9 mmol), DMSO (250 mL), and freshly prepared IBX (18.9 g, 67.4 mmol) were successively added to a 2 L flask. The mixture was stirred at room temperature for 2 h, at which point TLC indicated complete conversion. The mixture was diluted with 350 mL of EtOAc and washed with 150 mL of H2O. The aqueous layer was back-extracted with EtOAc (2 × 250 mL). The organic layers were combined and further washed with H2O (5 × 450 mL) and brine (250 mL). The organic matter was concentrated in vacuo, and the oil was then filtered through a pad of Celite and eluted with EtOAc. The eluate was concentrated to afford 8 (6.70 g, 90%) as a yellow oil, which was used directly in the next reaction. Note: Aldehyde 9 is prone to rearrangement when purified on unbuffered silica gel. In practice, this material is clean enough for subsequent reactions without chromatography; however, the crude 9 can be purified on neutral silica gel, eluting with a gradient of hexanes to 25% EtOAc / hexanes.

[0366] Aldehyde 9: 1 1H NMR (C6D6, 500 MHz) δ major isomer 9.26 (s, 1H), 7.44 (d, J = 8.7 Hz, 2H), 6.76 (d, J = 4.3 Hz, 2H), 5.80 (s, 1H), 5.67 (m, 1H), 5.25 (d, J = 12.9 Hz, 1H), 5.01 (d, J = 4.7 Hz, 1H), 3.99 (d, J = 6.6 Hz, 1H), 3.23 (s, 3H), 2.17 - 2.28 (m, 1H), 1.93 - 2.07 (m, 2H), 1.80 - 1.87 (m, 1H), 1.34 - 1.41 (m, 1H), 1.20 - 1.25 (m, 1H), 0.96 (s, 3H); minor isomer 9.36 (s, 1H), 7.42 (d, J = 8.7 Hz, 2H), 6.78 (d, J = 4.3 Hz, 2H), 6.00 (s, 1H), 5.62 (m, 1H), 5.21 (d, J = 12.9 Hz, 1H), 4.99 (d, J = 4.7 Hz, 1H), 4.07 (d, J = 6.6 Hz, 1H), 3.22 (s, 3H), 2.17 - 2.28 (m, 1H), 1.93 - 2.07 (m, 2H), 1.80 - 1.87 (m, 1H), 1.34 - 1.41 (m, 1H), 1.20 - 1.25 (m, 1H), 0.97 (s, 3H); 1313C NMR (C6D6, 500 MHz) δ 200.2, 200.0, 160.5, 160.2, 132.8, 132.7, 132.3, 130.4, 117.82, 117.78, 113.64, 113.56, 102.2, 101.9, 87.2, 85.3, 82.4, 81.0, 54.4, 38.5, 38.1, 28.9, 25.3, 22.2, 21.5.

[0367] Hundred-gram preparation of 2-iodoxybenzoic acid (IBX): Add solid potassium peroxymonosulfate and deionized H2O to a 5 L flask equipped with a magnetic stir bar. Stir the mixture and heat to 75 °C. After the potassium peroxymonosulfate is completely dissolved, add 2-iodobenzoic acid in solid form and stir the mixture vigorously at 75 °C for 4 h. After stirring stops, a white precipitate (product) precipitates at the bottom of the flask. Filter the mixture by vacuum filtration through a Buchner funnel and wash the separated white powder further with H2O (3 × 150 mL) and acetone (3 × 100 mL). Obtain IBX as a white crystalline powder and store at -20 °C. The characterization data match the literature values previously reported by Frigerio, M; et al.

[0368] (3R)-1-((R)-5-(tert-Butyl)-2-thioxothiazolidin-3-yl)-3-hydroxy-5-((4R,5S)-2-(4-methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)pentan-1-one (9). To a flame-dried 3 L flask equipped with a stir bar was added the auxiliary 6b (13.76 g, 63.3 mmol) in solid form and taken up in anhydrous toluene. The solution was concentrated by rotary evaporation to remove trace moisture. The flask was then purged with argon and taken up in anhydrous CH2Cl2 (600 mL). Dichlorophenylborane (8.22 mL, 63.3 mmol) was added at room temperature and the mixture was stirred for 15 minutes at room temperature. Pure (-)-sparteine (29.1 mL, 126.7 mmol) was added and the mixture became cloudy but cleared upon further stirring. After stirring for 30 minutes at room temperature, the mixture was cooled to -78 °C and a solution of aldehyde 8 (14.0 g, 50.7 mmol) in anhydrous DCM (75 mL) was added dropwise over 15 minutes. The mixture was stirred at -78 °C for 1 hour and slowly warmed to 0 °C over 3 hours, at which point NMR indicated complete consumption of the starting material. The mixture was quenched with saturated aqueous NaHCO3 (200 mL) and the organic layer was separated. The aqueous layer was washed with CH2Cl2 (200 mL) and the organic layers were combined, dried over Na2SO4, filtered, and concentrated in vacuo to afford crude 9 as a dark yellow oil. The material was then passed through a plug of neutral silica (DCVC) and eluted into a 3 L flask with a gradient of 50% EtOAc / hexanes. The mixture was concentrated and further dried by removing toluene and taken directly to the next step. A small aliquot was purified by preparative TLC for spectroscopic analysis. Note 1: The aldol reaction gave selectivity in a 10:1 ratio. Resolution of the unwanted diastereomer was achieved in a further step in saponification step 2. Note 2: The aldol adduct 9 is prone to hydrolysis when purified on untreated silica. Flash chromatography on neutral silica (Silicyce) eluting with a gradient from hexanes to 50% EtOAc / hexanes gave 9 in 95%+ purity. In practice, as described in this procedure, the material was clean enough after passing through the plug of neutral silica. Note 3: (-)-Sparteine can be recovered from the DCVC column.

[0369] Alcohol 9: TLC (25% EtOAc / Hex) R f = 0.23 11H NMR (C6D6, 500 MHz) δ for the major isomer: 7.49 (d, J = 8.6 Hz, 2H), 6.77 (d, J = 8.7 Hz, 2H), 6.23 (s, 1H), 5.83 - 5.76 (m, 1H), 5.31 - 5.25 (m, 1H), 5.08 - 5.00 (m, 2H), 4.18 (d, J = 6.7 Hz, 1H), 3.64 - 3.57 (m, 1H), 3.24 (s, 3H), 2.46 (m, 2H), 2.02 - 1.94 (m, 2H), 1.93 - 1.85 (m, 1H), 1.66 - 1.50 (m, 3H), 1.20 (s, 3H), 0.71 (s, 9H); for the minor isomer: δ 7.59 (d, J = 8.7 Hz, 2H), 6.83 (d, J = 8.8 Hz, 2H), 5.91 (s, 1H), 5.89 - 5.79 (m, 1H), 5.01 (dd, J = 9.2, 0.8 Hz, 1H), 4.09 (d, J = 6.8 Hz, 1H), 3.58 (m, 1H), 3.25 (s, 3H), 2.46 (m, 2H), 2.02 - 1.94 (m, 2H), 1.93 - 1.85 (m, 1H), 1.66 - 1.50 (m, 3H), 1.17 (s, 3H), 0.68 (s, 9H); 13 13C NMR (C6D6, 500 MHz) δ for the major isomer: 204.8, 172.6, 160.2, 133.5, 132.6, 128.0, 117.6, 113.5, 102.0, 86.0, 83.1, 71.6, 68.4, 54.4, 45.5, 30.5, 29.4, 29.1, 21.8; for the minor isomer: δ 204.8, 172.6, 160.4, 133.4, 130.8, 128.4, 117.7, 113.7, 102.4, 87.7, 81.9, 71.6, 68.4, 54.4, 45.5, 30.9, 29.4, 29.1, 22.4.

[0370] (3R)-1-((R)-5-(tert-Butyl)-2-thioxothiazolidin-3-yl)-3-((tert-butyldimethylsilyl)oxy)-5-((4R,5S)-2-(4-methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)pentan-1-one (10). To a 3 L flask charged with crude 9 were successively added CH2Cl2 (600 mL) and 2,6-lutidine (29.5 mL, 253.3 mmol). The mixture was purged with argon and cooled to 0 °C. TBSOTf (34.9 mL, 152.0 mmol) was added dropwise and the mixture was warmed to room temperature and stirred for 2 h, at which point NMR indicated complete consumption of the starting material. The solution was quenched by addition of solid sodium bicarbonate (20 g) and stirring for 15 min. The mixture was vacuum filtered through a pad of neutral silica (DCVC) eluting with CH2Cl2 (1.5 L) into a 3 L flask. The eluate was concentrated in vacuo to give 10 as a dark yellow crude oil and was used directly in the next reaction. A small aliquot was purified by preparative TLC for spectroscopic analysis.

[0371] Intermediate 10: TLC (100% CH2Cl2) R f = 0.40 11H NMR (C6D6, 500 MHz) δ For the major isomer: 7.61 (d, J = 8.6 Hz, 2H), 6.89 (d, J = 8.7 Hz, 2H), 5.94 (s, 1H), 5.85 (m, 1H), 5.34 (d, J = 17.1 Hz, 2H), 5.11 (d, J = 10.6 Hz, 2H), 5.03 (d, J = 8.3 Hz, 1H), 4.46 (m, 1H), 4.14 (d, J = 17.1, 1H), 3.85 - 3.55 (m, 2H), 3.31 (s, 3H), 2.57 (m, 2H), 2.03 (d, J = 11.8 Hz, 2H), 1.91 (m, 2H), 1.26 (s, 3H), 1.00 (s, 9H), 0.77 (s, 9H), 0.19 (s, 3H), 0.14 (s, 3H). For the minor isomer: 7.56 (d, J = 8.6 Hz, 2H), 6.81 (d, J = 8.7 Hz, 2H), 6.31 (s, 1H), 5.85 (m, 1H), 5.34 (d, J = 17.1 Hz, 2H), 5.11 (d, J = 10.6 Hz, 2H), 5.06 (d, J = 8.3 Hz, 1H), 4.54 (m, 1H), 4.23 (d, J = 17.1, 1H), 3.85 - 3.55 (m, 2H), 3.26 (s, 3H), 2.54 (m, 2H), 2.03 (d, J = 11.8 Hz, 2H), 1.91 (m, 2H), 1.9 (s, 3H), 1.03 (s, 9H), 0.78 (s, 9H), 0.22 (s, 3H), 0.19 (s, 3H); 13 13C NMR (C6D6, 500 MHz) δ 204.7, 204.6, 170.5, 170.4, 160.4, 160.2, 133.3, 133.2, 132.6, 130.9, 128.3, 128.0, 127.8, 127.6, 117.6, 117.5, 113.7, 113.6, 102.4, 102.0, 87.6, 85.7, 83.1, 82.1, 81.1, 71.7, 69.1, 69.0, 54.4, 46.0, 45.7, 37.5, 32.4, 31.7, 31.2, 29.4, 28.5, 26.1, 25.9, 25.8, 25.5, 22.4, 21.8, 18.0. Note 1: 10 can be further purified (95%+) by flash chromatography on neutral silica gel, eluting with a gradient of hexane to CH2Cl2. In practice, the material is clean enough to proceed to the next step without chromatography.

[0372] (3R)-3-((tert-Butyldimethylsilyl)oxy)-5-((4R,5S)-2-(4-methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)pentanoic acid (4)

[0373] Lithium hydroxide monohydrate (6.38 g, 0.152 mmol) was added to a 3 L flask containing a solution of crude 10 in 4:1 CH3CN / H2O (250 mL). The mixture was stirred overnight at room temperature, at which point the deep yellow color dissipated to a light brown. The mixture was diluted with 200 mL of H2O and 200 mL of diethyl ether. The aqueous layer was collected and the organic layer was back-extracted with H2O (2 × 100 mL). The aqueous layers were combined and carefully acidified to pH 6 with 1 M HCl. The mixture was extracted into EtOAc (3 × 500 mL), and the organic materials were combined, dried over Na2SO4, filtered, and concentrated in vacuo to give a clear brown oil. The material was purified on silica gel, eluting with a gradient of hexanes to 30% EtOAc / hexanes, to give acid 4 (5.5 g, 50% over four steps) as a light brown oil. Note 1: Minor diastereomers obtained from the oxy-acetal reaction were removed in this step after chromatography.

