MXPA00012534A - Macrocyclic analogs and methods of their use and preparation - Google Patents

Abstract

The invention provides halichondrin analogs having pharmaceutical activity, such as anticancer or antimitotic (mitosis-blocking) activity, and methods of identifying agents that induce a sustained mitotic block in a cell after transient exposure of the cell to the agents.

Description

MACROCYCLIC ANALOGS AND METHODS FOR USE AND PREPARATION BACKGROUND The invention relates to pharmaceutically active macrolides. Halicondrine B is a potent anticancer agent, which was originally isolated from the Halichondria okadai marine sponge, and was subsequently found in Axinella sp. , Phakellia carteri, and Lissondendryx sp. . In 1992 (Aicher, T. D. et al., J. Am. Chem.

Soc. 114: 3162-3164) a total synthesis of Halicondrine B was published. Halicondrine B has shown in vitro the inhibition of tubulin polymerization, the assembly of microtubules, the cross-linking of betas-tubulin, the fixation of GTP and vinblastine to tubulin, and hydrolysis of tubulin-dependent GTP, and has shown in vitro and in vivo anti-cancer properties. SUMMARY OF THE INVENTION The invention provides halicondrine analogues having pharmaceutical activity, such as anticancer or antimitotic activity (blocking of mitosis). These compounds are substantially smaller than the halicondrine B. The invention emphasizes a compound having the formula (I): Formula (I) In formula (I), A is a saturated hydrocarbon skeleton of 1 to 6 carbon atoms, or unsaturated of 2 to 6 carbon atoms, the skeleton being unsubstituted or having between 1 to 13 substituents, preferably between 1 and 10 substituents, for example, at least one substituent selected from cyano, halo, azido, Qi, and oxo. Each Qi is independently selected from ORi, SRi, S02R ?, OS02R ?, NR2, RX, NR2 (CO) Rlf NR2 (CO) (CO) Rl r NR4 (CO) NR2R ?, NR2 (CO) OR ?, (CO) OR ?, 0 (CO) R ?, (CO) NR2R ?, and 0 (CO) NR2R ?. The number of substituents can be, for example, between 1 and 6, 1 and 8, 2 and 5, or 1 and 4. Throughout the description it is understood that the numerical ranges are inclusive. Each of Ri, R2, R4, R5, and Re is independently selected from H, alkyl of 1 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms, hydroxyalkyl of 1 to 6 carbon atoms, aminoalkyl of 1 to 6 carbon atoms, aryl of 6 to 10 carbon atoms, haloaryl of 6 to 10 carbon atoms, (for example, p-fluorophenyl or p-chloro-phenyl), hydroxyaryl of 6 to 10 carbon atoms , C 1 -C 4 -alkoxy-C 6 -aryl alkoxy (e.g., p-methoxyphenyl, 3,4,5-trimethoxyphenyl, p-ethoxyphenol, or 3,5-diethoxyphenyl), aryl 6 10 carbon atoms-alkyl of 1 to 6 carbon atoms (for example, benzyl or phenethyl), alkyl of 1 to 6 carbon atoms-aryl of 6 to 10 carbon atoms, haloaryl of 6 to 10 carbon atoms-alkyl from 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms-haloaryl of 6 to 10 carbon atoms, (alkoxy of 1 to 3 carbon atoms-aryl of 6 carbon atoms) -alkyl of 1 to 3 atoms d e carbon, heterocyclic radical of 2 to 9 carbon atoms, heterocyclic radical of 2 to 9 carbon atoms-alkyl of 1 to 6 carbon atoms, heteroaryl of 2 to 9 carbon atoms, and heteroaryl of 2 to 9 carbon atoms -alkyl of 1 to 6 carbon atoms. There can be more than one Rl, for example, if A is replaced with two different alkoxy groups (ORÍ), such as butoxy and 2-aminoethoxy. Examples of A include 2,3-dihydroxypropyl, 2-hydroxyethyl, 3-hydroxy-4-perfluorobutyl, 2,4,5-trihydroxy-pentyl, 3-amino-2-hydroxypropyl, 1,2-dihydroxyethyl, 2,3 -dihydroxy-4-perfluorobutyl, 3-cyano-2-hydroxypropyl, 2-amino-1-hydroxyethyl, 3-azido-2-hydroxypropyl, 3, 3-difluoro-2,4-dihydroxybutyl, 2, -dihydroxybutyl, 2-hydroxy-2 (p-fluorophenyl) ethyl, -CH 2 (CO) (substituted or unsubstituted aryl), CH 2 (CO) (alkyl or substituted alkyl, such as haloalkyl or hydroxyalkyl) and 3, 3-difluoro-2-hydroxypent -4-enyl. Examples of Qi include -NH (CO) (CO) - (heterocyclic or heteroaryl radical), -0S02- (aryl or substituted aryl), -0 (CO) NH- (aryl or substituted aryl), aminoalkyl, hydroxyalkyl, - NH (C0) (C0) - (aryl or substituted aryl), NH (CO) (alkyl) (heteroaryl or heterocyclic radical), 0 (substituted or unsubstituted alkyl) (substituted or unsubstituted aryl), and -NH (CO) (alkyl) (aryl or substituted aryl). Each of D and D 'is independently selected from R3 and 0R3, wherein R3 is H, alkyl of 1 to 3 carbon atoms, or haloalkyl of 1 to 3 carbon atoms. Examples of D and D 'are methoxy, methyl, ethoxy, and ethyl. In some embodiments, one of D and D 'is H. The value for n is 1, or preferably 0, thereby forming a ring of either six members or five members. This ring may be unsubstituted or substituted, for example, wherein E is R5 and 0R5, and may be a heterocyclic radical or a cycloalkyl, for example, wherein G is S, CH2, NR6, or preferably 0. Each of J and J 'is independently H, alkoxy of 1 to 6 carbon atoms, or alkyl of 1 to 6 carbon atoms; or J and J 'taken together are = CH2, or -O- (straight or branched alkylene or alkylidene of 1 to 5 carbon atoms) -0-, such as exocyclic ethylidene, isopropylidene, methylene, or ethylene. Q is alkyl of 1 to 3 carbon atoms, and preferably it is methyl. T is ethylene or ethenylene, optionally substituted with (C0) 0R7, wherein R7 is H or alkyl of 1 to 6 carbon atoms. Each of U and U 'is independently H, alkoxy of 1 to 6 carbon atoms, or alkyl of 1 to 6 carbon atoms; or U and U 'taken together are = CH2, or -0- (alkylene or alkylidene of 1 to 5 straight or branched carbon atoms) -0-. X is H or alkoxy of 1 to 6 carbon atoms. Each of Y and Y 'is independently H or alkoxy of 1 to 6 carbon atoms; or Y and Y 'taken together are = 0, = CH2, or -0- (straight or branched alkylene or alkylidene of 1 to 5 carbon atoms) -0-. Each of Z and Z 'is independently H or alkoxy of 1 to 6 carbon atoms; or Z e Z 'taken together are = 0, = CH2, or -0- (straight or branched alkylene or alkylidene of 1 to 5 carbon atoms) -0-. The invention highlights compounds of sufficient stability to be suitable for pharmaceutical development. The invention also emphasizes pharmaceutically acceptable salts of the described compounds, novel synthetic intermediates described, pharmaceutical compositions containing one or more of the described compounds, methods for making the disclosed compounds or intermediates, and methods for using the compounds or the compositions described. Methods of use include methods for reversibly or irreversibly inhibiting mitosis in a cell, and for inhibiting cancer or tumor growth in vitro, in vivo, or in a patient. The invention also emphasizes methods for identifying an antimitotic or anticancer agent, such as a reversible agent or, preferably, an irreversible agent. Brief Description of the Figures Figure 1 is a graph showing the percentage of cells that have completed mitosis and returned to the Gi stage as a function of the concentration of compound B1939 in a mitotic block assay. The minimum concentration required to complete the mitotic block at time 0 is 10 nM. The minimum concentration required to complete the mitotic block at time 10 (after the washout) is also 10 nM. Therefore, the reversibility ratio is 1 for B1939. On this graph a curve showing the percentage of viable cells in 5 days is superimposed, as a function of the concentration of compound B1939. Viability falls to very low levels at the same concentration as the 10-hour mitotic block. Figure 2 is a graph showing the percentage of cells that have completed mitosis and returned to the Gi stage as a function of the concentration of compound B2042 in a mitotic block reversibility assay. The minimum concentration required to complete the mitotic block at time 0 is 3 nM. The minimum concentration required to complete the mitotic block at time 10 is 100 nM. Therefore, the reversibility ratio is 33. A curve showing the percentage of viable cells in 5 days is superimposed over this graph, as a function of the concentration of compound B2042. Viability falls to very low levels at the same concentration as the 10-hour mitotic block. Figure 3 is a graph showing the average volume of the tumor in microliters, as a function of time (days) in a growth inhibition assay of the melanoma xenograft LOX. This illustrates the antitumor activity of a compound of the formula (I), the compound B1939. Paclitaxel and a vehicle control were used. Figure 4 is a graph showing the average body weight per mouse, as a function of time (days) in the test described in Figure 3. Figure 5 is a graph showing the average tumor volume in microliters, as a function of time (days) in a human colon cancer xenograft growth inhibition assay COLÓ 205, which shows the antitumor activities of vinblastine and vincristine. Detailed Description of the Invention A. Definitions B. Halicondrine Analogs C. Synthesis of Halicondrine Analogs D. Pharmacological Activity E. Uses A. Definitions The following terms are described in part a below and for their use herein. The hydrocarbon skeletons contain carbon and hydrogen atoms, and can be linear, branched, or cyclic. Unsaturated hydrocarbons include one, two, three or more double bonds C-C (sp2) or triple bonds C-C (sp) Examples of unsaturated hydrocarbon radicals include ethynyl, 2-propynyl, 1-propenyl, 2-butenyl, 1,3-butadienyl, 2-pentenyl, vinyl (ethenyl), allyl, and isopropenyl. Examples of bivalent unsaturated hydrocarbon radicals include alkenylenes and alkylidenes such as methylidin, ethylidene, ethylidin, vinylidene, and isopropylidene. In general, the compounds of the invention have hydrocarbon backbones ("A" in the formula (I)) which are substituted, for example, with hydroxy, amino, cyano, azido, heteroaryl, aryl, and other fractions described herein . The hydrocarbon backbones can have two geminal hydrogen atoms replaced with oxo, a divalent carbonyl oxygen atom (= 0), or a ring-forming substituent, such as -O- (straight or branched alkylene or alkylidene) -0- to form an acetal or ketal. The alkyl of 1 to 6 carbon atoms includes linear, branched, and cyclic hydrocarbons, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, sec-pentyl, neopentyl, tert-pentyl, cyclopentyl, hexyl, isohexyl, sec-hexyl, cyclohexyl, 2-methylpentyl, tert-hexyl, 2,3-dimethylbuyl, 3, 3-dimethylbutyl, 1,3-dimethylbutyl, and 2,3-dimethylbutyl -2-ilo. Alkoxy (-0R), alkylthio (-SR), and other alkyl-derived moieties (substituted, unsaturated, or bivalent) are analogous to alkyl groups R. Alkyl groups, and alkyl-derived groups such as alkoxy groups haloalkyl, hydroxyalkyl, alkenyl, alkylidene, and alkylene, may be 2 to 6 carbon atoms, 3 to 6 carbon atoms, 1 to 3 carbon atoms, or 2 to 4 carbon atoms. The alkyls substituted with halo, hydroxy, amino, cyano, azido, etc., may have 1, 2, 3, 4, 5 or more substituents, which are independently selected (they may or may not be the same) and may or may not be in the same carbon atom. For example, haloalkyls are alkyl groups with at least one substituent selected from fluoro, chloro, bromo, and iodo. The haloalkyls may have two or more halo substituents, which may or may not be the same halogen, and may or may not be on the same carbon atom. Examples include chloromethyl, periodomethyl, 3,3-dichloropropyl, 1,3-difluorobutyl, and l-bromo-2-chloropropyl. Heterocyclic and heteroaryl radicals include furyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathienyl, 2H-pyrrolyl, pyrrolyl, imidazolyl (for example, 1-, 2- or 4-imidazolyl), pyrazolyl, isothiazolyl, isoxazolyl, pyridyl ( for example, 1-, 2-, or 3-pyridyl), pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl (for example, 1-, 2-, or 3-indolyl), indazolyl, purinyl, 4H-quinolixinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinolinyl, pteridinyl, pyrrolinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, indolinyl, isoindolinyl, and morpholinyl. The heterocyclic and heteroaryl radicals can be linked to the rest of the molecule at any position along the ring. The heterocyclic and heteroaryl radicals can be from 2 to 9 carbon atoms, or smaller, such as from 3 to 6 carbon atoms, from 2 to 5 carbon atoms, or from 3 to 7 carbon atoms. Aryl groups include phenyl, benzyl, naphthyl, tolyl, mesityl, xylyl, and cumenyl. It is understood that "heterocyclic radical", "aryl", and "heteroaryl" include those having 1, 2, 3, 4, or more substituents independently selected from lower alkyl, lower alkoxy, amino, halo, cyano, nitro, azido, and hydroxyl. The heterocyclic, heteroaryl, and aryl radicals can also be bivalent substituents of the hydrocarbon backbone "A" in the formula (I). B. Halicondrine Analogs Referring to formula (I) in the Summary section, embodiments of the invention include compounds wherein n is 0; wherein each of D and D 'is independently selected from R3, alkoxy of 1 to 3 carbon atoms, and haloalkyloxy of 1 to 3 carbon atoms; wherein Rs is selected from H, alkyl of 2 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms, hydroxyalkyl of 1 to 6 carbon atoms, aminoalkyl of 1 to 6 carbon atoms, aryl of 6 to 10 carbon atoms, haloaryl of 6 to 10 carbon atoms, hydroxyaryl of 6 to 10 carbon atoms, alkoxy of 1 to 3 carbon atoms-aryl of 6 carbon atoms, aryl of 6 to 10 carbon atoms-alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms-aryl of 6 to 10 carbon atoms, haloaryl of 6 to 10 carbon atoms-alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms carbon-haloaryl of 6 to 10 carbon atoms, (C 1 -C 3 alkoxy-C 6 aryl) -alkyl of 1 to 3 carbon atoms, heterocyclic radical of 2 to 9 carbon atoms, heterocyclic radical from 2 to 9 carbon atoms-alkyl of 1 to 6 carbon atoms, heteroaryl of 2 to 9 carbon atoms, and heteroaryl of 2 to 9 atoms carbon-alkyl of 1 to 6 carbon atoms; and combinations thereof. Another embodiment includes compounds having one or more of the following characteristics: (a) when A is a saturated hydrocarbon skeleton of 1 to 6 carbon atoms or unsaturated of 2 to 6 carbon atoms, the skeleton having at least one substituent selected from cyano, halo, azido, Qi, and oxo; (b) each Qi is independently selected from ORi, SRi, S02R ?, OS02R ?, NR2, RX, NR2 (CO) R ?, NR2 (CO) R ?, and 0 (CO) NR2R ?; (c) Z and Z 'taken together are = 0 or = CH2; (d) where each Qi is independently selected from ORi, SRX, S02R !, 0S02R ?, NH (CO) (CO) R ?, and 0 (C0) NHR ?; (e) each Ri is independently selected from alkyl of 1 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms, aryl of 6 carbon atoms, haloaryl of 6 carbon atoms, alkoxy of 1 to 3 atoms of carbon-aryl of 6 carbon atoms, aryl of 6 carbon atoms-alkyl of 1 to 3 carbon atoms, alkyl of 1 to 3 carbon atoms-aryl of 6 carbon atoms, haloaryl of 6 carbon atoms- alkyl of 1 to 3 carbon atoms, alkyl of 1 to 3 carbon atoms-haloaryl of 6 carbon atoms, (alkoxy of 1 to 3 carbon atoms-aryl of 6 carbon atoms) -alkyl of 1 to 3 atoms of carbon, heterocyclic radical of 2 to 9 carbon atoms, heteroaryl of 2 to 9 carbon atoms, and heteroaryl of 2 to 9 carbon atoms-alkyl of 1 to 6 carbon atoms; (f) one of D and D 'is methyl or methoxy and the other is H; (g) n is 0; (h) G is O; (i) J and J 'taken together are = CH2; (j) Q is methyl; (k) T is ethylene; (1) U and U 'taken together are = CH2; (m) X is H; (n) each of Y and Y 'is H; and (o) Z and Z 'taken together are = 0. Examples of combinations are the combination of (h) - (m), the combination of (a) and (b), the combination of (f) and (h), and the combination of (h) and where one of D and D 'is methyl and the other is H. The particularly preferred compounds are B1793 and B1939. 'Another embodiment includes compounds wherein Qi is independently selected from 0R ?, SRi, S02R ?, 0S02R ?; and each Ri is independently selected from alkyl of 1 to 6 carbon atoms, haloalkyl of 1 to 6 carbon atoms, aryl of 6 carbon atoms, haloaryl of 6 carbon atoms, alkoxy of 1 to 3 carbon atoms- aryl of 6 carbon atoms, aryl of 6 carbon atoms-alkyl of 1 to 3 carbon atoms, alkyl of 1 to 3 carbon atoms-aryl of 6 carbon atoms, haloaryl of 6 carbon atoms-alkyl of 1 to 3 carbon atoms, alkyl of 1 to 3 carbon-haloaryl atoms of 6 carbon atoms, and (C 1 -C 3 alkoxy-C 6 aryl) -alkyl of 1 to 3 carbon atoms. Other embodiments include compounds wherein: one of D and D 'is alkyl or alkoxy, wherein n is 1; (f) as above, where n is 1; E is alkoxy, wherein n is 1; n is 0, wherein one of D and D 'is hydroxy and the other is H; and (f) as above, wherein n is 1 and E is methyl. The invention also highlights compounds wherein: (1) A has at least one substituent selected from hydroxyl, amino, azido, halo, and oxo; (2) A is a saturated hydrocarbon skeleton having at least one substituent selected from hydroxyl, amino, and azido (e.g., B1793, B1939, B2042, B1794, and B1922); (3) A has at least two substituents independently selected from hydroxyl, amino, and azido (e.g., B2090 and B2136); (4) A has at least two substituents independently selected from hydroxyl and amino (eg, B2042 and B2090); (5) A has at least one hydroxyl substituent, and at least one amino substituent (eg, B1939 and B2136); (6) A has at least two hydroxyl substituents (e.g., B1793 and B1794); (7) A is a hydrocarbon skeleton of 2 to 4 carbon atoms, which is substituted (eg, B2004, B2037, B1920, B2039, B2070, B2090, and B2043); (8) A is a hydrocarbon skeleton of 3 carbon atoms, which is substituted (for example, B1793, B1920, B1984, B1988, B1939, B1940, B2014); (9) A has an (S) -hydroxyl alpha to the carbon atom linking A to the ring containing G (eg, B1793, B1939 or B1920) or an (R) -hydroxyl (eg, B2102, B2013, B2042 ); and (10) A is a saturated hydrocarbon skeleton of 1 to 6 carbon atoms, having at least one substituent selected from hydroxyl and cyano (eg, B2013, B2037, B2102, B2086, and B2091). With (S) -hydroxyl it is meant that the configuration of the carbon atom that 'has the hydroxyl group is (S). The embodiments of the invention also include compounds having at least two substituents on the carbon atoms (1) alpha and gamma, (2) beta and gamma, or preferably (3) alpha and beta to the carbon atom linking A to the ring containing G. The alpha, beta, and gamma carbon atoms may have a (R) or (S) configuration. The invention further provides preferred compounds having the formula (1) -A, which is shown below, wherein the substituents are identical to those defined above.