[0374] Acid 4: TLC (50% EtOAc / hexanes) R f = 0.54; 1 1H NMR (C6D6, 500 MHz) δ major isomer 7.51 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.7 Hz, 2H), 5.88 (s, 1H), 5.77 (m, 1H), 5.28 (d, J = 10.5, 1H), 5.06 (d, J = 10.5, 1H), 4.07 (m, 1H), 3.26 (s, 3H), 2.17 - 2.47 (m, 2H), 1.84 (m, 2H), 1.60 (m, 2H), 1.13 (s, 3H), 0.92 (s, 9H), 0.06 (s, 3H), 0.02 (s, 3H); minor isomer 7.50 (d, J = 8.7 Hz, 2H), 6.78 (d, J = 8.7 Hz, 2H), 6.16 (s, 1H), 5.77 (m, 1H), 5.28 (d, J = 10.5, 1H), 5.06 (d, J = 10.5, 1H), 4.15 (m, 1H), 3.23 (s, 3H), 2.17 - 2.47 (m, 2H), 1.84 (m, 2H), 1.60 (m, 2H), 1.16 (s, 3H), 0.95 (s, 9H), 0.10 (s, 3H), 0.07 (s, 3H); 13 13C NMR (C6D6, 500 MHz) δ.

[0375] (3S,4S,E)-1-Iodo-2,4-dimethylhexa-1,5-dien-3-yl (3R)-3-((tert-butyldimethylsilyl)oxy)-5-((4R,5S)-2-(4-methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)pentanoate (11)

[0376] The acid 4 (5.5 g, 12.2 mmol) and the alcohol 6c (3.23 g, 12.8 mmol) were combined in a 250 mL round bottom and dried by removing toluene prior to use. DMAP (0.150 g, 1.22 mmol) and pivalic anhydride (3.71 mL, 18.3 mmol) were added sequentially and the mixture was stirred neat at 50 °C for 5 h. The pivalic anhydride was then removed from the mixture overnight under a constant stream of air. The crude material was then loaded directly onto silica gel and eluted with a gradient of hexanes to 10% Et2O / hexanes to give the ester 11 (6.7 g, 80%) as a clear oil. Note 1: Pivalic anhydride tends to streak on silica gel and reduce resolution. Maximum purification resolution is achieved when little to no pivalic anhydride is present in the crude mixture prior to chromatography. Note 2: A thin 1 cm stir bar is most effective for this reaction as it allows for vigorous stirring without splashing up the sides of the flask.

[0377] Ester 11: 11H NMR (C6D6, 500 MHz) δ For the major isomer: 7.54 (d, J = 8.7 Hz, 2H), 6.83 (d, J = 8.6 Hz, 2H), 6.16 (s, 1H), 5.90 (s, 1H), 5.84 - 5.76 (m, 1H), 5.66 - 5.56 (m, 1H), 5.30 (d, J = 17.3 Hz, 1H), 5.13 (d, J = 8.1 Hz, 1H), 5.07 (d, J = 8.1 Hz, 2H), 4.99 - 4.87 (m, 2H), 4.09 (dt, J = 6.5, 1.3 Hz, 1H), 3.27 (s, 3H), 2.40 (dd, J = 15.1, 6.6 Hz, 1H), 2.19 (dd, J = 15.0, 5.7 Hz, 1H), 1.89 - 1.80 (m, 2H), 1.75 (dd, J = 12.9, 3.7 Hz, 1H), 1.66 (s, 3H), 1.65 (m, 2H), 1.19 (s, 3H), 0.95 (s, 9H), 0.66 (d, J = 6.9 Hz, 3H), 0.09 (s, 2H), 0.07 (s, 2H). For the minor isomer: δ 7.52 (d, J = 8.7 Hz, 2H), 6.79 (d, J = 8.6 Hz, 2H), 6.19 (s, 1H), 6.17 (s, 1H), 5.84 - 5.76 (m, 1H), 5.66 - 5.56 (m, 1H), 5.16 (d, J = 8.1 Hz, 1H), 5.07 (d, J = 8.1 Hz, 2H), 4.99 - 4.87 (m, 2H), 4.09 (dt, J = 6.5, 1.3 Hz, 1H), 3.23 (s, 3H), 2.47 (dd, J = 15.0, 6.3 Hz, 1H), 2.27 (m, 2H), 1.98 - 1.96 (m, 2H), 1.68 (s, 3H), 1.22 (s, 3H), 0.98 (s, 9H), 0.67 (d, J = 6.9 Hz, 3H), 0.12 (s, 3H), 0.11 (s, 3H); 1313C NMR (C6D6, 500 MHz) δ for the major isomer: 164.7, 160.4, 144.6, 139.3, 133.3, 130.9, 128.2, 128.0, 127.8, 127.6, 127.4, 127.2, 117.6, 115.4, 113.6, 102.3, 87.5, 81.7, 81.5, 80.0, 69.5, 54.4, 42.6, 42.3, 40.0, 32.5, 31.4, 25.7, 22.4, 20.0, 17.9, 16.0; for the minor isomer: 169.6, 160.2, 144.5, 139.5, 133.2, 132.5, 128.2, 128.0, 127.8, 127.6, 127.4, 127.2, 117.5, 115.4, 113.5, 120.0, 85.6, 83.0, 81.6, 80.0, 69.4, 54.4, 40.0, 30.0, 28.5, 25.7, 21.8, 20.0, 17.9, 16.1.

[0378] (3aS, 6S, 7S, 11R, 13aR, E)-11-((tert-Butyldimethylsilyl)oxy)-7-((E)-1-iodoprop-1-en-2-yl)-2-(4-methoxyphenyl)-6,13a-dimethyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f][1]oxacyclododecin-9-one (12). The ester 11 was dried by rotary evaporation of toluene in a 3 L flask and then charged with anhydrous toluene (700 mL). The mixture was purged with Ar and heated to reflux. A solution of Hoveyda-Grubbs 2nd generation catalyst (0.520 mg, 0.830 mmol) in anhydrous toluene (500 mL) was added dropwise via a 1 L addition funnel. After stirring for 20 minutes, the mixture changed from a clear green to a black solution and was stirred further at reflux for 5 hours. The mixture was then cooled to room temperature and concentrated. The crude black semi-solid was suspended in hexanes and filtered through a pad of Celite, eluting with hexanes. The eluate was concentrated to give a green oil which was purified on silica gel, eluting with a gradient of hexanes to 15% Et2O / hexanes to give the macrocycle 12 (3.25 g, 51%) as an off-white solid.

[0379] Macrocycle 12: 11H NMR (500 MHz, C6D6) major δ 7.57 (d, J = 8.7 Hz, 2H), 6.86 (d, J = 8.7 Hz, 2H), 6.24 (s, 1H), 5.93 (s, 1H), 5.88 - 5.75 (m, 1H), 5.72 - 5.56 (m, 1H), 5.33 (d, J = 17.2 Hz, 1H), 5.16 (d, J = 8.1 Hz, 1H), 5.03 - 4.90 (m, 2H), 4.25 - 4.10 (m, 1H), 3.30 (s, 3H), 2.45 (d, J = 6.6 Hz, 1H), 2.42 (d, J = 6.6 Hz, 1H), 2.34 - 2.20 (m, 3H), 2.04 - 1.75 (m, 2H), 1.69 (s, 3H), 1.22 (s, 3H), 0.98 (s, 9H), 0.69 (d, J = 6.9 Hz, 3H), 0.12 (s, 3H), 0.10 (s, 3H); minor δ 7.55 (d, J = 8.7 Hz, 2H), 6.83 (d, J = 8.7 Hz, 2H), 6.22 (s, 1H), 6.19 (s, 1H), 5.88 - 5.75 (m, 1H), 5.72 - 5.56 (m, 1H), 5.19 (d, J = 17.2 Hz, 1H), 5.10 (d, J = 8.1 Hz, 1H), 5.03 - 4.90 (m, 2H), 4.25 - 4.10 (m, 1H), 3.27 (s, 3H), 2.51 (d, J = 6.6 Hz, 1H), 2.49 (d, J = 6.6 Hz, 1H), 2.34 - 2.20 (m, 3H), 2.04 - 1.75 (m, 2H), 1.71 (s, 3H), 1.25 (s, 3H), 1.01 (s, 9H), 0.71 (d, J = 6.9 Hz, 3H), 0.15 (s, 3H), 0.14 (s, 3H); 13 13C NMR (500 MHz, C6D6) δ.

[0380] (4R, 7R, 8S, 11S, 12S, E)-4,7,8 - Trihydroxy - 12 - ((E)-1 - iodoprop - 1 - en - 2 - yl)-7,11 - dimethyloxacyclododec - 9 - en - 2 - one (13). Lactone 12 (3.25 g) was dissolved in 5:1 CH2Cl2 / MeOH (300 mL), and CSA in solid form (3.45 g, 14.9 mmol) was added. The mixture was stirred for 5 h, at which point TLC indicated complete conversion of the starting material. Saturated bicarbonate solution (50 mL) and the mixture were extracted into CH2Cl2. The organic layer was collected and concentrated to a crude oil, which was further purified on silica gel (CH2Cl2 to 35% acetone / CH2Cl2) to give pure 13 (1.10 g, 52%).

[0381] Triol 13: TLC (30% Ace / CH2Cl2) Rf = 0.25; 1 H NMR (CDCl3, 500 MHz) δ 6.49 (s, 1H), 5.76 (dd, J = 15.2, 9.7 Hz, 1H), 5.40 (dd, J = 15.2, 9.9 Hz, 1H), 5.31 (d, J = 10.7 Hz, 1H), 3.82 (d, J = 9.8 Hz, 1H), 3.77 (dt, J = 11.3, 3.6 Hz, 1H), 2.69 - 2.46 (m, 3H), 1.84 (s, 3H), 1.70 (tt, J = 13.1, 4.3 Hz, 1H), 1.52 - 1.36 (m, 2H), 1.32 (s, 3H), 1.25 (m, 1H), 0.93 (d, J = 6.7 Hz, 3H).

[0382] (2S, 3S, 6S, 7R, 10R, E)-7,10-Dihydroxy-2-((E)-1-iodoprop-1-en-2-yl)-3,7-dimethyl-12-oxoxacyclododec-4-en-6-yl acetate (2). Triol 13 (1.10 g, 2.6 mmol) and CSA (0.12 g, 0.52 mmol) were dissolved in CH2Cl2 (100 mL) and cooled to 0 °C. A solution of trimethyl orthoformate (0.40 mL, 3.1 mmol) in CH2Cl2 (20 mL) was added via a dropping funnel and the mixture was stirred at 0 °C for 1 h, at which point a saturated aqueous solution of disaccharide (5 mL) was added. The mixture was extracted into CH2Cl2 and the organic matter was concentrated to a crude oil, which was purified on silica gel (CH2Cl2 to 25% acetone / CH2Cl2) to give pure nucleus 2 (980 mg, 81%) as a pale yellow semi-solid.

[0383] Nucleus 2: TLC (3:1 hexane / EtOAc): Rf = 0.16; 1H NMR (CDCl3, 500 MHz) δ 6.47 (s, 1H), 5.67 (dd, J = 15.2, 9.5 Hz, 1H), 5.57 (dd, J = 15.2, 9.7 Hz, 1H), 5.29 (d, J = 10.5 Hz, 1H), 5.05 (d, J = 9.5 Hz, 1H), 3.75 (bs, 1H), 3.42 (d, J = 11.1 Hz, 1H), 2.66 - 2.44 (m, 3H), 2.09 (s, 3H), 1.82 (s, 3H), 1.62 - 1.31 (m, 4H), 1.20 (s, 3H), 0.90 (d, J = 6.7 Hz, 3H); 1313C NMR (CDCl3, 100 MHz) δ 172.0, 169.8, 143.5, 139.8, 126.3, 84.4, 80.4, 78.9, 73.5, 69.3, 41.1, 38.4, 35.3, 29.9, 24.8, 21.5, 19.2, 16.5; FTIR (film) νmax 3502, 3058, 2959, 2873, 1733, 1616, 1368, 1243, 1168, 1021 cm -1 ; C 18 H 27 IO6Na [M+Na] + HR-ESI-MS m / z calculated for

[0384] ((2R,3R)-3-((2R,3S)-3-methoxypentan-2-yl)oxiran-2-yl)methanol (14). A 5.5 M solution of tert-butyl hydroperoxide in decane (46.0 mL, 253 mmol) was added to a 1 L round-bottom flask containing Ti(O-iPr)4 (2.73 mL, 12.6 mmol), (-)-diethyl tartrate (2.2 mL, 12.6 mmol) and a stirred solution of powdered molecular sieve (2 g) in anhydrous CH2Cl2 (300 mL). The mixture was cooled to -20 °C. The resulting mixture was stirred at -20 °C for 30 min. A solution of alcohol 6d (20.0 g, 127 mmol) in CH2Cl2 (40 mL) was added dropwise. The reaction was warmed to -10 °C over 1 h and stirred at -10 °C for 2 h. The reaction was quenched by the addition of 10% aqueous NaOH (25 mL). MgSO4 (20 g) was added and the mixture was filtered through a pad of diatomaceous earth and the eluate was concentrated. The crude product was purified on silica gel (hexane to 50% EtOAc / hexane) to give the epoxy alcohol 14. Note: As determined by NMR, selectivity was obtained in a 11:1 ratio. The diastereomers were not separable and were carried directly to the oxidation step.

[0385] Epoxy alcohol 14: TLC (2:1 hexane / EtOAc): Rf = 0.10; 11H NMR (C6D6, 500 MHz) δ 3.56 - 3.48 (m, 1H), 3.33 - 3.26 (m, 1H), 3.17 (s, 3H), 3.07 - 3.03 (m, 1H), 2.86 (dd, J = 7.7, 2.3 Hz, 1H), 2.78 (dd, J = 7.2, 2.3 Hz, 1H), 2.59 (dt, J = 4.9, 2.6 Hz, 1H), 1.62 - 1.49 (m, 1H), 1.41 - 1.29 (m, 3H), 0.99 (d, J = 6.9 Hz, 1H), 0.84 - 0.79 (m, 3H). 13 13C NMR (C6D6, 500 MHz) δ 83.4, 61.9, 57.6, 57.5, 57.4, 38.4, 23.6, 10.0, 9.78; FTIR (film) νmax 3422, 2972, 2930, 2879, 1468, 1103 cm -1 ; C9H 18 O3 [M] + HR - ESI - MS m / z calcd for: 174.1250, found 174.1249; [α] 25 D = +4.0° (c = 0.075, CHCl3).

[0386] (2S, 3R)-3-((2R, 3S)-3-Methoxypentan-2-yl)oxirane-2-carbaldehyde (15). To a 1 L round-bottom flask equipped with a magnetic stir bar was added epoxy alcohol 14 and DMSO (200 mL). Freshly prepared solid IBX was added and the mixture was cooled to -20 °C in an ice-salt bath. The oxidation reaction was stirred and warmed to room temperature over 2 h. The mixture was then diluted with EtOAc (500 mL) and H2O (250 mL) and extracted. The aqueous layer was back-extracted with EtOAc (2 × 200 mL). The EtOAc layers were combined and washed with H2O (5 × 350 mL). The organic layer was then concentrated by rotary evaporation. The crude semi-solid was then vacuum filtered through a plug of Celite and the eluate concentrated. The crude product was purified by silica gel (hexanes to 30% EtOAc / hexanes) to afford aldehyde 15 as a clear oil. Note: The 10:1 ratio of diastereomers in this step was not separable and proceeded directly to the Marshall addition. Resolution was achieved after stannylation of the alkyne obtained in the next step.

[0387] Aldehyde 15: TLC (2:1 hexanes / EtOAc): R f = 0.55; 11H NMR (C6D6, 500 MHz) δ 8.68 (d, J = 6.4 Hz, 1H), 3.10 (s, 3H), 2.91 (td, J = 6.4, 4.0 Hz, 1H), 2.84 (dd, J = 7.5, 2.0 Hz, 1H), 2.79 (dd, J = 6.3, 2.0 Hz, 1H), 1.49 - 1.40 (m, 1H), 1.27 - 1.17 (m, 1H), 0.86 - 0.79 (m, 1H), 0.74 (t, J = 7.4 Hz, 3H), 0.64 (d, J = 7.0 Hz, 3H). 13 13C NMR (C6D6, 500 MHz) δ 197.3, 83.0, 59.0, 58.2, 57.4, 38.0, 23.4, 9.6, 9.4; FTIR (film) νmax 2972, 2930, 2879, 2828, 1732, 1468, 1103 cm -1 ; C9H 17 O3 [M + H] + The HR-ESI-MS m / z calculated value for: 173.1172, found 173.1174.

[0388] (1S, 2R)-1-((2R, 3R)-3-((2R, 3S)-3-Methoxypentan-2-yl)oxiran-2-yl)-2-methylbut-3-yn-1-ol (5). Toluene or benzene was removed by azeotroping under vacuum prior to the reaction. The aldehyde 15 (7.01 g, 40.8 mmol) and the allenylstannane 6e (21.0 g, 61.0 mmol) were dried in a 500 mL round-bottom flask. Anhydrous CH2Cl2 (200 mL) was added to the flask and cooled to -78 °C. BF3 etherate (7.53 mL, 61.0 mmol) was added dropwise over 5 minutes. The reaction was stirred at -78 °C for 1 hour. A mixture of MeOH (50 mL) and saturated NaHCO3 (10 mL) was added and the mixture was warmed to room temperature. The layers were separated and the aqueous layer was extracted with ether (3 × 20 mL). The organic layers were combined, washed with brine and dried over Na2SO4 and concentrated. Flash chromatography was performed with a gradient of hexanes to 4:1 hexanes / EtOAc to afford the alkyne 5 (80%) as a clear oil. Note: Any minor diastereomers obtained from the Marshall addition were removed after chromatography. The remaining diastereomers from the Sharpless epoxidation were separated after further purification in the next step.

[0389] Alkyne 5: TLC (2:1 hexanes / EtOAc); R f = 0.50; CAM staining; single spot; 11H NMR (CDCl3, 500 MHz) δ 3.58 (dd, J = 4.4 Hz, 1H), 3.41 (s, 3H), 3.20 (td, J = 6.4, 4.1 Hz, 1H), 3.06 (dd, J = 8.1, 2.3 Hz, 1H), 2.91 (dd, J = 4.5, 2.3 Hz, 1H), 2.81 (ddd, J = 7.0, 4.3, 2.6 Hz, 1H), 2.17 (d, J = 2.5 Hz, 1H), 2.05 (d, J = 4.8 Hz, 1H), 1.67 (ddd, J = 14.2, 7.6, 6.7 Hz, 1H), 1.53 - 1.44 (m, 2H), 1.31 (dd, J = 7.1, 0.7 Hz, 3H), 0.97 (d, J = 7.1 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H); 13 13C NMR (CDCl3, 500 MHz) δ 84.4, 83.8, 72.3, 71.4, 59.0, 58.3, 38.9, 30.4, 23.9, 17.0, 10.6, 10.1.

[0390] (1S, 2R, E)-1-((2R, 3R)-3-((2R, 3S)-3-Methoxypentan-2-yl)oxiran-2-yl)-2-methyl-4-(tributylstannyl)but-3-en-1-ol (3). To a solution of alkyne 5 in a 500 mL round-bottom flask equipped with a magnetic stir bar was added freshly distilled THF (200 mL) via Na benzophenone and PdCl2(PPh3)2. The mixture was cooled to -20 °C in an ice-salt bath. Tributyltin hydride was added dropwise, at which point the mixture gradually turned into a black solution. After the mixture was stirred at -20 °C for 45 minutes, the black solution was concentrated to give a black crude oil. The material was taken up in hexane, filtered through a pad of Celite and the eluate was concentrated. This process was repeated again to remove as much palladium catalyst as possible. The crude orange-yellow oil was purified twice on silica gel (hexane to 5% Et2O / hexane) to give vinyltin 3 as a single diastereomer.

[0391] Vinyltin 3: TLC (10∶1 hexane / Et2O): R f =; 11H NMR (C6D6, 500 MHz) δ 6.24 (dd, J = 19.1, 6.8 Hz, 1H), 6.16 (d, J = 6.8 Hz, 1H), 3.42 (td, J = 4.9, 1.8 Hz, 1H), 3.20 (s, 3H), 3.13 (td, J = 6.3, 4.2 Hz, 1H), 3.04 (dd, J = 8.0, 2.3 Hz, 1H), 2.70 (dd, J = 4.3, 2.3 Hz, 1H), 2.48 (td, J = 6.9, 5.2 Hz, 1H), 1.58 (m, 6H), 1.45 - 1.29 (m, 7H), 1.16 (d, J = 6.9 Hz, 3H), 0.99 - 0.89 (m, 19H), 0.83 (t, J = 7.4 Hz, 3H); 13 13C NMR (C6D6, 500 MHz) δ 154.5, 150.5, 150.4, 150.4, 150.3, 83.8, 83.3, 72.8, 59.0, 57.5, 57.3, 57.2, 39.0, 39.0, 29.3, 27.4, 23.5, 15.9, 15.8, 13.4, 10.5, 9.63, 9.41.

[0392] Convergent Stille coupling to 17S-FD-895 (1). Vinyl stannane 5 (1.33 g, 2.57 mmol) and nuclear macrolide 2 (1.00 g, 2.14 mmol) were combined in a 100 mL flask and dried by rotary evaporation of benzene. Then CuCl (0.425 g, 4.29 mmol), KF (0.249 g, 4.29 mmol), XPhos Pd G2 (0.169 g, 0.214 mmol), and anhydrous tert-butanol (25 mL) were added to the mixture in sequence. The reaction vessel was purged with Ar, heated to 50 °C and stirred overnight, at which time the solution became a gray turbid mixture. Then the mixture was filtered directly through a plug of diatomaceous earth and the plug was washed with acetone. The eluate was concentrated to give a crude brown semi-solid, which was then purified by neutral silica gel, eluting with a gradient of hexane to 30% acetone / hexane to give 17S-FD-895 as a white semi-solid.

[0393] 17S-FD-895 (1): Isomer 1SR: 11H NMR (C6D6, 400 MHz) δ 171.9, 168.7, 140.4, 138.1, 131.4, 131.3, 126.2, 125.8, 83.5, 82.4, 79.0, 73.1, 71.9, 69.1, 58.9, 57.4, 57.0, 41.6, 40.9, 38.9, 38.3, 35.6, 29.9, 24.5, 23.7, 20.5, 16.2, 16.1, 11.6, 10.6, 9.8; 13 13C NMR (C6D6, 100 MHz) δ 172.1, 169.0, 140.7, 137.9, 132.5, 132.4, 131.7, 131.3, 126.4, 126.4, 83.7, 82.6, 79.2, 73.3, 72.9, 69.3, 59.6, 57.7, 57.7, 41.5, 41.1, 39.3, 38.5, 35.8, 32.4, 30.3, 30.1, 29.8, 24.8, 23.9, 23.1, 20.7, 17.2, 16.4, 14.4, 11.9, 10.8, 10.0; FTIR (film) νmax 3447, 2963, 2930, 2875, 1739, 1457, 1374, 1239, 1176, 1089, 1021 cm -1 ; [M+Na] + ; C 31 H 50 O9Na1 [M+Na] + HR-ESI-MS m / z calculated for C29H37O9Na1 [M+Na]: 589.3345, found 589.3347.

[0394] Example 3. Additional synthetic work

[0395] The compound numbers used in Examples 3, 4, and 6 correspond to the compounds described in these examples, and Figures 3A - 3B , Figures 4A - 4F , Figures 5A - 5H , Figure 6 , Figures 7A - 7C , Scheme A1 ( Figure 8 ) and Scheme A2 ( Figure 9 ), Scheme AS1 ( Figure 10 ), Scheme AS2, Scheme AS3 ( Figure 11 ), Scheme AS4 ( Figure 12 ), Scheme AS5 and the compounds described in Tables S1 - S3.

[0396] Since their first discovery in the mid-1990s, the natural product family of polyketides, including FD-895, pladienolides, spliceostatin, herboxidiene, and thiostrepton, has attracted interest due to their selective anti-tumor activity (1-5). In recent years, two lead candidate drugs, E7107 (6) and H3B-8800 (7), have entered phase I clinical trials for solid tumors and leukemia. Studies on the mode of action have shown that they share a similar ability to regulate splicing through interactions within the SF3B component of the spliceosome (11) (8-10). First proposed as a common motif (12) and subsequently verified by structural analysis (13), these small molecules uniquely position themselves at the interface between SF3B1, PHF5A, and SF3B3 (14), which is the hinge region involved in regulating the branch site adenosine binding pocket (15, 16). These splicing regulators all have a similar structural backbone that contains a macrolactone ring connected by a diene to a side chain (17, 18). Here, the importance and positioning of the stereochemical centers in these molecules clearly demonstrate unique geometric requirements for activity.

[0397] Although many of these splicing regulators show the necessary functional spatiality to be able to bind readily to the SF3B pocket in vitro, the high density of their functional groups results in low stability in biological media, leading to short half-lives (t 1 / 2 ≤ 30 minutes) (19). Recent studies have shown that synthetic modifications along the side chain are not only tolerable but also allow for the acquisition of three-dimensional arrangements that reduce the degradation rate (19). These studies have also shown that synthetic analogs meet the requirements for active binding to the spliceosome pocket in vivo (13, 14). This ultimately led to the identification of 17S-FD-895 (1) as a therapeutic lead (20).