Formula 1-A The invention also highlights the following monosaccharide intermediate having the formula (II): Formula (II) wherein R is methyl or methoxy, and each of Pl, P2, and P3 is independently selected from H and primary alcohol protecting groups. Preferably, the side chain of God is below the page plane and 0P2 is on the plane of the page. Primary alcohol protecting groups include esters, ethers, silyl ethers, alkyl ethers, and alkoxyalkyl ethers. Examples of esters include formats, acetates, carbonates, and sulfonates. Specific examples include format, benzoyl formate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4, 4- (ethylendithio) -pentanoate, pivaloate, crotonate, 4-methoxy- crotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate, carbonates such as methyl, 4-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2- (trimethylsilyl) ethyl, 2- (phenylsulfonyl) -ethyl, vinyl, allyl, and p-nitrobenzyl. Examples of silyl ethers include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, allyl, and ethers or allyloxycarbonyl derivatives. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy) methyl, benzyloxymethyl, beta- (rimethylsilyl) ethoxymethyl, and tetrahydropyranyl ethers. Examples of benzyl ethers include p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-picolyl. Preferably, each of Pl and P2 are TBS and P3 is MPM (see alcohol 19 below). In one aspect, formula (II) can be modified such that the hydroxyethyl side chain can also be a protected hydroxyl, -CH2CH20-P4, wherein P4 is independently selected from the values for Pl. A related intermediate is alcohol 17, wherein the hydroxyethyl side chain is a hydroxymethyl side chain. Similarly, a hydroxypropyl side chain, or corresponding aminoalkyl side chain, can be prepared. Pl and P2, taken together, can be a diol protecting group, such as cyclic acetals and ketals (methylene, ethylidene, benzylidene, isopropylidene, cyclohexylidene, and cyclopentylidene), silylene derivatives such as di-t-butylsilylene and 1, 1 , 3, 3-tetra-isopropyl disiloxanilidene, cyclic carbonates, and cyclic boronates. Methods for adding and removing those hydroxyl protecting groups, and additional protecting groups, are well known in the art and are available, for example, in P.J. Kocienski, Protecting Groups, Thieme, 1994, and in T. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2nd edition, John Wiley & amp; amp;; Sons, 1992. The following section provides the representative syntheses of the intermediates of the formula (II) and the halicondrine analogs of the formula (I). C. Synthesis of Halicondrine Analogs The following is an overview, followed by synthetic schemes 1-16, and many detailed protocols. The compounds of the general formula 4 can be prepared by the route delineated in Scheme 1. The key fragment F-2 exemplified by the vinyl iodide compound X2 can be prepared according to the procedure of Kishi, et al. (Total Synthesis of halicondrine B and norhalicondrine B. Aicher, TD, Buszek, KR, Fang, FG, Forsyth, CJ, Jung, SH, Kishi, Y., Matelich, MC, Scola, M., Spero, DM, Yoon, SKJ Am. Chem. Soc. 1992, 114, 3162-4).

Vinyl Iodide X2 The key fragment F-3 can be obtained by reduction by DIBALH of the corresponding methyl ester, XF3, prepared according to the procedure of Stamos, et al. (Scheme 2). [Synthetic studies on halichondrins: a practical synthesis of the C.1-C.13 segmeht. Stamos, D.P .; Kishi, Y. Tetrahedron Lett. 1996, 31, 8643-8646]. The synthesis of key fragment F-1 exemplified by compound 20 can be synthesized as described in Scheme 3 or Scheme 4. Using B1793 as a representative example, the coupling of the three key fragments proceeded as outlined in the Scheme 5: Nozaki-Hiy-ama-Kishi coupling of fragments 20 and X2, followed by the formation of intramolecular Williamson's ether, provided tetrahydropyran B2318. The modification of the protecting group as described in Scheme 5, or alternatively in Scheme 6, provided the primary iodide B2313. The halogen-metal exchange reaction and the coupling with the key fragment F-3 provided a mixture of diastereomeric alcohols B2308. The manipulation and oxidation of the additional protective group, followed by an intramolecular Nozaki-Hiyama-Kishi reaction, provided an intermediate, which, when oxidized and treated with TBAF, experienced the closure of the intramolecular hetero-Michael ring. Acetal formation mediated by PPTs provided B1793. As exemplified in Scheme 7, aryl groups may be incorporated within the C32 side chain (e.g., B2043). Intermediate B2318 was deprotected and the resulting diol was oxidatively dissociated to the corresponding aldehyde. Treatment with a Grignard agent (e.g., pF-PhMgBr), separation of the resulting diastereomers and silylation yielded 204, which was converted to the final product in a manner similar to that described in Scheme 6. prepare ether analogs from B1793, by treatment with an appropriate alkylating agent (e.g., Scheme 8). Similarly, sulfonates, esters, carbamates, etc. can be prepared from B1793 by treatment with an activated carbonyl component. Oxidative dissociation of the diol and reduction or selective oxidation of the hydroxyl group can provide derivatives such as B2037 and B1934, respectively. Alternatively, one or more hydroxyl groups can be converted to the corresponding amino groups, with subsequent coupling with an activated carbonyl component (Scheme 9). The displacement of the sulfonyl intermediate (eg, B1920) by carbon or heteroatom nucleophiles can also be performed rapidly (Scheme 10). C31 methyl analogs can be prepared as outlined in Scheme 11. Indium-mediated coupling of an allyl bromide ester, with 2,3-di- (3-isopropylidine) -D-glyceraldehyde provided lactone 103. hetero-Michael addition, lactone reduction, Wittig coupling and intramolecular Michael addition, provided tetrahydrofuran 107. Pummerer reconfiguration, protective group adjustment and reduction by DIBALH provided key fragment 1 (e.g. , 114), which was converted to the final compound in a manner analogous to that described in Scheme 6. Fluoro atoms can be introduced as described in Schemes 12-14. Starting with the appropriate tetrahydrofuran intermediate, the fluorinated key fragment 1 was obtained, and the final compound was taken in a manner analogous to that illustrated in Scheme 6. Similarly, triol derivatives can be prepared from the intermediary of tetrahydrofuran. For example, as outlined in Scheme 15, the addition of allyltributylstannane to aldehyde X32 yielded homoallyl alcohol 33, which was brought to the final compound in a manner similar to that described in Scheme 6. These triols can be further modified as described. exemplifies in Scheme 6. The 1,3-diol derivatives can be prepared from the intermediates previously described. For example, B2086 can be oxidatively dissociated, and reduced to provide the 1,3-diol B2091 (Scheme 16).

?? Summary of Key Fragment F-3: XF-3 F-3 Fragment Key F-3. DIBALH (1M in toluene, 3.86 milliliters) was added to a solution of XF-3 (1.46 grams, 1.93 mmol) in toluene (37 milliliters) at -78 ° C.

After stirring for 10 minutes, the reaction was cooled by careful addition of MeOH (0.46 milliliters) and H20 (0.21 milliliters), was warmed to room temperature and stirred for 15 minutes. The white suspension was filtered through Celite with CH2C12 / Et20 1: 1. The filtrate was concentrated and purified by column chromatography (10 percent EtOAc - hexanes) to give the key fragment F-3 (1.34 grams, 96 percent) as an oil. Synthesis of B1793: L-arabinose Triol 1. A solution of TBDPSCI (444 milliliters, 1.7 moles) in DMF (0.5 liters) was added in three portions, to a suspension of L-arabinose (250.0 grams, 1.66 moles), imidazole (231.4 grams, 3.40 moles) and DMF (2.5 liters). The addition of each serving took 1.5 hours with 30 minutes, and an interval of 15 hours separating the second and third portions, respectively. The resulting solution was stirred for 3 hours, concentrated and purified by flash chromatography (5 percent to 33 percent EtOAc-hexanes) to provide triol 1 (394 grams, 61 percent).

Triacetate 2. Acetic anhydride (6.06 moles) was added over 1.5 hours to triol 1 (1.01 moles) in pyridine. (1.0 liter) at 15 ° C. The solution was stirred for 1 hour, concentrated and purified by flash chromatography (15 percent to 25 percent EtOAc-hexanes) to give triacetate 2 (518 grams, 97 percent).

Diacetates 3. Allytrimethylsilane (1.11 mol) was added followed by BF3-OEt2 (1.11 mmol), for 1.5 hours, to triacetate 2 (164 grams, 0.32 mol) in toluene (1.5 liters) at 0 ° C. The orange solution was stirred for 1 hour at 0 ° C, and for 2 hours at room temperature. The mixture was poured slowly into saturated aqueous NaHC03 (1.7 liters) at 0 ° C, and stirred for 30 minutes. The separated aqueous layer was extracted with EtOAc (3 x 600 milliliters), and the combined organic layers were dried over Na 2 SO 4, concentrated and purified by flash chromatography (5 percent to 10 percent EtOAc-hexanes) to provide a mixture. of diacetates 3 (108 grams, 69 percent).

Diol 4a. K2C03 (72 mmol) was added to the diacetates 3 (108 grams, 218 mol) in MeOH '(0.5 liters) at room temperature. The suspension was stirred for 2.5 hours and then concentrated. The orange residue was suspended in saturated aqueous NH4C1 (150 milliliters), extracted with EtOAc (3 x 150 milliliters), and the combined organic layers were dried over Na2SO4, concentrated and purified by flash chromatography (15% EtOAc a). 50 percent - hexanes) to provide alpha isomer 4a (33.86 grams, 37 percent), and beta-isomer 4b (58 grams, 63 percent). 4a 5 Alcohol 5. Imidazole (16.75 grams, 246 mmoles) and TBSC1 (16.08 grams, 107 mmoles) were added to a solution of diol 4a (33.86 grams, 82 mmoles) in CH2C12 (250 milliliters) at 0 ° C. After 18 hours at 0 ° C and 5 hours at room temperature, the reaction mixture was diluted with saturated aqueous NaHCO3 (250 milliliters), stirred for 30 minutes and the layers allowed to separate. The aqueous layer was extracted with EtOAc (3 x 250 milliliters), and the combined organic layers were dried over Na 2 SO 4, concentrated and purified by flash chromatography (2 percent to 50 percent EtOAc-hexanes) to provide the alcohol. (36.0 grams, 83 percent). 6 Methyl ether 6. Iodomethane (16.5 milliliters, 265 mmol) and NaH (60 percent in mineral oil, 5.28 grams, 132 mmol) were added to a solution of the alcohol (34.93 grams, 66 mmol), THF (320 milliliters) and DMF (80 milliliters) at 0 ° C. After 19 hours at 0 ° C the reaction was quenched with saturated aqueous NHC1 and saturated aqueous Na2S203.

The resulting mixture was stirred for 20 minutes and the layers allowed to separate. The aqueous phase was extracted with EtOAc (3 x 200 milliliters), and the combined organic layers were dried over Na 2 SO 4, concentrated and purified by flash chromatography (3 percent EtOAc-hexanes) to give the methyl ether. (34.23 grams, 96 percent).

MeQ OTBS HC | MeQ OH ^ \ Q ../OTBDPS "" ^ \ 0 ... / OH Diol 7. HCl (37 percent aqueous solution, 12.75 milliliters, 153 mmol) was added to a solution of methyl ether 6 (32.93 grams, 61 mmol) in MeOH (110 milliliters) at room temperature. After 17 hours, NaHCO3 (17 grams) was added to the reaction mixture. The mixture was stirred for 30 minutes, concentrated, suspended in EtOAc and filtered. The filtrate was concentrated and purified by flash chromatography (50 percent HertOAc-hexanes to EtOAc) to give diol 7 (10.0 grams, 87 percent). 8 Alcohol 8. A solution of pivaloyl chloride (8.4 milliliters, 67 mmol) in pyridine (50 milliliters), for 1.5 hours, was added to a solution of diol 7 (12.24 grams, 65 mmol) in pyridine (100 milliliters) at 0 ° C.

After 1 hour at 0 ° C and 18 hours at room temperature, the mixture is diluted with saturated aqueous NHC1 and extracted with EtOAc (3 x 800 milliliters). The combined organic layers were dried over Na 2 SO 4, concentrated and purified by flash chromatography (50 percent EtOAc-hexanes) to provide alcohol 8 (16.9 grams, 96 percent). 9 Olefin 9. Benzyl bromide (62 milliliters, 521 mmol) and BuNHS04 (10.6 grams, 31 mmol) were added to a solution of alcohol 8 (16.9 grams, 62 mmol) in CH2C12 (100 milliliters) at 0 ° C. A solution of NaOH (9.95 grams, 248 mmol) in H20 (10 milliliters) was added to the reaction mixture, for 15 minutes. After 30 minutes at 0 ° C and 18 hours at room temperature, the reaction mixture was diluted with saturated aqueous NH4C1, and extracted with CH2C12 (3 x 100 milliliters). The combined organic layers were dried over Na 2 SO, concentrated and purified by flash chromatography (hexanes to 30 percent EtOAc-hexanes) to provide olefin 9 (22.1 grams, 98 percent).

Diol 10. Os04 (0.1 M solution in toluene, 7.3 milliliters, 0.73 mmol) and a solution of olefin 9 (24.9 grams, 69 mmol) in t-BuOH (165 milliliters) were added to a solution of K2C03 (31.2 grams, 161 mmole), K3Fe (CN) 6 (74.4 grams, 161 mmol), (DHQ) 2PYR (1.33 grams, 1.50 mmol), H20 (500 milliliters) and t-BuOH (330 milliliters) at 0 ° C. After 3 hours at 0 ° C, Na2S205-5 H20 (37.3 grams, 150 mmol) was added. The reaction mixture was warmed to room temperature, stirred for 1 hour, and extracted with EtOAc • (3 x 300 milliliters). The combined organic layers were dried over NaSO4, concentrated and purified by flash chromatography (5 percent isopropanol-CH2Cl2) to give diol 10 (17.98 grams, 75 percent). 11 Silyl Ether 11. Imidazole (21 grams, 308 mmol) and TBSC1 (26.5 grams, 176 mmol) were added to a solution of diol 10 (17.4 grams, 44 mmol) in DMF (90 milliliters) at room temperature. After 18 hours, the reaction mixture was diluted with saturated aqueous NaHCO3 (250 milliliters), stirred for 1 hour and extracted with CH2C12. (3 x 100 milliliters). The combined organic layers were dried over Na 2 SO 4, concentrated and purified by flash chromatography (5 percent EtOAc-hexanes) to provide silyl ether 11 (25.7 grams, 94 percent). 11 12 Alcohol 12. A mixture of silyl ether 11 was stirred. (21.2 grams, 33.8 mmol), Pd (OH) 2 (20 percent, 4.7 grams, 33.8 mmol) and EtOAc (200 milliliters) at room temperature, under an atmosphere of H2, for 3 hours. The mixture was filtered through Celite, concentrated and purified by flash chromatography (10 percent to 20 percent EtOAc-hexanes) to yield alcohol 12 (17.4 grams, 96 percent). 12 13 Olefin 13. 4-Methylmorpholine N-oxide (7.66 grams, 65 mmol) and TPAP (1.15 grams, 3.26 mmol) were added in four portions, over 20 minutes, to a solution of alcohol 12 (17.4 grams, 32.5 mmol) in CH2C12 (145 milliliters) at 0 ° C. After 20 minutes, the reaction mixture was diluted with Et20 (50 milli-liters) and saturated aqueous Na2S20s (50 milliliters), and filtered through Celite. The organic layer was separated, washed sequentially with CuSO4-saturated aqueous (1: 1) saline and saline, dried over Na2SO4, filtered through Celite, and concentrated to yield the desired crude ketone. The Tebbe reagent was prepared by stirring bis (cyclopentadienyl) titanium (11.36 grams, 45.6 mmol) and Me3Al (2.0 M in toluene, 45.6 milliliters, 91.2 mmol) for 4 days at room temperature. This material was cooled to -25 ° C and a crude ketone solution in THF (150 milliliters) was added. The reaction mixture was heated to 0 ° C, stirred for 30 minutes, cooled by the slow addition of 0.1 N NaOH (3.5 milliliters), and then stirred for an additional 20 minutes at room temperature. The mixture was diluted with Et20, filtered through Celite and concentrated. The residue was dissolved in CH2C12, filtered through basic A1203, concentrated and purified by flash chromatography (5 percent EtOAc-hexanes) to give olefin 13 (12.8 grams, 74 percent for two steps). 13 14 Alcohol 14. 9-BBN (0.5 M in THF, 165 milliliters, 83 mmol) was added to a solution of olefin 13 (12.78 grams, 24 mmol) in THF (280 milliliters) at 0 ° C. After stirring for 5 hours at room temperature, the reaction mixture was again cooled to 0 ° C, at which time H20 (200 milliliters), THF (100 milliliters) and NaB03-4 H20 (75 grams) were added. . The mixture was warmed to room temperature, stirred for 16 hours and then concentrated. The aqueous residue was extracted with EtOAc (4 x 300 milliliters), and the combined organic layers were dried over Na 2 SO 4. Concentration and purification by flash chromatography (20 percent to 35 percent EtOAc-hexanes) yielded alcohol 14 (12.05 grams, 91 percent). 14 15 Alcohol 15. DMSO (9 milliliters, 127 mmol) was added to a solution of oxalyl chloride (5.6 milliliters, 64 mmol) in CH2C12 (350 milliliters) at -78 ° C. After stirring for 15 minutes, a solution of alcohol 14 (11.7 grams, 0.021 mmol) in CH2C12 (50 milliliters) was added and stirring was continued for 1 hour, after which Et3N (26.7 milliliters, 192 mmol) was added. . The reaction mixture was warmed to 0 ° C, stirred for 15 minutes, diluted with saturated aqueous NHC1, and extracted with CH2C12 (3 x 200 milliliters). The combined organic layers were dried over Na 2 SO, and concentrated to provide the desired crude aldehyde. This material was dissolved in CH2C12 (200 milliliters) and treated with Et3N (20 milliliters) at room temperature. After stirring overnight, the reaction mixture was diluted with saturated aqueous NH4C1, and extracted with CH2C12 (3 x 200 milliliters). The combined organic layers were dried over Na 2 SO 4, concentrated and filtered through a short SiO 2 column (20 percent EtOAc-hexanes) to provide the crude epimerized product. The aldehyde was dissolved in Et20-EtOH (1: 1, 100 milliliters), cooled to 0 ° C and treated with sodium borohydride (1.21 grams, 32 mmol). The mixture was stirred for 20 minutes, carefully diluted with saturated aqueous NH4C1, stirred for 30 minutes at room temperature, and extracted with CH2C12 (3 x 150 milliliters). The combined extracts were dried over Na 2 SO 4, concentrated and purified by flash chromatography (20 percent EtOAc-hexanes) to provide alcohol 15 (9.95 grams, 85 percent for three steps). 16 MPM-ether 16. BF3-OEt2 (0.1 M in CH2C12 was added, 1.8 milliliters, 0.18 mmol) to a solution of alcohol 15 (9.87 grams, 18 mmol), MPM-trichloroimidate (4.9 milliliters, 27 mmol) and CH2C12 (175 milliliters) at 0 ° C. After 40 minutes, a second portion of BF3-OEt2 (0.1M in CH2C12, 0.9 milliliters, 0.09 mmol) was added to the reaction mixture. After 20 minutes, the reaction was quenched with saturated aqueous NHC1, stirred for 1 hour at room temperature, and diluted with Et20 (600 milliliters). The organic layer was separated and the aqueous layer was extracted with Et20 (150 milliliters). The combined organic extracts were washed sequentially with 0.1 N aqueous NaOH, NaHCO3, saline, dried over Na2SO4, concentrated and purified by flash chromatography (20 percent EtOAc-hexanes) to give MPM-ether 16 (10.20. grams, 85 percent). 16 17 Alcohol 17. LAH (1 M in THF, 22.5 milliliters, 22.5 mmol) was added to a solution of MPM-ether 16 (10.05 grams, 15 mmol) in Et20 (1.0 liter) at 0 ° C. After 30 minutes the reaction was cautiously cooled with H20 (1.3 milliliters), and 1 N of aqueous NaOH (1.3 milliliters). After stirring for 1 hour at room temperature, the suspension was filtered through Celite, concentrated and purified by flash chromatography (20 percent EtOAc-hexanes) to provide alcohol 17 (B.18 grams, 93%). hundred) . 17 18 Olefin 18. DMSO (5.8 milliliters, 82.4 mmol) was added to a solution of oxalyl chloride (3.6 milliliters, 41.2 mmol) in CH2C12 (100 milliliters) at -78 ° C. After 15 minutes, a solution of alcohol 17 (7.94 grams, 13. 5 mmoles) in CH2C12 (35 milliliters) to the reaction mixture. After stirring for 1 hour, Et3N was added (17 milliliters, 122 mmol), the mixture was warmed to 0 ° C, stirred for 20 minutes, diluted with saturated aqueous NH4C1, and then extracted with CH2C12 (3 x 100 milliliters). The combined organic extracts were dried over Na 2 SO 4, concentrated and filtered through a short SiO 2 column (20 percent EtOAc-hexanes) to provide the desired crude aldehyde. N-BuLi (1.6 M, 20 milliliters, 30 mmol) was added dropwise to a solution of CH3PPh3Br (10.1 grams, 30 mmol) in THF (350 milliliters) and DMSO (100 milliliters) at 0 ° C. After 1 hour, a solution of the crude aldehyde in THF (50 milliliters) was added. The reaction mixture was warmed to room temperature and stirred for 3 hours. Saturated aqueous NH4C1 was added, and the mixture was extracted with EtOAc (3 x 500 milliliters). The combined extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (7 percent EtOAc-hexanes) to provide the olefin 18 (5.57 grams, 71 percent yield for two steps). 18 19 Alcohol 19. 9-BBN (0.5 M in THF, 65 milliliters, 33 mmol) was added to a solution of olefin 18 (5.56 grams, 9.6 mmol) in THF (85 milliliters) at 0 ° C. The mixture was stirred for 5 hours at room temperature, and then re-cooled to 0 ° C, H20 was added sequentially (200 milliliters), THF (100 milliliters), and NaB03-4 H20 (30 grams). After stirring overnight at room temperature, the organic volatiles were removed under reduced pressure. The aqueous residue was extracted with EtOAc (3 x 200 milliliters), and the combined organic layers were dried over Na 2 SO 4. Concentration and purification by flash chromatography (30 percent EtOAc-hexanes) provided alcohol 19 (12.05 grams, 92 percent) 19 20 Aldehyde 20. DMSO (1.36 milliliters, 19.2 mmol) was added dropwise to a solution of oxalyl chloride (1.26 milliliters, 14.4 mmol) in CH2C12 (120 milliliters) at -78 ° C. After stirring for 10 minutes, a solution of alcohol 19 (5.76 grams, 9.61 mmol) in CH2C12 (20 milliliters) was added via a cannula. The transfer was completed by means of rinsing with additional CH2C12 (2 x 5 milliliters). After stirring for 20 minutes, the mixture was treated with Et2N (5.36 milliliters, 38.4 mmol), and stirred for 10 minutes at -78 ° C, 30 minutes at 0 ° C and 10 minutes at room temperature. The reaction mixture was poured into saturated aqueous NaHCO3 (200 milliliters) and the separated aqueous layer was extracted with CH2C12 (3x), followed by EtOAc (100 milliliters). The combined organic phases were dried over Na 2 SO, concentrated and purified by column chromatography (10 percent to 20 percent EtOAc-hexanes) to provide the aldehyde compound 20 (5.28 grams, 92 percent) as an oil.