[0398] Although efforts have been made to obtain gram quantities of pladienolides by fermentation (21), these methods are limited to the production of natural substances. To obtain the unnatural C17 stereocenter in 17S-FD-895, we focused on synthetic methods. To date, the reported gram-scale syntheses have only been able to obtain the less complex herboxidiene (22). The synthetic challenges faced in the gram-scale preparation of 17S-FD-895 (1, Figure 3A ) include: a total of 11 stereocenters (6 adjacent), a substituted diene, distal functional groups, quaternary carbons, and a 12-membered lactone. Our method ( Figure 3A ) builds on previous milligram-scale efforts ( Figure 3B ) (23-28), which established the importance of group assembly. Due to the high biological activity of 1, at an estimated dose of 4 mg / m 2At the maximum tolerated dose (MTD) of (6), we chose an approach that avoids the generation of the active substance until the last step. In general, our goal was to develop a method that can withstand large-scale synthesis by reducing the handling and chromatography requirements.

[0399] We began the development of a method for preparing 20 g (0.039 mol) of side chain 2 (Scheme A1, Figure 8 ) to ensure more than 15 g (0.027 mol) of 1. This started with the optimization and preparation of the kilogram-scale Crimmins auxiliary 7 (29). Diastereoselective aldol addition, followed by ammonolysis and subsequent methylation, enabled the successful conversion of 235 g (0.94 mol) of 7 (23) into 155 g (0.82 mol) of Weinreb amide 10 per batch. Fortunately, we were able to recover 65% ± 5% of 6. At this point, we encountered our first challenge: the high volatility of aldehyde 11. This was circumvented by changing the solvent to 2-methyltetrahydrofuran, enabling the reduction and homologation of 10 to 12 without the isolation of 11. Next, reduction with DIBAL-H gave alcohol 13, which could be stored at 4 °C for over 2 years. Sharpless epoxidation of 13 gave 14 in 6:1 dr (diastereomeric ratio), which was oxidized to 15 using TEMPO. As shown in Scheme A1 ( Figure 8 ), condensation of aldehyde 15 with Marshall allenylstannane 16 (30) gave alkyne 17.

[0400] The next problem arose in the hydrostannylation of 17, where the use of a palladium catalyst gave only 1:5 α:β regioselectivity. This led to contamination with trace amounts of the unwanted α-vinyltin, which was reduced to 1:10 dr in favor of the desired β-stannane by using the Figueroa molybdenum catalyst (31) (inset, Scheme A1). Finally, effective chromatography conditions facilitated the obtainment of 2 with a purity of 95+% by LC / MS analysis. To date, we have stockpiled over 200 g (1.3 mol) of 13. After several repetitions, we were able to synthesize 6.5 ± 0.5 g (0.013 mol) of 2 from 25 g (0.16 mol) of 13 in one week.

[0401] Parallel work was also initiated to produce 20 g (0.043 mol) of 3. We developed a scalable method for the preparation of intermediate 22 (23) from the mono-protected 18 in 300 g batches. To achieve this, TEMPO oxidation enabled the scalable conversion of 18 to 19 and 20 to 21 without chromatography. Lowering the reaction temperature (-78 °C to -94 °C) improved the dr of the allylboration of aldehyde 19 to 20 (85% to 95%). Solvent change (THF to Et2O) and reaction temperature optimization (-78 °C to -94 °C) increased the selectivity of the Grignard addition of 21 to give 22 (85% to 90% dr). The process currently requires a single chromatography step (20, Scheme A2( Figure 9 ))). With over 4 years of stability at -20 °C, compound 22 provides an ideal storage point for the batch preparation of nucleus 3.

[0402] The conversion of 22 to 3 presented the most significant challenge. The previously established method for the conversion of 22 to 23 (23) relied on extremely pure ZnBr2, the hygroscopicity of which increased the complexity upon scale-up. After reaction screening, we observed that the in situ decomposition of CBr4 in i-PrOH (32) reproducibly returned 65% ± 5% of 23, enabling three conversions in one step. The next challenge arose in the installation of the C1-C3 fragment. After oxidation to 24, we used a chiral tert-leucine-derived thiazolidinethione auxiliary (29) to install the distal C3 stereocenter with 9:1 dr. Subsequent protection and saponification gave acid 27, which was esterified with alcohol 33 (34) in neat pivalic anhydride (35) to give 34. This 6-step sequence could be carried out in 3 days to obtain a 10 g (0.015 mol) batch of 34 from 25 g (0.069 mol) of 22. At this point, we had installed the remaining 5 stereocenters required for 1 in 34, with a purity of 95+%.

[0403] Next, we turned our attention to the challenging ring-closing metathesis (Scheme A2( Figure 9 ))). Previously, despite the use of additives (36), the reaction was carried out at a maximum of 1 g (28) and allylic isomerization occurred. After screening catalysts and reaction conditions, we found that reversing the order of addition (solution of the 2nd generation Hoveyda-Grubbs catalyst in toluene to 34 in refluxing toluene) provided an acceptable yield of 35 on the 5-10 g scale. Subsequent complete deprotection of 35 with a weak acid, followed by selective acetylation of C7 in 36 via orthoester formation, gave nucleus 3. After optimization, we are now able to convert 30 g (0.083 mol) of 22 to 1.8 ± 0.2 g (0.0039 mol) of 3 (purity by LC / MS of 95+%) in less than 2 weeks.

[0404] At this stage, we are ready to perform the final step ( Figure 4A ). We chose to perform olefin cross-coupling at C13-C14 because alternating installation of the C14-C15 olefin via cross-metathesis or Julia-Kocienski olefination ( Figure 4B )(24, 28, 38) can be complicated by the formation of unwanted cis-olefins. After parallel reaction screening, we decided to perform a Stille coupling (39) using Buchwald XPhos Pd G2 catalyst with CuCl and KF in anhydrous t-BuOH. Under Class III safety conditions, we prepared 1 in 80% ± 2% yield with a worker exposure time of less than 3 hours per 5 g batch. Fortunately, we were able to recover 16% ± 3% of 3, which can be recycled, thus providing an effective mass balance in the 3 to 1 transformation. The side chain 2 is not recoverable.

[0405] To further evaluate this route, we independently introduced 13 C labels at C1 and C30 into 1 ( Figure 4B ). By preparing the Sammakia auxiliary with 1- 13 C acetyl chloride (Scheme AS1( Figure 10 )), relaying it to the corresponding 13 C1-labeled nucleus 3, and coupling it with side chain 2, the 13 C isotope label was installed at C1 to give 1 g of 13 C1-17S-FD-895. The 13 C label at C30 was introduced by selective acetylation of 36 with 1- 13 C acetic anhydride (Scheme S2). The resulting 13 C30-labeled 3 was coupled with 2 to prepare 100 mg of 13 C30-17S-FD-895. 13 C-NMR spectroscopy ( Figure 4B ) confirmed that the 13 C1-17S-FD-895 and 13 C30-17S-FD-895 batches were single compounds with a purity of 98%. In summary, this improved route produced over 17 g of 17S-FD-895 (1) with all 11 stereocenters installed with high selectivity and reproducibility. Additionally, the ability to produce gram-scale batches of stable isotope-labeled material is particularly advantageous for in vivo pharmacological evaluation.

[0406] Next, we wished to expand the structure-activity relationship (SAR) profile of 1 ( Figure 4C )(2, 40-42) by obtaining unnatural analogs from late-stage intermediates using this route. The C3-isomer 1a ( Figure 4D), C7-isomer 1b( Figure 4E ) and C18-C19 epoxide isomer 1c( Figure 4F ) were synthesized by varying the chiral reagent (1a, Scheme AS3( Figure 11 ) and 1b, Scheme AS4( Figure 12 )) or by collecting the minor isomeric byproduct (1c) generated during the synthesis of 1. Screening of 1a-1c in human colorectal tumor HCT-116 cells showed that inversion of the C3 or C7 stereocenter in 1a and 1b impaired activity, while the epoxide isomer 1c retained potency compared to 1.

[0407] These results are consistent with the established x-ray crystal structure of the SF3B core complexed with pladienolide B (14) (Figure 5). In this and related structures (18), inversion of the C3 hydroxyl in 1a eliminated its interaction with K1071 of the SF3B1 subunit( Figures 5A - 5F ). The lack of activity of the C7 isomer follows a similar rationale, as inversion of the C7 acetate in 1b disrupted its interaction with R38 in PHF5A. These findings support a strict SAR within the 12-membered core as it bridges the interface between SF3B1 and PHF5A. Structurally, it also supports tolerance to inversion of the C18-C19 epoxide in 1c, an isomer with activity comparable to 1 in HCT-116 cells. The rotational freedom within the side chain( Figures 5G - 5H ) allows pladienolide B and related analogues to adopt different conformations to access the same binding pocket. Overall, this synthesis facilitated the complete preclinical evaluation of the substance, provided isotopically labeled material, filled gaps in SAR data, and contributed to understanding the structural features required for small molecule splicing modulation.

[0408] Example 4. General Experimental Methods

[0409] Chemical reagents were purchased from Acros Organics, Alfa Aesar, Chem-Impex Int., CreoSalus, Fischer Scientific, Fluka, Oakwood Chemical, Sigma-Aldrich, Spectrum Chemical Mfg. Corp. or TCI Chemicals. Deuterated NMR solvents were obtained from Cambridge Isotope Laboratories. All reactions were carried out using rigorously dried anhydrous solvents, which were obtained by passing through columns composed of activated Al alumina or purchased in anhydrous form. By active Trace amounts of dimethylamine were removed by zeolite and subsequent NaOCN column to obtain anhydrous N,N-dimethylformamide. Triethylamine (Et3N) was dried over Na and freshly distilled. Ethyl-N,N-diisopropylamine (EtNiPr2) was distilled from ninhydrin and then from KOH. Anhydrous CH3CN was obtained by distillation from CaH2. All reactions were carried out under a positive pressure of Ar in oven-dried glassware sealed with septa and stirred with a Teflon-coated stir bar using an IKAMAG RCT-basic stirrer (IKA GmbH). The solution was heated on the adapter of the IKAMAG RCT-basic stirrer. Analytical thin-layer chromatography (TLC) was performed on silica gel 60 F254 pre-coated glass plates (EMSciences). Preparative TLC (pTLC) was carried out on silica gel 60 plates (EM Sciences). Visualization was achieved with UV light and / or appropriate staining agents (I2, KMnO4, bromocresol green, 2,4-dinitrophenylhydrazine, ninhydrin, and ammonium cerium molybdate on SiO2). Flash chromatography was performed on 230 - 400 mesh 60 grade silica gel from Fischer Scientific or 40 - 63 μm mesh 60 grade SiliaFlash irregular silica gel P60. Yields correspond to isolated, chromatographically and spectroscopically homogeneous materials. 1 1H NMR and 13 13C NMR spectra were recorded on a Varian VX500 spectrometer equipped with an Xsens Cold probe. 1 1H and 13 13C spectral chemical shift δ values were reported in parts per million (ppm), and multiplicities were abbreviated as s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad. All 13 13C NMR spectra were recorded with complete proton decoupling. FID files were processed using MestraNova 12.0.3 (MestreLab Research). Electrospray (ESI) mass spectrometry was performed using a ThermoFinnigan LCQ Deca spectrometer, and high-resolution analysis was carried out using a ThermoFinnigan MAT900XL mass spectrometer with electron impact (EI) ionization. High-resolution electrospray ionization mass spectrometry (HR-ESI-MS) was performed using a Thermo Scientific LTQ Orbitrap XL mass spectrometer. FTIR spectra were obtained as thin films on KBr or NaCl disks on a Nicolet magna 550 II series spectrometer, and the peaks were reported in wave numbers (cm -1 -1). The specific rotation [α] D was measured using a Perkin-Elmer model 241 polarimeter with the specified solvent and concentration and reported in deg cm2 g -1 in units. Spectral data and procedures for all new compounds were provided, and copies of selected spectra were provided.

[0410] Example 5. Experimental data for additional synthetic work

[0411] Synthetic procedure for side chain 2 ( Figure 8 , Scheme A1). An eleven-step sequence was developed to prepare 20 g of component 2 starting from auxiliary 6.

[0412] This procedure was partially optimized from published methods (19). Although compound 9 has previously been synthesized in ten-gram quantities (33), large amounts of toxic AlMe3 were required to hydrolyze the oxazolidinone auxiliary. Switching to the more labile thiazolidinethione auxiliary allowed for mild hydrolysis and facilitated the ten-gram production of ethanol 13 and subsequent gram-scale production of vinyltin 2. Each batch of 25 g of 13 provided 6.5 g of 2 with 95% purity, and a total of 20 g of 2 has been produced to date.

[0413] Synthesis of auxiliary 7

[0414]

[0415] Reagents: Et3N, 98% (Fischer Scientific): Redistilled before use. DMAP, 98% (CreoSalus): Can be used without further purification. Propionyl chloride, 98% (Sigma-Aldrich): Freshly distilled before use.