B2318 B2318. 0.1% (weight / weight, 3.21 grams) NiCl2 / CrCl2 and 1% NiCl2 / CrCl2 (weight / weight, 4.31 grams) were added to a solution of aldehyde 20 (3.73 grams)., 6.25 mmole), the F-2 fragment exemplified by vinyl iodide X2 (5.10 grams, 9.16 mmol), THF (85 milliliters) and DMF (21 milliliters) at room temperature in a glove box. The reaction mixture was stirred for 24 hours, removed from the glove box, cooled to 0 ° C, diluted with EtOAc (100 milliliters), cooled with saturated NHC1 (200 milliliters) and stirred for 30 minutes. The separated aqueous phase was extracted with EtOAc (6x), and the combined organic layers were dried over Na 2 SO, concentrated and purified by column chromatography (20 percent to 30 percent) to give B2318 (~ 3 grams), contaminated with impurities that ran closely and the intermediary not made cyclic (4.61 grams). The latter (4.61 grams, 4.48 mmol) in THF (150 milliliters) was dissolved, cooled to 0 ° C, and treated with KHMDS (0.5 M in toluene, 14 milliliters, 7.0 mmol) over a period of 2 minutes. After stirring at 0 ° C for 15 minutes, the reaction was quenched with saturated aqueous NH 4 Cl (150 milliliters) and warmed to room temperature. The separated aqueous layer was extracted with EtOAc (3x) and the combined organic phases were dried over Na 2 SO, concentrated and combined with the above partially purified product. Column chromatography (10 percent EtOAc-hexanes) yielded B2318 (3.17 grams, 55 percent) as a inseparable 3: 1 mixture of C27 diastereomers.

B2318 B2317 B2317. DDQ (1.45 grams, 6.42 mmol) was added per portion for 30 minutes to a stirred solution of B2318 (3.12 grams, 3.35 mmol) in CH2C12 (50 milliliters) and phosphate pH regulator of pH 7 (5 milliliters) to the room temperature. The reaction was quenched with saturated aqueous NaHCO3 (50 milliliters), stirred for 5 minutes, diluted with additional aqueous saturated NaHCO3 (100 milliliters), H20 (200 milliliters) and extracted with Et20 (5x). The combined organic phases were dried over Na2SO4, concentrated and purified by column chromatography (15 to 30 percent EtOAc-hexanes) to give recovered B2318 (1.40 grams) and a mixture of the C27 isomeric products. The recovered B2318 was re-subjected to the reaction conditions described above, to produce additional product. The recovered starting material was cycled again, through the deprotection conditions. All desired material was combined and separated by MPLC to produce B2317 (1.65 grams, 61 percent).

B2317 B2316 B2316. TsCl (0.63 grams, 3.30 mmol) was added to a solution of B2317 (1.60 grams, 1.97 mmol) in CH2C12 (8 milliliters) and pyridine (2 milliliters) at room temperature. After stirring for 29 hours, the reaction mixture was cooled with saturated aqueous NaHCO3 (30 milliliters), and H20 (10 milliliters). The separated aqueous layer was extracted with Et20, and the combined organic layers were dried over Na2SO4, concentrated and purified by column chromatography (15 to 30 percent EtOAc-hexanes) to give B2316 (2.01 grams, 92%). percent) as an oil, along with the recovered B2317 (92 milligrams, 5.8 percent).

B2316 B2315 B2315 LAH (1 M in THF, 2.61 milliliters, 2.61 mmol) was added, over 1 minute, to a solution of B2316 (1.68 grams, 1.74 mmol) in Et20 (80 milliliters) at 0 ° C. After stirring for 7 minutes, the reaction was quenched by careful addition of MeOH (0.42 milliliters, 10.4 mmol) and H20 (0.19 milliliters, 10 mmol), warmed to room temperature, and stirred for 20 minutes. Filtration through Celite with 1: 1 CH2Cl2-Et2 ?, concentration and purification by column chromatography (30 percent EtOAc at 40 percent - hexanes) gave B2315 (1.38 grams, 90 percent) as an oil.

B2315 B2314 B2314. MMTrCl (0.70 grams, 2.26 mmol) was added to a solution of B2315 (1.33 grams, 1.51 mmol) in CH2C12 (25 milliliters) and iPr2Net (0.79 milliliters, 4.53 mmol) at room temperature. The resulting mixture was stirred for 1 hour, and then poured into a mixture of NaHCO3 (20 milliliters), H20 (10 milliliters) and Et20 (50 milliliters). The separated aqueous layer was extracted with Et20 (3x). The combined organic phases were dried over Na2SO, concentrated and purified by column chromatography (CH2C12 followed by 15 to 30 percent EtOAc-hexanes) to give B2314 (1.65 grams, 95 percent) as a solid foam. .

B2314 B2313 B2313. A mixture of B2314 (1.60 grams, 1.39 mmoles) and Nal (3.10 grams, 20.8 mmoles) in acetone (50 milliliters) was heated under reflux for 13 hours. After cooling to room temperature, the reaction mixture was diluted with EtOAc, and concentrated. H20 (5 milliliters), saline (20 milliliters) and Na2S203 (200 milligrams) were added, and the resulting mixture was extracted with Et20 (4x). The combined extracts were dried over Na 2 SO, concentrated and purified by column chromatography (10 percent EtOAc-hexanes) to give B2313 (1.50 grams, 97 percent) as an oil.

B2308. Tert-BuLi (1.7 M in pentane, 1.00 milliliter, 1.7 mmol) was added over 1 minute to a solution of B2313 (0.90 grams, 0.81 mmol) in Et20 (14 milliliters) at -78 ° C. After stirring for 9 minutes, the mixture was transferred through a cannula, for 4 minutes, to a solution of the key fragment F-3 (0.83 grams, 1.14 mmoles) in Et20 (4 milliliters) at -78 ° C. The transfer was completed by means of rinsing with additional Et20 (2 milliliters). The resulting mixture was stirred at -78 ° C, for 5 minutes-, and then at 0 ° C for 10 minutes, cooled with saturated aqueous NaHCO 3 (30 milliliters) and warmed to room temperature. The separated aqueous layer was extracted with Et20 (3xN, and the combined organic phases were dried over Na2SO4, and concentrated.The residue was combined with those of two other batches (corresponding to 0.11 grams and 0.44 grams of B2313), and purified by column chromatography (10 percent to 20 percent EtOAc-hexanes) to give a mixture of B2307 and B2308 (1.86 grams, 83 percent) as a solid spur., Although the isomers could be separated by TLC preparation (EtOAc at 20 percent hexanes), these were carried forward as a mixture.

B2308 B2305, B2306 B2305 and B2306. The mixture of B2307 / B2308 was dissolved 4 (1.80 grams, 1.05 mmol) in EtOH (20 milliliters), treated with PPTS (10.0 milligrams, 0.04 mmol), stirred at room temperature for 11 hours, and then cooled with NaHCO3 (20.0 milligrams, 0.24 mmol). After stirring for 15 minutes, the mixture was concentrated, azeotroped with toluene (15 milliliters), and purified by column chromatography (20 to 30 percent EtOAc-hexanes), to give a mixture of B2305 and B2306 (1.22 grams, 81 percent) as a solid foam. Although the isomers could be separated by TLC preparation (30 percent EtOAc-hexanes), these were carried out as a mixture.

B2305, B2306 B2304 > 0 B2304. A mixture of B2305 / B2306 '(1.16 grams, 0.68 mmol), and Dess-Martin periodinone (0.61 grams, 1.44 mmol) in CH2Cl2 (35 milliliters), was stirred at room temperature for 1 hour. Dess-Martin periodinana (0.54 grams, 1.27 mmoles) was added to the mixture, and stirring was continued for an additional hour. The mixture was diluted with Et20 (100 milliliters), stirred for 20 minutes and filtered through Celite with Et20. The colorless filtrate was washed with saturated aqueous NaHCO3 (100 milliliters), and the separated aqueous layer was extracted with Et20 (3x). The combined organic phases were dried over Na 2 SO 4, concentrated and purified by column chromatography (10 percent to 15 percent EtOAc-hexanes) to give B2304 (0.98 grams, 84 percent) as a solid foam. Alternatively, B2304 can be prepared as follows, and in fact, the synthesis described below is superior to that given above.

B2317 ER804025 To a solution of the alcohol, 2.4 g mg, in methylene chloride, 29 milliliters, triflic anhydride, 770 milligrams was added. The mixture was stirred for 15 minutes, extracted with saturated sodium bicarbonate, dried and chromatographed to give 2.737 grams, 100 percent.

ER804025 ER804026 To a solution of the mesylate, 405 milligrams, in DMF, 0.06 milliliters, di-isopropylethylamine, 0.130 milliliters was added, followed by bencentiol, 0.061 milliliters. After 4 hours and after 22 hours, additional amine, 0.03 milliliters, and bencentiol, 0.015 milliliters were added. After 24 hours the mixture was diluted with 5 percent ethyl acetate / hexane, 1 milliliter, and chromatographed to give 409 milligrams.

ER804026 ER804027 To a solution of sulfur, 1.97 grams, in acetonitrile, 16 milliliters, was added N-methylmorpholine oxide (NMO), and then a solution of 1.02 grams, perruthenate (VII) of tetrapropylammonium, (TPAP), 38 milligrams, in acetonitrile , 1 milliliter. After 3.5 hours at room temperature, the mixture was heated at 40 ° C for 1 hour. The mixture was cooled and saturated aqueous sodium thiosulfate was added, and the mixture was partitioned between water and ethyl acetate. The usual preparation gave 1,567 grams of a brown oil.

ER804027 ER804028 To a solution of the pivaloate ester, 1567 grams, in methylene chloride, 11.2 milliliters, at -78 ° C, DIBAL, 2.5 milliliters of a 1M solution in toluene was added. After 15 minutes, additional DIBAL, 0.8 milliliters was added. After about 5 additional minutes, methanol, 0.46 milliliters, was slowly added, followed by water, 0.2 milliliters. The mixture was filtered through Celite and chromatographed to give 1.386 grams of an oil.

To a solution of the sulfone, 36 milligrams, in DME, milliliter, -40 ° C, n-butylithium, 2.8 equivalents, was added. After 35 minutes a solution of the aldehyde, 42 milligrams, in DME, 0.5 milliliters was added. After 40 minutes, saturated aqueous ammonium chloride was added, and the mixture was extracted with ethyl acetate. The usual preparation, followed by chromatography gave 52 milligrams of an oil.

ER804029 ER804030 To a solution of the alcohol, 42 milligrams, in methylene chloride, 2 milliliters, was added the Dess-Martin reagent, 36.4 milligrams. The mixture was stirred for 30 minutes and ether was added. The mixture was filtered through Celite, washed with saturated sodium bicarbonate, saturated sodium thiosulfate, prepared in the usual manner and chromatographed to give 38 milligrams of an oil.

ER804030 B2304 Preparation of the Sml2 Solution A solution of 1,2-di-iodoethane in 10 milliliters of THF was added to a suspension of Sm, 0.16 grams, in THF, 1 milliliter. The mixture was stirred for 1 hour. An aliquot of this solution, 0.03 milliliters, was added to a solution of the sulfone in THF at -78 ° C. After 5 minutes, additional Sml reagent, 0.05 milliliter was added. After a few additional minutes, more reagent was added, 0.25 milliliters. The cooling bath was removed and saturated aqueous sodium bicarbonate, 3 milliliters was added. The mixture was divided between ether and water, and the usual preparation gave 9.1 milligrams, 81 percent of an oil. Synthesis of B1794: Arabinose B1794 B1794- Except for the stereochemical and protective group differences (Schemes 3 and 5), arabinose was converted to B1794 in a manner similar to that described for B1793 (see Schemes 4 and 5). HRMS: calculated for C40H56O12 + Na 753.3826. Found: 753.3856. Synthesis of B1793 Analogs Representative: 81793 B1M0. R = H B1921- »T * B1920 and B1921. TsCl (9.9 milligrams, 0.052 mmol) was added to a solution of B1793 diol (7.6 milligrams, 0.010 mmol) in CH2C12 (1 milliliter) and pyridine (0.1 milliliter) at room temperature. After 48 hours the reaction was cooled with a 1: 4 mixture of saturated aqueous NaHC03 -salt solution, and extracted with CH2C12 (4x). The combined extracts were dried over Na 2 SO, and concentrated.

B2302, B2303 B2301 B2301. A mixture of B2302 / B2303 (0.79 grams, 0.60 mmol) and Dess-Martin periodinone (0.26 grams, 0.60 mmol) in CH2C12 (30 milliliters) was stirred at room temperature for 30 minutes. Additional Dess-Martin periodinnan (0.26 grams, 0.60 mmol) was added to the mixture, and stirring was continued for an additional 1.5 hours. The mixture was then diluted with Et20 (100 milliliters), stirred for 15 minutes and filtered through Celite. The filtrate was washed with saturated aqueous NaHCO3 (100 milliliters), and the separated aqueous layer was extracted with Et20 (3x). The combined organic phases were dried over Na 2 SO, were concentrated and purified by column chromatography (10 percent to 15 percent EtOAc. hundred - hexanes) to give the B2301 (0.67 grams, i5 percent) as an oil.

B2301 B1793 B1793 TBAF (1 M in THF containing 0.5 M imidazole HCl, 4.60 milliliters, 4.60 mmol) was added, over 2 minutes, to a solution of B2301 (0.62 grams, 0.48 mmol), in THF (29 milliliters) at temperature environment, and the resulting mixture was stirred for 18 hours. After dilution with hexanes (10 milliliters), the reaction mixture was directly loaded onto a Si02 column packed with 50 percent EtOAc-hexanes and leached with 50 percent EtOAc-hexanes (1 liter), followed by 10% MeOH / EtOAc to collect a mixture of intermediates. After removal of the solvent, the residue was dissolved in CH2C12 (15 milliliters), and treated with PPTS (645 milligrams). After stirring for 1 hour at room temperature, additional PPTS (414 milligrams) was added, and the resulting white suspension was stirred for 4.5 hours. The reaction mixture was then loaded directly onto a Si02 column packed with 70 percent EtOAc-hexanes and leached with 70 percent EtOAc / hexanes (0.5 liters), EtOAc (1 liter). Leaching with 5 percent to 10 percent MeOH / EtOAc yielded pure B1793 (181 milligrams), and leaching with 15 percent MeOH / EtOAc gave additional semi-hard product, which, after purification by TLC preparation ( 10 percent MeOH-EtOAc) provided additional pure B1793 (42 milligrams). B1793 (223 milligrams total, 64 percent) was obtained as a white solid. HRMS: calculated for C40H58Oi2 + Na 753.3826. Found: 753.3808. Synthesis of B1794: Arabinose B1794 B1794. Except for the stereochemical and protective group differences (Schemes 3 and 5), the arabinose was converted to B1794 in a manner similar to that described for B1793 (see Schemes 4 and 5). HRMS: calculated for C0H58Oi2 + Na 753.3826. Found: 753.3856. Synthesis of B1793 Analogs Representative: B1793 B1920: R = H B1921: R = Ts B1920 and B1921. TsCl (9.9 milligrams, 0.052 mmol) was added to a solution of B1793 diol (7.6 milligrams, 0.010 mmol) in CH2C12 (1 milliliter) and pyridine (0.1 milliliter) at room temperature. After 48 hours, the reaction was cooled with a 1: 4 mixture of saturated aqueous NaHC03 -salt solution, and extracted with CH2C12 (4x). The combined extracts were dried over Na2SO4, and concentrated. Purification by TLC preparation (80 percent EtOAc-hexanes) provided the monosylate B1920 (6.0 milligrams, 67 percent), and the ditosylate B1921 (1.8 milligram, 18 percent).