[0416] (R)-1-(4-Benzyl-2-thioxothiazolidin-3-yl)propan-1-one (7). Et3N (700 mL, 5.20 mol) and DMAP (105 g, 862 mol) were added to a 20 L reaction vessel at room temperature, which contained a solution of 6 (892 g, 4.26 mol) in anhydrous CH2Cl2 (9 L). The mixture was cooled to 0 °C, and propionyl chloride (490 mL, 5.61 mol) dissolved in CH2Cl2 (2.3 L) was added dropwise over 1.5 h while maintaining the temperature at 0 °C. The mixture was then stirred at room temperature. After 18 h, the mixture was cooled to 0 °C, and saturated NH4Cl (5.8 L) was added dropwise while keeping the temperature below 0 °C. The mixture was extracted with CH2Cl2 (3 × 2 L). The combined organic phases were washed with saturated NaHCO3 (4 L) and brine (4 L), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Pure auxiliary 7 (950 g, 83%) was obtained by crystallization from CH3CN. The characterization data matched the literature values (43). 11H NMR (500 MHz, CDCl3) δ 7.30 (m, 3H), 7.24 (m, 2H), 5.34 (ddd, J = 10.9, 7.2, 3.8 Hz, 1H), 3.36 (m, 2H), 3.17 (dd, J = 13.2, 3.8 Hz, 1H), 3.05 (m, 2H), 2.84 (d, J = 11.6, 1H), 1.15 (t, J = 7.2 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 201.2, 175.0, 136.7, 129.6, 129.0, 127.3, 68.8, 36.8, 32.5, 32.0, 8.9; C 12 H 13 NOS2 [M+1] + The LCMS (ES-API) m / z calculated value for

[0417] Synthesis of Adduct 8

[0418]

[0419] Reagents: Propionaldehyde, 98% (Alfa Aesar): Redistilled before use. EtN(i-Pr)2, 97% (Fisher Scientific): Redistilled before use. TiCl4, 98% (Alfa Aesar): Can be used without further purification.

[0420] (2R,3S)-1-((S)-4-Benzyl-2-thiazolidinyl)-3-hydroxy-2-methylpentan-1-one (8). (S)-1-(4-Benzyl-2-thiazolidinyl)propan-1-one (7) (235 g, 887 mmol) was added to a 20 L reaction flask and dissolved in CH2Cl2 (7 L) with mechanical stirring. The mixture was cooled to below 0 °C. TiCl4 (1 M solution in CH2Cl2, 922 mL, 922 mmol) was added dropwise over 1 h while maintaining the temperature below 0 °C, at which point the mixture turned orange. EtN(i-Pr)2 (168 mL, 966 mmol) was added dropwise over 30 min, and the resulting black mixture was stirred at 0 °C for 15 min. After cooling the reaction to -94 °C, a solution of propionaldehyde (71.0 mL, 984 mmol) in anhydrous CH2Cl2 (350 mL) was added dropwise over 6 h. The mixture was stirred at -94 °C for 30 min and then slowly warmed to room temperature overnight. The mixture was cooled to 0 °C and saturated NaHCO3 (1.7 L) was added slowly. Note rapid heating. The phases were separated, and the aqueous phase was extracted with CH2Cl2 (3 × 1 L). The combined organic phases were washed with brine (2 L), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Pure adduct 8 (250 g, 88%) was obtained by flash chromatography, eluting with a gradient of heptane to 1:3 EtOAc / heptane with a 9.5:1 dr.

[0421] Adduct 8: TLC (1:3 EtOAc / heptane): R f = 0.63 (CAM staining); 1 H NMR (500 MHz, CDCl3) δ 7.34 (m, 2H), 7.29 (m, 3H), 5.37 (ddd, J = 11.2, 7.1, 4.4 Hz, 1H), 4.73 (qd, J = 7.1, 2.3 Hz, 1H), 3.97 (ddd, J = 8.1, 5.3, 2.2 Hz, 1H), 3.38 (ddd, J = 11.5, 7.2, 1.1 Hz, 1H), 3.25 (dd, J = 13.2, 4.1 Hz, 1H), 3.05 (dd, J = 13.2, 10.5 Hz, 1H), 2.89 (dd, J = 11.6, 0.8 Hz, 1H), 2.77 (bs, 1H), 1.61 (m, 1H), 1.45 (m, 1H) 1.18 (d, J = 7.1 Hz, 3H), 0.98 (t, J = 7.5 Hz, 3H); 1313C NMR (125 MHz, CDCl3) δ 201.7, 178.7, 136.5, 129.6, 129.1, 129.1, 127.4, 72.6, 69.1, 42.3, 37.1, 31.9, 26.7, 10.6, 10.5; FTIR (film) ν max 3444, 3027, 2964, 2937, 2876, 1689, 1455, 1352, 1258, 1191, 1164, 1041, 1029, 960 cm -1 ; C 15 H 19 NO2S2 [M+1] + of LCMS (ES-API) m / z calculated value: 324.40; [α] 25 D = 199.5° (c = 1.0 CH2Cl2).

[0422] Conversion of alcohol 8 to Weinreb amide 9

[0423]

[0424] Reagents: N,O-dimethylhydroxylamine hydrochloride, 99% (Alfa Aesar): can be used without further purification. Imidazole, 99% (Sigma-Aldrich): can be used without further purification.

[0425] (2R,3S)-3-Hydroxy-N-methoxy-N,2-dimethylpentanamide (9). N,O-Dimethylhydroxylamine hydrochloride (174 g, 1.78 mol) and imidazole (182 g, 2.68 mol) were sequentially added to a solution of 8 (288 g, 892 mmol) in CH2Cl2 (13 L) in a 20 L reaction vessel at room temperature. The mixture was stirred at room temperature for 16 h. H2O (3 L) was added, and the mixture was separated, and then the aqueous phase was extracted with CH2Cl2 (3 × 2.5 L). The combined organic phases were washed with brine (5 L), dried over Na2SO4, filtered and concentrated on a rotary evaporator to give a yellow oil. Pure amide 9 (131 g, 80%) was obtained by flash chromatography, eluting with a gradient of heptane to 3:1 EtOAc / heptane. Note 1: 65% ± 5% of the auxiliary agent 6 was recovered after chromatography. Note 2: Rotational isomers were observed by NMR.

[0426] Amide 9: TLC (3:1 EtOAc / heptane): R f = 0.17 (KMnO4); 11H NMR (500 MHz, CDCl3) δ 3.79 (bs, 1H), 3.76 (td, J = 5.4, 2.6 Hz, 1H), 3.69 (s, 3H), 3.17 (s, 3H), 2.90 (bs, 1H), 1.77 (bs, 1H), 1.57 (m, 1H), 1.39 (m, 1H), 1.15 (d, J = 7.1 Hz, 3H), 0.95 (t, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 178.5, 73.1, 61.7, 38.1, 32.0, 26.8, 10.5, 10.1; FTIR (film) ν max 2969, 2917, 2855, 1719, 1449, 1265, 1178, 1108, 1020, 715 cm -1 ; C8H 17 NO3 [M+1] + of LCMS (ES-API) m / z calculated value: 176.40; [α] 25 D = -11.3° (c = 1.0, CH2Cl2).

[0427] Methylation of Amides 9 to 10

[0428]

[0429] Reagents: NaH, 60% in mineral oil (Alfa Aesar): Can be used without further purification. MeI, 98% (Sigma-Aldrich): Can be used without further purification.

[0430] (2R,3S)-N,3-Dimethoxy-N,2-dimethylpentanamide (10). In a 20 L reaction vessel at room temperature, MeI (1.12 L, 18.0 mol) was added to a solution of amide 9 (155 g, 886 mmol) in a mixture of anhydrous THF (6 L) and anhydrous DMF (1.5 L). The mixture was cooled to 0 °C and NaH (60% in mineral oil, 88.5 g, 2.21 mol) was added in portions, ensuring that the mixture remained at 0 °C. The mixture was slowly warmed to room temperature and stirred for 16 h. After cooling the mixture to 0 °C, pH 7 phosphate buffered saline (1.5 L) was added dropwise. The volatiles were concentrated on a rotary evaporator. H2O (4.5 L) was added to the residue and the resulting mixture was extracted with tert-butyl methyl ether (3 × 3 L). The combined organic phases were washed with brine (3 L), dried over Na2SO4, filtered and concentrated on a rotary evaporator. Pure amide 10 (129 g, 77%) was obtained as a colorless oil by flash chromatography, eluting with a gradient of heptane to 1:1 EtOAc / heptane. Note 1: Rotational isomers were observed by NMR.

[0431] Amide 10: TLC (3:1 EtOAc / heptane): R f = 0.27 (KMnO4); 1 1H NMR (500 MHz, CDCl3) δ 3.68 (s, 3H), 3.41 (s, 3H), 3.30 (tdd, J = 7.0, 4.0, 1.0 Hz, 1H), 3.18 (s, 3H), 3.03 (bs, 1H), 1.58 (dqd, J = 14.9, 7.5, 3.9 Hz, 1H), 1.42 (dt, J = 14.4, 7.2 Hz, 1H), 1.21 (d, J = 6.9 Hz, 3H), 0.93 (t, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 176.5, 83.9, 61.6, 58.7, 39.6, 32.2, 25.3, 14.5, 9.6; FTIR (film) ν max 3581, 3502, 2969, 2934, 2882, 2820, 1658, 1457, 1379 cm -1 ; C9H 19 NO3 [M+1] + of LCMS (ES-API) m / z calcd for: 190.40; [α] 25 D = -13.0° (c = 1.0 CHCl3).

[0432] Conversion of 10 to ester 12

[0433]

[0434] Reagents: DIBAL-H, 1.0 M solution in hexanes (Sigma-Aldrich): Can be used without further purification. NaH, 60% in mineral oil (Alfa Aesar): Can be used without further purification. Triethyl phosphonoacetate, 99% (Oakwood Chemical): Can be used without further purification.

[0435] (4S,5S,E)-Ethyl 5-methoxy-4-methylhept-2-enoate (12) In a 5 L flask, dissolve amide 10 (107 g, 565 mmol) in anhydrous CH2Cl2 (2 L). Cool the mixture to -78 °C. Add DIBAL-H (1.0 M, 880 mL, 886 mmol) dropwise over 45 min at -78 °C and stir for 15 min. Add acetone (100 mL) dropwise over 10 min and warm the mixture to 0 °C. Add saturated Rochelle’s salt (2 L) over 30 min and stir the mixture at room temperature for 1.5 h. Separate the phases and extract the aqueous phase with CH2Cl2 (3 × 500 mL). Dry the combined organic phases over Na2SO4, filter and concentrate on a rotary evaporator. Then dry the residue by azeotropic removal of toluene to afford aldehyde 11, which is used immediately after preparation. A solution of triethyl phosphonoacetate (572 mL, 2.88 mol) in anhydrous 2-methyltetrahydrofuran (400 mL) is added dropwise over 30 min to a 5 L reaction flask containing a suspension of NaH (60% in mineral oil, 97.4 g, 2.44 mol) in anhydrous 2-methyltetrahydrofuran (1 L) cooled to 0 °C. Caution rapid evolution of H2. Stir the mixture at 0 °C for 15 min and add a solution of 11 in anhydrous 2-methyltetrahydrofuran (1 L) dropwise over 30 min. Stir the mixture at room temperature for 16 h, cool to 0 °C and quench with saturated NH4Cl (1.6 L). Concentrate the organics on a rotary evaporator. Extract the mixture with EtOAc (2 × 1 L) and dry the combined organic phases over Na2SO4, filter and concentrate on a rotary evaporator. Purify by flash chromatography, eluting with a gradient of CH2Cl2 to 1:10 EtOAc / CH2Cl2 to afford pure ester 12 (88.3 g, 78% over two steps) as a colorless oil.

[0436] Ester 12: TLC (CH2Cl2): R f = 0.14 (CAM staining); 11H NMR (500 MHz, CDCl3) δ 6.95 (dd, J = 15.8, 7.7 Hz, 1H), 5.82 (dd, J = 15.8, 1.3 Hz, 1H), 4.18 (q, J = 7.1 Hz, 2H), 3.36 (s, 3H), 3.00 (ddd, J = 7.4, 5.6, 4.4 Hz, 1H), 2.57 (m, 1H), 1.51 (m, 1H), 1.41 (m, 1H), 1.28 (t, J = 7.1 Hz, 3H), 1.07 (d, J = 6.8 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 166.8, 151.3, 121.1, 85.6, 60.4, 58.0, 39.3, 20.0, 14.9, 14.4, 10.0; FTIR (film) ν max 2978, 2934, 2882, 2820, 1719, 1650, 1466 cm -1 ; C 11 H 20 O3 [M+NH4] + of LCMS (ES-API) m / z calculated value: 218.6; [α] 25 D = -45.4° (c = 1.0, CH2Cl2).