B1793 B2294 B2294. MsCl (0.3 M in CH2C12, 98 μL, 0.030 mmol) was added dropwise, over 40 minutes, to a mixture of collidine (7 μL, 0.054 mmol), B1793 (20.8 milligrams, 0.028 mmol) and CH2C12 (1 milliliter) to 0 ° C. After 76 hours at 4 ° C, the reaction was cooled with a 1: 4 mixture of saturated aqueous NaHC03-saline, and extracted with CH2C12 (4x). The combined extracts were dried over Na2SO4, and concentrated. The crude product was dissolved in toluene (3 milliliters), concentrated and purified by preparative TLC (1.5 percent MeOH-EtOAc) to produce mesylate B2294 (21.4 milligrams, 95 percent).

B2014 B2015 B2014 and B2015. A solution of 0.016 M of 4-fluorobenzyl bromide in Et2 was added. (800 μL, 13 μmol) and Ag2Ü (10 milligrams, 43 μmol), in three portions at 1 hour intervals, at room temperature, to a solution of B1793 (1.7 milligrams, 2.3 μmol) in Et20 (1.2 milliliters). The mixture was protected from light, stirred for 7 hours, and then filtered through Celite. Concentration and purification by TLC preparation (EtOAc) provided the primary ether B2014 (1.1 milligrams, 56 percent), and the secondary ether B2015 (0.6 milligrams, 31 percent). HRMS (FAB): calculated for C47H63FO? 2 + Na 861.4201. Found: for B2014 861.4178, for B2015 861.4160.

General. A mixture of B1793 (1 milligram, 1.37 micromoles), Et3N (10 microliters, 72 micromoles) and ArNCO (2 to 4 equivalents) in CH2C12 (0.2 milliliters) was stirred at room temperature for 4 hours at night, until it was considered, by TLC, that the reaction was complete. The reaction mixture was diluted with NaHCO3 (3 milliliters), extracted with CH2C12 (3x) and EtOAc (2x), dried over Na2SO4 and purified by preparative TLC (5 percent MeOH-CH2C12) to give the products : B1984. (1.0 milligram, 86 percent) HRMS (FAB): calculated for C47H63N013 + Na 872.4197. Found: 872.4214. B1990 (1.1 milligrams, 92 percent) HRMS (FAB): calculated for C47H62CIN0? 3 + Na 906.3807. Found: 906.3826. B1992 (1.0 milligram, 83 percent) HRMS (FAB): calculated for C48H65NO? 4 + Na 902.4303. Found: 902.4269.

B2042 B1793 B2042. DEAD (0.4 M in ether, 50 μL, 19 μmol) was added to a solution of B1793 (2.0 milligrams, 2.7 umoles), triphenylphosphine (5 milligrams, 19 μmoles), 4-nitrobenzoic acid (3.2 milligrams, 19 μmoles) and Et20 (500 μL) at room temperature. After 22 hours, the reaction mixture was directly loaded onto a preparation TLC plate, and was leached with 60 percent EtOAc-hexanes, to give the intermediate diester (3.0 milligrams). This material was taken in MeOH (300 μL) and treated with K2CO3 (approximately 1 milligram). After stirring at room temperature for 30 minutes, the reaction mixture was diluted with saline, and extracted with CH2Cl2 (5x). The combined extracts were dried over Na 2 SO 4, concentrated and purified by preparative TLC (5 percent MeOH-EtOAc) to give B2042 (1.2 milligrams, 60 percent for two steps). HRMS (FAB): calculated for C4oH58 ?? 2 + Na 753.3826. Found: 753.3810.

B1793 B1896, B1897 B1896 and B1897. NaBH4 (3 milligrams, 0.08 mmol) was added to a solution of B1793 (2.30 milligrams, 3.15 umoles) in 1: 1 CH2C12-MeOH (0.2 milliliters) at room temperature. Concentration of the reaction mixture and purification by TLC preparation (8 percent MeOH-EtOAc) yielded B1896 (0.80 milligrams, 35 percent) and B1897 (2: 1 mixture, 0.15 milligrams, 6.5 percent) ). HRMS (FAB): for B1896: calculated for C40H60O? 2 + Na 755.3983. Found: 753.3969.

B1793 B1918 B1918 A mixture of B1793 (2.0 milligrams, 2.74 μmoles), NaI04 (35 milligrams, 0.16 mmol), MeOH (0.8 milliliters) and H20 (0.2 milliliters) was stirred at room temperature for 40 minutes. The reaction mixture was diluted with H20 (1 milliliter), extracted with CH2C12 (6x), dried over Na2SO4, concentrated and purified by column chromatography (5 percent MeOH-CH2C12) to give B1918 (1.9 milligrams, 100 percent).

B1918 B2037 B2037 A solution of 0.034 M NaBH (0.1 milliliter, 3.4 μmol) in EtOH was added portionwise to a solution of B1918 (1.9 milligrams, 2.72 μmol) in MeOH (0.8 milliliters) and CH2C12 (0.2 milliliters) at -78 ° C. at room temperature, until it was considered by TLC that the reaction was complete. The reaction was quenched with NH4C1 (2 milliliters) at -78 ° C, warmed to room temperature, extracted with CH2C12 (6x), dried over Na2SO4 and purified by preparative TLC (5% MeOH - CH2C12 ) to provide B2037 (1.7 milligrams, 89 percent), HRMS (FAB): calculated for C39H56? n + Na 723.3720. Found: 723.3749.

B1793 B2035 B2035 NaI0 (35 milligrams, 0.16 mmol) was added to a solution of B1793 (1.7 milligrams, 0.0023 mmol), MeOH (800 μL) and H20 (200 μL), and after 15 minutes, the mixture was diluted with H2O, and it was extracted with CH2C12 (5x). The organic extracts were dried over Na 2 SO 4, concentrated, and the intermediate aldehyde was immediately dissolved in DMF (300 μL). 3-Bromo-3, 3-difluoropropene (3 μL, 0.023 mmol) and indium powder (3 milligrams, 0.23 mmol) were added, and after 3 hours 3-bromo-3,3-difluoropropene (1 μL, 0.008 mmoles) additional. After 18 hours, H20 was added, and the mixture was extracted with EtOAc (3x). The combined organic extracts were washed successively with H20 and saline, dried over Na2SO4 and purified by preparative TLC (80 percent EtOAc hexanes) to give B2035 (0.74 milligram, 41 percent for two steps) as a mixture. of isomers. HRMS (FAB): calculated for C42H58F20n + Na 799.3845. Found: 799.3883.

B 1793 B2011: X = H B2008: X = F B2008, B2011 NaBH4 (2 milligrams, 0.05 mmol) was added to a solution of B1793 (2.2 milligrams, 0.003 mmol) in 1: 1 of CH2C12-MeOH (200 μL) at room temperature. After 15 minutes saturated aqueous NH4C1 and H20 were added, and the mixture was extracted with CH2C12 (6x) and EtOAc (2x). The combined organic extracts were dried over Na2SO4, concentrated and purified by column chromatography (10 percent MeOH-EtOAc), to provide an intermediate triol, which was dissolved in MeOH (800 μL) and H20 (200 μL) . NaI04 (35 milligrams, 0.16 mmol) was added, and after 20 minutes the mixture was diluted with H20 and extracted with CH2C12 (6x). The organic extracts were dried over Na 2 SO 4, concentrated and the intermediate aldehyde dissolved immediately in THF (500 μL). 4-Fluorophenylmagnesium bromide (2 M in Et20, 12 μL, 0.024 mmol) was added, and after 20 minutes the reaction was quenched with saturated aqueous NH4C1. The mixture was extracted with CH2C12 (6x), and combined organic extracts were dried over Na2SO4, and concentrated. Purification by TLC preparation (EtOAc) provided the desired product as a mixture of 4 isomers (1.32 milligrams, 55 percent for 3 steps). Dess-Martin periodinane (~ 3 milligrams, 0. 007 immoles) to a solution of the above product (0.95 milligrams, 0.0012 mmol) in CH2C12 (300 μL), and the mixture was stirred at room temperature for 20 minutes. Dess-Martin periodinane (~3 milligrams, 0.007 mmol) and CH2C12 (300 μL) was added, and after a further 40 minutes Et20, saturated aqueous NaHCO3 (4 milliliters), and saturated aqueous Na2S203 (1 milliliter) were added. The mixture was extracted with Et20 (3x), and the combined extracts were washed with saline, dried over Na2SO4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes) to give B2008 (0.58 milligrams). , 61 percent). HRMS (FAB): calculated for C45H57FOu + Na 815.3783. Found: 815.3758. B2011. In an analogous manner, B1793 (1.9 milligrams, 0.003 mmol) was converted to B2011 (0.87 milligrams, 42 percent for 4 steps). HRMS (FAB): calculated for C45H580n + Na 797.3877. Found: 797.3877.

B2013 A solution of B1920 (1.4 milligrams, 0.0016 mmol), KCN (1 milligram, 0.016 mmol) and DMSO (500 μL) was heated at 60 ° C for 8 hours. After cooling to room temperature, H20 was added and the mixture was extracted with EtOAc (3x). The combined organic extracts were washed successively with H20 and saline, dried over Na2SO4, concentrated and purified by preparative TLC (80 percent EtOAc-hexanes) to give B2013 (0.78 milligram, 67 percent). HRMS (FAB): calculated for C4? H57NOu + Na 762.3829. Found: 762.3848.

B1920 X1920 X1920 A mixture of B1920 (1.3 milligrams, 1.47 umoles), Nal (30 milligrams, excess) and acetone (1 milliliter) was stirred at 60 ° C for 3.5 hours. After cooling to room temperature, the reaction mixture was diluted with saturated aqueous NaHCO3 (3 milliliters), extracted with CH2C12 (5x) and EtOAc, dried over Na2SO4, and purified by column chromatography (50% EtOAc). percent - CH2C12 to 80 percent EtOAc - hexanes) to give the iodide X1920 (1.3 milligrams, 100 percent). twenty B1998: Ar = p-Cl-Ph X1920 B2010: Ar = p-MeO-Ph B2019: Ar = 2-imidazole General. A mixture of the iodide XI920 (1.0 equivalent), iPr2EtN (11 to 22 equivalents), ArSH (9 to 46 equivalents), and DMF (0.3 milliliters) were stirred at room temperature, until it was considered, by TLC, that the reaction It was complete (typically 24 to 48 hours). The reaction mixture was diluted with saturated aqueous NaHCO3 (2 milliliters), extracted with CH2C12 and EtOAc, dried over Na2SO4 and purified by preparative TLC (80 percent EtOAc-hexanes, or 5 percent MeOH-CH2Cl2 ) to provide sulfur products: B1998. (1.3 milligrams gave 1.1 milligrams, 85 percent) HRMS (FAB): calculated for C46H6? C10uS + Na 897.3521. Found: 897.3533. B2010 (1.1 milligrams gave 0.7 milligrams, 59 percent) HRMS (FAB): calculated for C47H640? 2S + Na 875.4016. Found: 875.4057. B2019. (1.1 milligrams gave 0.7 milligrams, 61 percent) MS (FAB): M + Na. mCPBA B1998: Ar = p-Cl-Ph B2016: Ar = p-Cl-Ph B2010: Ar = p-MeO-Ph B2030: Ar = p-MeO-Ph General. A solution of 0.01 M mCPBA (1.2 equivalents) in CH2C12 was added to a solution of a sulfide (1.0 equivalent) in CH2C12 (0.5 milliliters) at 0 ° C, for 30 minutes. The reaction mixture was diluted with saturated NaHCO3 (2 milliliters), extracted with CH2C12 and EtOAc, dried over Na2SO and purified by TLC preparation (80 percent EtOAc-hexanes, or EtOAc) to give the products: B2016 . (0.9 milligrams gave 0.7 milligrams, 74 percent). B2030. (1.0 milligram gave 0.6 milligrams, 61 percent) HRMS (FAB): calculated for C47H60? 4S + Na 907.3914. Found: 907.3950. 1) TBDPSCI 2) Dess-Martin 3) TBAF B1793 B1934 B1934. TBDPSCI (3.0 μL, 12 μmol) was added to a solution of B1793 (1.3 milligrams, 1.78 umoles), imidazole (10 milligrams, 166 μmol) and DMF (0.10 milliliters) at room temperature. After stirring for 1 hour, the reaction mixture was diluted with saturated aqueous NaHCO3 (2 milliliters), extracted with CH2C12 (3x) and EtOAc (2x), dried over Na2SO4 and purified by preparative TLC (5% MeOH). percent - CH2Cl2) to give the intermediate silyl ether (1.3 milligrams, 77 percent). This material was dissolved in CH2C12 (0.5 milliliters) and treated with Dess-Martin periodinnan (10 milligrams, 24 μmol), for 1.5 hours at room temperature, diluted with Et20, and filtered through Celite. The filtrate was concentrated and purified by preparative TLC (50 percent EtOAc-hexanes) to provide the intermediate diketone (1.0 milligram, 77 percent), which was dissolved in THF (0.5 milliliters) and treated with 0.02M. of TBAF containing 0.01 M imidazole hydrochloride (THF solution, 75 μL, 1.5 μmol) at room temperature, for 15 minutes. The reaction mixture was leached through a Si02 column (50 percent EtOAc-hexanes to 5 percent MeOH-CH2C12), and the desired product was further purified by preparative TLC (5 percent MeOH - CH2C12 ) to provide B1934 (0.75 milligrams, 100 percent). HRMS (FAB): calculated for C40H56O? 2 + Na 751.3669. Found: 751.3690. Synthesis of B1939: B1922 B2294 B1922. Tetra-n-butylammonium azide (0.2 M in DMF, 0.5 milliliter, 0.10 mmol) was added to a solution of mesylate B2294 (21.4 milligrams, 0.026 mmol) in DMF (2 milliliters) at room temperature. After stirring at 83 ° C for 3.5 hours, the reaction mixture was cooled to room temperature, diluted with toluene, concentrated and purified by preparative TLC (80 percent ethyl acetate-hexanes) to provide the B1922 (18 milligrams, 92 percent).

B1922 B1939 B1939. Me3P was added sequentially (1 M in THF) and H20 (0.8 milliliters) to a solution of the azide B1922 (24.6 milligrams, 0.032 mmol) in THF (3.2 milliliters) at room temperature. The mixture was stirred for 22 hours, diluted with toluene, concentrated and purified by flash chromatography [step gradient, 10 percent MeOH-EtOAc followed by MeOH-EtOAc-30 percent aqueous NH 4 OH (9: 86: 5)] to provide the desired primary amine (23.3 milligrams), which contained according to XH -NMR-1 percent trimethylphosphine oxide. Lyophilization from benzene and remaining under high vacuum for 2 days, provided B1939 (20.3 milligrams, 87 percent). Synthesis of B1939 Analogs Representative: B1930, B1940, B1973, B1987, B1988, B1991, B2003, B2004 B1930. Me3P (1 M in THF, 13 μL, 0.013 mmol) was added to a solution of B1922 (1.6 milligrams, 2.1 μmoles), THF (400 μL) and H20 (100 μL) at room temperature. The mixture was stirred for 22 hours, diluted with toluene, concentrated, and dried azeotropically with toluene (2x) to give the crude amine, which was used directly in the next step. EDC (0.06 M in CH2C12, 100 μL, 11 μmol) was added to a solution of the crude amine, benzoylmorphic acid (0.8 milligram, 5.3 μmol) and CH2C12 (200 μL) at room temperature. After 30 minutes, the reaction was cooled with a mixture of 1: 4. of saturated aqueous NaHC03 - saline, and extracted with CH2C12 (5x). The combined extracts were dried over Na 2 SO 4, and concentrated and purified by preparative TLC (EtOAc) to give B1930 (1.5 milligrams, 83 percent for two steps). HRMS (FAB): calculated for C48H63NO? 3 + Na 884.4197. Found: 884.4166. B1940 Using the procedure described above for B1930, B1922 was reduced, coupled with 3-pyridylacetic acid hydrochloride, and purified by TLC preparation [(MeOH-EtOAc-30% aqueous NH 4 OH (9:86: 5)] to provide B1940 (0.8 milligrams, 67 percent for two steps) HRMS (FAB): calculated for C47H64N2O? 2 + Na 871.4357. Found: 871.4362.

B1973. Using the procedure described above, B1922 (0.9 milligram, 1.2 μmol) was reduced, coupled with phenylacetic acid, and purified by TLC preparation (5 percent MeOH-EtOAc) to provide B1973 (0.44 milligrams, 44 ppm). one hundred for two steps). HRMS (FAB): calculated for C48H65NO? 2 + Na 870.4404. Found: 870.4447. B1987 Using the procedure described above, B1922 was reduced (0.9 milligrams, 1.2 μmoles), coupled with 3-indole-glyoxylic acid, and purified by preparative TLC (3 percent MeOH -EtOAc) to provide B1987 (0.8 milligrams, 75 percent for two steps). HRMS (FAB): calculated for C5oH64N2O? 2 + Na 923.4306. Found: 923.4338. B1991 Using the procedure described above, B1922 (1 milligram, 1.3 pmol) was reduced, coupled with 4-chlorobenzoic acid, and purified by preparative TLC (3 percent MeOH-EtOAc) to provide B1991 (0.8 milligrams, 70 percent for two steps). HRMS (FAB): calculated for C47H62ClNO? 2 + Na 890.3858. Found: 890.3843. B2003 Using the procedure described above, B1922 (1 milligram, 1.3 μmol) was reduced, coupled with 3, 4, 5-trimethoxybenzoylmorphic acid, and purified by preparative TLC (EtOAc) to provide B2003 (0.7 milligrams, 56 percent for two steps). HRMS (FAB): calculated for C5? H69NO? 6 + Na 974.4514. Found: 974.4525. B2004 Using the procedure described above, B1922 (1 milligram, 1.3 μmol) was reduced, coupled with 3, 4, 5-trimethoxybenzoic acid, and purified by preparative TLC (5 percent MeOH-EtOAc) to give B2004 (0.7 milligrams, 58 percent for two steps). HRMS (FAB): calculated for C49H65NO? 3 + Na 946.4565. Found: 946.4599.

B1988 B1930 B1988. Dess-Martin periodinane (1 milligram, 2.3 umoles) was added to a solution of B1930 (0.80 milligrams, 0.93 μmoles) in CH2C12 (500 μL) at room temperature. After one hour the reaction was diluted with Et20 and filtered through Celite. The filtrate was sequentially washed with a 1: 9 mixture of saturated aqueous NaHC03-Na2S203 and saline, dried over Na2SO4, concentrated and purified by preparative TLC (80 percent EtOAc-hexanes) to give B1988 ( 0.45 milligrams, 56 percent). HRMS (FAB): calculated for C48H6? NO? 3 + Na 882.4041. Found: 884.4012. Synthesis of B2090: l or i 102 103 Compound 103. Indium powder (1.35 grams, 11.8 mmol) was added to a solution of 102 (3.38 grams, 17.6 mmol) in DMF (20 milliliters) at room temperature. After stirring for 30 minutes the reaction mixture was cooled to 0 ° C. Then the pure aldehyde 101 (3.72 grams, 28.6 mmol) was added, and the mixture was stirred overnight, while allowing the temperature to warm to room temperature. The reaction mixture was again cooled to 0 ° C and then carefully cooled with saturated aqueous NH 4 Cl (100 milliliters). After stirring for 30 minutes, the resulting mixture was extracted with Et20 (3x), dried over Na 2 SO, concentrated and purified by column chromatography (10 percent EtOAc a percent - hexanes) to give the pure crystalline 103 (2.20 grams, 59 percent). 103 104 105 Compound 104. Et3N (72 μL, 0.51 μmoles) was added to a solution of 103 (1.09 grams, 5.13 mmol) and thiophenol (0.63 milliliters, 7.16 mmol) in CH2C12, and the resulting mixture was stirred at 0 ° C for 1 hour. . Filtration through Si02 gave a mixture of 104 and 105, which after MPLC (15 percent to 20 percent EtOAc - hexanes) yielded 104 (0.53 grams, 32 percent) and 105 (0.92 grams, 56 percent) . 104 106 Compound 106. DIBALH (1 M in toluene, 3.28 milliliters, 3.28 mmol) was added to a solution of 104 (0.53 grams, 1.64 mmol) in toluene (10 milliliters) at -78 ° C, and the mixture was stirred at -78. ° C for 10 minutes. The reaction was cooled by careful addition of MeOH (0.40 milliliters, 9.84 mmol) and H20 (0.17 milliliters, 9.84 mmol), warmed to room temperature, and stirred for 20 minutes. The white suspension was filtered through a mixture of Celite and Si02 with 1: 1 CH2C12-Et20 and concentrated to give 106 (0.53 grams, 100 percent) as an oil. 106 107 Compound 107. A mixture of 106 (0.53 grams, 1.64 mmol) and ethyl (triphenylphosphoranylidene) acetate (1.15 grams, 3.29 mmol) in toluene (10 milliliters) was heated at 80 ° C, for 15 hours. The mixture was cooled to room temperature and DBU (25 μL, 0.16 mmol) was introduced. The mixture was heated at 80 ° C for 1.5 hours, cooled to room temperature, concentrated and purified by column chromatography (10 to 20 percent EtOAc-hexanes) to give 107 (0.54 grams, 83%). cent) as an oil (ratio of 3: 1 of the a isomers: ß). 107 108 Compound 108. A solution of mCPBA (~ 55 percent, 450 milligrams in 4.5 milliliters of CH2C12, 1.44 mmol) was added, to a solution of 107 (0.54 grams, 1.36 mmol) in CH2C12 (10 milliliters) at -78 ° C. The reaction mixture was diluted with saturated aqueous NaHCO3 (50 milliliters), H20 (10 milliliters), and Et20 (60 milliliters), and then warmed to room temperature. The separated aqueous layer was extracted with EtOAc (4x), and the combined organic phases were dried over Na 2 SO, concentrated and purified by column chromatography (50 percent EtOAc-hexanes) to give 108 (0.51 grams, 92 percent ) as an oil. 108 109 Compound 109. A mixture of 108 (0.51 grams, 1.24 mmol) and NaOAc (1.00 gram, 12.4 mmol) in Ac20 (10 milliliters) was stirred at 140 ° C for 12 hours, cooled to room temperature and then heated to room temperature. concentrated. The residue was partitioned between NaHCO3 (20 milliliters), and Et20 (30 milliliters), and stirred vigorously at room temperature for 30 minutes. The separated aqueous layer was extracted with Et20 (2x), and the combined organic phases were dried over Na2SO, concentrated and purified by column chromatography (5 percent to 15 percent EtOAc-hexanes) to give 109 (0.41 grams , 73 percent) as an oil. 109 110 Compound 110. A mixture of 109 (0.41 grams, 0.91 mmol) and K2C03 (44.3 milligrams, 0.32 mmol) in EtOH (5 milliliters) was heated at 60-70 ° C for 1 day. After cooling to room temperature, the reaction mixture was concentrated and leached through a SiO2 column (10 percent to 20 percent EtOAc-hexanes) to give the partially purified aldehyde intermediate. This material was dissolved in EtOH (2.5 milliliters), treated with NaBH (50 milligrams, 1.32 mmol), and stirred at room temperature for 30 minutes. The mixture was concentrated and purified by column chromatography (40 percent EtOAc-hexanes) to give 110 (181 milligrams, 66 percent). 110 111 Compound 111. BF3-OEt2 (0.05 M in CH2C12, 175 μL, 8.75 μmol) was added to a solution of 110 (181 milligrams, 0.60 mmol) and 2, 2, 2-p-methoxybenzyl trichloroacetimidate (0.50 milliliters, 1.80 mmoles) in CH2C12 (5 milliliters) at 0 ° C. The resulting mixture was stirred for 1.5 hours at 0 ° C, and for 2 hours at room temperature, until the reaction was complete. The mixture was cooled with saturated aqueous NaHCO3 (25 milliliters) and extracted with Et20 (5x). The combined organic phases were dried over a2SO4, concentrated and purified by column chromatography (CH2C12 and then 20 percent EtOAc-hexanes) to give the semi-pure 111 (0.37 grams,> 100 percent) as an oil. 112 111 Compound 112. A mixture of 111 (0.37 grams, max = 0.60 mmol) and TsOH-H20 (36 milligrams) in EtOH (5 milliliters) was stirred, initially at room temperature overnight, and then at 60 ° C during the night. 1 hour. Additional TsOH * H20 (31 milligrams) was added at room temperature and the reaction mixture was stirred for 1 hour at room temperature. The mixture was then concentrated, cooled with saturated aqueous NaHCO3 and extracted with EtOAc (5x). The combined organic phases were dried over Na 2 SO 4, concentrated and purified by column chromatography (20 percent to 50 percent EtOAc-hexanes and then 5 percent MeOH-CH 2 C 12) to give 112 (121 milligrams, 53 percent). cent) as an oil, along with the recovered 111 (49 milligrams, 21 percent). 112 113 Compound 113. TBSOTf (250 μL, 1.09 mmol) was added to a solution of 112 (121 milligrams, 0.32 mmol) and Et3N (176 μL, 1.26 mmol) in CH2C12 at 0 ° C, and the resulting mixture was stirred for 25 minutes. . The reaction was quenched with saturated aqueous NaHCO3 (15 milliliters) and the separated aqueous layer was extracted with ether (3x). The combined organic phases were dried over Na 2 SO 4, concentrated and purified by column chromatography (5 percent to 10 percent EtOAc / hexanes) to give 113 (165 milligrams, 85 percent) as an oil. 113 114 Compound 114. DIBALH (1M in toluene, 0.54 milliliters, 0.54 mmol) was added to a solution of 113 (165 milligrams, 0.27 mmol) in toluene (5 milliliters) at -78 ° C, and the resulting mixture was stirred at -78 ° C for '10 minutes. The reaction was cooled by careful addition of MeOH (65 μL, 0.81 mmol) and H20 (29 μL, 0.81 mmol), warmed to room temperature and stirred for 25 minutes. The white suspension was filtered through Celite with 1: 1 CH2C12-Et20. Concentration and purification by column chromatography (10 percent to 20 percent EtOAc-hexanes) gave 114 (153 milligrams, 100 percent) as an oil. 114 B2090 B2090. In a manner similar to that described in Scheme 6 for the synthesis of B1794, intermediate 114 was converted to B2090. HRMS (FAB): calculated for C39H56? N + Na 723.3720. Found: 723.3731.

B2136 B2090 B2136. In a manner analogous to that of B1939, B2090 was converted to B2136. HRMS (FAB): calculated for C39H57NO? Or + Na 722.3880. Found: 722.3907. Synthesis of B2039 / B2043: X2318 201 Diol 201. TBAF (1 M in THF, 383 μL, 0.383 mmol) was added to a solution of X2318 (350-LS-218) (80.8 milligrams, 0.0765 mmol) in THF (7 milliliters), and stirred at the temperature environment for 16 hours. After partial concentration the residue was directly loaded onto a packed Si02 column using 30 percent EtOAc-hexanes. Gradient leaching (30 percent EtOAc-hexanes to EtOAc) provided 201 diol (49.7 milligrams, 92 percent). 201 202 Aldehyde 202. A mixture of diol 201 (49.7 milligrams, 0.0707 mmol), NaI04 (100 milligrams, 0.47 mmol), MeOH (10 milliliters) and H20 (2.5 milliliters) was stirred at room temperature for 30 minutes. H20 was added and the mixture was extracted with CH2C12 (4x). The combined organic extracts were dried over Na 2 SO 4, concentrated and purified by column chromatography (30 percent EtOAc-hexanes) to provide aldehyde 202 (41.7 milligrams, 88 percent). 202 203 Alcohol 203. 4-Fluorophenylmagnesium bromide (2 M in Et20, 155 μL, 0.31 mmol) was added to a solution of aldehyde 202 (41.7 milligrams, 0.062 mmol) in THF (6 milliliters). After 15 minutes at room temperature, the reaction was quenched with saturated aqueous NH4C1, and extracted with CH2C12 (4x). The combined organic extracts were dried over Na2SO4, concentrated and purified by preparative TLC (40 percent EtOAc-hexanes) to provide alcohol 203 (32.4 milligrams, 68 percent) as a mixture of 1: 1 C34 isomers . In this step the undesired minor isomer C27 was separated, and furthermore it was isolated as a mixture of 1: 1 C34 isomers (8.4 milligrams, 18 percent). 203 204 Ether 204. Et3N (18 μL, 0.13 mmol) and TBSOTf (15 μL, 0.063 mmol) were added to a solution of 203 alcohol (32.4 milligrams, 0.042 mmol) in CH2C12 (5 milliliters) at 0 ° C. After 20 minutes the reaction was quenched by the addition of saturated aqueous NH4C1, and extracted with CH2C12 (3x). The combined organic extracts were dried over Na 2 SO 4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes) to provide ether 204 (33.1 milligrams, 89 percent). 204 205 Alcohol 205. LAH (1M in THF, 113 μL, 0.113 mmol) was added dropwise to a solution of ether 204 (33.1 milligrams, 0.0375 mol) in Et20 (10 milliliters) at 0 ° C. After 20 minutes, H20 and 1 M NaOH were added, and the mixture was stirred at room temperature for 10 minutes. Filtration through Celite, concentration and purification by column chromatography (40 percent EtOAc-hexanes) provided alcohol 205 (28.4 milligrams, 95 percent). 205 206 Ether 206. Diisopropylethylamine (31 μL, 0.18 mmol) and MMTrCl (22 milligrams, 0.071 mmol) were added to a solution of alcohol 205 (28.4 milligrams, 0.0356 mmol) in CH2C12 (4 milliliters) at 0 ° C. After 15 hours at room temperature, H20 was added, and the mixture was extracted with CH2C12 (3x). The combined extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by preparative TLC (40 percent EtOAc-hexanes) to provide ether 206 as a mixture of ~ 1.5: 1 C34 epimer (45). milligrams, cant), which contained a small amount of impurities that ran closely. 206 207 Alcohol 207. DDQ (40 milligrams, 0.18 mmol) was added to a solution of ether 206 (37 milligrams, 0.034 mol) in CH2C12 (4 milliliters) and a 1:10 mixture of rBuOH: phosphate pH regulator with pH 7 (2 milliliters) at 0 ° C. The mixture was stirred vigorously in the dark for 15 minutes. Three additional portions of DDQ (40 milligrams, 0.18 mmol) were added at 10 minute intervals, then the reaction was diluted with saturated aqueous NaHCO3 and extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by preparative TLC (30 percent EtOAc-hexanes) to provide alcohol 207 (19.2 milligrams, 59 percent), as well as ether 206 recovered (9.7 milligrams, 26 percent). 207 208A and 208B Mesylates 208A and 208B. They were added sequentially Et3N (19 μL, 0.13 mmol) and Ms20 (10 milligrams, 0.056 mmol) to a solution of 207 alcohol (21.3 milligrams, 0.022 mmol) in CH2C12 (6 milliliters) at 0 ° C. After 30 minutes, saturated aqueous NaHCO3 was added, and the mixture was extracted with CH2C12 (3x). The combined extracts were washed with saline, dried over Na2SO4, concentrated and purified by preparative TLC (30 percent EtOAc-hexanes) to provide the mesylates 208A (11.7 milligrams, 51 percent) and 208B (6.5 milligrams) , 28 percent) as individual C34 isomers. 208A and 208B B2039, B2043 B2039 and B2043. In a manner similar to that described in Scheme 6 for the synthesis of B1794, both diastereomers 208A and 208B were independently converted to B2039 and B2043. HRMS (FAB): calculated for C45H59FO ?? + Na 817.3939. Found: for B2039 817.3896, for B2043 817.3910. Synthesis of B2086, B2088, B2091 X- 20 10a, 10b Alcohol X-20. NaI04 (1.16 grams, 5.4 ICO Immoles) to a solution of diols 10 a, b (1.19 grams, 3.0 mmol) in MeOH-H20 (4: 1, 75 milliliters) at 0 ° C. The reaction mixture was allowed to warm to room temperature. After stirring for 40 minutes, the mixture was diluted with EtOAc, filtered through Celite, concentrated and partitioned between saline and CH2C12. The separated aqueous layer was extracted with CH2Ci2 (2x). The combined organic layers were dried over Na2SO4, and concentrated to provide the crude aldehyde intermediate. NaBH (228 milligrams, 6.0 mmol) was added to a solution of the aldehyde in MeOH-Et20 (1: 1, 40 milliliters) at 0 ° C. The mixture was stirred for 30 minutes, carefully quenched with saturated aqueous NH4C1, stirred for 20 minutes at room temperature, and extracted with CH2C12 (3x). The combined extracts were dried over NaSO, concentrated and purified by flash chromatography (40 percent to 50 percent EtOAc-hexanes) to provide the X-20 alcohol (1.02 grams, 93 percent for two steps). 21 Silyl ether 21. Imidazole (0.94 grams, 13.9 mmol) and TBSC1 (0.59 grams, 3.89 mmol) were sequentially added to a solution of the alcohol X-20 (1.02 grams, 2.78 mmol) in DMF (10 milliliters) at room temperature. After 14 hours, the reaction mixture was diluted with saturated aqueous NH4C1 and extracted with EtOAc (3x). The combined organic extracts were washed with H20, saline, dried over Na2SO4, concentrated and purified by flash chromatography (5 percent to 15 percent EtOAc-hexanes) to give the silyl ether 21 (1.3 grams, 98%). percent). 21 22 Alcohol 22. A mixture of Pd (OH) 2 (20 percent, 0.8 grams), silyl ether 21 (1.3 grams, 2.70 mmol) and EtOAc (30 milliliters) was stirred for 1 hour under 1 atmosphere of H2 to the Room temperature was filtered through Celite, concentrated and purified by flash chromatography (20 percent to 40 percent EtOAc-hexanes) to provide alcohol 22 (0.96 grams, 91 percent). 22 25 Alcohol 25. 4-Methylmorpholine N-oxide (980 milligrams, 8.4 mmol) and TPAP (131 milligrams, 3.26 mmol) were added sequentially to a solution of alcohol 22 (1.78 grams, 4.6 mmol) in CH2C12 (45 milliliters) to the room temperature. A cold bath was necessary to control the exotherm. After 20 minutes the reaction mixture was diluted with hexanes, filtered through a short SiO2 column (15 percent EtOAc-hexanes) and concentrated to give the crude ketone. Tebbe reagent (14.9 milliliters, 9.0 mmol) was added over 10 minutes to a solution of the crude ketone in THF (60 milliliters) at 0 ° C. After 20 minutes the reaction mixture was poured into Et20 (100 milliliters) which had been previously cooled to -78 ° C, cooled by the slow addition of H20 (30 milliliters), warmed to room temperature, stirred for 30 minutes and extracted with Et20 (4x). The combined extracts were washed with saline, dried over Na2SO4, concentrated and purified by flash chromatography (10 percent EtOAc-hexanes) to provide the desired olefin contaminated by the gem-dimethyl product (1.07 grams). This mixture was used directly in the next step. 9-BBN (0.5 M in THF, 11.6 milliliters, 5.8 mmol) was added to a solution of the olefin in THF (15 milliliters) at 0 ° C. The reaction mixture was allowed to warm to room temperature, it was stirred for 5 hours, and then it was cooled again to 0 ° C. H20 (60 milliliters), THF (60 milliliters) and NaB03-4 H20 (5.7 grams) were added. After stirring for 5 hours at room temperature, the THF was removed under reduced pressure and the aqueous residue was extracted with EtOAc (4x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (20 to 40 percent EtOAc-hexanes) to provide alcohol 25 (605 milligrams, 18 percent for three Steps) . 26 Alcohol 26. Using the procedure previously described, alcohol was sequentially oxidized (604 milligrams, 1.49 mmol), made isomeric, and reduced. The 1C4 purification by flash chromatography (20 percent EtOAc to 40 percent hexanes) provided the alcohol 26 (550 milligrams, 91 percent for three steps). 26 27 MPM-ether 27. BF3-OEt2 (0.05 M in CH2C12, 270 μL, 0.013 mmol) was added to a solution of alcohol 26 (545 milligrams, 1.35 mmol) and MPM-trichloroimidate (1.14 grams, 4.0 mmol) in CH2C12 (40 μM). milliliters) at 0 ° C. After 1 hour, the reaction was cooled with saturated aqueous NaHCO3, extracted with CH2C12, dried over Na2SO4, concentrated and purified by flash chromatography (10 percent to 15 percent EtOAc-hexanes) to provide the MPM- ether 27 (580 milligrams, 82 percent). 28 27 Alcohol 28. LAH (1 M in THF, 1.9 milliliters, 1.9 mmol) was added to a solution of MPM-ether 27 (580 milligrams, 1.11 mmol) in Et20 (100 milliliters) at 0 ° C. After 30 minutes the reaction was carefully cooled with H20 (0.5 milliliters), and 1 N aqueous iuOH (0.5 milliliters), stirred for 1 hour at room temperature, filtered through Celite, concentrated and purified by flash chromatography (30 percent to 50 percent EtOAc-hexanes) to provide alcohol 28 (460 milligrams, 95 percent). 29 28 Olefin 29. DMSO (441 μL, 6.23 mmol) was added to a solution of oxalyl chloride (272 μL, 3.12 mmol) in CH2C12 (30 milliliters) at -78 ° C. After 15 minutes a solution of the alcohol 28 (458 milligrams, 1.04 mmol) in CH2C12 (15 milliliters) was added to the reaction mixture. After stirring for 1 hour at -78 ° C, Et3N (1.3 milliliters, 9.35 mmol) was added. The reaction mixture was warmed to 0 ° C, stirred for 10 minutes, diluted with saturated aqueous NH4C1 and extracted with CH2C12 (3x). The combined organic extracts were dried over Na 2 SO 4, concentrated and filtered through a short SiO2 column (20 percent to 30 percent EtOAc-hexanes) to provide the crude aldehyde. N-BuLi (1.63 M, 1.4 milliliters, 2.28 mmol) was added dropwise to a solution of CH3PPh3Br (815 milligrams, 2.28 mmol), THF (20 milliliters) and DMSO (7.5 milliliters) at 0 ° C.