[0437] Reduction of 12 to alcohol 13

[0438]

[0439] Reagent: DIBAL-H, 1.0 M hexane solution (Sigma-Aldrich): Can be used without further purification.

[0440] (4S, 5S, E)-5-Methoxy-4-methylhept-2-en-1-ol (13). DIBAL-H (1.0 M, 700 mL, 0.85 mol) was added dropwise to a 5 L reaction flask containing a solution of ester 12 (56.5 g, 282 mmol) in anhydrous CH2Cl2 (1.5 L) cooled to -78 °C over 60 minutes. The mixture was stirred at -78 °C for 1 hour. Then acetone (100 mL) was added dropwise over 25 minutes. The mixture was warmed to 0 °C, saturated Rochelle salt (1 L) was added, and the mixture was stirred at room temperature for 2 hours. The phases were separated, and the aqueous phase was extracted with CH2Cl2 (3 × 500 mL). The combined organic phases were washed with brine (250 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Pure alcohol 13 (36.5 g, 82%) was obtained by flash chromatography, eluting with a gradient of heptane to 1:1 EtOAc / heptane.

[0441] Alcohol 13: TLC (1:3 EtOAc / heptane): R f = 0.26 (CAM staining); 1 1H NMR (500 MHz, CDCl3) δ 5.65 (m, 2H), 4.10 (bs, 2H), 3.36 (s, 3H), 2.92 (ddd, J = 7.5, 5.7, 4.2 Hz, 1H), 2.44 (m, 1H), 1.52 (m, 1H), 1.40 (m, 1H), 1.01 (d, J = 6.9 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 135.2, 129.0, 86.4, 64.0, 57.7, 38.9, 23.5, 16.0, 10.0; FTIR (film) ν max 3388, 2968, 2932, 2876, 2826, 1460, 1375 cm -1 ; C9H 18 O2 [M+1] + of LCMS (ES-API) m / z calcd for: 158.20; [α] 25 D = -34.5° (c = 0.2, CHCl3).

[0442] Epoxidation of alcohol 13 to epoxide 14

[0443]

[0444] Reagents: Ti(Oi-Pr)4, 97% (Sigma-Aldrich): Vacuum distilled at 90 °C and 5 mbar. Diethyl (-)-tartrate, 99% (Alfa Aesar): Used without further purification. tert-Butyl hydroperoxide, 3.3 M solution in toluene: Dried from the 70% aqueous solution according to the method developed in the Sharpless laboratory (44).

[0445] ((2R,3R)-3-((2R,3S)-3-Methoxypentan-2-yl)oxiran-2-yl)methanol (14). tert-Butyl hydroperoxide (3.3 M, 76.6 mL, 253 mmol) was added to a 1 L flask containing Ti(Oi-Pr)4 (2.73 mL, 12.6 mmol), diethyl (-)-tartrate (2.21 mL, 12.6 mmol), and powdered molecular sieves (2 g) in a stirred solution of anhydrous CH2Cl2 (400 mL). The mixture was cooled to -20 °C and stirred for 30 minutes. A solution of alcohol 13 (20.0 g, 127 mmol) in CH2Cl2 (50 mL) was added dropwise. The reaction was stirred at -20 °C for 4 hours. The reaction was quenched by the addition of 10% NaOH (25 mL). The mixture was then extracted into CH2Cl2 and concentrated on a rotary evaporator. Pure epoxy alcohol 14 (22.1 g, 88%) was obtained as a 6:1 mixture of diastereomers by flash chromatography, eluting with a gradient of hexanes to 1:1 EtOAc / hexanes. Note 1: The diastereomers were not separable and were carried directly to the next step.

[0446] Epoxy alcohol 14: TLC (1:2 EtOAc / hexanes): R f = 0.10 (CAM staining); 1 1H NMR (500 MHz, C6D6) δ 3.55 (m, 1H), 3.33 (m, 1H), 3.20 (s, 3H), 3.08 (rd, J = 6.3, 4.5 Hz, 1H), 2.89 (dd, J = 7.6, 2.3 Hz, 1H), 2.63 (dt, J = 4.9, 2.6 Hz, 1H), 1.59 (tt, J = 13.9, 7.4 Hz, 1H), 1.41 (m, 1H), 1.35 (m, 1H), 1.02 (d, J = 6.9 Hz, 1H), 0.85 (t, J = 7.4 Hz, 3H), 0.84 (d, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, C6D6) δ 83.8, 62.2, 58.0, 57.9, 57.7, 38.8, 24.0, 10.4, 10.1; FTIR (film) ν max3422, 2972, 2930, 2879, 1468, 1103 cm -1 ; C9H 18 O3[M] + HR-ESI-MS m / z calculated value for : 174.1250, found 174.1249; [α] 25 D = +182.4° (c = 1.0, CHCl3).

[0447] Oxidation of epoxy alcohol 14 to epoxy aldehyde 15

[0448]

[0449] Reagents: TEMPO, 99% (Oakwood Chemical): used without further purification. KBr, (Spectrum Chemical Mfg. Corp.): used without further purification. NaOCl, 2 M, 10% - 15% available chlorine (Spectrum Chemical Mfg. Corp.): used without further purification.

[0450] (2S,3R)-3-((2R,3S)-3-methoxypentan-2-yl)oxirane-2-carbaldehyde (15). A solution of KBr (1.21 g, 10.2 mmol) in H2O (50 mL), saturated NaHCO3 (100 mL), and TEMPO (1.33 g, 8.50 mmol) were added successively to a 2 L flask containing a solution of epoxy alcohol 14 (22.1 g, 127 mmol) in CH2Cl2 (600 mL). The mixture was cooled to 0 °C and a solution of NaOCl (2 M, 85 mL, 170 mmol) and saturated NaHCO3 (100 mL) was added dropwise via an addition funnel. The mixture was warmed to room temperature and stirred for 2 h. The phases were separated and the aqueous phase was extracted with CH2Cl2 (3 × 300 mL). The combined organic phases were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Aldehyde 15 (21.8 g, 99%) was obtained without further purification and used directly in the next step. Note 1: The diastereomers obtained from the epoxidation reaction were not separable in this step and thus carried on.

[0451] Aldehyde 15: TLC (1∶2 EtOAc / hexane): R f = 0.55 (CAM staining); 11H NMR (500 MHz, C6D6) δ 8.67 (d, J = 6.4 Hz, 1H), 3.10 (s, 3H), 2.90 (td, J = 6.4, 4.0 Hz, 1H), 2.84 (dd, J = 7.5, 2.0 Hz, 1H), 2.79 (dd, J = 6.4, 2.0 Hz, 1H), 1.44 (m, 1H), 1.21 (m, 1H), 0.82 (m, 1H), 0.74 (t, J = 7.4 Hz, 3H), 0.63 (d, J = 7.0 Hz, 3H); 13 13C NMR (125 MHz, C6D6) δ 197.7, 83.4, 58.6, 58.5, 57.7, 38.3, 23.7, 10.0, 9.8; FTIR (film) ν max 2972, 2930, 2879, 2828, 1732, 1468, 1103 cm -1 ; C9H 16 O3 [M+H] + HR-ESI-MS m / z calcd for: 173.1172, found 173.1174; [α] 25 D = -89.0° (c = 1.0, CH2Cl2).

[0452] Synthesis of allenylic stannane 16. Starting from commercially available (R)-but-3-yn-2-ol (29), a two-step sequence was used to prepare gram-scale allenylic stannane 16.

[0453]

[0454] Reagents: Et3N, 98% (Fischer Scientific): redistilled over CaH2 before use. MsCl, 98% (Alfa Aesar): used without further purification.

[0455] (R)-But-3-yn-2-yl methanesulfonate. Et3N (198 mL, 1.43 mol) was added dropwise to a 3 L three-necked flask containing a solution of (R)-but-3-yn-2-ol (50.0 g, 713 mmol) in CH2Cl2 (750 mL) cooled to -78 °C over 15 minutes. After 10 minutes, MsCl (83.4 mL, 1.07 mol) was added dropwise over 2 hours. The mixture was stirred at -78 °C for 1 hour, at which point saturated NaHCO3 (500 mL) was added slowly. The mixture was warmed to room temperature and the phases were separated. The aqueous phase was extracted with CH2Cl2 (3 × 500 mL). The combined organic phases were washed with brine (250 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator. The crude product was passed through a SiO2 plug and the eluate was concentrated. (R)-But-3-yn-2-yl methanesulfonate (99%, 107.5 g) was obtained without further purification and used directly in the next step. The characterization data matched the literature values.

[0456] (R)-But-3-yn-2-yl methanesulfonate: 1 1H NMR (500 MHz, CDCl3) δ 5.27 (qd, J = 6.7, 2.1 Hz, 1H), 3.11 (s, 3H), 2.71 (d, J = 2.2 Hz, 1H), 1.65 (d, J = 6.7 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 80.2, 76.4, 67.6, 39.2, 22.5; C6H8O3S [M+1] + Calculated LCMS (ES-API) m / z for: 148.08.

[0457] (R)-But-3-yn-2-yl methanesulfonate to allenylic tin 16 conversion

[0458]

[0459] Reagents: n-BuLi, 2.5 M in hexanes (Acros Organics): used without further purification. iPr2NH, 98% (Alfa Aesar): distilled over CaH2. n-Bu3SnH, 97%, containing 0.05% BHT as stabilizer (Acros Organics): used without further purification. CuBr·DMS, 99% (Acros Organics): used without further purification.

[0460] (S)-Buta-1,2-dien-1-yltributylstannane (16). In a 5 L flask, n-BuLi (2.5 M, 172 mL, 429 mmol) was added dropwise to a solution of iPr2NH (60.7 mL, 429 mmol) in anhydrous THF (800 mL) at 0 °C over 10 min. After 15 min, n-Bu3SnH (135 mL, 501 mmol) was added dropwise over 10 min, and the mixture was stirred at 0 °C for 2.5 h. After cooling the mixture to -85 °C, CuBr·DMS (88.2 g, 429 mmol) was added in portions over 40 min. The mixture was stirred at -85 °C for 30 min. (R)-But-3-yn-2-yl methanesulfonate (53.0 g, 358 mmol) was added dropwise, and the mixture was stirred for 10 min. The mixture was poured into a mixture of tert-butyl methyl ether (2 L), 25% aqueous NH3 (260 mL), and saturated NH4Cl (2 L) and stirred vigorously for 1 h. The phases were separated, and the organic layer was dried over Na2SO4, filtered, and concentrated on a rotary evaporator. The allenylic stannane 16 (77.2 g, 63%) was obtained with 96% ee by vacuum distillation (1 mbar, 150 °C). The characterization data matched the literature values. Note 1: This procedure was repeated to provide a total of over 500 g of 16.

[0461] Allenylic stannane 16: 1 H NMR (500 MHz, CDCl3) δ 5.20 (dq, J = 6.9, 4.0 Hz, 1H), 4.68 (p, J = 6.9 Hz, 1H), 1.64 (dd, J = 6.9, 1.4 Hz, 3H), 1.60 (m, 12H), 1.37 (m, 6H), 0.93 (t, J = 7.4 Hz, 9H); 13 C NMR (125 MHz, CDCl3) δ 210.0, 75.6, 74.9, 29.4, 27.6, 14.0, 10.6; C 13 H 32 Sn [M+1] + The LCMS (ES-API) m / z calculated for: 345.15.

[0462] Derivatization of 16 for determination of enantiomeric excess.

[0463] Reagents: Isobutyraldehyde (Alfa Aesar): Can be used without further purification. BF3·Et2O, 46.5% BF3 (Alfa Aesar): Can be used without further purification.

[0464] A solution of isobutyraldehyde (40 μL, 0.44 mmol) in CH2Cl2 (4 mL) was added dropwise to a solution of allenylic tin 16 (200 mg, 583 μmol) and BF3·OEt2 (210 μL, 1.66 mmol) cooled to -78 °C. After stirring for 1 h at -78 °C, the reaction was quenched with saturated NaHCO3 (4 mL). The mixture was warmed to room temperature and the phases were separated. The organic phase was stirred with KF on Celite (50 w%, 100 mg) and Na2SO4 (100 mg). The solids were removed by filtration and an aliquot of the filtrate was used for chiral GC analysis, indicating 96% ee.

[0465] Marshall addition of allenylic tin 16 to aldehyde 15

[0466]

[0467] Reagents: BF3·Et2O, 46.5% BF3 (Alfa Aesar): can be used without further purification.