After 1 hour, a solution of the aldehyde in THF was added (10 milliliters). The reaction mixture was warmed to room temperature and stirred for 3 hours. Saturated aqueous NH4C1 was added, and the mixture was extracted with EtOAc (4x). The combined extracts were washed with H20, saline, dried over Na2SO4, concentrated and purified by flash chromatography (10 percent to 15 percent EtOAc-hexanes) to provide the olefin 29 (380 milligrams, 95% yield). one hundred for two steps). 29 31 Compound 31. 9-BBN (0.5 M in THF, 6 milliliters, 3 mmol) was added to a solution of olefin 29 (370 milligrams, 0.85 mmol) in THF (7 milliliters) at 0 ° C. The mixture was allowed to warm to room temperature and was stirred for 1 hour. After re-cooling to 0 ° C, H20 (30 milliliters), THF (20 milliliters) and NaB03-4 H20 (2.8 grams) were added. After stirring for 3 hours at room temperature, the THF was removed under reduced pressure. The aqueous residue was extracted with EtOAc (4x), dried over Na 2 SO 4, concentrated and purified by flash chromatography (25 percent to 50 percent EtOAc-hexanes) to provide alcohol 30, which was used directly in the next step. Pivaloyl chloride (157 μL, 1.27 mmol) was added to a solution of alcohol 30 in CH2C12-pyridine (1: 1 mixture, 10 milliliters) at room temperature. After 18 hours, additional pivaloyl chloride (100 μL, 0.81 mmol) was added. After 1 hour the reaction mixture was cooled to 0 ° C, cooled with MeOH (0.5 milliliters), concentrated, diluted with saline, and extracted with CH2C12 (4x). The combined organic extracts were dried over Na 2 SO 4, concentrated and purified by flash chromatography (10 percent to 15 percent EtOAc-hexanes) to provide compound 31 (410 milligrams, 90 percent for two steps). 31 2 Alcohol 32. TBAF (1M in THF, 1.14 milliliters, 1.14 mmol) was added to a solution of 31 (410 milligrams, 0.761 mmol) in THF (5 milliliters) at room temperature. After 1.5 hours, the reaction mixture was concentrated and purified by flash chromatography (40 percent EtOAc-hexanes to 100 percent EtOAc) to provide alcohol 32 (320 milligrams, 100 percent). 32 33a: C34 a-OH 33b: C34ß-OH Alcohols 33a and 33b. Dess-Martin periodinane (925 milligrams, 2.18 mmol) was added to a solution of alcohol 32 (309 milligrams, 0.727 mmol) in CH2C12 (19 milliliters) at room temperature. After 1 hour the reaction was diluted with Et20 and filtered through Celite. The filtrate was sequentially washed with a 1: 9 mixture of saturated aqueous NaHC03-Na2S203 and saline, dried over Na2SO4, concentrated and purified by flash chromatography (20 percent to 30 percent EtOAc-hexanes) to provide the desired aldehyde, which was taken immediately to the next step. BF3-OEt2 (135 μL, 1.1 mmol) was added to a solution of the crude aldehyde, tri-n-butylaliptin (337 μL, 1.08 mmol) and CH2C12 (16 milliliters) at -78 ° C. After 1 hour, the reaction was cooled with saturated aqueous NaHCO3, extracted with CH2C12 (3x). The combined organic extracts were dried over Na 2 SO, concentrated and purified by MPLC (25 percent to 30 percent EtOAc-hexanes) to provide the largest, most polar alcohol 33a (165 milligrams, 49 percent for two steps) and the least polar minor product 33b (90 milligrams, 27 percent for two steps).

MeQ / -OM TBSOTf 33a 34 Compound 34. TBSOTf (163 μL, 0.710 mmol) was added to a solution of alcohol 33a (165 milligrams, 0.355 mmol), Et3N (247 μL, 1.78 mmol) and CH2C12 (5 milliliters) at 0 ° C. After 25 minutes the reaction was quenched with saturated aqueous NaHCO3, extracted with CH2C12 (3x), dried over Na2SO4, concentrated and purified by flash chromatography (15 to 20 percent EtOAc-hexanes) to provide the compound 34 (200 milligrams, 98 percent). 34 35a and 35b Diols 35a and 35b. Os04 (0.1 M solution in toluene, 32 μL, 3.2 μmol) was added to a solution of K2C03 (168 milligrams, 1.22 mmol), K3Fe (CN) 6 (400 milligrams, 1.22 mmol), (DHQ) 2PYR (11 milligrams , 12 μmol), H20 (3.2 milliliters) and t-BuOH (2.2 milliliters) at 0 ° C. Then a solution of olefin 34 (200 milligrams, 0.345 mmol) in t-BuOH (1 milliliter) was added to the reaction mixture. After 5 hours at 0 ° C, Na2S205-5 H20 (200 milligrams) was added. The reaction mixture was warmed to room temperature, stirred for 30 minutes, and extracted with extracted with CH2C12 (5x). The combined extracts were washed with saline, dried over Na 2 SO, concentrated and purified by preparative TLC (70 percent EtOAc-hexanes) to provide the major, less polar diol 35a (118 milligrams, 56 percent), and the minor, more polar 35s diastarreomeric product (74 milligrams, 35 percent). Each of the individual diastereomers were carried out separately. 35a 36 Compound 36. TBSOTf (177 μL, 0.77 mmol) was added to a solution of diol 35a (118 milligrams, 0.912 mmol), Et3N (267 μL, 1.92 mmol) and CH2C12 (5 milliliters) at 0 ° C. After 25 minutes, the reaction was quenched with saturated aqueous NaHCO3, extracted with CH2C12 (3x), dried over Na2SO4, concentrated and purified by flash chromatography (10 percent to 15 percent EtOAc-hexanes) to provide compound 36 (161 milligrams, 100 percent). 36 37. Alcohol 37. Using the procedure previously described for the preparation of alcohol 28, compound 36 (161 milligrams, 0.192 mmol) yielded alcohol 37 (135 milligrams, 93 percent) after purification by flash chromatography (20% EtOAc). percent to 40 percent - hexanes). 37 38 Aldehyde 38. Dess-Martin periodinane (227 milligrams, 0.535 mmol) was added to a solution of alcohol37 (135 milligrams, 0.178 mmol) in CH2C12 (5 milliliters), at room temperature. After 1 hour the reaction mixture was diluted with Et20 and filtered through Celite. The filtrate was washed sequentially with a mixture of 1: 9 saturated aqueous NaHC03 - Na2S203 and saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (10 percent to 20 percent EtOAc-hexanes) to provide aldehyde 38 (127 milligrams, 95 percent). 38 B2086, B2102 B2086, B2102. Each of the diastereomers obtained above were separately taken to the final product in a manner similar to that described in Scheme 6 for B1794. The diastereomer 35a provided B2086. The diastereomer 35b provided B2102.

B2088 B2086 B2088. NaI04 was added to a solution of B2086 (1 milligram, 1.29 pmol) in MeOH-H20 (4: 1, 1 milliliter) at room temperature. After 30 minutes the reaction mixture was diluted with H20, extracted with CH2C12 (6x), dried over Na2SO4, and concentrated to give B2088 (1.2 milligrams).

B2088 B2091 B2091. NaBH4 (0.013 M in EtOH, 20 μL, 0.27 μmoles) was added to a solution of B2088 (1 milligram, 1.29 umoles) in MeOH-CH2C12 (4: 1, 0.5 milliliters) at -78 ° C. Additional NaBH4 was added periodically with accurate monitoring of the reaction by TLC (a total of 220 μL of the NaBH solution was required). The reaction mixture was cooled to 0 ° C with saturated aqueous NHC1, stirred for 20 minutes at room temperature, and extracted with CH2C12 (6x). The combined extracts were dried over Na 2 SO 4, concentrated and purified by preparative TLC (7 percent MeOH-EtOAc) to provide B2091 (0.40 milligrams, 50 percent). Synthesis of B1933: 302 303 Alcohol 303. 9-BBN (0.5 M in THF, 23 milliliters, 0.012 moles) was added dropwise, during 30 minutes, to a solution of alkene 302 (1.51 grams, 0.00386 moles) in THF (40 milliliters) at 0 ° C. . After stirring for 80 minutes at room temperature, the mixture was cooled to 0 ° C, and H20 (80 milliliters) was carefully added, followed by NaB03-4 H20 (4.2 grams, 0.027 moles). The mixture was vigorously stirred at room temperature for 2.3 hours, then extracted with EtOAc (3x). The combined organic extracts were washed with saline, dried over Na2SO4, concentrated and purified by column chromatography (50 percent EtOAc-hexanes) to provide alcohol 303 (1.37 grams, 87 percent). 303 304 Aldehyde 304. Oxalyl chloride was added by dripping (88 μL, 1.00 mmol) to a solution of DMSO (142 μL, 2.00 mmol) in CH2C12 (20 milliliters) at -78 ° C. After 30 minutes a solution of the alcohol 303 (137 milligrams, 0.335 mmol) in CH2C12 (5 milliliters) was added, and it was stirred at -78 ° C for 1 hour. Et3N (420 μL, 3.01 mmol) was added, and after 10 minutes the reaction was stirred for 10 minutes at 0 ° C, at which point saturated aqueous NH4C1 was added, and the resulting mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (50 percent EtOAc-hexanes) to provide the intermediate aldehyde 304 (0.114 grams, 84 percent) which was used immediately in the next step. 304 305 Alcohol 305. TBAF (1 M in THF, 5 μL, 0.005 mmol) was added to a solution of aldehyde 304 (0.114 grams, 0.27 mmol) in CF3TMS (0.5 M in THF, 1.1 milliliters, 0.54 mmol) at 0 °. C. After 20 minutes, a second portion of TBAF (1 M in THF, 100 μL, 0.1 mmol) was added, and the mixture was stirred for 10 minutes, at which point excess TBAF was added dropwise (1 M in THF, 270 μL, 0.27 mmole), to dissociate the intermediate silyl ether. After 30 minutes, the mixture was diluted with H20 and extracted with EtOAc (3x). The organic extracts were washed with H20, saline, dried over Na2SO4, concentrated and purified by column chromatography (50 percent EtOAc-hexanes) to provide alcohol 305 (123 milligrams, 95 percent) as a mixture. inseparable from 1: 1 isomers. 306 305 Silyl ether 306. TBSOTf (265 μL, 1.16 mmol) was added to a solution of alcohol 305 (123 milligrams, 0.257 mmol), Et3N (430 μL, 3.08 mmol) in CH2C12 (8 milliliters) a 0 ° C. After stirring at room temperature for 20 hours, saturated aqueous NaHCO3 was added, and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes) to give silyl ether 306 (148 milligrams, 97 percent). 306 307 Alcohol 307. LAH (1 M in THF, 220 μL, 0.22 mmol) was added dropwise to a solution of silyl ether 306 (131 milligrams, 0.22 mmol) in Et20 (5 milliliters) at 0 ° C. After 20 minutes H20 and 1 M NaOH were carefully added. The mixture was stirred at room temperature for 30 minutes, filtered through glass wool, concentrated and purified by column chromatography (50 percent EtOAc-hexanes) to provide alcohol 307 (112 milligrams, cant. ). 307 309 Alkene 309. Oxalyl chloride (58 μL, 0.66 mmol) was added dropwise to a solution of DMSO (94 μL, 1.3 mmol) in CH2C12 (10 milliliters) at -78 ° C. After 30 minutes a solution of the alcohol 307 (112 milligrams, 0.22 mmol) in CH2C12 (3 milliliters) was added. After 1 hour Et3N (276 μL, 1.98 mmol) was added, and after 10 minutes at -78 ° C the reaction was stirred at 0 ° C for 10 minutes. Saturated aqueous NH 4 Cl was added, and the mixture was extracted with CH 2 Cl 2 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (50 percent EtOAc-hexanes) to provide aldehyde 308 (101 milligrams, 91 percent) which was used immediately in the next step. N-BuLi (1.63 M, in THF, 200 μL, was added dropwise, 0. 33 mmol) was added to a solution of CH3PPh3Br (118 milligrams, 0.33 mmol) in THF (3 milliliters) and DMSO (1.2 milliliters) at 0 ° C.

After 70 minutes a solution of the aldehyde 308 was added (101 milligrams, 0.20 mmol) in THF (3 milliliters), and after 10 minutes at 0 ° C the reaction was stirred at room temperature for 1 hour. Saturated aqueous NH4C1 was added, and the mixture was extracted with EtOAc (3x). The combined organic extracts were washed with saline, dried over Na 2 SO, concentrated and purified by column chromatography (20 percent EtOAc-hexanes) to provide the alkene 309 (90.9 milligrams, 90 percent). 309 310 Alcohol 310. 9-BBN (0.5 M in THF, 17 milliliters, 8.45 mmol) was added dropwise to a solution of alkene 309 (1.06 grams, 2.11 mmol) in THF (30 milliliters) at 0 ° C. After stirring for 2.5 hours at room temperature, the reaction was cooled to 0 ° C, and H20 (60 milliliters) was carefully added, followed by NaB03-4 H20 (3.25 grams, 21.1 mmol). The mixture was vigorously stirred at room temperature for 2 hours, then diluted with H20 and extracted with EtOAc (3x). The combined organic extracts were washed with saline, dried over Na 2 SO, concentrated and purified by column chromatography (20 to 30 percent EtOAc-hexanes) to provide 310 alcohol (0.920 grams, 84 percent). . 311 310 Pivaloate 311. A mixture of 310 alcohol was stirred (65.8 milligrams, 0.0126 mmol), pyridine (61 μL, 0.76 mmol) and PvCl (23 μL, 0.189 mmol) in CH2C12 (3 milliliters), at room temperature, for 5 hours. A second reaction was run using alcohol 310 (0.92 grams, 1.76 mmole) under similar conditions, and both reactions were combined during the preparation; saturated aqueous NHC1 was added, and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes) to provide the pivaloate 311 (1.08 grams, cant.) 311 312 Alcohol 312. A mixture of ether 311 (0.811 grams, 1.33 mmoles), DDQ (6.1 grams, 27 mmoles) and 10: 1 of tBuOH: phosphate pH regulator was vigorously stirred with pH 7 (42 milliliters) in CH2C12 (84 milliliters) in the dark, at room temperature, for 1.5 hours, at which point additional DDQ (1.0 gram, 4.4 mmol) was added. After 1 hour saturated aqueous NaHCO3 was added and the mixture was extracted with CH2C12 (4x). The combined organic extracts were washed successively with saturated aqueous NaHCO3 and saline, dried over Na2SO4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes) to provide alcohol 312 (0.56 grams, 87 percent ), as well as the recovered starting material 311 (97 milligrams, 12 percent) » Ketone 313. Oxalyl chloride was added dropwise (21 μL, 0.12 mmol) to a solution of DMSO (34 μL, 0.48 mmol) in CH2C12 (3 milliliters) at -78 ° C. After 1 hour a solution of the alcohol 312 (39.4 milligrams, 0.081 mol) in CH2C12 (1.5 milliliters) was added and the mixture was stirred for 1.5 hours. Et3N (100 μL, 0.73 mmol) was added, and after 10 minutes the mixture was heated to 0 ° C. Saturated aqueous NH4C1 was added, and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (30 percent EtOAc-hexanes) to provide ketone 313 (36.6 milligrams, 93 percent) which was used immediately in the next step. 313 314 Alqueno 314. Tebbe reactant was added (~0.65 M in toluene, 720 μL, 0.47 mmole) to a solution of ketone 313 (151 milligrams, 0.31 mmol) in THF (5 milliliters) at 0 ° C. After 15 minutes H20 was carefully added and the mixture was extracted with EtOAc (3x). The combined organic extracts were washed with saline, dried over Na 2 SO, concentrated and purified by column chromatography (10 percent EtOAc-hexanes) to provide the alkene 314 (139 milligrams, 93 percent). 314 315 Alcohol 315. 9-BBN (0.5 M in THF, 6.0 milliliters, 2.9 mmol) was added dropwise to a solution of alkene 314 (468 milligrams, 0.97 mmol) in THF (10 milliliters) at 0 ° C. The mixture was stirred at room temperature for 2 hours, at which point additional 9-BBN (0.5 M in THF, 500 μL, 0.25 mmol) was added. After 2.5 hours the mixture was cooled to 0 ° C, and H20 (10 milliliters) was carefully added, followed by NaB03-4 H20 (1.5 grams, 9.7 mmol). The mixture was vigorously stirred at room temperature for 5 hours, diluted with H20 and extracted with EtOAc. (3x) The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by column chromatography (gradient percent to 30 percent EtOAc - hexanes) to provide alcohol 315 (0.47 grams, 97 percent). 315 316 Alcohol 316. Oxalyl chloride (246 μL, 2.82 mmol) was added dropwise to a solution of DMSO (400 μL, 5.64 mmol) in CH2C12 (40 milliliters) at -78 ° C. After 1 hour a solution of alcohol 315 (0.47 grams, 0.94 mmoles) in CH2C12 (10 milliliters) was added and the mixture was stirred for 1 hour. Et3N (1.2 milliliters, 8.5 mmol) was added, and after 10 minutes the mixture was warmed to 0 ° C, and stirred for 10 minutes. Saturated aqueous NHC1 was added, and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na2SO4, and concentrated. The crude aldehyde was stirred in CH2Cl2 (20 milliliters) and Et3N (2 milliliters) at room temperature, overnight. Saturated aqueous NH4C1 was added, and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by flash chromatography (30% EtOAc-hexanes), providing the epimerized aldehyde which was immediately dissolved in 1: 1 Et20: EtOH (10 milliliters) and cooled to 0 ° C. NaBH 4 (35 milligrams, 0.94 mmole) was added, and after 10 minutes the reaction was quenched with saturated aqueous NHC 1. The mixture was extracted with EtOAc (3x), and the combined organic extracts were washed with saline, dried over Na 2 SO 4, concentrated and purified by column chromatography (30 percent EtOAc-hexanes) to provide alcohol 316 ( 0.410 grams, 87 percent yield for 3 steps). 316 317 Ether 317. Alcohol 316 (60.7 milligrams, 0.12 mmol) and MPMOTCl (0.10 grams, 0.36 mmol) were combined, made azeotropic from toluene (3x), and dried under high vacuum overnight. CH2C12 was added (3 milliliters), and the mixture was cooled to 0 ° C. It was added BF3-OEt2 (approximately 1 μL, 0.01 mmol), and after stirring for 10 minutes the reaction was quenched with saturated aqueous NH4C1. The mixture was extracted with CH2C12 (3x), and the combined extracts were washed with saline, dried over Na2SO, concentrated and purified by preparative TLC (30 percent EtOAc-hexanes) to provide ether 317 (55.4 milligrams, 74 percent). 317 318 Alcohol 318. LAH (1 M in THF, 104 μL, 0.104 mmol) was added dropwise to a solution of ether 317 (54 milligrams, 0.087 mmol) in Et20 (5 milliliters) at 0 ° C. After 30 minutes, H20 and 1 M NaOH were carefully added. The mixture was stirred at room temperature for 10 minutes, filtered through glass wool, concentrated and purified by column chromatography (30 percent to 50 percent EtOAc-hexanes) to provide alcohol 318 (45.5 milligrams, 98 percent). 318 319 Aldehyde 319. Oxalyl chloride (11 μL, 0.13 mmol) was added dropwise to a solution of DMSO (18 μL, 0.25 mmol) in CH2C12 (2 milliliters) at -78 ° C. After 1.8 hours, a solution of alcohol 318 (22.6 milligrams, 0.042 mmol) in CH2C12 (1 milliliter) was added and the mixture was stirred for 1 hour. Et2N (53 μL, 0.38 mmol) was added and after 10 minutes, the reaction was warmed to 0 ° C and stirred 10 minutes. Saturated aqueous NHC1 was added and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by flash chromatography (20 percent EtOAc-hexanes) to provide the aldehyde 319 (21.7 milligrams, 97 percent). 319 B1933 B1933. In a manner similar to that described in Scheme 6 for the synthesis of B1794, the 319 intermediary was converted to B1933. HRMS (FAB): calculated for C? H5F3On + H 783.3931. Found: 783.3940.