[0468] (1S,2R)-1-((2R,3R)-3-((2R,3S)-3-methoxypentan-2-yl)oxiran-2-yl)-2-methylbut-3-yn-1-ol (5). Aldehyde 15 (7.01 g, 40.8 mmol) and allenylic tin 16 (21.0 g, 61.0 mmol) were dissolved in anhydrous CH2Cl2 (400 mL) in a 1 L flask and purged with an Ar atmosphere. The mixture was cooled to -78 °C and BF3·Et2O (7.53 mL, 61.0 mmol) was added dropwise over 5 min. The reaction was stirred at -78 °C for 1 h. A mixture of MeOH (50 mL) and saturated NaHCO3 (10 mL) was added and the solution was warmed to room temperature. The phases were separated and the aqueous phase was extracted with Et2O (3 × 400 mL). The organic phases were combined, dried over Na2SO4 and concentrated on a rotary evaporator. By flash chromatography, eluting with a gradient of hexanes to 1:3 Et2O / hexanes, the alkyne 17 (6.92 g, 75%) was obtained as a colorless oil with 10:1 dr. Note 1: The minor C16-C17 Marshall diastereomer was removed by chromatography. Note 2: The remaining C18-C19 epoxide diastereomers from the Sharpless epoxidation were separated after the next purification step.

[0469] Alkyne 17: TLC (1:2 EtOAc / hexanes); R f = 0.50 (CAM staining); 11H NMR (500 MHz, CDCl3) δ 3.58 (dd, J = 4.4, 4.4 Hz, 1H), 3.41 (s, 3H), 3.20 (td, J = 6.5, 4.1 Hz, 1H), 3.06 (dd, J = 8.1, 2.3 Hz, 1H), 2.91 (dd, J = 4.5, 2.3 Hz, 1H), 2.81 (qdd, J = 7.0, 4.7, 2.4 Hz, 1H), 2.17 (d, J = 2.6 Hz, 1H), 2.05 (d, J = 4.8 Hz, 1H), 1.67 (ddd, J = 14.2, 7.6, 6.7 Hz, 1H), 1.48 (m, 2H), 1.31 (dd, J = 7.2, 0.7 Hz, 3H), 0.97 (d, J = 7.1 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, CDCl3) δ 84.4, 83.8, 72.3, 71.4, 58.9, 58.3, 38.9, 30.4, 23.9, 17.1, 10.6, 10.1; FTIR (film) ν max 3438, 3310, 2973, 2937, 2879, 1457, 1090 cm -1 ; C 13 H 22 O3 [M + H] + The calculated value of HR-ESI-MS m / z for is 226.1642, the measured value 226.1641; [α] 25 D = +45.4° (c = 1.0, CH2Cl2).

[0470] Hydrostannylation of 17

[0471]

[0472] Reagents: n-Bu3SnH, 97%, containing 0.05% BHT as stabilizer (Acros Organics): can be used without further purification. PdCl2(PPh3)2 (Oakwood Chemical): dried by azeotropic distillation with benzene.

[0473] (1S,2R,E)-1-((2R,3R)-3-((2R,3S)-3-methoxypentan-2-yl)oxiran-2-yl)-2-methyl-4-(tributylstannyl)but-3-en-1-ol (2). PdCl2(PPh3)2 (1.55 g, 2.21 mmol) was added to a solution of alkyne 17 (5.01 g, 22.1 mmol) in anhydrous THF (200 mL) in a 500 mL flask. The mixture was cooled to 0 °C and n-Bu3SnH (17.9 mL, 66.3 mmol) was added dropwise. The mixture was stirred at 0 °C for 45 minutes, at which point the resulting mixture was concentrated to give a black crude oil. The material was extracted into hexanes, filtered through a pad of Celite and eluted with hexanes. The eluate was concentrated on a rotary evaporator and the process was repeated twice until a clear black solution was obtained. Pure vinyl stannane 2 (5.72 g, 50%) was obtained by flash chromatography eluting with a gradient of hexanes to CH2Cl2 to 1:20 Et2O / CH2Cl2 as a 1:5 α:β regioisomer mixture. The desired regioisomer of 95+% purity was obtained by additional flash chromatography eluting with a gradient of hexanes to CH2Cl2 to 1:20 Et2O / CH2Cl2.

[0474] Alternative procedure using the Figueroa catalyst.

[0475]

[0476] Alkyne 17 (5.01 g, 22.1 mmol) was dissolved in benzene (200 mL) in a 500 mL flask and cooled to -78 °C. n-Bu3SnH (17.9 mL, 66.3 mmol) was added dropwise. The Figueroa catalyst (MoI2(CO)2(CNAr Dipp2 )2) (31) was added as a solid. The resulting frozen red mixture was slowly thawed to room temperature with stirring over 4 hours. The mixture was concentrated on a rotary evaporator. Pure vinyl stannane 2 (11.3 g, 55%) was obtained by flash chromatography eluting with a gradient of hexanes to CH2Cl2 to 1:20 Et2O / CH2Cl2 as a 1:10 α:β regioisomer. Note 1: The unwanted epoxide diastereomeric by-products were also removed by chromatography.

[0477] Vinyl stannane 2: TLC (1:10 Et2O / hexanes): R f = 0.28 (CAM stain); 11H NMR (500 MHz, C6D6) δ 6.27 (dd, J = 19.1, 6.8 Hz, 1H), 6.19 (d, J = 19.1 Hz, 1H), 3.45 (m, 1H), 3.23 (s, 3H), 3.16 (m, 1H), 3.07 (dd, J = 8.0, 2.3 Hz, 1H), 2.73 (dd, J = 4.4, 2.3 Hz, 1H), 2.51 (td, J = 6.9, 5.2 Hz, 1H), 1.61 (m, 8H), 1.39 (m, 8H), 1.19 (d, J = 6.9 Hz, 3H), 1.01 (d, J = 7.1 Hz, 3H), 1.00 (d, J = 8.1 Hz, 3H), 0.95 (t, J = 7.4 Hz, 12H), 0.86 (t, J = 7.4 Hz, 3H); 13 13C NMR (125 MHz, C6D6) δ 150.8, 129.0, 83.7, 73.1, 59.3, 57.8, 57.7, 46.1, 39.3, 29.6, 27.7, 23.9, 16.2, 14.0, 10.9, 10.0, 9.8; FTIR (film) ν max 3454, 3310, 2973, 2937, 2890, 1459, 1101, 840 cm -1 ; C 25 H 50 O3Sn[M + H] + The HR-ESI-MS m / z calculated value for is 519.2843, found 519.2839; [α] 25 D = +12.3° (c = 1.0, CH2Cl2).

[0478] Synthesis procedure for nucleus 3. An optimized twelve-step sequence from the published method (1) was developed for the gram-scale preparation of 3, starting from the commercially available 18 (Scheme A2( Figure 9 )) and as follows.

[0479] The alcohol 22 was prepared in hectogram quantities. Each batch of 20 g of alcohol 22 yielded 6 g of 27, and a total of 90 g of 27 has been synthesized to date. Then each batch of 6 g of acid 27 yielded 1.1 g of nucleus 3, and a total of 18 g of 3 has been synthesized to date.

[0480] Oxidation of 18 to aldehyde 19

[0481]

[0482] Reagents: TEMPO, 99% (Oakwood Chemical): Can be used without further purification. KBr (Spectrum Chemical Mfg. Corp.): Can be used without further purification. NaOCl 2M, 10% - 15% available chlorine (Spectrum Chemical Mfg. Corp.): Can be used without further purification.

[0483] 4 - ((tert - Butyldimethylsilyl)oxy)butanal (19). A solution of KBr (6.99 g, 58.7 mmol) in H2O (60 mL) was added to a 3 L flask containing a solution of 18 (100 g, 489 mmol) in CH2Cl2 (1 L), followed by the addition of saturated NaHCO3 (100 mL) and TEMPO (2.29 g, 14.7 mmol). The mixture was cooled to 0 °C and a mixture of NaOCl (2M, 318 mL, 636 mmol) and saturated NaHCO3 (300 mL) was added dropwise in portions via a dropping funnel. The mixture was allowed to warm to room temperature and stirred for 3 h. The mixture was extracted with CH2Cl2 (3 × 250 mL). The combined organic phases were washed with H2O (500 mL), brine (500 mL), dried over Na2SO4, filtered and concentrated on a rotary evaporator. Aldehyde 19 (100 g, 99%) was obtained as a clear oil without further purification. The characterization data matched the literature values.

[0484] Aldehyde 19: TLC (1∶10 EtOAc / hexane): R f = 0.20 (KMnO4); 1 1H NMR (500 MHz, CDCl3) δ 9.79 (t, J = 1.7 Hz, 1H), 3.65 (t, J = 6.0 Hz, 2H), 2.50 (td, J = 7.1, 1.7 Hz, 2H), 1.86 (tt, J = 7.1, 5.9 Hz, 2H), 0.90 (m, 9H), 0.04 (s, 6H); 13 13C NMR (125 MHz, CDCl3) δ 202.5, 62.1, 40.8, 25.9, 25.5, 18.2, -5.4; C 10 H 22 O2Si [M + 1] + The LCMS (ES - API) m / z calculated value for C

[0485] The Brown addition of aldehyde 19

[0486]

[0487] Reagents: s-BuLi, 1.4 M solution in cyclohexane (Sigma-Aldrich): can be used without further purification. (+)-B-Methoxydiisopinocampheylborane, 99% (Sigma-Aldrich): can be used without further purification. BF3·Et2O, 46.5% BF3 (Alfa Aesar): can be used without further purification.

[0488] (8S,9S)-14,14,15,15-Tetramethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadecane-9-ol (20). A solution of s-BuLi (1.4 M, 353 mL, 494 mmol) was added dropwise to a 3 L three-necked flask containing a solution of MEM-protected allyl alcohol (86.7 g, 593 mmol) in anhydrous THF (1 L) cooled to -78 °C over 30 minutes. The resulting solution was stirred at -78 °C for 1 hour, then a solution of (+)-B-methoxydiisopinocampheylborane (156 g, 494 mmol) in anhydrous THF (500 mL) was added. The resulting clear mixture was stirred at -78 °C for 1 hour again. BF3·Et2O (79.3 mL, 642 mmol) was added, followed by a solution of 4-((tert-butyldimethylsilyl)oxy)butanal (19) (100 g, 494 mmol) in anhydrous THF (200 mL). The mixture was stirred at -78 °C for 3 hours. After cooling to 0 °C, saturated NH4Cl (500 mL) was added to the mixture, and it was extracted with CH2Cl2 (3 × 250 mL). The combined organic phases were washed with H2O (500 mL), brine (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. The pure alcohol 20 (134 g, 78%) was obtained with 90.5% dr as determined by chiral HPLC by flash chromatography eluting with a gradient of heptane to 1:1 EtOAc / heptane.

[0489] Alcohol 20: TLC (1:5 EtOAc / hexane): R f = 0.25 (CAM staining); 11H NMR (500 MHz, CDCl3) δ 5.68 (ddd, J = 17.3, 10.5, 8.0 Hz, 1H), 5.32 (m, 2H), 4.79 (d, J = 7.0 Hz, 1H), 4.70 (d, J = 7.0 Hz, 1H), 3.91 (t, J = 7.9 Hz, 1H), 3.83 (ddd, J = 10.9, 5.3, 3.5 Hz, 1H), 3.64 (m, 3H), 3.55 (ddd, J = 5.3, 3.6, 1.9 Hz, 2H), 3.39 (s, 3H), 2.98 (bs, J = 3.5 Hz, 1H), 1.71 (m, 1H), 1.63 (m, 2H), 1.40 (m, 1H), 0.88 (s, 9H), 0.04 (s, 6H); 13 13C NMR (125 MHz, CDCl3) δ 134.9, 120.0, 93.1, 81.6, 73.3, 71.7, 67.5, 63.3, 59.2, 29.5, 29.0, 26.1, 18.5, -5.2; FTIR (film) ν max 3347, 2927, 2856, 1616, 1250, 1021 cm -1 ; C 17 H 36 O5SiNa [M+Na] + HR-ESI-MS m / z calculated for C 25 D H

[0490] Oxidation of ketone 20 to ketone 21

[0491]

[0492] Reagents: TEMPO, 99% (Oakwood Chemical): can be used without further purification; KBr (Spectrum Chemical Mfg. Corp.): can be used without further purification; NaOCl 2 M, 10% - 15% available chlorine (Spectrum Chemical Mfg. Corp.): can be used without further purification.

[0493] (S)-14,14,15,15-Tetramethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadec-9-one (21). A solution of KBr (3.65 g, 30.6 mmol) in H2O (100 mL), saturated NaHCO3 (250 mL), and TEMPO (3.99 g, 25.5 mmol) were added successively to a 2 L flask containing a solution of 20 (89.0 g, 255 mmol) in CH2Cl2 (400 mL). The mixture was cooled to 0 °C and a solution of NaOCl (2 M, 255 mL, 511 mmol) and saturated NaHCO3 (300 mL) was added in portions (20 mL at a time) while maintaining the temperature below 0 °C. The mixture was warmed to room temperature and stirred for 2 h. The phases were separated and the aqueous phase was extracted with CH2Cl2 (2 × 200 mL). The combined organic phases were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. The ketone 21 (88.0 g, 99%) was obtained without further purification.

[0494] Ketone 21: TLC (1:3 EtOAc / hexanes): R f = 0.40 (CAM staining); 1 1H NMR (500 MHz, CDCl3) δ 5.77 (ddd, J = 17.2, 10.4, 6.8 Hz, 1H), 5.46 (dt, J = 17.2, 1.3 Hz, 1H), 5-36 (dt, J = 10.4, 1.0 Hz, 1H), 4.80 (d, J = 7.0 Hz, 1H), 4.74 (d, J = 7.0 Hz, 1H), 4.62 (dt, J = 6.7, 1.2 Hz, 1H), 3.76 (dt, J = 11.0, 4.4 Hz, 1H), 3.67 (m, 1H), 3.59 (t, J = 6.1 Hz, 2H), 3.52 (t, J = 4.6 Hz, 2H), 3.37 (s, 3H), 2.62 (m, 2H), 1.76 (m, 2H), 0.87 (s, 9H), 0.02 (s, 6H); 13 13C NMR (125 MHz, CDCl3) δ 208.2, 132.6, 120.2, 93.7, 82.7, 71.8, 67.5, 62.1, 59.2, 34.8, 26.4, 26.0, 18.4, -5.2; FTIR (film) ν max 2954, 2929, 2857, 1720, 1472, 1256, 1101 cm -1 ; C 17 H 34 O5SiNa [M+Na] +Calculated HR-ESI-MS m / z: 369.2068, found 369.2067; [α] 25 D = +22.0° (c = 1.0, CH2Cl2).

[0495] Stereoselective Grignard addition of ketone 21

[0496]

[0497] Reagent: MeMgBr, 3 M solution in Et2O (Sigma-Aldrich): used without further purification.

[0498] (8S,9R)-9,14,14,15,15-Pentamethyl-8-vinyl-2,5,7,13-tetraoxa-14-silahexadecan-9-ol (22). At -94 °C, MeMgBr (3 M, 462 mL, 1.39 mmol) was added dropwise to a 5 L reaction flask containing a solution of ketone 21 (160 g, 462 mmol) in anhydrous THF (1.5 L). The mixture was stirred at -94 °C for 2 h, warmed to room temperature, and then stirred for an additional 16 h. After recooling to -78 °C, saturated NH4Cl (500 mL) was added dropwise to the mixture. The mixture was diluted with H2O (1 L) and extracted with tert-butyl methyl ether (2 × 500 mL). The combined organic phases were washed with H2O (500 mL) and brine (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. The crude product was filtered through a pad of Celite, eluted with EtOAc, and the eluate was concentrated on a rotary evaporator. Alcohol 22 (155 g, 88%) was obtained in 90% dr without further purification as determined by chiral HPLC. Note 1: The average batch of crude 22 contained <5% of starting material 21. Note 2: The solution of MeMgBr in Et2O gave better yields and selectivities (≤70% yield, ≤90% de) compared to the solution in THF.

[0499] Alcohol 22: TLC (1∶5 EtOAc / hexane): R f = 0 - 30 (CAM staining); 11H NMR (500 MHz, CDCl3) δ 5.73 (ddd, J = 17.2, 10.5, 8.1 Hz, 1H), 5.29 (ddd, J = 14.7, 1.9, 0.8 Hz, 1H), 5.26 (ddd, J = 21.6, 1.9, 0.8 Hz, 1H), 4.75 (d, J = 7.0 Hz, 1H), 4.70 (d, J = 7.0 Hz, 1H), 3.86 (d, J = 8.0 Hz, 1H), 3.82 (dd J = 5.2, 3.7 Hz, 1H), 3.80 (dd, J = 5.5, 3.4 Hz, 1H), 3.61 (m, 3H), 3.53 (dd, J = 3.3, 2 - 3 Hz 1H), 3.52 (dd, J = 3.3, 1.9 Hz, 1H), 3 - 36 (s, 3H), 2.69 (s, 1H), 1.64 (m, 1H), 1.59 (m, 2H), 1.42 (m, 1H), 1.14 (s, 3H), 0.87 (s, 9H), 0.02 (s, 6H); 13 13C NMR (125 MHz, CDCl3) δ 134.3, 120.3, 93.3, 87.5, 73.4, 71.8, 67.5, 63.9, 59.1, 33.9, 26.6, 26.1, 23.6, 18.5, -5.2; FTIR (film) ν max 2954, 2929, 2857, 2359, 1472, 1255, 1097, 1037 cm -1 ; C 18 H 38 O5SiNa [M + Na] + HR - ESI - MS m / z calculated for C 25 D H

[0500] The conversion of 22 to 23

[0501]

[0502] Reagents: CBr4, 99% (TCI Chemicals): Can be used without purification. Imidazole, 99% (Sigma - Aldrich): Can be used without purification.

[0503] p - Anisaldehyde dimethyl acetal, 98% (Acros Organics): Can be used without further purification. i - PrOH, 99% (Fischer Scientific): Used as received without further drying.

[0504] 3-((4R,5S)-2-(4-Methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)propan-1-ol (23). CBr4 (27.7 g, 63.8 mmol) and imidazole (500 mg, 7.34 mmol) were added to a solution of alcohol 22 (20.0 g, 55.2 mmol) in i-PrOH (2 L). The mixture was heated to reflux and stirred at 100 °C overnight, at which point an orange color appeared and NMR analysis indicated complete consumption of the starting material. The mixture was cooled to room temperature and concentrated in vacuo. The resulting brown crude oil was immediately taken up in anhydrous CH2Cl2 (700 mL) and purged with Ar. Anisaldehyde dimethyl acetal (20.0 mL, 117 mmol) was added in one aliquot, and the mixture turned purple after stirring at room temperature for 10 minutes. The reaction was stirred overnight. Saturated NaHCO3 (100 mL) was added, and the mixture was extracted with CH2Cl2 (2 × 500 mL). The organics were combined and concentrated on a rotary evaporator to give a brown oil. Pure alcohol 23 (9.21 g, 60%) was obtained by flash chromatography, eluting with a gradient of hexanes to 1:3 EtOAc / hexanes. Note 1: Batches of 23 were obtained in an unruly mixture of acetal diastereomers, as shown in its structure.

[0505] Alcohol 23: TLC (1:1 EtOAc / hexanes): R f = 0.37 (CAM staining); 1 1H NMR (500 MHz, C6D6) δ 7.55 (d, J = 8.6 Hz, 2H), 7.50 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.7 Hz, 2H), 6.81 (d, J = 8.6 Hz, 2H), 6.16 (s, 1H), 5.91 (s, 1H), 5.79 (m, 1H), 5.71 (m, 1H), 5-30 (dt, J = 3.5, 1.6 Hz, 1H), 5.27 (dt, J = 3.5, 1.6 Hz, 1H), 5.07 (dd, J = 1.7, 1.7 Hz, 1H), 5.05 (dd, J = 1.7, 1.7 Hz, 1H), 4.17 (dt, J = 6.7, 1.2 Hz, 1H), 4.09 (dt, J = 6.7, 1.2 Hz, 1H), 3.42 (m, 2H), 3.38 (m, 2H), 3.27 (s, 3H), 3.26 (s, 3H), 1.73 (m, 2H)1.53 (m, 2H), 1.33 (m, 1H)1.19 (s, 3H)1.17 (s, 3H); 1313C NMR (125 MHz, C6D6) δ 160.8, 160.6, 133.9, 133.8, 133.8, 133.7, 132.8, 131.1, 128.5, 128.3, 117.9, 117.9, 117.8, 117.6, 114.0, 113.9, 107.7, 102.5, 102.2, 96.3, 88.0, 86.5, 86.2, 86.2, 83.6, 82.5, 82.4, 81.8, 63.1, 63.0, 63.0, 58.4, 58.4, 33.8, 32.7, 31.3, 29.9, 28.6, 27.5, 27.3, 27.2, 27.1, 22.9, 22.5, 22.0, 21.8; FTIR (film) ν max 3421, 3080, 2938, 1718, 1614, 1516, 1932, 1303, 1249, 1170, 1032 cm -1 ; C 16 H 22 O4Na [M+Na] + HR-ESI-MS m / z calculated for C15H15O4Na [M+Na]: 301.1410, found 301.1411; [α] 25 D = +14.8° (c = 0.4, CH2Cl2).

[0506] Oxidation of Aldehyde 23 to Aldehyde 24

[0507]

[0508] Reagents: TEMPO, 99% (Oakwood Chemical): used without further purification; KBr (Spectrum Chemical Mfg. Corp.): used without further purification; NaOCl, 2 M, 10% - 15% available chlorine (Spectrum Chemical Mfg. Corp.): used without further purification.

[0509] 3-((4R,5S)-2-(4-Methoxyphenyl)-4-methyl-5-vinyl-1,3-dioxolan-4-yl)propanal (24). A solution of KBr (0.699 g, 5.87 mmol) in H2O (60.0 mL) was added to a 2 L flask containing a solution of alcohol 23 (11.2 g, 40.2 mmol) in CH2Cl2 (750 mL), followed by the addition of NaHCO3 (75 mL) and TEMPO (229 mg, 1.47 mmol). The mixture was cooled to 0 °C, and a mixture of NaOCl (2 M, 32.0 mL, 63.6 mmol) and saturated NaHCO3 (50 mL) was added in portions (20 mL). The mixture was warmed to room temperature. After stirring for 3 h at room temperature, the mixture was extracted with CH2Cl2 (3 × 250 mL). The combined organic phases were washed with H2O (500 mL) and brine (500 mL), dried over Na2SO4, filtered, and concentrated on a rotary evaporator. Aldehyde 24 (11 g, 99%) was used without further purification. Note 1: Aldehyde 24 readily undergoes rearrangement when purified on unbuffered silica gel.

[0510] Aldehyde 24: TLC (1:1 EtOAc / hexane): R f = 0.70 (CAM staining); 1 1H NMR (500 MHz, C6D6) δ 9.39 (s, 1H), 9.29 (s, 1H), 7.47 (d, J = 8.7 Hz, 2H), 7.45 (d, J = 8.7 Hz, 2H), 6.81 (d, J = 4.3 Hz, 2H), 6.79 (d, J = 4.3 Hz, 2H), 6.02 (s, 1H), 5.83 (s, 1H), 5.70 (m, 1H), 5.65 (m, 1H), 5.28 (dt, J = 13.0, 1.6 Hz, 1H), 5.24 (dt, J = 12.8, 1.8 Hz, 1H), 5.04 (dt, J = 4.7, 1.5 Hz, 1H), 5.02 (dt, J = 4.6, 1.4 Hz, 1H), 4.10 (dt, J = 6...

Claims

1. A method for preparing a compound having the following formula: wherein the method comprises reacting a compound having the formula with an acetylating agent in the presence of a strong acid and one or more organic solvents; wherein the acetylating agent is 1,1,1-trimethoxyethane or acetic anhydride, and the strong acid is a sulfonic acid.

2. A method for preparing a compound having the following formula: wherein said method comprises reacting a compound having the formula with an acetylating agent in the presence of a strong acid and one or more organic solvents; wherein said acetylating agent is 1,1,1-trimethoxyethane or acetic anhydride, and said strong acid is p-toluenesulfonic acid.

3. A method for preparing a compound having the following formula: wherein the method comprises reacting a compound having the formula with an acetylating agent in the presence of a strong acid and one or more organic solvents; wherein the acetylating agent is 1,1,1-trimethoxyethane or acetic anhydride, and the strong acid is camphorsulfonic acid.

4. The method according to claim 1, wherein the organic solvent is dichloromethane or chloroform.

5. The method according to claim 2, wherein the organic solvent is dichloromethane or chloroform.

6. The method according to claim 3, wherein the organic solvent is dichloromethane or chloroform.

7. A method for preparing a compound having the following formula: The method comprises the method according to any one of claims 1-6.

8. The method according to claim 7, wherein the compound is at least 95% enantiomerically pure.

9. A compound having the following formula: Wherein the compound is prepared by a method as described in any one of claims 1-6.

10. The compound according to claim 9, wherein the compound is at least 95% enantiomerically pure.

11. Use of the compound according to claim 9 or 10 in a method for preparing a compound having the following formula:

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