B1897 B1942 B1942. A mixture of B1897 (2 milligrams, 2.73 μL), NaI04 (35 milligrams, 0.16 mmol), MeOH (0.8 milliliters) and H20 (0.2 milliliters) was stirred at room temperature for 30 minutes. The reaction mixture was then diluted with H20 (3 milliliters) and extracted with CH2C12 (6x) and EtOAc (2x) The combined organic phases were dried over Na2SO4 and purified by column chromatography (5 percent MeOH-CH2Cl2) to give the desired aldehyde. This material was dissolved in THF (0.1 milliliter), cooled to 0 ° C and treated with 0.5 M CF3TMS (30 μL, 15 mmol), followed by 0.05 M TBAF in THF (5 milliliters, 0.025 mmol). After stirring for 30 minutes, the reaction mixture was diluted with saturated aqueous NaHCO3 (2 milliliters) and H20 (1 milliliter) with EtOAc (6x), dried over Na2SO4, filtered and concentrated to give the bis-ether. Raw TMS This material was dissolved in THF (0.5 milliliters) with 1 M TBAF in THF containing 0.5 imidazole hydrochloride (8 μL, 8 μmol) at room temperature for 30 minutes. The reaction mixture was levigated through a Si02 column (50 percent EtOAc-hexanes to EtOAc) to provide the intermediate diol. A mixture of this product and Dess-Martin periodinnan (10 milligrams, 24 mmol) in CH2C12 (0.5 milliliters) was stirred at room temperature for 1 hour, diluted with. Et20 (5 milliliters) and filtered through Celite. The filtrate was concentrated and purified by preparative TLC (50 percent EtOAc-hexanes) to deliver B1942 (1.5 milligrams, 72 percent for the 5 steps). HRMS (FAB): calculated for C0H55F3O11 + H 767.3516. Found: 767.3542.

Synthesis of B2070 / B2073: MeQ OMPM 1) Nal04 MeQ OMPM HC H \, A "T0 ?? ../OPv 2) CH2 = CHCF2Br ../OPv In F F X400 401 Alcohol 401. A mixture of NaI04 (375 milligrams, 1.74 mmol), X400 (674 milligrams, 1.58 mmol), MeOH (16 milliliters) and H20 (4 milliliters) was stirred at room temperature for 1 hour. After dilution with H20, the mixture was extracted with CH2Cl2 (4x) and the combined organic extracts were dried over Na2SO4, concentrated and purified by flash chromatography (30 percent EtOAc-hexanes), to provide the intermediate aldehyde (570 milligrams), which was dissolved immediately in DMF (15 milliliters). Indium (275 milligrams, 2.4 mmol) and 3-bromo-3, 3-difluoropropene (240 μL, 2.4 mmol) were added and after stirring at room temperature for 17 hours, H20 and 0.1 M HCl were added. The mixture was extracted with EtOAc (3x) and the combined organic extracts were washed successively with H20 and brine, dried over Na2SO4, concentrated and purified by column chromatography. (20 percent to 30 percent EtOAc-hexanes), to provide alcohol 401 as a 1: 1 mixture of the C34 isomers (605 milligrams, 81 percent for the 2 steps). 401 402 Diol 402. A mixture of 0s04 (1 xtal), alcohol 401 (605 milligrams, 1.28 mmol), 4-methylmorpholine N-oxide (0.45 milligrams, 3.84 mmol), acetone (30 milligrams) and H20 was stirred. (6 milliliters) at room temperature for 29 hours. Os 04 (3 xtals) and N-oxid-or 4-methyl-morpholine (0.1 grams, 0.8 mmol) were added and after 2 days? A2S203 was added. The mixture was extracted with CH2C12 (6x) and the combined organic extracts were dried over? A2S04 and concentrated. The crude intermediate triol was immediately dissolved in 4: 1:: MeOH: H20 (25 milliliters) and added? AI0 (0.41 grams, 1.9 mmol). After vigorous stirring at room temperature for 2 hours, the mixture was diluted with H20, extracted with CHC12 (3x) and the combined organic extracts were dried over? A2S04 and concentrated to provide the intermediate aldehyde, which was dissolved Immediate 1: 1 EtOH-Et20 (30 milliliters) and cooled to 0 ° C. Add? ABH4 (48 milligrams, 1.3 mmol) and, after 20 minutes, quench the reaction with H20 and extract with CH2C12 (4x). The combined organic extracts were dried over α2S04, concentrated and purified by column chromatography (50 percent EtOAc-hexanes), to provide the diol 402 (485 milligrams, 80 percent for the 3 steps). 402 403 Silyl Ether 403. TBSOTf (2.3 milliliters, 10 mmol) was added dropwise to a mixture of diol 402 (485 milligrams, 1.0 mmol), Et3N (2.8 milliliters, 20 mmol) and CH2C12 (30 milliliters) at 0 ° C. After stirring for 1 hour at room temperature, saturated aqueous NH2C1 was added and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes), to provide silyl ether 403 (668 milligrams, 95 percent). 403 04 Alcohol 404. LAH (1 M in THF, 2.8 milliliters, 2.8 mmol) was added dropwise to a solution of 403 silyl ether (668 milligrams, 0.948 mmol) in Et20 (60 milliliters) at 0 ° C. After 15 minutes, they carefully added H20 and NaOH. The mixture was stirred at room temperature for 20 minutes, filtered through glass cloth, concentrated and purified by column chromatography (30 percent EtOAc-hexanes), to provide 404 alcohol (500 milligrams, 85%). hundred) .

Aldehyde 405. Oxalyl chloride (210 μL, 2.42 mmol) was added dropwise to a solution of DMSO (345 μL, 4.84 mmol) in CH2C12 (30 milliliters) at -78 ° C. After 1 hour, a solution of 404 alcohol (500 milligrams, 0.806 mmol) in CH2C12 (10 milliliters) was added. After 40 minutes, Et3N (1.0 milliliter, 7.2 mmol) was added. After stirring at -78 ° C for 10 minutes, the reaction mixture was warmed to 0 ° C and stirred for an additional 10 minutes. Saturated aqueous NH4C1 was added and the mixture was extracted with CH2C12 (3x).

The combined organic extracts were washed successively with H20, brine, dried over Na2SO4 and concentrated. Purification by flash chromatography (30 percent EtOAc-hexanes) provided aldehyde 405 (486 milligrams, 98 percent) which was used immediately in the next step. 405 406 Alkene 406. nBuLi (1.63 M, 860 μL, 1.4 mmol) was added dropwise to a solution of CH3PPh3Br (500 milligrams, 1.4 mmol) in THF (15 milliliters) and DMSO (6 milliliters) at 0 ° C. After 1 hour, a solution of aldehyde 405 (486 milligrams) in THF (15 milliliters) was added. The reaction mixture was warmed to room temperature and stirred for 30 minutes. Saturated aqueous NH4C1 was added, the mixture was extracted with EtOAc (3x) and the combined extracts were washed successively with H2O and brine, dried over Na2SO4, concentrated and purified by column chromatography (20% EtOAc- hexanes), to provide the alkene 406 (450 milligrams, 93 percent). 406 407 Ester 407. 9-BBN (0.5 M in THF, 9.0 milliliters, 4.5 mmol) was added dropwise to a solution of alkene 406 (0.460 grams, 0.746 mmol) in THF (10 milliliters) at 0 ° C. After warming to room temperature, the mixture was stirred for 3 hours and two additional portions of 9-BBN (0.5 M in THF, 3.0 milliliters, 1.5 mmol) were added at 30 minute intervals. The reaction mixture was again cooled to 0 ° C, after which THF (10 milliliters), H20 (10 milliliters) and NaB03 «4 H20 (1.72 grams, 11.2 moles) were carefully added. The mixture was stirred vigorously at room temperature for 1.5 hours, and additional NaB03 »4 H20 (1.0 grams, 6.5 mmol) was added. After 2 hours, the mixture was diluted with H20 and extracted with EtOAc (3x). The combined extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 to 30 percent EtOAc-hexanes) to provide the intermediate alcohol (509 milligrams), which was dissolved immediately. in CH2C12 (10 5 milliliters) and treated with pyridine (600 μL, 7.5 mmol) and PvCl (275 μL, 2.2 mmol). After 6 hours saturated aqueous NH4C1 was added and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 to 30 percent EtOAc-hexanes) to give the ester 407 (473 milligrams, 79 percent for the 2 steps). 407 408 Alcohol 408. A mixture of ester 407 (11 milligrams, 0.015 mmol) and Pd (OH) 2 / C (10 milligrams) in EtOAc 500 μL) was stirred vigorously under an atmosphere of H2 at room temperature for 6 hours. The mixture was filtered Through Celite, concentrate and purify by column chromatography (30 percent EtOAc-hexanes) to provide alcohol 408 (9.4 milligrams, quant). 408 409 Alkene 409. Oxalyl chloride (7 μL, 0.075 mmol) was added dropwise to a solution of DMSO (11 μL, 0.15 mmol) in CH2C12 (2 milliliters) at -78 ° C under N2. After 40 minutes, a solution of 408 alcohol (15.2 milligrams, 0.025 mmol) in CH2C12 (1 milliliter) was added and the reaction was stirred at -78 ° C for 1 hour. Et3N (31 μL, 0.22 mmol) was added and after stirring for 10 minutes, the mixture was heated to 0 ° C. After 10 minutes, the mixture was cooled with saturated aqueous NH4C1 and extracted with CH2C12 (3x). The combined extracts were washed successively with H20 and brine, dried over Na2SO4 and concentrated. After flash chromatography (30 percent EtOAc-hexanes), the intermediate acetone (13 milligrams) was dissolved immediately in THF (500 μL) and treated with Tebbe reagent (~ 0.65 M in toluene, 62 μL, 0.040 mmole) at 0 ° C. After 1.5 hours, additional Tebbe reagent (~ 0.65 M in toluene, 62 μL, 0.040 mmol) was added and after 10 minutes H20 and brine were carefully added. The mixture was extracted with EtOAc (3x) and the combined organic extracts were washed with brine, dried over Na 2 SO, concentrated and purified by column chromatography (10 percent EtOAc-hexanes) to provide the alkene 409 (11.9 milligrams, 80 percent for the 2 steps). 409 410 Alcohol 410. 9-BBN (0.5 M in THF, 1.5 milliliters, 0.72 mmol) was added dropwise to a solution of alkene 409 (0.144 grams, 0.242 mmol) in THF (2 milliliters) at 0 ° C. After warming to room temperature, the mixture was stirred for 3 hours. The reaction mixture was again cooled to 0 ° C, after which THF (2 milliliters), H20 (2 milliliters) and NaB03 «4 H20 (0.38 grams, 2.4 moles) were carefully added. The mixture was stirred vigorously at room temperature for 4 hours, diluted with H2O and extracted with EtOAc (3x). The combined extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 percent EtOAc-hexanes), to provide alcohol 410 (0.140 grams, 94 percent). 410 411 Alcohol 411. Oxalyl chloride (26 μL, 0.30 mmol) was added dropwise to a solution of DMSO (43 μL, 0.60 mmol) in CH2C12 (4 milliliters) at -78 ° C. After 1 hour, a solution of alcohol 410 (57 milligrams, 0.093 mmol) in CH2C12 (2 milliliters) was added. After 45 minutes, Et3N (125 μL, 0.90 mmol) was added. After stirring at -78 ° C for 10 minutes, the reaction mixture was warmed to 0 ° C and stirred for an additional 10 minutes. Saturated aqueous NH4C1 was added and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with brine, dried over Na 2 SO and concentrated. The crude product was dissolved in CH2C12 (4 milliliters), treated with Et3N (400 μL) and stirred at room temperature for 15 hours. Saturated aqueous NHC1 was added and the mixture was extracted with CH2C12 (3x). The combined organic extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by flash chromatography (30 percent EtOAc-hexanes), to provide the intermediate aldehyde (48 milligrams), which was immediately dissolved in 1: 1 Et20-EtOH (4 milliliters), cooled to 0 ° C and treated with solid NaBH (~4 milligrams, 0.09 mmol). After stirring for 15 minutes, saturated aqueous NH4C1 was carefully added and the mixture was extracted with EtOAc (3x). The combined extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by column chromatography (20 to 30 percent EtOAc-hexanes), to provide alcohol 411 (45.6 milligrams, 80 percent for the 3 steps ). 4U 4 * 2A, 412B 412A and 412B. Alcohol 411 (120 milligrams, 0.196 mmol) and MPMOTCI (0.17 grams, 0.59 mmol) were added, made azeotropic from toluene (3x) and dried under high vacuum for 1 hour. CH2C12 (9-milliliters) was added and the mixture was cooled to 0 ° C. BF3 * Oet2 (0.016 M in CH2Cl2, 125 μL, 0.002 mmol) was added dropwise and after stirring for 20 minutes, the reaction was quenched with saturated aqueous NH4C1. The mixture was extracted with CH2C12 (3x) and the combined extracts were washed with brine, dried over Na2SO4, concentrated and purified by preparative TLC (20 percent EtOAc-hexanes) to provide the intermediate MPM ether, which it contained some impurities that ran closely. This material was immediately dissolved in Et20 (10 milliliters) and treated with LAH (1M in THF, 300 μL, 0.300 mmol) at 0 ° C. After 10 minutes, H20 and 1M NaOH were added, and after stirring for 10 minutes at room temperature, the mixture was filtered through Celite, concentrated and purified by preparative TLC (35 percent EtOAc-hexanes) , to provide the 412A (49 milligrams, 39 percent for the 2 steps) as a single C34 isomer and the 412B (46 milligrams, 36 percent for the 2 steps) as a ~ 9: 1 mixture of C34 isomers.

B2070, B2073 412A, 412B B2070 and B2073. Intermediates 412A and 412B were converted to B2070 and B2073, respectively, in a manner similar to that described in Schemes 4 and 6. For B2070: HRMS (FAB): calculated for C? H58F20? 2 + Na 803.3794. Found: 803.3781.

Synthesis of B1963: 501 Diol 501 (64). NaHCO3 (21 milliliters) and KBr (89 milligrams, 0.75 mmol) were added to a solution of diol 10 (1.35 grams, 3.4 mmol) in CH2C12 (34 milliliters). The mixture was cooled to 0 ° C, and 4-methoxy-2,2,6-tetramethyl-1-piperidinyloxy (0.05 M in CH2C12, 7.45 milliliters, 0.37 mmol) and NaOCl (0.07 M in H20, 5.6 milliliters, were added). 0.39 mmole), sequentially. After 1 hour, the reaction mixture was cooled with Na2S203, diluted with saturated aqueous NaHCO3, and extracted with CH2C12 (3x). The combined extracts were dried over Na2SO4, concentrated and dissolved in THF (21 milliliters). After cooling to 0 ° C, CF3TMS (1.5 grams, 10.5 mmol) and TBAF (0.1 M in THF, 680 μL, 0.068 mmol) were added sequentially. After stirring for 40 minutes, additional TBAF (1 M in THF, 8.3 milliliters, 8.3 mmol) was added. After 30 minutes, the reaction was quenched with H20 and extracted with EtOAc (3x). The combined organic extracts were washed with brine, dried over Na 2 SO 4, concentrated and purified by flash chromatography (30 percent EtOAc, 40 percent, 50 percent-hexanes, followed by EtOAc), to provide a mixture of 2%. : 1 diols (553 milligrams, 35 percent). Separation by MPLC (1.5 percent, MeOH-CH2C12) gave the larger, more polar isomer 501 (64) (340 milligrams, 22 percent) and the smaller, less polar isomer (152 milligrams, 10 percent).

B1963 B1963. The intermediate 501 was converted to B1963, in a manner similar to that described in Schemes 4 and 6 for the synthesis of B1794.

D. Synthesis of B2320 and Related Analogs B2294 These compounds are manufactured by treating B2294 with an appropriate amine in a solvent such as methanol, for a period of from a few hours to several days. The progress of the reaction can be monitored by thin layer chromatography. A standard preparation procedure, well known to those of skill in the art, provides the desired compounds. The procedure below is to prepare ER803868; however, this procedure is general and can be used to prepare any desired analogue.

E. Synthesis of ER803868 To a solution of B2294, 1.2 milligrams in methanol, 0.5 milliliters, morpholine, 0.012 milliliters was added. The mixture was stirred for 10 days with additional morpholine, 0.012 milliliters, being added on days 1, 2, 3, 4 and 8. The mixture was then chromatographed, to give 1.4 milligrams of the desired compound.

B2320 R = N, N-dimethylamino B2330 R = N-isopropylamino B2336 R = N-methylamino B2339 R = Nt-butylamino B2417 R = N-2-hydroxyethylamino B2418 R = N-piperazinyl B2489 R = N, N-bis- ( 2-hydroxyethyl) amino B2490 R = N-1, 3-dihydroxy-2-propylamino B2491 R = N-benzylamino ER803834 R = N-piperidinyl ER803835 R = N-pyrrolidinyl ER803836 R = N-3- (R) -hydroxypyrrolidinyl ER803843 R = N-homopiperidinyl ER803845 R = N-para-methoxybenzylamino ER803846 R = N-phenethylamino ER803851 R = N-2- (S-hydroxymethyl) pyrrolidinyl ER803852 R = N-2- (R-hydroxymethyl) pyrrolidinyl ER803868 R = N-morpholinyl ER803869 R = N-ethylamino ER803870 R = N-imidazoyl ER803883 R = N, N-diethylamino ER803884 R = N-para-chlorobenzylamino D. Pharmacological Activity Many of the drugs that were individually described for their in vitro and in vivo activity were tested (see Table 1, below). The scanning methods included an inhibition assay of cell growth in vitro, using human colon cancer cells DLD-1 (accession number CCL 221 of the ATCC), in a microtiter plate format (Finlay, GJ and contributors, Analytical Biochemistry 139: 272-277, 1984), a mitral-block reversibility test U937 (accession number CRL 1593 of the ATTC) (described below), and in some cases, an in vivo growth inhibition assay of xenograft of LOX human melanoma tumor (see Table 1). The chemical stability to degradation of esterase was also examined. U937 Mitotic Block Reversibility Assay U937 human histiocytic lymphoma cells were added to 75 square centimeter tissue culture flasks as 2.5 x 106 cells in 22.5 milliliters of RPMI 1640 Medium containing 10 percent Fetal Bovine Serum. The cells were allowed to adapt to the culture for 36 hours of incubation at 37 ° C in a humidified atmosphere containing 5 percent C02. Then each test drug was added to a flask as 2.5 milliliters of 10x final concentration. The final concentrations that were obtained were from 0.1 - 1000 nM, in average log increments, for a total of 10 concentration steps that include a drug-free control flask that received 2.5 milliliters of medium. The cells were incubated with the drug for a period of 12 hours prior to treatment, at 37 ° C in a humidified atmosphere containing 5 percent C02. The contents of each flask were stirred and centrifuged at 300 x gram for 10 minutes at room temperature, after which the medium containing the drug was removed from the cell pellet. The cells were resuspended in 25 milliliters of drug-free hot medium and centrifuged at 300 x gram for 10 minutes at room temperature. After removing the medium from the cell pellet, the cells were resuspended in 35 milliliters of drug-free hot medium, transferred to fresh flasks, and a 10-milliliter sample of the cells was immediately removed from each cell. flask, were processed immediately as described below and stored for a subsequent cell cycle analysis (0 hours of drug wash). Incubation of the remaining 25 milliliters of cells was continued in the drug-free medium for another 10 hours. A 10 milliliter sample was removed from each flask, processed immediately and stored for subsequent cell cycle analysis (10 hours of drug wash) and a new 10 milliliter replacement medium was added to each incubation flask . Incubation of the cells in the drug-free medium continued for 5 days. After day two, 20 milliliters of medium and cells were removed from each flask and replaced with 20 milliliters of fresh medium. The viability of the cells was quantified after 5 days by trypan blue exclusion techniques, using hemacytometer counting. The cells were processed for cell cycle analysis using modifications of the method published in the source book of Becton Dickinson Immunocytometry Systems, section 1.11 (Preparation of Fixed Complete Cells by Alcohol Based on Suspensions for DNA Analysis). Briefly, each 10 milliliter sample that was removed from the flasks at the 0 and 10 hours of drug wash, was centrifuged separately at 300 x gram for 10 minutes. After removing the medium from the cell pellet, the cells were resuspended in 3 milliliters of cold saline. Seven milliliters of 100 percent cold ethanol were added slowly with vigorous vortex motion. The cell samples that were treated with ethanol from the periods of 0 hours to 10 hours of washing the compound were stored overnight at 4 ° C. The cells that were treated with ethanol were centrifuged at 300 x gram for 10 minutes, the ethanol was removed and then the cells were washed in 10 milliliters of Phosphate Regulated Serum (PBS). The cells were resuspended in 0.5 milliliters of 0.2 milligram / milliliter of Ribonuclease A (Sigma Number R-5503) in phosphate-buffered serum at a7 ° C water bath for 30 minutes. The cells were transferred to appropriate flow cytometry tubes and 0.5 milliliters of 10 milligrams / milliliters of propidium iodide (Pl) (Sigma Number P4170) in phosphate-buffered serum was added to each tube. The cells were incubated with Pl at room temperature in the dark for at least 15 minutes before analysis with a flow cytometer (Becton Dickinson FACScan flow cytometer or its equivalent). The cells should have been analyzed within an hour and kept in the dark at 4 ° C until they were ready. Cycle analysis of the cells was performed in cells of 0 hours and 10 hours using the flow cytometric measurement of the intensity of the cellular fluorescence. The fluorescence intensity of propidium iodide was measured for each cell on a linear amplification scale, ignoring even events using pair discrimination. The results that were obtained from the analysis of 15,000 cells were presented as a histogram with the increasing fluorescence intensity on the x axis and the number of cells at a particular intensity level, on the y axis. The intensity of spotting with Pl depends on the amount of DNA in the cell, so that it is possible to identify cells in different phases of the cell cycle, such as cells that have not yet synthesized DNA since the last mitosis (Gi stage), cells that are intermediate stages of DNA synthesis (S phase), and cells that have doubled their DNA complement and are ready to divide (phase G2). Cells that are blocked in the mitosis phase of the cell cycle have also doubled the amount of DNA compared to cells in the Gi phase. If all the cells are blocked in mitosis, there are no cells of the Gi phase, but if the block is removed when the compound is removed, the cells complete the mitosis and reappear in the Gi phase. In this way, the number of cells that reappear in this way in Gi or S phase, is a measure of the number of cells that have recently completed mitosis. For each sample at hours 0 and 10 after the compound was removed, the percentage of cells that terminate mitosis (such as the number of cells that reappear in the Gi phase) was quantified and tested as a function of the concentration initial of the compound that was used during the 12-hour pre-treatment period. The percentage of cells that were still viable 5 days after washing the drug on the same graph was superimposed, for example, see Figure 1 and Figure 2. A ratio between the concentration of the compound that is required to block by complete all cells in mitosis at hour 0 and the concentration that is required to maintain block 10 after the compound is removed. This was taken as a measure of the reversibility of a compound, indicating the proportions close to, or equal to one, the probably potent anti-tumor compounds in vivo (see Table 1, columns 4-6, and Figures 3 and 4). ). Table 1 In Vitro Inhibition and Reversibility Data Compound B1793 B1994 B1994 B1919 B1918 B1919 B1919 B1919 B1919 B2004 B2019 B2010 B2010 B2013 B2014 B2014 B2019 B2019 B2034 B2035 B2037 B2039 B2042 B2043 B2070 B2073 B2086 B2088 B2090 B2091 B2102 B2136 B2294 N2320 B2330 B2336 B2339 B2417 B2418 B2489 B2489 B2490 B2491 ER803834 ER803835 ER803836 ER803843 ER803845 ER803846 ER803846 ER803851 ER803852 ER803868 ER803869 ER803870 ER803883 ER803884 * = inhibition of cells in vitro ** = before washing # = after washing The invention also presents a method for identifying an agent that induces a Mitotic block held in a cell, after transient exposure of the cell to the agent. The invention presents the determination of the relative reversibility of the test compound, by relating the measurement of step (d) and the measurement of step (f), as described below. This determination can be a ratio, or an arithmetic difference, for example. In one aspect, the method includes: (a) incubating a first cell sample with a predetermined concentration of a test compound for a time between that sufficient to empty the Gi population and that equivalent to a cell cycle (eg, typically, 8-16 hours, or approximately 12 hours). (b) substantially removing the test compound from the first cellular sample (eg, by washing or changing the medium); (c) incubating the first sample in free medium of the test compound for a sufficient time interval to allow at least 80 percent (eg, 85 percent, 90 percent, and preferably 95 percent, 98 per one hundred or 99 percent) of the cells that were released from the mitotic block that was induced by a highly mitotic inhibitor, complete mitosis and return to the Gi phase (eg, typically 6-14 hours, or approximately 10 hours after from step (b) of separation); and (d) measuring the percentage of cells transiently exposed from step (c) that have completed mitosis and returned to Gi stage (e.g., by measuring a cell cycle marker, such as Pl-dependent fluorescence of DNA). One aspect of this scanning method includes the additional steps of: (e) incubating a second sample of cells with a concentration of the test compound that is less than or equal to that which was used in step (a) for a period of time between enough to empty the Gi population and that equivalent to a cell cycle; (f) measuring the percentage of cells from step (e) that has completed mitosis and that have returned to phase Gi; Y (g) determining a rate of reversibility of the test compound. In one embodiment of the method, the first and second cell samples are suspension culture cells that are selected from, for example, human leukemia, human lymphoma, murine leukemia, and murine lymphoma cells. The first and second cell samples can be incubated simultaneously (steps (a) and (e)) or in separate portions. Other embodiments further include, before step (a), step (i) of calculating a desirable time interval for incubating the first cell sample with a reversible mitotic blocking agent (or, alternatively, the test compound), to provide a satisfactory majority of cells that were harvested in the mitotic block; and wherein the incubation of step (a) is for the time interval that is calculated in step (i). Another embodiment of the method further includes, before step (c), step (ii) of calculating a desirable time interval for the free incubation of the test compound of step (c), step (ii) comprising the determination of the time interval after which at least 80% of the cells were previously treated with a complete mitosis of the highly reversible antimitotic agent and re-entered the Gi stage.; and wherein the incubation of step (c) is for the time interval that was determined in step (ii). Another embodiment of the method uses non-suspension culture cells from, for example, adherent murine or human cancer cells, which were cultured by any means to detach them from tissue culture flasks. One aspect of the method further includes repeating steps (a) - (f), using a range of relative concentrations of the test compound to determine which two substantially minimal concentrations of the test compound substantially provide the complete mitotic block in step ( d) and in step (f), respectively. The ratio of these sufficient minimum concentrations is an index of reversibility (see detailed U937 protocol for the preparation of exemplary dose response curves). These concentrations can be determined by extrapolating the percent cell curves (from steps (d) and (f)) as a function of concentration (for example, by testing only a few concentrations, such as 3 or less), or by empirically testing a full range of concentrations. The above methods are useful for identifying an agent (test compound) that inhibits mitosis, for identifying a mitotic blocking agent that substantially retained its effectiveness in blocking mitosis after its removal, and for predicting, for example, the IC50. or the IC95 of a mitotic blocking agent. When compared to relatively reversible antimitotic agents, substantially irreversible antimitotic agents, in other words, agents that continue to block mitosis in a cell, which has only been transiently exposed to the agent, are probably more effective in vivo in where natural processes, including multidrug resistance (MDR) pumps and metabolic trajectories or other degrading trajectories, prevent prolonged exposure. The effectiveness of relatively reversible antimitotic agents may depend on a period of sustained exposure.

In view of the cost to develop the pharmaceutical products, the economic advantages of determining the proportions of reversibility, as described above, are considerable. The above methods can be used, for example, to predict whether a test compound with good in vi tro activity will be effective in vivo, such as in a clinical trial. Relatively reversible agents would not be expected to perform as well as irreversible agents. This is shown, for example, by contrasting the data for the two known compounds, the vincristine of the relatively irreversible antimitotic agent and the vinblastine of the highly reversible antimitotic agent.

Table 2 Reversibility Characteristics of Vinblastine and Vincristine Concentration of the required matrix for the complete mitotic block, nM Compound Time 0 Hour 10 Interpretation Rate (before washing) (after washing) Reversibility Vinblastine 10 600 60 Highly Reversible Vincristine 10 10 1 Irreversible The analyzes of the antimitotic drugs, vinblastine and vincristine, in the Reversibility Test of the Mitotic Block U937, indicate that in spite of the identical potencies to induce the initial mitotic blocks (values of hour 0), the capacity of the two drugs to inducing the mitotic blocks that are sustained 10 hours after the drug wash (10 o'clock values), are very different: vincristine induces irreversible mitotic blocks, whereas those induced by vinblastine are highly reversible. Analyzes of in vivo anticancer activities of the antimitotic drugs, vinblastine and ia vincristine against human colon cancer xenografts COLÓ 205 that were cultured subcutaneously in immunocompromised mice (without hair), indicate that at equivalent doses of 1 milligram / kilogram, vincristine shows substantial cancer growth inhibitory activity, whereas vinblastine is inactive (Figure 5). At the lowest dose of 0.3 milligrams / kilogram, vincristine still produces moderate growth inhibition, while vinblastine is again inactive. The greater in vivo activity of vincristine correlates with its irreversibility in relation to the high reversibility of vinblastine. E. Use The compounds that are described have pharmaceutical activity, which includes anti-tumor and anti-mitotic activity, as demonstrated in section D above. Examples of tumors include melanoma, fibrosarcoma, monocytic leukemia, colon carcinoma, ovarian carcinoma, breast carcinoma, osteosarcoma, prostatic carcinoma, lung carcinoma, and ras-transformed fibroblasts. The invention features pharmaceutical compositions that include a compound of the formula (I) and a pharmaceutically acceptable carrier, the compositions may also include a combination of the compounds that were described, or a combination of one or more of the compounds that were described and others. pharmaceutically active agents, such as an anti-tumor agent, an immune stimulating agent, an interferon, a cytokine, an anti-MDR agent or an anti-angiogenesis agent. The compositions may be formulated for oral, topical, parenteral, intravenous or intramuscular administration, or administration by injection or inhalation. The formulations can also be prepared for controlled release, which includes transdermal patches. A method for inhibiting the growth of tumors in a patient includes the step of administering to the patient an effective anti-tumor amount of a compound or composition described. The invention also contemplates combination therapies, which include methods for the co-administration of a compound of formula (I) before, during, or after administering another pharmaceutically active agent. The methods of administration may be the same or different. Inhibition of tumor growth includes a growth of the cell or tissue that is exposed to the test compound that is at least 20 percent lower, and preferably 30 percent, 50 percent, or 75 percent less than the control growth (absence of known inhibitor or test compound). Other Modes The essential features of the invention can be readily discerned from the foregoing description and the claims below. On the basis of the above description, variations of the described compounds and methods of the invention can be designed and adapted without departing from the spirit and scope of the claims and the description. The references and publications described herein are incorporated herein in their entirety.

Claims (21)

1. CLAIMS 1. A compound that has the formula where A is a skeleton of saturated C1-6 hydrocarbons or C2.6 unsaturated, said skeleton being unsubstituted or having between 1 and 10 substituents, inclusive, independently selected from cyano, halo, azido, oxo and Qx; each Q1 is independently selected from OR ^ SR1 (S02Rlf OSOaRi, NR2Rlf NR2 (CO) R1, NR2 (CO) (CO) R1 # NR4 (CO) NR2RX, NR2. (CO) OR1 # (CO) OR1 # 0 (C0) ) R1 ((CO) NR2R1 (and 0 (CO) NR2R ?; each Rx, R2, R4, R5 and R6 is independently selected from H, C1_e haloalkyl, hydroxyalkyl aminoalkyl C6.10 aryl, C6.10 haloaryl, C6 hydroxyaryl .10, alkoxy Cj ^ .j-aryl C6.10, aryl C6.10-alkyl C ^ g, alkyl C6.10-aryl C6.10, haloaryl C6.10, (C6-C6-alkoxy) -alkyl Cx.z, C2.9 heterocyclic radical, C2.9 heterocyclic radical C2_9 heteroaryl alkyl, and C2.9-heteroaryl C ^ g alkyl, - each of D and D 'is independently selected from R3 and 0R3, wherein R3 is H, C1_3 alkyl, or C3 haloalkyl; n is 0 or 1; E is R5 or 0R5; G is 0, S, CH2 or NR6; each of J and J 'is independently H, C- ^ g alkoxy, or straight or branched alkyl) -0-; Q is C1-3 alkyl; T is ethylene or ethenylene, optionally substituted with (CO) OR7, wherein R7 is H or C ^ g alkyl, - each of U and U 'is independently H, C- ^ g alkoxy, or C- ^ g alkyl, - or U and U' taken together are = CH2 or -O- (straight or branched Cx_5 alkylene) -0-; X is H or C ^ g alkoxy, - each of Y and Y 'is independently H or C 1-6 alkoxy; or Y and Y 'taken together with = 0, = CH2, or -0- (straight or branched Cx.5 alkylene) -O-; and each of Z and Z 'is independently H or C 1-6 alkoxy; or Z and Z 'taken together with = 0, = CH2, or -O- (straight or branched Cx.5 alkylene) -O-; or a pharmaceutically acceptable salt thereof.
2. 2. The compound of claim 1, wherein n is 0.
3. 3. The compound of claim 1, wherein each of D and D 'is independently selected from R3, C4 alkoxy, and haloalkyloxy C-, ^.
4. 4. The compound of claim 1, wherein R5 is selected from H, C1-6alkyl, haloalkyl C ^ g, hydroxyalkyl aminoalkyl C1-6, aryl C6-10, haloarylC6.10, hydroxyarylC6. 10, C1-3 alkoxy-C6_10 aryl, C6.10 aryl-C6.10 alkyl-C6.10 aryl, C1.6 haloaryl, C1.s-haloaryl C6.10 alkyl, (C1-3 alkoxy-C6 aryl) C1-3 alkyl, C2_9 heterocyclic radical, C2.9 heterocyclic radical, C1-6 alkyl, C2.9 heteroaryl, and C2.9 heteroaryl-C1-6 alkyl. The compound of claim 1, wherein A comprises a skeleton of saturated or C2.6 unsaturated hydrocarbons, said backbone having at least one substituent selected from cyano, halo, azido, oxo and Q1 # - each Q1 is independently selected from OR ^ SRX, S02Rx, OS02R1 # NR2Rlf NR- (CO) R1 (NR2 (CO) (CO) R1 (NR4 (CO) NR2R1 # NR2. (CO) OR1 ((COJOR ^ 0 (CO) Rlf (CO) NR2R1, and 0 ( CO) NR2R ?; n is 0; G is O; J and J 'taken together are = CH2, Q is methyl, T is ethylene, U and U' taken together are = CH2, X is H, each of Y and Y 'is H; and Z and Z' taken together are = 0 or = CH2 6. The compound of claim 1, wherein each Qx is independently selected from OR !, SR17 SO ^, OSO ^, NH (C0) Rlf NH (CO) (C0) R1 # and 0 (C0) NHR ?; each Rl is independently selected from CX_6 alkyl, C1_6 haloalkyl, hydroxy alkyl amino C6-10 aryl, C6.10 haloaryl, C6.10 hydroxyaryl, alkoxy C1.3-C6.10 aryl, C6.10-aryl, C1.6-alkyl, C6.10-C6.10 aryl, C1-6 haloaryl-C1-6 alkyl, C6 g -haloaryl C6.10 alkyl, ( C.sub.3-alkoxy C6) -C.sub.4 alkyl, C2.9 heterocyclic radical, C2_9 heterocyclic radical C2-9 heteroaryl alkyl, and C2.9 heteroaryl-alkyl one of D and D 'is methyl or methoxy, and the other is H; n is 0; G is O; J and J 'taken together are = CH2; Q is methyl; T is ethylene; U and U 'taken together are = CH2; X is H; each of Y and Y 'is H; and Z and Z 'taken together are = 0. The compound of claim 6, wherein A has at least one substituent selected from hydroxyl, amino, azido, halo and oxo. 8. The compound of claim 7, wherein A comprises a skeleton of saturated hydrocarbons having at least one substituent selected from hydroxyl, amino and azido. 9. The compound of claim 8, wherein A has at least two substituents independently selected from hydroxyl, amino and azido. The compound of claim 8, wherein A has at least two substituents independently selected from hydroxyl and amino. The compound of claim 8, wherein A has at least one hydroxyl substituent and at least one amino substituent. The compound of claim 8, wherein A has at least two hydroxyl substituents. The compound of claim 8, wherein A comprises a C2_4 hydrocarbon backbone. The compound of claim 8, wherein A comprises a C3 hydrocarbon skeleton. 15. The compound of claim 13, wherein A has a (S) -hydroxyl at the alpha carbon atom to the carbon atom linking A with the ring containing G. The compound of claim 6, wherein A comprises a skeleton of saturated hydrocarbons having at least one substituent selected from hydroxyl and cyano. 17. The compound of claim 6, wherein Qx is independently selected from OR ^ SR1 (SO ^ and OSO ^, where each Rx is independently selected from C x g alkyl, C 1 β haloalkyl, C 6 aryl, C 6 haloaryl, C 1 alkoxy C6-aryl, Cg-C6-alkyl, C6-alkyl, C6-haloaryl, C6-alkyl, C6-haloalkyl, C6-alkyl, and (C6-alkoxy) C3-alkyl. composed of the following structure: 19. The compound of the following structure: and their pharmaceutically acceptable salts. 20. A method for identifying an agent that induces a mitotic block sustained in a cell after transient exposure of said cell to said agent, said method comprising the steps of: (a) incubating a first cellular sample with a predetermined concentration of a compound test for a time interval between enough to empty the Gx population and the equivalent to a cell cycle; (b) substantially separating said test compound from said first cellular sample; (c) incubating said first sample in a medium free of said test compound for a sufficient time interval to allow at least 80% of the cells released from the mitotic block induced by a highly reversible mitotic inhibitor to complete mitosis and return to phase G1; Y (d) measure the percentage of cells exposed transiently from step (c) that have completed mitosis and returned to phase G1. The method of claim 20, further comprising the steps of: (e) incubating a second sample of cells with a concentration of said test compound less than or equal to that used in step (a) for a time interval between enough to empty the Gx population and the equivalent to a cell cycle; (f) measuring the percentage of cells from step (e) that have completed mitosis and have returned to the Gx phase; and (g) determining the relative reversibility of said test compound by relating the measurement of step (d) and the measurement of step (f).